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Search Results (339)

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23 pages, 2901 KiB  
Article
Wood Gasification Biochar as an Effective Biosorbent for the Remediation of Organic Soil Pollutants
by Elisabetta Loffredo, Nicola Denora, Danilo Vona, Antonio Gelsomino, Carlo Porfido and Nicola Colatorti
Soil Syst. 2025, 9(1), 18; https://doi.org/10.3390/soilsystems9010018 - 24 Feb 2025
Abstract
A biochar (BC) generated by the pyrogasification of wood chips from authorized forestry cuts was extensively characterized and evaluated for its efficacy in retaining/releasing two agrochemicals, namely the fungicide penconazole (PEN), the herbicide S-metolachlor (S-MET), and the xenoestrogen bisphenol A (BPA) widely present [...] Read more.
A biochar (BC) generated by the pyrogasification of wood chips from authorized forestry cuts was extensively characterized and evaluated for its efficacy in retaining/releasing two agrochemicals, namely the fungicide penconazole (PEN), the herbicide S-metolachlor (S-MET), and the xenoestrogen bisphenol A (BPA) widely present in industrial effluents. The elemental composition of BC was evaluated using CN elemental analysis and total reflection X-ray fluorescence (TXRF) spectroscopy which showed the abundance of elements typically found in BCs (Ca, K, P) along with essential trace elements such as Fe and Mn. Scanning electron microscopy coupled with energy-dispersive X-ray analysis (SEM-EDX) described the surface features of BC along with the major surface elements, while Brunauer–Emmett–Teller (BET) analysis revealed, as expected, a large specific surface area (366 m2 g−1). High porosity (0.07 cm3 g−1) was demonstrated by the density functional theory (DFT) method, while Fourier transform infrared (FT-IR) spectroscopy highlighted the presence of a prominent aromatic structure and the abundance of reactive functional groups responsible for the binding of the compounds. The sorption/desorption capacity of BC was studied by means of sorption kinetics and isotherms in batch trials, and by modeling the experimental data with various theoretical equations. All compounds reached sorption equilibrium on BC very rapidly, following preferentially pseudo-second-order kinetics. Freundlich adsorption constants of PEN, S-MET, and BPA were 37.3, 13.2, and 11.6 L g−1, respectively, thus demonstrating the great affinity of BC for hydrophobic pollutants. The adsorption process was hysteretic as only a small fraction of each compound was slowly desorbed from BC. The overall results obtained highlighted the great potential of BC of acting as a biosorbent of contaminants, which is of great importance for the containment of pollution in agricultural soils and for limiting the entry of toxic compounds into the human and animal food chain. Full article
(This article belongs to the Special Issue Adsorption Processes in Soils and Sediments)
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<p>Representative TXRF spectrum of the BC sample.</p>
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<p>SEM images at magnifications of 1500 (<b>above left</b>) and 10,000 (<b>above right</b>) and EDX spectrum (<b>below</b>) of BC.</p>
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<p>FTIR-ATR spectra of BC (<b>A</b>) and of BC-compound interaction product (<b>B</b>). Highlighted areas (pink boxes) indicate the main signals (<b>A</b>). Highlighted areas (<b>B</b>): aromatic overtones signals (green box), specific -C-C-/-CH peaking (blue box), specific -C-O/-Si-O: -C-O/-C-N cluster signals (pink box).</p>
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<p>Effects of solution/BC ratio on the concentration of the adsorbed compounds at equilibrium. Data were statistically treated by ANOVA analysis and Duncan’s new multiple range test. For each compound, different letters indicate statistically significant differences at <span class="html-italic">p</span> ≤ 0.05 (<span class="html-italic">n</span> = 3).</p>
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<p>Adsorption kinetics of the compounds on BC. Points indicate experimental data, while the solid lines are plots of the pseudo-second-order model (PSO). The vertical bar on each point indicates the standard error (<span class="html-italic">n</span> = 3). The initial concentration of each compound was 2 mg L<sup>−1</sup>, and the pH of the solution/BC (ratio 10,000, <span class="html-italic">v</span>/<span class="html-italic">w</span>) suspension was 8.2.</p>
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<p>Adsorption/desorption isotherms of the compounds on/from BC. Points indicate experimental data, while solid lines and dashed lines are plots of the Freundlich model for adsorption and desorption, respectively. The vertical bar on each point indicates the standard error (<span class="html-italic">n</span> = 3). The equilibrium time was 16 h.</p>
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17 pages, 3763 KiB  
Article
Bio-Based and Solvent-Free Epoxy Vitrimers Based on Dynamic Imine Bonds with High Mechanical Performance
by Lei Chen, Na Ning, Gang Zhou, Yan Li, Shicheng Feng, Zhengyan Guo and Yi Wei
Polymers 2025, 17(5), 571; https://doi.org/10.3390/polym17050571 - 21 Feb 2025
Abstract
Conventional epoxy thermosets, with irreversible crosslinking networks, cannot be reprocessed and recycled. Furthermore, the utilization of petroleum-based materials accelerates the depletion of non-renewable resources. The introduction of dynamic covalent bonds and the use of bio-based materials for thermosets can effectively address the above [...] Read more.
Conventional epoxy thermosets, with irreversible crosslinking networks, cannot be reprocessed and recycled. Furthermore, the utilization of petroleum-based materials accelerates the depletion of non-renewable resources. The introduction of dynamic covalent bonds and the use of bio-based materials for thermosets can effectively address the above issues. Herein, a series of bio-based epoxy vitrimers with dynamic covalent imine bonds were synthesized via a simple solvent-free, one-pot method using vanillin-derived aldehyde monomers, 4,4-diaminodiphenylsulfone (DDS) and bisphenol F diglycidyl ether (BFDGE) as raw materials. The effect of crosslinking density, crosslinking structure and imine bond content on the resulting bio-based vitrimers was studied, demonstrating their excellent thermal properties, UV shielding and solvent resistance, as well as outstanding mechanical properties compared to those of the previously reported vitrimers. In particular, the cured neat resin of vitrimer had a maximum tensile strength of 109 MPa and Young’s modulus of 6257 MPa, which are higher than those of previously reported imine-based vitrimers. The dynamic imine bonds endow these vitrimers with good reprocessability upon heating (over 70% recovery) and degradation under acidic conditions, enabling recycling by physical routes and gentle degradation by chemical routes. This study demonstrates a simple and effective process to prepare high-performance bio-based and recycled epoxy thermosets. Full article
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<p>(<b>a</b>) DSC curves and (<b>c</b>) storage modulus of the control and <span class="html-italic">tri</span>-PHV with different imine bond contents; (<b>b</b>) DSC curves and (<b>d</b>) storage modulus of the control, <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20 and <span class="html-italic">di</span>-CHV-20; (<b>e</b>) TGA curvesof the control, <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20 and <span class="html-italic">di</span>-CHV-20 with different imine bond contents; (<b>f</b>) comparison of thermal stability of <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20, and <span class="html-italic">di</span>-CHV-20 (in dash circle) with reference [<a href="#B31-polymers-17-00571" class="html-bibr">31</a>,<a href="#B33-polymers-17-00571" class="html-bibr">33</a>,<a href="#B34-polymers-17-00571" class="html-bibr">34</a>,<a href="#B37-polymers-17-00571" class="html-bibr">37</a>,<a href="#B40-polymers-17-00571" class="html-bibr">40</a>,<a href="#B43-polymers-17-00571" class="html-bibr">43</a>,<a href="#B46-polymers-17-00571" class="html-bibr">46</a>,<a href="#B49-polymers-17-00571" class="html-bibr">49</a>,<a href="#B50-polymers-17-00571" class="html-bibr">50</a>,<a href="#B51-polymers-17-00571" class="html-bibr">51</a>].</p>
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<p>(<b>a</b>) Stress relaxation curves of <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20, <span class="html-italic">di</span>-CHV-20 at 210 °C; (<b>b</b>) stress relaxation curves of <span class="html-italic">tri</span>-PHV-20 at temperatures of 200–230 °C; (<b>c</b>) linear fitting line of <span class="html-italic">tri</span>-PHV-20; (<b>d</b>) activation energy (<span class="html-italic">E<sub>a</sub></span>) and topological freezing transition temperature (<span class="html-italic">T<sub>v</sub></span>) of <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20, <span class="html-italic">di</span>-CHV-20.</p>
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<p>(<b>a</b>) Typical stress–strain curves and (<b>b</b>) the detailed mechanical properties for the control, <span class="html-italic">tri</span>-PHV, <span class="html-italic">di</span>-PHV and <span class="html-italic">di</span>-CHV; (<b>c</b>) comparison of tensile properties and Young’s modulus of <span class="html-italic">tri</span>-PHV, <span class="html-italic">di</span>-PHV and <span class="html-italic">di</span>-CHV with those of other imine vitrimers from the references [<a href="#B29-polymers-17-00571" class="html-bibr">29</a>,<a href="#B30-polymers-17-00571" class="html-bibr">30</a>,<a href="#B31-polymers-17-00571" class="html-bibr">31</a>,<a href="#B34-polymers-17-00571" class="html-bibr">34</a>,<a href="#B37-polymers-17-00571" class="html-bibr">37</a>,<a href="#B38-polymers-17-00571" class="html-bibr">38</a>,<a href="#B39-polymers-17-00571" class="html-bibr">39</a>,<a href="#B41-polymers-17-00571" class="html-bibr">41</a>,<a href="#B42-polymers-17-00571" class="html-bibr">42</a>,<a href="#B46-polymers-17-00571" class="html-bibr">46</a>,<a href="#B49-polymers-17-00571" class="html-bibr">49</a>,<a href="#B50-polymers-17-00571" class="html-bibr">50</a>]; (<b>d</b>) UV spectra of the control, <span class="html-italic">tri</span>-PHV, <span class="html-italic">di</span>-PHV and <span class="html-italic">di</span>-CHV.</p>
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<p>(<b>a</b>) Digital photo of degradation process of <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20 and <span class="html-italic">di</span>-CHV-20 in <span class="html-italic">x</span> M HCl + EDA + DMSO (<span class="html-italic">x</span> = 0.1, 0.2, 0.5, 1, V<sub>HCl</sub>:V<sub>EDA</sub>:V<sub>DMSO</sub> = 1:1:8); (<b>b</b>) degradation mechanism of vitrimers with HCl and EDA.</p>
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<p>(<b>a</b>) Illustration of thermal reprocessing progress; (<b>b</b>) imine metathesis reaction of <span class="html-italic">tri</span>-PHV-20; (<b>c</b>) stress–strain curves for original and recycled <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20 and <span class="html-italic">di</span>-CHV-20.</p>
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<p>(<b>a</b>) Photographs of the control, <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20 and <span class="html-italic">di</span>-CHV-20 samples dispersed in DMSO solution before and after 24 hat RT; (<b>b</b>) gel fraction and swelling ratio of the Control, <span class="html-italic">tri</span>-PHV-20, <span class="html-italic">di</span>-PHV-20 and <span class="html-italic">di</span>-CHV-20 in DMSO solution.</p>
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<p>Comparison of overall performance of <span class="html-italic">tri</span>-PHV with that of other imine vitrimers from the references [<a href="#B42-polymers-17-00571" class="html-bibr">42</a>,<a href="#B44-polymers-17-00571" class="html-bibr">44</a>,<a href="#B46-polymers-17-00571" class="html-bibr">46</a>,<a href="#B49-polymers-17-00571" class="html-bibr">49</a>,<a href="#B50-polymers-17-00571" class="html-bibr">50</a>,<a href="#B52-polymers-17-00571" class="html-bibr">52</a>].</p>
Full article ">Scheme 1
<p>Synthetic route of (<b>a</b>) vanillin-based aldehyde monomers (<span class="html-italic">di</span>-Ali, <span class="html-italic">di</span>-Aro, and <span class="html-italic">tri</span>-Aro) and (<b>b</b>) imine vitrimer (<span class="html-italic">di</span>-CHV, <span class="html-italic">di</span>-PHV, <span class="html-italic">tri</span>-PHV); (<b>c</b>) dynamic crosslink of imine epoxy vitrimers.</p>
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16 pages, 3488 KiB  
Article
Toxic Effects of Bisphenol A on L. variegatus and A. punctulata Sea Urchin Embryos
by Jacob D. Kunsman, Maya C. Schlesinger and Elizabeth R. McCain
Hydrobiology 2025, 4(1), 5; https://doi.org/10.3390/hydrobiology4010005 - 19 Feb 2025
Abstract
Bisphenol A, BPA, is a small molecule frequently used in large-scale plastic production. The chemical has garnered a reputation for its association with harmful human health effects, and numerous animal studies have contributed to its classification as an endocrine disruptor. Prior research has [...] Read more.
Bisphenol A, BPA, is a small molecule frequently used in large-scale plastic production. The chemical has garnered a reputation for its association with harmful human health effects, and numerous animal studies have contributed to its classification as an endocrine disruptor. Prior research has investigated the impact of the chemical on echinoderms, including seven species of sea urchin. Our project investigated the toxic effects of this chemical on two uninvestigated species: Lytechinus variegatus and Arbacia punctulata. We exposed embryos to a range of environmentally relevant BPA concentrations (1 µg/L, 10 µg/L, 100 µg/L, and 1000 µg/L) for 48 h, until the pluteus stage. Larvae were classified according to the type of abnormality they exhibited, using a light microscope, and the EC50 was determined through probit analysis and dose–response curves. We also examined isolated plutei skeletons under a scanning electron microscope to assess changes to the skeletal structure under increasing concentrations of BPA. Our results suggest BPA induces embryotoxicity and soft tissue abnormalities more severely in L. variegatus, whereas A. punctulata exhibits more resistance to these effects. The EC50 values, over 1000 µg/L for A. punctulata and approximately 260 µg/L for L. variegatus, support this. These relative values also agree with our hypothesis that sea urchin embryos in a single genus have a similar level of BPA embryotoxicity. Interestingly, under SEM examination, the A. punctulata skeletal microstructure appears to be altered as a result of BPA exposure. While the EC50s are below what has been documented in many, but not all, marine environments, longer and consistent exposure may have a more deleterious impact. These findings suggest BPA’s effects on echinoderms should be further explored with multiple forms of analysis and over the long term. Full article
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<p>Observed embryonic malformations in <span class="html-italic">Lytechinus variegatus</span> 48 h post-fertilization. (<b>A</b>) Normal pluteus, (<b>B</b>) shortened arms, (<b>C</b>) incomplete/missing arm(s), (<b>D</b>) underdeveloped pluteus, (<b>E</b>) deteriorated epithelium of a pluteus, (<b>F</b>) prism, and (<b>G</b>) ball of cells. Scale bar represents 10 µm.</p>
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<p>Observed embryonic malformations in <span class="html-italic">Arbacia punctulata</span> 48 h post-fertilization. Asterisk indicates pluteus of interest. (<b>A</b>) Normal pluteus, (<b>B</b>) shortened arms, (<b>C</b>) incomplete/missing arm(s), (<b>D</b>) underdeveloped pluteus, (<b>E</b>) deteriorated epithelium of a pluteus, (<b>F</b>) prism, and (<b>G</b>) ball of cells. Scale bar represents 10 µm.</p>
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<p>Percentage of total normal (dots) and abnormal (horizontal lines) embryos in different BPA concentrations within (<b>A</b>) <span class="html-italic">Arbacia punctulata</span> (n = approximately 200). (<b>B</b>) Dose–response curve for <span class="html-italic">L. variegatus</span> embryos/larvae exposed to seawater (control) or one of four concentrations of BPA for 48 h. (<b>C</b>) The total prevalence of each abnormality across all BPA samples combined, not including controls. Blue bars indicate <span class="html-italic">A. punctulata</span> abnormalities while orange bars indicate <span class="html-italic">L. variegatus</span> abnormalities.</p>
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<p>Depictions of measurements performed on sea urchin plutei and visual differences in sea urchin basket structure between <span class="html-italic">Arbacia punctulata</span> and <span class="html-italic">Lytechinus variegatus</span>. (<b>A</b>) Demonstration of the measurement parameters for full length (FL), arm length (AL), bridge length (BL), arm width (AW), and bridge width (BW) on an <span class="html-italic">A. punctulata</span> pluteus. (<b>A</b>) Scale bar represents 33.3 µm. Differences in basket structure can be observed between <span class="html-italic">A. punctulata</span> (<b>B</b>) and <span class="html-italic">L. variegatus</span> (<b>C</b>). (<b>B</b>,<b>C</b>) scale bars are 5 µm.</p>
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<p>Average length (µm) of each skeletal measurement performed in each treatment group (n = 30) of <span class="html-italic">Arbacia punctulata</span>. Horizontal brackets indicate which pairs are being statistically compared. One star (*) denotes <span class="html-italic">p</span>-value &lt; 0.05, two stars (**) denotes <span class="html-italic">p</span>-value &lt; 0.01, and three stars (***) denotes <span class="html-italic">p</span>-value &lt; 0.001.</p>
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<p>Average length (µm) of each skeletal measurement performed in each treatment group (n = 30) of <span class="html-italic">Lytechinus variegatus</span>. Horizontal brackets indicate which pairs are being statistically compared. One star (*) denotes <span class="html-italic">p</span>-value &lt; 0.05, two stars (**) denotes <span class="html-italic">p</span>-value &lt; 0.01, and three stars (***) denotes <span class="html-italic">p</span>-value &lt; 0.001.</p>
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17 pages, 2312 KiB  
Article
Green Chemistry Method for Analyzing Bisphenol A in Milk
by Angela M. Encerrado Manriquez and Wen-Yee Lee
Separations 2025, 12(2), 25; https://doi.org/10.3390/separations12020025 - 25 Jan 2025
Viewed by 181
Abstract
A simple, fast, green, and sensitive method for determining Bisphenol A (BPA) levels in commercial milk was developed using a solventless sample preparation technique known as stir bar sorptive extraction, coupled with thermal desorption–gas chromatography/mass spectrometry. BPA was selected due to its ubiquitous [...] Read more.
A simple, fast, green, and sensitive method for determining Bisphenol A (BPA) levels in commercial milk was developed using a solventless sample preparation technique known as stir bar sorptive extraction, coupled with thermal desorption–gas chromatography/mass spectrometry. BPA was selected due to its ubiquitous presence in the environment and its classification as an endocrine-disrupting chemical of concern (i.e., its ability to mimic hormone functions). Studies have reported that BPA can leach into various food sources, including milk, a dietary staple for infants. It is critical to have an effective and efficient process for monitoring the presence of BPA in milk to protect children’s health. Current detection methods for BPA in milk are lengthy and tedious and tend to require the use of organic solvents for the extraction of BPA. This optimized “green” method provides an effective alternative for BPA detection in a challenging matrix, e.g., milk. Factors such as pH (1.5, 6, and 13), temperature (70–80 °C), and sonication (1 h, 2 h, and 3 h) were studied with a BPA-spiked whole milk sample (final concentration of 8 ppb) to optimize the extraction efficiency without the use of solvents. The developed methodology improves BPA recovery from whole milk by over 50%, with a detection limit in the parts per trillion range (45 ng/L). The sample preparation developed in this report rendered a more sensitive option for analyzing BPA in milk, with a limit of detection in the parts per trillion range (compared to low ppb) even though the recovery performance is not as good as in reported studies (54% vs. >85%); nonetheless, it provides a green alternative for future studies assessing BPA exposure through dairy products. Full article
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<p>Diagrams showing the experimental design. (<b>A</b>) The sample preparation specifics and pre-extraction conditions that were studied. (<b>B</b>) The different extraction conditions that were studied to improve extraction efficiency during SBSE.</p>
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<p>BPA recovery (based on instrument response) under neutral (MN), basic (MB), and acidic (MA) conditions. MA and MB results were normalized and compared to MN; therefore, the standard deviation was excluded from the dataset (<span class="html-italic">n</span> = 8). Statistical analysis revealed that the data did not follow a normal distribution, and a non-parametric test revealed no statistically significant difference across treatments (<span class="html-italic">p</span>.adj-value &gt; 0.05). The same letter above each treatment indicates no significant differences at Tukey’s test (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>The effects of sonication time and temperature on BPA recovery from MN, MB, and MA. (<b>A</b>) Effects of 0–3 h of continuous sonication starting at 25 °C; (<b>B</b>) effects of temperature maintained at 60–80 °C for 3 h; and (<b>C</b>) combined effects of sonication (0–3 h) and temperature (69 °C). Responses were normalized against the control condition, as there was no pH adjustment or sonication. The standard error is not shown since all samples were normalized. A complete statistical analysis can be found in the <a href="#app1-separations-12-00025" class="html-app">Supplementary Section (Table S1)</a>.</p>
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<p>Recovery of BPA from different solvents using SBSE: (<b>A</b>) BPA recovered from water and milk samples with 0%, 10%, and 30% of ACN, and (<b>B</b>) BPA recovered from water and milk samples with 0%, 10%, and 30% of MeOH. BPA recovery was based on the instrument’s response, i.e., BPA peak area in the chromatograms. Data are means of three replicates ± standard error (error bar).</p>
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<p>Recovery of BPA during SBSE through the addition of EDTA. Data are means of three replicates ± standard error (shown as error bars).</p>
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14 pages, 391 KiB  
Article
Distribution of Environmental Phenols into Follicular Fluid and Urine of Women Attending Infertility Clinic
by Anna Klimowska, Joanna Jurewicz, Michał Radwan, Paweł Radwan, Paweł Pol and Bartosz Wielgomas
J. Xenobiot. 2025, 15(1), 17; https://doi.org/10.3390/jox15010017 - 21 Jan 2025
Viewed by 370
Abstract
Infertility and environmental pollution are two globally prevalent and related issues. To explore women’s reproductive health, the composition of follicular fluid (FF) has been studied and it was found that changes to its composition, including the presence of exogenous chemicals, can adversely affect [...] Read more.
Infertility and environmental pollution are two globally prevalent and related issues. To explore women’s reproductive health, the composition of follicular fluid (FF) has been studied and it was found that changes to its composition, including the presence of exogenous chemicals, can adversely affect the fertilization process. Two groups of women (idiopathic infertility and controls) who were patients at a fertility clinic were recruited for this study. Samples of urine and FF were gathered from each participant to determine the concentration of 14 common phenols (four parabens, six bisphenols, two benzophenones, and two naphthols). Associations between phenol concentrations (free and total) in both matrices were described using Spearman’s correlation coefficient and were compared between two groups by the Mann–Whitney U test. Eight phenols were quantified in more than 50% of the urine samples, while only three parabens were quantified in hydrolyzed FF samples, and only methylparaben was quantified in non-hydrolyzed FF samples. Conjugates were the predominant form in FF samples. However, a significant correlation of 0.533 (p < 0.0001) was observed between free and total methylparaben concentrations in FF. Differences in concentrations between cases and controls in both matrices were not statistically significant, except for benzophenone-3 in urine, with a higher median observed in the control group (p = 0.04). The total paraben concentrations in urine and FF samples were rather weakly correlated (r = 0.232–0.473), implying that urine concentrations may not be appropriate for predicting their concentration in FF. Full article
(This article belongs to the Section Emerging Chemicals)
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<p>Spearman’s correlation plots for total paraben concentrations in urine and follicular fluid samples. Correlation between (<b>a</b>) free and total concentrations of methylparaben in follicular fluid, (<b>b</b>) total concentrations of parabens in the follicular fluid and (<b>c</b>) between the matrices. Plots include only samples with concentrations &gt;LOD.</p>
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<p>Box plots of (<b>A</b>) total urinary phenolic concentrations and (<b>B</b>) total and (<b>C</b>) free concentrations of parabens in follicular fluid. Analytes with detection &gt;50% were only plotted. Box plots’ description: line—median, box—25th and 75th percentile, whiskers—5th and 95th percentile, dots—results below 5th and above 95th percentile. Dotted line—LOD in FF samples (0.2 ng/mL). *—<span class="html-italic">p</span> &lt; 0.05 (Mann–Whitney U test).</p>
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30 pages, 6320 KiB  
Article
Environmental Exposure to Bisphenol A Enhances Invasiveness in Papillary Thyroid Cancer
by Chien-Yu Huang, Ren-Hao Xie, Pin-Hsuan Li, Chong-You Chen, Bo-Hong You, Yuan-Chin Sun, Chen-Kai Chou, Yen-Hsiang Chang, Wei-Che Lin and Guan-Yu Chen
Int. J. Mol. Sci. 2025, 26(2), 814; https://doi.org/10.3390/ijms26020814 - 19 Jan 2025
Viewed by 328
Abstract
Bisphenol A (BPA) is a prevalent environmental contaminant found in plastics and known for its endocrine-disrupting properties, posing risks to both human health and the environment. Despite its widespread presence, the impact of BPA on papillary thyroid cancer (PTC) progression, especially under realistic [...] Read more.
Bisphenol A (BPA) is a prevalent environmental contaminant found in plastics and known for its endocrine-disrupting properties, posing risks to both human health and the environment. Despite its widespread presence, the impact of BPA on papillary thyroid cancer (PTC) progression, especially under realistic environmental conditions, is not well understood. This study examined the effects of BPA on PTC using a 3D thyroid papillary tumor spheroid model, which better mimicked the complex interactions within human tissues compared to traditional 2D models. Our findings demonstrated that BPA, at environmentally relevant concentrations, could induce significant changes in PTC cells, including a decrease in E-cadherin expression, an increase in vimentin expression, and reduced thyroglobulin (TG) secretion. These changes suggest that BPA exposure may promote epithelial–mesenchymal transition (EMT), enhance invasiveness, and reduce cell differentiation, potentially complicating treatment, including by increasing resistance to radioiodine therapy. This research highlights BPA’s hazardous nature as an environmental contaminant and emphasizes the need for advanced in vitro models, like 3D tumor spheroids, to better assess the risks posed by such chemicals. It provides valuable insights into the environmental implications of BPA and its role in thyroid cancer progression, enhancing our understanding of endocrine-disrupting chemicals. Full article
(This article belongs to the Special Issue Design, Synthesis, and Bioapplications of Multifunctional Materials)
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<p>Spheroid formation and cell viability testing of TPC-1 and BCPAP spheroids. (<b>a</b>) Bright-field images of TPC-1 spheroids formed with cell densities of 200 and 1000 cells/well on days 1, 4, and 7. Scale bar: 100 µm. (<b>b</b>) Growth curve of TPC-1; the results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>c</b>) Bright-field images of BCPAP spheroids formed with cell densities of 200 and 1000 cells/well on days 1, 4, and 7. Scale bar: 100 µm. (<b>d</b>) Growth curve of BCPAP; the results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>e</b>) Cell viability was examined by Calcein-AM/PI staining. Fluorescence images show live (green) and dead (red) TPC-1 spheroid cells after 4 and 7 days of culture. Scale bar: 100 µm. (<b>f</b>) Fluorescence images show live (green) and dead (red) BCPAP spheroid cells after 4 and 7 days of culture. Scale bar: 100 µm.</p>
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<p>Cell proliferation of TPC-1 after treatment with BPA. (<b>a</b>) Bright-field images show TPC-1 spheroids treated with various concentrations (0~10<sup>−4</sup> M) of BPA for 24 h and 48 h. Scale bar: 200 µm. (<b>b</b>) Line graph showing the relative growth area (%) of TPC-1 spheroids with a cell density of 200 cells/well after BPA treatment for 24 h and 48 h. The results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>c</b>) Line graph showing the relative growth area (%) of TPC-1 spheroids with the cell density of 1000 cells/well after BPA treatment for 24 h and 48 h. The results were the means from three independent experiments, and the error bars indicate the standard errors.</p>
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<p>Cell proliferation of BCPAP after treatment with BPA. (<b>a</b>) Bright-field images show BCPAP spheroids treated with 0 to 10<sup>−4</sup> M BPA for 24 h and 48 h. Scale bar: 200 µm. (<b>b</b>) Line graph showing the relative growth area (%) of BCPAP spheroids with a cell density of 200 cells/well after BPA treatment for 24 h and 48 h. The results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>c</b>) Line graph showing the relative growth area (%) of BCPAP spheroids with t a cell density of 1000 cells/well after BPA treatment for 24 h and 48 h. The results were the means from three independent experiments, and the error bars indicate the standard errors.</p>
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<p>Cell viability of thyroid cancer cell line after treatment with BPA. (<b>a</b>) Fluorescence images showing live (green) and dead (red) TPC-1 spheroid cells after treatment with 0 to 10<sup>−4</sup> M BPA for 24 h and 48 h. Scale bar: 100 µm. (<b>b</b>) Fluorescence images show live (green) and dead (red) BCPAP spheroid cells after treatment with 0 to 10<sup>−4</sup> M BPA for 24 h and 48 h. Scale bar: 100 µm.</p>
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<p>Immunofluorescence images showing the cytoskeletal arrangement in BCPAP spheroids after 48 h treatment with 10<sup>−4</sup> M BPA. Spheroids were stained with Hoechst (blue) for nuclei and phalloidin (green) for F-actin. Scale bar: 50 µm.</p>
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<p>PTC expression of thyroglobulin (TG). (<b>a</b>) Immunofluorescence images showing TG expression (green) and DAPI-stained nuclei (blue) in TPC-1 and BCPAP cells cultured in monolayer. Scale bar: 250 µm. (<b>b</b>) Immunofluorescence images showing TG expression (green) and DAPI-stained nuclei (blue) in TPC-1 spheroids before and after 48 h treatment with 10<sup>−4</sup> M BPA. Scale bar: 100 µm. (<b>c</b>) The fluorescence intensity of TG per unit area in TPC-1 spheroids, compared before and after treatment with 10<sup>−4</sup> M BPA. The results presented in these histograms were the means from three independent experiments, and the error bars indicate the standard errors (** <span class="html-italic">p</span> &lt; 0.01). (<b>d</b>) Immunofluorescence images showing TG expression (green) and DAPI-stained nuclei (blue) in BCPAP spheroids before and after 48 h treatment with 10<sup>−4</sup> M BPA. Scale bar: 100 µm. (<b>e</b>) The fluorescence intensity of TG per unit area in BCPAP spheroids, compared before and after treatment with 10<sup>−4</sup> M BPA. The results presented in these histograms are the means from three independent experiments, and the error bars indicate the standard errors (** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>PTC expression of E-cadherin. (<b>a</b>) Immunofluorescence images showing E-cadherin expression (red) and DAPI-stained nuclei (blue) in TPC-1 and BCPAP cells cultured in monolayer. Scale bar: 250 µm. (<b>b</b>) Immunofluorescence images showing E-cadherin expression (red) and DAPI-stained nuclei (blue) in TPC-1 spheroids before and after 48 h treatment with 10<sup>−4</sup> M BPA. Scale bar: 100 µm. (<b>c</b>) The fluorescence intensity of E-cadherin per unit area in TPC-1 spheroids, compared before and after treatment with 10<sup>−4</sup> M BPA. The results presented in these histograms are the means from three independent experiments, and the error bars indicate the standard errors (**** <span class="html-italic">p</span> &lt; 0.0001). (<b>d</b>) Immunofluorescence images showing E-cadherin expression (red) and DAPI-stained nuclei (blue) in BCPAP spheroids before and after 48 h treatment with 10<sup>−4</sup> M BPA. Scale bar: 100 µm. (<b>e</b>) The fluorescence intensity of E-cadherin per unit area in BCPAP spheroids, compared before and after treatment with 10<sup>−4</sup> M BPA. The results presented in these histograms were the means from three independent experiments, and the error bars indicate the standard errors (** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>PTC expression of vimentin. (<b>a</b>) Immunofluorescence images showing vimentin expression (red) and DAPI-stained nuclei (blue) in TPC-1 and BCPAP cells cultured in monolayer. Scale bar: 250 µm. (<b>b</b>) Immunofluorescence images showing vimentin expression (red) and DAPI-stained nuclei (blue) in TPC-1 spheroids before and after 48 h treatment with 10<sup>−4</sup> M BPA. Scale bar: 100 µm. (<b>c</b>) The fluorescence intensity of vimentin per unit area in TPC-1 spheroids, compared before and after treatment with 10<sup>−4</sup> M BPA. The results presented in these histograms were the means from three independent experiments, and the error bars indicate the standard errors (* <span class="html-italic">p</span> &lt; 0.05). (<b>d</b>) Immunofluorescence images showing vimentin expression (red) and DAPI-stained nuclei (blue) in BCPAP spheroids before and after 48 h treatment with 10<sup>−4</sup> M BPA. Scale bar: 100 µm. (<b>e</b>) The fluorescence intensity of vimentin per unit area in BCPAP spheroids, compared before and after treatment with 10<sup>−4</sup> M BPA. The results presented in these histograms were the means from three independent experiments, and the error bars indicate the standard errors (**** <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>Concentration of Matrigel required for maintaining TPC-1 3D spheroid structure and invasion assay. (<b>a</b>) Bright-field images of spheroids with cell densities of 200 cells/well, embedded in 3 and 5 mg/mL Matrigel for 7 days. Scale bar: 200 µm. (<b>b</b>) Bright-field images of spheroids with cell densities of 1000 cells/well, embedded in 3 and 5 mg/mL Matrigel for 7 days. Scale bar: 200 µm. (<b>c</b>) Quantitative analysis of the invasion area (%) for cell densities of 200 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>d</b>) Quantitative analysis of the invasion area (%) for cell densities of 1000 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors.</p>
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<p>Concentration of Matrigel required for maintaining BCPAP 3D spheroid structure and invasion assay. (<b>a</b>) Bright-field images of spheroids with cell densities of 200 cells/well, embedded in 3 and 5 mg/mL Matrigel for 7 days. Scale bar: 200 µm. (<b>b</b>) Bright-field images of spheroids with cell densities of 1000 cells/well, embedded in 3 and 5 mg/mL Matrigel for 7 days. Scale bar: 200 µm. (<b>c</b>) Quantitative analysis of the invasion area (%) for cell densities of 200 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>d</b>) Quantitative analysis of the invasion area (%) for cell densities of 1000 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors.</p>
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<p>TPC-1 invasion assay across 0 to 10<sup>−4</sup> M BPA treatment groups. (<b>a</b>) Bright-field images of spheroids with a cell density of 200 cells/well after 7 days. Scale bar: 200 µm. (<b>b</b>) Bright-field images of spheroids with cell densities of 1000 cells/well in 7 days. Scale bar: 200 µm. (<b>c</b>) Quantitative analysis of the invasion area (%) for cell densities of 200 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>d</b>) Quantitative analysis of the invasion area (%) for cell densities of 1000 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors.</p>
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<p>BCPAP invasion assay across 0 to 10<sup>−4</sup> M BPA treatment groups. (<b>a</b>) Bright-field images of spheroids with a cell density of 200 cells/well after 7 days. Scale bar: 200 µm. (<b>b</b>) Bright-field images of spheroids with cell densities of 1000 cells/well in 7 days. Scale bar: 200 µm. (<b>c</b>) Quantitative analysis of the invasion area (%) for cell densities of 200 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors. (<b>d</b>) Quantitative analysis of the invasion area (%) for cell densities of 1000 cells/well relative to day 0. Results were the means from three independent experiments, and the error bars indicate the standard errors.</p>
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<p>This study established a 3D in vitro thyroid cancer model to evaluate the effects of BPA exposure. The results revealed that BPA exposure promotes dedifferentiation of thyroid cancer spheroids, induces EMT, and enhances invasive behavior.</p>
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<p>Conclusive image illustrating the impact of BPA on the malignant progression of thyroid cancer.</p>
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20 pages, 4798 KiB  
Article
Impact of Ex Vivo Bisphenol A Exposure on Gut Microbiota Dysbiosis and Its Association with Childhood Obesity
by Gracia Luque, Pilar Ortiz, Alfonso Torres-Sánchez, Alicia Ruiz-Rodríguez, Ana López-Moreno and Margarita Aguilera
J. Xenobiot. 2025, 15(1), 14; https://doi.org/10.3390/jox15010014 - 17 Jan 2025
Viewed by 526
Abstract
Dietary exposure to the plasticiser bisphenol A (BPA), an obesogenic and endocrine disruptor from plastic and epoxy resin industries, remains prevalent despite regulatory restriction and food safety efforts. BPA can be accumulated in humans and animals, potentially exerting differential health effects based on [...] Read more.
Dietary exposure to the plasticiser bisphenol A (BPA), an obesogenic and endocrine disruptor from plastic and epoxy resin industries, remains prevalent despite regulatory restriction and food safety efforts. BPA can be accumulated in humans and animals, potentially exerting differential health effects based on individual metabolic capacity. This pilot study examines the impact of direct ex vivo BPA exposure on the gut microbiota of obese and normal-weight children, using 16S rRNA amplicon sequencing and anaerobic culturing combined methods. Results showed that direct xenobiotic exposure induced modifications in microbial taxa relative abundance, community structure, and diversity. Specifically, BPA reduced the abundance of bacteria belonging to the phylum Bacteroidota, while taxa from the phylum Actinomycetota were promoted. Consistently, Bacteroides species were classified as sensitive to BPA, whereas bacteria belonging to the class Clostridia were identified as resistant to BPA in our culturomics analysis. Some of the altered bacterial abundance patterns were common for both the BPA-exposed groups and the obese non-exposed group in our pilot study. These findings were also corroborated in a larger cohort of children. Future research will be essential to evaluate these microbial taxa as potential biomarkers for biomonitoring the effect of BPA and its role as an obesogenic substance in children. Full article
(This article belongs to the Special Issue The Role of Endocrine-Disrupting Chemicals in the Human Health)
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Graphical abstract
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<p>Description of children’s gut microbiota before and after ex vivo BPA exposure according to the study groups, results based on 16S rRNA gene amplicon sequencing. (<b>a</b>) Box plots of the alpha diversity indices; (<b>b</b>) beta diversity: nMDS plot based on Bray–Curtis distance, with samples as points and ellipses coloured by study groups. PERMANOVA test results (R<sup>2</sup>, F, <span class="html-italic">p</span>-value) are indicated in the plot. NW, green; OB, blue; NW10, pink; OB10, orange. Mean relative abundance of indicated phyla (<b>c</b>) and ASVs (<b>d</b>).</p>
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<p>Differentially abundant ASVs between exposed and non-exposed groups. To identify taxa that were differentially abundant due to BPA exposure within each BMI group, ANCOM-BC2 analysis was performed: (<b>a</b>) green bars indicate taxa that were significantly more abundant in the NW group, while pink represents taxa that were significantly more abundant in the NW10 group; (<b>b</b>) blue bars indicate taxa that were significantly more abundant in the OB group, while red represents taxa that were significantly more abundant in the OB10 group. * features not sensitive to pseudo-count addition. ASVs with a log fold change between—1.99 and 0 are not shown here to simplify plot size.</p>
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<p>A heatmap displaying the top 50 ASVs with significant changes in relative abundance between the study groups and NW group (used as the reference), detected by MaAsLin2s with default parameters. An increase in relative abundance is shown in red (+) and a decrease in this is shown in blue (−).</p>
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<p>Box plot representing CFU/g from each study group after 1, 15, and 30 days of anaerobic culture in recovery enrichment media. NW, green; OB, blue; NW10, pink; OB10, orange.</p>
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<p>Heatmap of the relative abundance of identified isolates in each group, accompanied by a phylogenetic tree. Stars highlight species identified by sequencing after MALDI TOF failed to identify them. Circles indicate species sensitive to BPA, cultured in control samples (NW, OB) but absent after being exposed to BPA (NW10, OB10). Triangles indicate species showing resistance to BPA, only cultured after exposure to BPA or those whose relative abundance was at least double that of non-exposed samples.</p>
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<p>Comparison of 16S rRNA sequencing and culturing results. (<b>a</b>) Venn diagram depicting overlap between genera identified by 16S rRNA sequencing (blue) and culturing (yellow); (<b>b</b>,<b>c</b>) heatmaps representing the relative abundance of the common genera identified by 16S rRNA sequencing and culturing. “g_<span class="html-italic">Clostridium</span>” represents the relative abundance of “g_<span class="html-italic">Clostridium sensu stricto</span> 1”.</p>
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<p>Box plots showing the relative abundance distribution of significantly different bacterial genera and phyla and their specific ratios between study groups: (<b>a</b>) <span class="html-italic">Bifidobacterium</span>; (<b>b</b>) <span class="html-italic">Bacteroides</span>; (<b>c</b>) <span class="html-italic">Clostridium</span>; (<b>d</b>) <span class="html-italic">Bacteroides</span>/<span class="html-italic">Bifidobacterium</span> ratio; (<b>e</b>) <span class="html-italic">Bacteroides</span>/<span class="html-italic">Clostridium</span> ratio; (<b>f</b>) <span class="html-italic">Actinomycetota</span>; (<b>g</b>) <span class="html-italic">Bacteroidota</span>; (<b>h</b>) <span class="html-italic">Bacillota</span>; (<b>i</b>) <span class="html-italic">Bacteroidota</span>/<span class="html-italic">Actinomycetota</span> ratio; (<b>j</b>) <span class="html-italic">Bacteroidota/Bacillota</span> ratio. NW, green; OB, blue.</p>
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<p>Stool sampling and ex vivo BPA exposure procedure.</p>
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<p>Boxplots showing the relative abundance distribution of significantly different bacterial genera and phyla and their specific ratios between study groups (<b>a</b>) <span class="html-italic">Bifidobacterium</span>; (<b>b</b>) <span class="html-italic">Bacteroides</span>; (<b>c</b>) <span class="html-italic">Clostridium</span>; (<b>d</b>) Ratio <span class="html-italic">Bacteroides</span>/<span class="html-italic">Bifidobacterium</span>; (<b>e</b>) Ratio <span class="html-italic">Bacteroides</span>/<span class="html-italic">Clostridium</span>; (<b>f</b>) <span class="html-italic">Actinomycetota</span>; (<b>g</b>) <span class="html-italic">Bacteroidota</span>; (<b>h</b>) <span class="html-italic">Bacillota</span>; (<b>i</b>) Ratio <span class="html-italic">Bacteroidota</span>/<span class="html-italic">Actinomycetota</span>; (<b>j</b>) Ratio <span class="html-italic">Bacteroidota/Bacillota</span>. NW, green; OB, blue; NW10, pink; OB10, orange.</p>
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11 pages, 5003 KiB  
Article
SERS Detection of Hydrophobic Molecules: Thio-β-Cyclodextrin-Driven Rapid Self-Assembly of Uniform Silver Nanoparticle Monolayers and Analyte Trapping
by Qi Yuan and Yunqing Wang
Biosensors 2025, 15(1), 52; https://doi.org/10.3390/bios15010052 - 15 Jan 2025
Cited by 1 | Viewed by 455
Abstract
High-sensitivity and repeatable detection of hydrophobic molecules through the surface-enhanced Raman scattering (SERS) technique is a tough challenge because of their weak adsorption and non-uniform distribution on SERS substrates. In this research, we present a simple self-assembly protocol for monolayer SERS mediated by [...] Read more.
High-sensitivity and repeatable detection of hydrophobic molecules through the surface-enhanced Raman scattering (SERS) technique is a tough challenge because of their weak adsorption and non-uniform distribution on SERS substrates. In this research, we present a simple self-assembly protocol for monolayer SERS mediated by 6-deoxy-6-thio-β-cyclodextrin (β-CD-SH). This protocol allows for the rapid assembly of a compact silver nanoparticle (Ag NP) monolayer at the oil/water interface within 40 s, while entrapping analyte molecules within hotspots. The proposed method shows general applicability for detecting hydrophobic molecules, exemplified as Nile blue, Nile red, fluconazole, carbendazim, benz[a]anthracene, and bisphenol A. The detection limits range from 10−6to 10−9 M, and the relative standard deviations (RSDs) of signal intensity are less than 10%. Moreover, this method was used to investigate the release behaviors of a hydrophobic pollutant (Nile blue) adsorbed on the nanoplastic surface in the water environment. The results suggest that elevated temperatures, increased salinities, and the coexistence of fulvic acid promote the release of Nile blue. This simple and fast protocol overcomes the difficulties related to hotspot accessibility and detection repeatability for hydrophobic analytes, holding out extensive application prospects in environmental monitoring and chemical analysis. Full article
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<p>Schematic diagram of SERS detection utilizing the synergistic interaction between substrate self-assembly and target analyte capture.</p>
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<p>(<b>a</b>) Images of the self-assembly process of Ag NPs assisted by β-CD-SH, (<b>b</b>) SEM image of the film formed by Ag NPs with β-CD-SH modification, (<b>c</b>) SEM image of the film formed by Ag NPs without β-CD-SH modification, (<b>d</b>) SERS spectra of NB at different concentrations collected on a monolayer film (10<sup>−5</sup> M β-CD-SH), (<b>e</b>) SERS spectra of 10<sup>−7</sup> M NB collected from 10 random points on a monolayer film, (<b>f</b>) Intensity of the 593 cm<sup>−1</sup> peak at 10 random points on the substrate, (<b>g</b>) SERS spectra of NB (10<sup>−7</sup> M) detected on 10 batches of monolayer films prepared under the same conditions, and (<b>h</b>) Statistical distribution of SERS intensity at 593 cm<sup>−1</sup> (NB, 10<sup>−7</sup> M) corresponding to 10 batches of co-assembled films.</p>
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<p>(<b>a</b>) SERS spectra of 10<sup>−7</sup> M NB collected from 10 random points on a monolayer film (without β-CD-SH modification), (<b>b</b>) Intensity of the 593 cm<sup>−1</sup> peak at 10 random points on the substrate (without β-CD-SH modification), (<b>c</b>) SERS spectra of NB (10<sup>−7</sup> M) at 10 random points on a β-CD-SH-Ag film detected by droplet application, (<b>d</b>) Intensity of the 593 cm<sup>−1</sup> peak at 10 random points on a β-CD-SH-Ag film detected by droplet application, (<b>e</b>) SERS spectra of NB (10<sup>−7</sup> M) at 10 random points on a β-CD-SH-Ag film detected by soaking, (<b>f</b>) Intensity of the 593 cm<sup>−1</sup> peak at 10 random points on a β-CD-SH-Ag film detected by soaking, (<b>g</b>) SERS spectra of NB (10<sup>−7</sup> M) at 10 random points on a PVP-Ag film detected by droplet application, and (<b>h</b>) Intensity of the 593 cm<sup>−1</sup> peak at 10 random points on a PVP-Ag film detected by droplet application.</p>
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<p>SERS spectra of (<b>a</b>) Nile red, (<b>b</b>) fluconazole, (<b>c</b>) carbendazim, (<b>d</b>) benz[a]anthracene, and (<b>e</b>) bisphenol A at different concentrations.</p>
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<p>SERS spectra of NB released from PMMA under the influence of different (<b>a</b>) temperatures, (<b>b</b>) salinities, (<b>c</b>) pH values, and (<b>d</b>) concentrations of fulvic acid.</p>
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30 pages, 1492 KiB  
Review
Maternal–Foetal Effects of Exposure to Bisphenol A: Outcomes and Long-Term Consequences
by Henrique Eloi Costa, Ines Medeiros, Melissa Mariana and Elisa Cairrao
Appl. Sci. 2025, 15(2), 697; https://doi.org/10.3390/app15020697 - 12 Jan 2025
Viewed by 732
Abstract
Exposure to bisphenol A (BPA), one of the most widely produced plasticisers, can have a major effect on the growing embryo and the mother during pregnancy; as this is the most vulnerable period, the cutoff established in the legislation does not take this [...] Read more.
Exposure to bisphenol A (BPA), one of the most widely produced plasticisers, can have a major effect on the growing embryo and the mother during pregnancy; as this is the most vulnerable period, the cutoff established in the legislation does not take this factor into account. Thus, this narrative review aims to highlight the consequences for the foetus and the pregnant woman of maternal and foetal exposure to BPA by analysing epidemiological and experimental studies on humans. Extensive research has examined the effects of BPA on several systems outcomes. Specifically, BPA exposure affects the immune system of the offspring and promotes the development of respiratory diseases, including asthma and wheezing. Moreover, BPA has been negatively associated with children’s neurodevelopment, leading to behavioural changes; autism; and reproductive changes, mainly deviations in anogenital distance, sexual hormone levels and sexual maturation, which can result in infertility. Furthermore, in mothers, BPA exposure may be linked to pre-eclampsia and gestational diabetes mellitus and affects birth parameters, leading to a higher risk of preterm delivery, shorter birth lengths and lower birth weights, although the results were not always consistent. These results demonstrate the urgent need for stricter legislation banning the use of BPA during pregnancy to reduce the hazards to the health and development of the foetus and the unborn child. Full article
(This article belongs to the Special Issue Exposure Pathways and Health Implications of Environmental Chemicals)
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<p>Flow diagram of the literature review process.</p>
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<p>Sources, routes of exposure, maternal biological samples of BPA exposome and consequent metabolism. The figure was created on PowerPoint version 2204 using pictures from BioRender.</p>
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<p>Main changes caused by exposure to BPA in pregnant women and in foetal and post-natal development. The figure was created on PowerPoint version 2204 using pictures from BioRender.</p>
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18 pages, 2894 KiB  
Article
Ficus lindsayana Leaf Extract Protects C2C12 Mouse Myoblasts Against the Suppressive Effects of Bisphenol-A on Myogenic Differentiation
by Pornsiri Pitchakarn, Jirarat Karinchai, Pensiri Buacheen, Arisa Imsumran, Ariyaphong Wongnoppavich, Kongsak Boonyapranai and Sakaewan Ounjaijean
Int. J. Mol. Sci. 2025, 26(2), 476; https://doi.org/10.3390/ijms26020476 - 8 Jan 2025
Viewed by 423
Abstract
Recently, toxicological and epidemiological research has provided strong support for the unfavorable effects of bisphenol-A (BPA, 2,2′-bis(4-hydroxyphenyl) propane) on myogenesis and its underlying mechanisms. Researchers have therefore been looking for new strategies to prevent or mitigate these injurious effects of BPA on the [...] Read more.
Recently, toxicological and epidemiological research has provided strong support for the unfavorable effects of bisphenol-A (BPA, 2,2′-bis(4-hydroxyphenyl) propane) on myogenesis and its underlying mechanisms. Researchers have therefore been looking for new strategies to prevent or mitigate these injurious effects of BPA on the human body. It has been found that plant extracts may act as potential therapeutic agents or functional foods, preventing human diseases caused by BPA. We previously reported that Ficus lindsayana (FL) extract exhibits anti-inflammation activity in macrophages via suppressing the expression of inflammation-related molecules and anti-insulin resistance in inflammation-treated adipocytes. In this study, we investigated whether Ficus lindsayana leaf extract (FLLE) protects C2C12 mouse myoblasts against the suppressive effects of BPA on myogenic differentiation. The viability of BPA-stimulated C2C12 myoblasts was significantly increased when co-treated with FLLE (200 µg/mL), suggesting that the extract may lessen the inhibitory effects of BPA on cell division. We also found that FLLE significantly increased neo-myotube formation by inducing the fusion of myoblasts into multinucleated myotubes when compared to the BPA-treated control cells, without impacting cell viability. In addition, the levels of myogenin and myocyte enhancer factor 2A (MEF2A), which are crucial markers and regulators of myogenesis, were markedly increased by the addition of FLLE (50 µg/mL) to the BPA-treated C2C12 cells. This finding suggests that FLLE effectively improved myogenic differentiation in BPA-exposed myoblasts. FLLE treatment (50 µg/mL) significantly raised total Akt protein levels in the BPA-treated C2C12 cells, enhancing protein phosphorylation. In addition, FLLE (50 µg/mL) obviously increased the phosphorylation levels of p70S6K and 4E-BP1, key downstream targets of the Akt/mTOR signaling cascade, by elevating total p70S6K and 4E-BP1 levels. These results suggest that FLLE diminishes the decline in myogenic differentiation induced by BPA via the regulation of the myocyte differentiation-related signaling pathway. The information obtained from this study demonstrates the health benefits of this plant, which warrants further investigation as an alternative medicine, functional ingredient, or food supplement that can prevent the negative health effects of BPA or other toxicants. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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<p>HPLC chromatogram of the standard mixture containing chlorogenic acid, catechin, vanillic acid, and rutin (<b>A</b>). Phytochemical profile of FLLE analyzed by HPLC (<b>B</b>).</p>
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<p>Effect of FLLE on the viability of C2C12 myoblasts. The cells were treated with various concentrations of the extracts (0–800 µg/mL) for 48 h. Cell viability was determined via MTT assay. Each value represents mean ± SD (n = 3) ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001 vs. non-treated control.</p>
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<p>Effect of BPA and FLLE co-treatment on the viability of C2C12 myoblasts under non-differentiated (<b>A</b>) and differentiated conditions (<b>B</b>). (<b>A</b>) The nondifferentiated C2C12 myoblasts were treated with various concentrations of the extracts (0–200 µg/mL) in the presence or absence of 50 µM BPA for 72 h. (<b>B</b>) The cells were treated with the extracts (0–200 µg/mL) in the presence or absence of 50 µM BPA during differentiation for 6 days. At the indicated time, the cell viability was determined via MTT assay. Each value represents mean ± SD (n = 3) * <span class="html-italic">p</span> &lt; 0.05 vs. BPA-treated control, <sup>###</sup> <span class="html-italic">p</span> &lt; 0.001 vs. non-treated control.</p>
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<p>Effects of FLLE on the myogenesis of BPA-treated C2C12 myoblasts. The cells were treated with the extracts (0–50 µg/mL) in the presence of 50 µM BPA during differentiation for 6 days. (<b>A</b>) Morphological changes in C2C12 cells under microscopic observation on the sixth day of differentiation. (<b>B</b>) The effects of FLLE on myogenic differentiation, determined by measuring the fraction of nuclei incorporated into myotubes on the sixth day of myogenic differentiation. Each value represents mean ± SD (n = 3) ** <span class="html-italic">p</span> &lt; 0.01 vs. BPA-treated control, <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 vs. non-treated control.</p>
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<p>Effect of FLLE on the protein level of MEF2A and myogenin myogenesis markers in BPA-treated C2C12 myoblasts. The cells were treated with the extracts (0–50 µg/mL) in the absence or presence of 50 µM BPA during differentiation for 6 days. After the treatment, the protein samples were collected and myogenin and MEF2A levels were determined via Western blotting (normalized with β-actin level). (<b>A</b>) The expression of MEF2A and myogenin in non-differentiated myoblasts compared to non-treated control myocytes. (<b>B</b>) A representative result of three independent experiments. (<b>C</b>,<b>D</b>) Relative band density of MEF2A (<b>C</b>) and myogenin (<b>D</b>) normalized with β-actin level. Each value in represents mean ± SD (n = 3) ** <span class="html-italic">p</span> &lt; 0.01 vs. BPA-treated control, <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 vs. non-treated control.</p>
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<p>Effects of FLLE on the Akt/p70S6K/4EBP1 pathway in C2C12 myotubes. After the treatment, the protein samples were collected and used to determine Akt/p70S6K/4EBP1 phosphorylation levels via Western blotting. The relative phosphorylation levels of Akt, p70S6K, and 4EBP1 were calculated by dividing the amount of protein detected by the phosphorylated antibody by the amount detected by the antibody to determine total protein levels. (<b>A</b>) The level of the phospho- and total forms of Akt, p70S6K, and 4EBP1 in non-differentiated myoblasts compared to non-treated control myocytes. (<b>B</b>) A representative Western blotting result of three independent experiments. (<b>C</b>,<b>E</b>,<b>G</b>) Relative phosphorylation levels of Akt (<b>C</b>), p70S6K (<b>E</b>), and 4EBP1 (<b>G</b>). (<b>D</b>,<b>F</b>,<b>H</b>) Band density of phospho- and total forms of Akt (<b>D</b>), p70S6K (<b>F</b>), and 4EBP1 (<b>H</b>), normalized with β-actin levels. Each value in (<b>C</b>–<b>H</b>) represents mean ± SD (n = 3) * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 vs. BPA-treated control, <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>##</sup> <span class="html-italic">p</span> &lt; 0.01 vs. non-treated control.</p>
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16 pages, 2193 KiB  
Article
Comprehensive Analysis of the Proteome of S. cerevisiae Wild-Type and pdr5Δ Cells in Response to Bisphenol A (BPA) Exposure
by Valentina Rossio and Joao A. Paulo
Microorganisms 2025, 13(1), 114; https://doi.org/10.3390/microorganisms13010114 - 8 Jan 2025
Viewed by 490
Abstract
Bisphenol A, an endocrine-disrupting compound, is widely used in the industrial production of plastic products. Despite increasing concerns about its harmful effects on human health, animals, and the environment, the use of BPA has been banned only in infant products, and its effects [...] Read more.
Bisphenol A, an endocrine-disrupting compound, is widely used in the industrial production of plastic products. Despite increasing concerns about its harmful effects on human health, animals, and the environment, the use of BPA has been banned only in infant products, and its effects on cellular processes are not fully understood. To investigate the impact of BPA on eukaryotic cells, we analyzed the proteome changes of wild-type and PDR5-deleted S. cerevisiae strains exposed to different doses of BPA using sample multiplexing-based proteomics. We found that the ABC multidrug transporter Pdr5 plays an important role in protecting yeast cells from BPA toxicity, with its absence significantly sensitizing cells to BPA. BPA inhibited yeast growth in a dose-dependent manner, with a more pronounced effect in PDR5-deleted cells. Proteomic analysis revealed that BPA induces widespread dose-dependent changes in protein abundance, including the upregulation of metabolic pathways such as arginine biosynthesis and the downregulation of mitochondrial proteins. Additionally, we observed markers of cellular stress induced by BPA by identifying multiple stress-induced proteins that were upregulated by this compound. As cellular processes affected by BPA have been shown to be evolutionarily conserved, these insights can advance our understanding of BPA’s cellular impact and its broader effects on human health. Full article
(This article belongs to the Section Molecular Microbiology and Immunology)
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<p>Experimental workflow, dataset summary, and the effect of bisphenol A (BPA) on cellular growth after six-hour treatment. (<b>A</b>) Wild-type and <span class="html-italic">pdr5</span>∆ <span class="html-italic">S. cerevisiae</span> cells were grown in duplicate to exponential phase (24 °C) and treated with the indicated BPA concentrations or ethanol (EtOH) as a control for six hours. (<b>B</b>) Cells were harvested and processed for mass spectrometry analysis. In brief, yeast cells were lysed, and total protein was extracted and digested. The subsequent peptides were labeled with tandem mass tag (TMTpro) reagents, as indicated, pooled 1:1, and fractionated by basic pH reversed-phase (BPRP) HPLC prior to mass spectrometry analysis. This panel was assembled, in part, using Biorender.com. (<b>C</b>) Percentage of cells at 6 h treated with the indicated BPA concentration compared to EtOH-treated cells. (<b>D</b>) Dataset summary.</p>
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<p>Principal component analysis (PCA), hierarchical clustering analysis (HCA), and differentially abundant proteins (DAPs) in wild-type and <span class="html-italic">pdr5</span>Δ cells treated with BPA. (<b>A</b>) PCA of the dataset illustrates the clustering of the replicates. (<b>B</b>) HCA of the TMT relative abundance (TMT RA) for the 4687 proteins quantified across the 16 TMT channels. Duplicates of each condition are indicated as A and B. (<b>C</b>) The table summarizes the differentially abundant proteins (DAPs) in the two yeast strains, wt and <span class="html-italic">pdr5</span>Δ, treated with the indicated BPA concentrations compared to the control (EtOH-treated) strains.</p>
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<p>Proteome-wide profiling of differentially abundant proteins in wild-type cells after treatment with 300 mg/mL BPA. (<b>A</b>) The volcano plot illustrates differentially abundant proteins (i.e., |log<sub>2</sub> ratio| &gt; 0.5, and <span class="html-italic">p</span>-value &lt; 0.05) in wild-type cells after treatment with 300 mg/mL BPA. Proteins highlighted in (<b>C</b>,<b>E</b>) are labeled. (<b>B</b>) The top gene ontology (GO) biological processes (BP) terms associated with the proteins that are increasing in (<b>A</b>). (<b>C</b>) Bar graphs illustrate the TMT relative abundance (RA) of the classes of proteins in (<b>B</b>). (<b>D</b>) The top GO cellular component (CC) terms associated with proteins with decreased abundance after BPA treatment. (<b>E</b>) TMT relative abundance measurements of mitochondrial proteins with decreased abundance in (<b>A</b>).</p>
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<p>Proteins changing with lower doses of BPA in wild-type cells and proteome-wide profiling of differentially abundant proteins in <span class="html-italic">pdr5</span>∆ cells treated with 150 mg/mL BPA. (<b>A</b>) The table summarizes the differentially abundant proteins in wild-type cells treated with 50 and 150 mg/mL of BPA compared to the EtOH condition. (<b>B</b>) The volcano plot illustrates differentially abundant proteins (i.e., |log<sub>2</sub> ratio| &gt; 0.5, and <span class="html-italic">p</span>-value &lt; 0.05) in <span class="html-italic">pdr5</span>Δ cells after treatment with 150 mg/mL BPA compared to <span class="html-italic">pdr5</span>Δ cells treated with EtOH. (<b>C</b>) The top gene ontology (GO) biological processes (BP) terms associated with the proteins increasing after BPA treatment in <span class="html-italic">pdr5</span>Δ cells. Bar graphs illustrate the TMT relative abundance (RA) of proteins increasing in <span class="html-italic">pdr5</span>Δ cells involved in (<b>D</b>) methionine metabolism and of (<b>E</b>) the protein Ykl071 (Osi1, oxidative stress-induced protein 1).</p>
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15 pages, 3568 KiB  
Article
Bisphenol S Induces Lipid Metabolism Disorders in HepG2 and SK-Hep-1 Cells via Oxidative Stress
by Kai-Xing Lin, Zi-Yao Wu, Mei-Lin Qin and Huai-Cai Zeng
Toxics 2025, 13(1), 44; https://doi.org/10.3390/toxics13010044 - 8 Jan 2025
Viewed by 529
Abstract
Bisphenol S (BPS) is a typical endocrine disruptor associated with obesity. To observe BPS effects on lipid metabolism in HepG2 and SK-Hep-1 human HCC cells, a CCK-8 assay was used to assess cell proliferation in response to BPS, and the optimal concentration of [...] Read more.
Bisphenol S (BPS) is a typical endocrine disruptor associated with obesity. To observe BPS effects on lipid metabolism in HepG2 and SK-Hep-1 human HCC cells, a CCK-8 assay was used to assess cell proliferation in response to BPS, and the optimal concentration of BPS was selected. Biochemical indices such as triglyceride (TG) and total cholesterol (T-CHO), and oxidative stress indices such as malondialdehyde (MDA) and catalase (CAT) were measured. ROS and MDA levels were significantly increased after BPS treatment for 24 h and 48 h (p < 0.05), indicating an oxidative stress response. Alanine aminotransferase (ALT), T-CHO, and low-density lipoprotein cholesterol (LDL-C) levels also increased significantly after 24 or 48 h BPS treatments (p < 0.05). RT-PCR and Western blot analyses detected mRNA or protein expression levels of peroxisome proliferator-activated receptor α (PPARα) and sterol regulatory element-binding protein 1c (SREBP1C). The results indicated that BPS could inhibit the mRNA expression of PPARα and carnitine palmitoyl transferase 1B (CPT1B), reduce lipid metabolism, promote mRNA or protein expression of SREBP1C and fatty acid synthase (FASN), and increase lipid synthesis. Increased lipid droplets were observed using morphological Oil Red O staining. Our study demonstrates that BPS may cause lipid accumulation by increasing oxidative stress and perturbing cellular lipid metabolism. Full article
(This article belongs to the Special Issue Drug Metabolism and Toxicological Mechanisms)
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<p>Effect of different concentrations of BPS on the viability of HepG2 and SK-Hep-1 cells. Note: (<b>A</b>,<b>B</b>) presents the alterations in cell viability following treatment with varying concentrations of BPS. “*” represent cell viability following a 24 h exposure to BPS relative to the 0 μmol/L group, <span class="html-italic">p</span> &lt; 0.05. “#” represents cell viability following a 48 h exposure to BPS relative to the 0 μmol/L group, <span class="html-italic">p</span> &lt; 0.05. n = 3, the same below.</p>
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<p>Effect of BPS treatment on ROS levels in HepG2 and SK-Hep-1 cells. Note: (<b>A</b>,<b>B</b>) shows the results of reactive oxygen species detection in HepG2 and SK-Hep-1 cells exposed to BPS for 24 h or 48 h conditions with a microscopic scale of 100 μm. (<b>C</b>) indicates reactive oxygen species fluorescence intensity quantification, and “#” represents the oxidative stress of cells after exposure compared to that of the control group, <span class="html-italic">p</span> &lt; 0.05. n = 3.</p>
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<p>BPS-induced lipid droplet deposition in HepG2 cells. Note: (<b>A</b>–<b>D</b>) shows Oil Red O staining in HepG2 cells following BPS exposure for 24 h or 48 h with a microscopic scale of 50 μm. (<b>E</b>–<b>H</b>) presents the proportionally enlarged “□” window in (<b>A</b>–<b>D</b>), while “↑” in (<b>F</b>,<b>H</b>) refers to red fat droplets.</p>
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<p>BPS-induced lipid droplet deposition in SK-Hep-1 cells. Note: (<b>A</b>–<b>D</b>) presents Oil Red O staining of SK-Hep-1 cells after exposure to BPS for 24 h or 48 h with a microscopic scale of 50 μm. (<b>E</b>–<b>H</b>) presents the proportionally enlarged “□” window in (<b>A</b>–<b>D</b>), while “↑” in (<b>F</b>,<b>H</b>) refers to the red fat droplets.</p>
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<p>Effect of BPS on oxidative stress levels in HepG2 cells and SK-Hep-1 cells. Note: (<b>A</b>) shows the MDA levels within SK-Hep-1 and HepG2 cells following BPS treatments for 24 h and 48 h. (<b>B</b>) shows the CAT levels within SK-Hep-1 and HepG2 cells following BPS treatments for 24 h and 48 h; “#” represents BPS compared to the control group, <span class="html-italic">p</span> &lt; 0.05. n = 3.</p>
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<p>Effects of BPS on HepG2 and SK-Hep-1 cell damage and metabolic-related indicators. Note: (<b>A</b>–<b>D</b>) presents the results of TG, T-CHO, ALT, and LDL-C analyses after BPS exposure in SK-Hep-1 and HepG2 cells for 24 h and 48 h, respectively. “#” represents BPS compared to the control group, <span class="html-italic">p</span> &lt; 0.05. n = 3.</p>
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<p>Effect of BPS on the expression levels of related mRNAs in HepG2 and SK-Hep-1 cells. Note: (<b>A</b>–<b>D</b>) indicates the mRNA expression results of PPARα, CPT1B, CD36, SREBP1C, and FAFSN in HepG2 and SK-Hep-1 cells following 24 h and 48 h of exposure, respectively. “#” represents BPS compared to the control group, <span class="html-italic">p</span> &lt; 0.05. n = 3.</p>
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<p>Effects of BPS on the expression levels of the lipid synthesis proteins SREBP1C and FASN in HepG2 and SK-Hep-1 cells. Note: (<b>A</b>–<b>H</b>) indicates the relative protein expression of SREBP1C and FASN in SK-Hep-1 and HepG2 cells after 24 h and 48 h of exposure, respectively. “#” represents BPS compared to the control group, <span class="html-italic">p</span> &lt; 0.05. n = 3.</p>
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17 pages, 7619 KiB  
Article
The Impact of an MDP-Containing Primer on the Properties of Zinc Oxide Networks Infiltrated with BisGMA-TEGDMA and UDMA-TEGDMA Polymers
by Benjamin Wellhäußer, Lena Marie Saure, Fabian Schütt, Franziska Scherer, Sebastian Wille and Matthias Kern
Materials 2025, 18(1), 137; https://doi.org/10.3390/ma18010137 - 31 Dec 2024
Viewed by 486
Abstract
This study was conducted to evaluate the material properties of polymer-infiltrated zinc oxide networks (PICN) and the effect of using a phosphate monomer-containing primer applied before polymer infiltration. A total of 148 ZnO-network (zinc oxide) specimens were produced: n = 74 were treated [...] Read more.
This study was conducted to evaluate the material properties of polymer-infiltrated zinc oxide networks (PICN) and the effect of using a phosphate monomer-containing primer applied before polymer infiltration. A total of 148 ZnO-network (zinc oxide) specimens were produced: n = 74 were treated with a primer before polymer infiltration and light curing, while the remaining specimens were untreated. Each group was divided into two subgroups (n = 37) based on the infiltrating polymer: UDMA (aliphatic urethane-dimethacrylates)-TEGDMA (triethylene glycol-dimethacrylate) or BisGMA (bisphenol A-glycidyl-methacrylate)-TEGDMA. Additionally, n = 7 specimens of each polymer type were prepared for comparison. Then, biaxial flexural strength was measured before and after 150 days of water storage at 37 °C, including 37,500 thermal cycles (5 °C to 55 °C). The Vickers hardness, surface roughness, and water absorption at 37 °C were also tested. The initial biaxial flexural strength was reduced in the ZnO network specimens compared to in the pure polymers. Primer application improved the flexural strength, though the strength of BisGMA-TEGDMA significantly decreased after water storage. The ZnO network increased hardness, and the polymer-infiltrated networks showed higher roughness post-grinding and absorbed less water than the pure polymer groups. The ZnO networks did not improve the flexural strength over that of the pure polymers. However, the primer’s positive impact and the network’s long-term stability suggest potential if the network structure can be modified to contain thicker, more stable branches. Full article
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<p>SEM image of the ZnO powder before sintering to form a network.</p>
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<p>Exemplary cross-section SEM image of the ZnO network before polymer infiltration.</p>
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<p>Exemplary cross-section SEM image of the PICN network after polymer infiltration.</p>
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<p>PICN specimen placed on three steel balls; the piston applies pressure to the specimen to determine the flexural strength (2-column fitting image).</p>
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<p>Different hydrolytic effects on BisGMA-TEGDMA specimens after 150 days of water storage with thermocycling.</p>
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<p>Topography images of different surface roughnesses obtained from confocal laser scanning microscopy of the BisGMA-TEGDMA specimens before and after polishing, marked with lowercase letters: (<b>a</b>) BT before; (<b>b</b>) BT after; (<b>c</b>) Z-BT before; (<b>d</b>) Z-BT after; (<b>e</b>) Z-BT-P before; (<b>f</b>) Z-BT-P after.</p>
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<p>Topography images of different surface roughnesses obtained from confocal laser scanning microscopy of the UDMA-TEGDMA specimens before and after polishing, marked with lowercase letters: (<b>a</b>) UT before; (<b>b</b>) UT after; (<b>c</b>) Z-UT before; (<b>d</b>) Z-UT after; (<b>e</b>) Z-UT-P before; (<b>f</b>) Z-UT-P after.</p>
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<p>Relative water absorption in all test groups from 0 to 456 h (19 days).</p>
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13 pages, 2097 KiB  
Article
Taxonomic and Functional Dynamics of Bacterial Communities During Drift Seaweed Vermicomposting
by Manuel Aira, Ana Gómez-Roel and Jorge Domínguez
Microorganisms 2025, 13(1), 30; https://doi.org/10.3390/microorganisms13010030 - 27 Dec 2024
Viewed by 498
Abstract
Seaweed is a valuable natural resource, but drift or beach-cast seaweed is considered a waste product. Although seaweed is traditionally used as an organic amendment, vermicomposting has the potential to transform the material into valuable organic fertilizer, thereby enhancing its microbial properties. This [...] Read more.
Seaweed is a valuable natural resource, but drift or beach-cast seaweed is considered a waste product. Although seaweed is traditionally used as an organic amendment, vermicomposting has the potential to transform the material into valuable organic fertilizer, thereby enhancing its microbial properties. This study aimed to investigate the dynamics of the taxonomic and functional bacterial communities in seaweed during the vermicomposting process by high-throughput sequencing of 16S rRNA gene amplicons. Vermicomposting changed the composition of the bacterial communities, as indicated by the low proportion of bacterial taxa common to the bacterial communities in the raw seaweed and vermicompost (21 to 56 ASVs from more than 900 ASVs per sample type). The observed increase in taxonomic diversity (32% mean increase across sampling times) also affected the functionality of the bacterial communities present in the vermicompost. The diverse bacterial community showed enriched functional pathways related to soil health and plant growth, including the synthesis of antibiotics, amino acids, and phytohormones, as well as the degradation of bisphenol. In conclusion, in terms of microbial load and diversity, vermicompost derived from seaweed is a more valuable organic fertiliser than seaweed itself. Full article
(This article belongs to the Section Environmental Microbiology)
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<p>Relative abundance of bacterial communities at phylum and genus level during seaweed vermicomposting. The least abundant bacterial taxa (relative abundance &lt; 1 and 2% for bacterial phyla and genera, respectively) were collapsed into ‘others’.</p>
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<p>UpSet plot showing the unique and shared bacterial ASVs during seaweed vermicomposting. The mean relative abundances of unique and shared ASVs at the phylum level are shown. Black dots represent sampling times and lines indicate the intersections between sampling times.</p>
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<p>Changes in bacterial alpha diversity during seaweed vermicomposting, represented as (<b>A</b>) taxonomic amplicon sequence variant (ASV) and (<b>B</b>) phylogenetic diversity (Faith index). Different letters indicate significant differences between time points (paired <span class="html-italic">t</span>-test, FDR corrected). Changes in bacterial taxonomic and phylogenetic beta diversity during seaweed vermicomposting were determined by principal coordinate analysis of Bray-Curtis (<b>C</b>) and weighted UniFrac (<b>D</b>) distances, respectively.</p>
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<p>Heatmap of predicted bacterial community pathways and corresponding changes in contribution (i.e., bacterial ASVs with these functions), alpha diversity (inverse Simpson index), and beta diversity (Bray-Curtis distance) during seaweed vermicomposting. KEGG orthologue (KO) abundance data were z-score transformed. Different letters indicate significant differences between time points (paired <span class="html-italic">t</span>-test, FDR corrected).</p>
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18 pages, 8968 KiB  
Article
Integrated Metagenomic and Metabolomic Analysis of In Vitro Murine Gut Microbial Cultures upon Bisphenol S Exposure
by Amon Cox, Farrhin Nowshad, Evelyn Callaway and Arul Jayaraman
Metabolites 2024, 14(12), 713; https://doi.org/10.3390/metabo14120713 - 18 Dec 2024
Viewed by 531
Abstract
Background: The gut microbiota are an important interface between the host and the environment, mediating the host’s interactions with nutritive and non-nutritive substances. Dietary contaminants like Bisphenol A (BPA) may disrupt the microbial community, leaving the host susceptible to additional exposures and pathogens. [...] Read more.
Background: The gut microbiota are an important interface between the host and the environment, mediating the host’s interactions with nutritive and non-nutritive substances. Dietary contaminants like Bisphenol A (BPA) may disrupt the microbial community, leaving the host susceptible to additional exposures and pathogens. BPA has long been a controversial and well-studied contaminant, so its structural analogues like Bisphenol S (BPS) are replacing it in consumer products, but have not been well studied. Methods: This study aimed to determine the impact of BPS on C57BL/6 murine gut microbiota using shotgun metagenomic sequencing and the metabolomic profiling of in vitro anaerobic cultures. Results: The results demonstrated that a supraphysiologic BPS dose did not overtly distort the metagenomic or metabolomic profiles of exposed cultures compared to controls. A distinct BPS-associated metabolite profile was not observed, but several metabolites, including saturated fatty acids, were enriched in the BPS-exposed cultures. In the absence of a BPS-associated enterotype, Lactobacillus species specifically were associated with BPS exposure in a discriminant model. Conclusions: Our study provides evidence contrasting the effects of BPS in the gut microbiome to its predecessor, BPA, but also emphasizes the role of inter-animal variation in microbiome composition, indicating that further study is needed to characterize BPS in this context. Full article
(This article belongs to the Special Issue Effects of Environmental Exposure on Host and Microbial Metabolism)
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<p>Overview of multi-omics analyses. PCA score plots of metabolomic (<b>A</b>) and metagenomic (<b>C</b>) profiles with results from MANOVA on BPS-treated cultures vs. controls (n = 5; excluding inoculum). PCA scores by first six components over time for metabolomic (<b>B</b>) and metagenomic (<b>D</b>) profiles.</p>
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<p>Diversity measures of metagenomic profiles at species level. (<b>A</b>) Violin plot of alpha diversity indices (Shannon and Inverse Simpson) for 0 h, 24 h, and 48 h profiles by group. (<b>B</b>) Beta diversity of metagenomic profiles as a Principal Coordinates Analysis scores plot, with PERMANOVA on 24 h and 48 h profiles (n = 5). (<b>C</b>) Relative abundance of species means per group.</p>
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<p>Discriminant analysis of microbial species. (<b>A</b>) sPLS-DA scores for species relative abundance by time point. (<b>B</b>) sPLS-DA loadings for bacterial species by time point. Loadings signs for 24 h were flipped to align with 48 h. (<b>C</b>) Heatmap displaying relative abundances by group per biological replicate at the species level, including species with mean relative abundances &lt; 1%.</p>
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<p>Discriminant analysis of microbial metabolic pathways. (<b>A</b>) sPLS-DA scores for metabolic pathway relative abundance by time point. sPLS-DA loadings for differential metabolic pathways retained in the 24 h hour model (<b>B</b>) and 48 h model (<b>C</b>).</p>
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<p>Differentially altered metabolomic features. Log2 fold changes (BPS-treated/control) against negative log 10 transformed adjusted <span class="html-italic">p</span>-values from empirical Bayes moderated <span class="html-italic">t</span>-test (n = 5), for 24 h (<b>A</b>) and 48 h (<b>B</b>) profiles. (<b>C</b>) Scores plot of sPLS-DA, considering only the 24 h and 48 h profiles and respective top 50 metabolomic features contributing the greatest variance. (<b>D</b>) Heatmap of metabolomic feature z-scores with hierarchical clustering. Heatmap row names represent culture replicates; “A-E” denote mouse, “s” or “v” denote BPS-exposed and control, respectively; 0–2 denote culture time points 0 h, 24 h, and 48 h.</p>
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<p>Canonical correlations between differential ‘omics features. Sparse Canonical Correlations Analysis (sCCA) scores plots of differential annotated metabolites with color scaled by species component 1 (<b>A</b>) and of sPLS-DA-selected species colored by metabolites component 1 (<b>B</b>). sCCA loadings for metabolites (<b>C</b>) and species (<b>D</b>); metabolomic features with non-zero loadings are denoted by their annotation, whereas features with loadings equal to zero are only listed by their analysis code.</p>
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