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13 pages, 1346 KiB  
Review
Beta-Thalassemia and Male Infertility: Unraveling the Oxidative Stress Connection—An Up-to-Date Review
by Christos Roidos, Christos-Alexandros Batakoias, Evangelos N. Symeonidis, Aris Kaltsas, Vasileios Tzikoulis, Georgios Tsampoukas, Chara Tsiampali, Natalia Palapela, Athanasios Zachariou, Nikolaos Sofikitis and Fotios Dimitriadis
Diagnostics 2024, 14(24), 2789; https://doi.org/10.3390/diagnostics14242789 (registering DOI) - 12 Dec 2024
Abstract
Background/Objectives: Beta-thalassemia (BTH), a genetic disorder resulting from beta-globin gene mutations, affects over 1.5 million people globally. The disorder’s multifactorial impact on male fertility, particularly through oxidative stress (OS), warrants focused study. This review examines the mechanisms of OS in TM, its implications [...] Read more.
Background/Objectives: Beta-thalassemia (BTH), a genetic disorder resulting from beta-globin gene mutations, affects over 1.5 million people globally. The disorder’s multifactorial impact on male fertility, particularly through oxidative stress (OS), warrants focused study. This review examines the mechanisms of OS in TM, its implications for male infertility, and the potential of antioxidant therapies to mitigate fertility challenges. Methods: A non-systematic review was conducted using the PubMed, Cochrane, and Medscape databases, focusing on studies on beta-thalassemia (BTH), erectile dysfunction (ED), hormonal alterations, and OS. Studies were screened based on relevance, language, and topic, with 71 articles meeting the inclusion criteria after removing duplicates. Results: The findings reveal that OS, exacerbated by iron overload from regular blood transfusions, is significantly associated with impaired sperm quality and fertility in patients with TM. Iron toxicity affects gonadotropin levels, reduces sperm quality, and contributes to hypogonadism. Additionally, antioxidant therapies show promise in reducing OS-induced sperm damage, though efficacy is limited by a lack of robust clinical trials. Conclusions: OS plays a considerable role in male infertility among patients with TM, primarily through iron-induced sperm damage and hormonal disruptions. While antioxidant therapies may offer a partial remedy, further research is necessary to understand OS’s mechanisms in TM and develop effective fertility treatments. This review highlights the need for personalized antioxidant approaches to improve reproductive outcomes in this population. Full article
(This article belongs to the Special Issue Advances in the Diagnosis and Management of Male Infertility)
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<p>A flow diagram of the studies included in the narrative review.</p>
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<p>Oxidative stress and antioxidant therapy in beta-thalassemia-related infertility.</p>
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16 pages, 3568 KiB  
Article
Antibiofilm Activity of Epinecidin-1 and Its Variants Against Drug-Resistant Candida krusei and Candida tropicalis Isolates from Vaginal Candidiasis Patients
by Sivakumar Jeyarajan, Sukumar Ranjith, Raja Veerapandian, Kalimuthusamy Natarajaseenivasan, Prahalathan Chidambaram and Anbarasu Kumarasamy
Infect. Dis. Rep. 2024, 16(6), 1214-1229; https://doi.org/10.3390/idr16060096 (registering DOI) - 12 Dec 2024
Abstract
Background/Objective: Indwelling intrauterine contraceptive devices (IUDs) have surfaces that facilitate the attachment of Candida spp., creating a suitable environment for biofilm formation. Due to this, vulvovaginal candidiasis (VVC) is frequently linked to IUD usage, necessitating the prompt removal of these devices for effective [...] Read more.
Background/Objective: Indwelling intrauterine contraceptive devices (IUDs) have surfaces that facilitate the attachment of Candida spp., creating a suitable environment for biofilm formation. Due to this, vulvovaginal candidiasis (VVC) is frequently linked to IUD usage, necessitating the prompt removal of these devices for effective treatment. In this study, we evaluated the susceptibility of antimicrobial peptides in vitro against biofilm forming, Amphotericin B (MIC50 > 2 mg L−1) resistant Candida krusei and Candida tropicalis isolated from IUD users who had signs of vaginal candidiasis (hemorrhage, pelvic pain, inflammation, itching, and vaginal discharge). Three antimicrobial peptides, namely, epinecidin-1 (epi-1) and its two variants, namely, variant-1 (Var-1) and variant-2 (Var-2), which were reported to have enhanced antibacterial activity were tested against IUD isolates (C. krusei and C. tropicalis) with pathogenic form of Candida albicans as control. Variants of epi-1, namely, Var-1 and Var-2 were created by substituting lysine in the place of histidine and alanine. Methods: The antimicrobial activity was measured using the microbroth dilution method to determine the minimum inhibitory concentration (MIC) of peptides against C. albicans, C. krusei and C. tropicalis. The MIC of each peptide was used for biofilm assay by Crystal violet staining, Scanning Electron Microscopy, and Reactive Oxygen Species (ROS) assay. To find the possible mechanism of anti-biofilm activity by the peptides, their ability to interact with Candida spp. cell membrane proteins such as Exo-β-(1,3)-Glucanase, Secreted Aspartic Proteinase (Sap) 1, and N-terminal Domain Adhesin: Als 9-2 were determined through PatchDock. Results: The MIC values of peptides: epi-1, var-1 and var-2 against C. albicans are 128 μg mL−1, 64 μg mL−1 and 32 μg mL−1, C. tropicalis are 256 μg mL−1, 64 μg mL−1, and 32 μg mL−1 and C. krusei are 128 µg mL−1, 128 µg mL−1 and 64 µg mL−1, respectively. Both the variants outperformed epi-1. Specifically for tested Candida spp., var-1 showed two- to four-fold enhancements and var-2 showed two- to eight-fold enhancements compared to epi-1. Electron microscopy confirmed that the mechanism of action involves pore formation thus inducing reactive oxygen species in Candida spp. cell membrane. Computational analysis showed that the peptides have a high tendency to interact with Candida spp. cell membrane proteins such as Exo-β-(1,3)-Glucanase, Secreted Aspartic Proteinase (Sap) 1, and N-terminal Domain Adhesin: Als 9-2, thereby preventing biofilm formation. Conclusions: The in vitro evidence supports the potential use of epi-1 and its variants to be used as an anti-biofilm agent to coat IUDs in the future for therapeutic purposes. Full article
(This article belongs to the Special Issue New Advances in Drugs/Vaccines against Infectious Diseases)
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<p>Susceptibility of the peptides against (<b>a</b>) <span class="html-italic">C. albicans</span> (MTCC 227), (<b>b</b>) <span class="html-italic">C. tropicalis</span> (CA4) and (<b>c</b>) <span class="html-italic">C. krusei</span> (CA54). Epi-1, var-1, and var-2 inhibited growth of <span class="html-italic">C. albicans</span> (MTCC 227) at 128 μg mL<sup>−1</sup>, 64 μg mL<sup>−1</sup> and 32 μg mL<sup>−1</sup>. For <span class="html-italic">C. tropicalis;</span> epi-1, Var-1 and Var-2 showed MIC of 256 μg mL<sup>−1</sup>, 64 μg mL<sup>−1</sup>, and 32 μg mL<sup>−1</sup>, respectively. For <span class="html-italic">C. krusei</span>; epi-1 and Var-1 showed MIC of 128 μg mL<sup>−1</sup> and variant-2 showed MIC of 64 μg mL<sup>−1</sup>. Cell density was measured at 595 nm after 24 h. Growth of the cells measured without peptides is used as a control with 100% growth. The level of significance was measured for variants compared to epi-1 at their respective concentrations tested. Statistical value representations are * <span class="html-italic">p</span>  &lt;  0.5, ** <span class="html-italic">p</span>  &lt;  0.1, *** <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>Light microscopic image of crystal violet stained <span class="html-italic">C. albicans</span> (MTCC 227), <span class="html-italic">C. tropicalis</span> (CA4), and <span class="html-italic">C. Krusei</span> (CA54) are shown in individual rows at 40× magnification. The columns represent (<b>a</b>) control, (<b>b</b>) epinecidin-1, (<b>c</b>) variant-1, (<b>d</b>) variant-2. The quantitative plots for each row is provided in the right column, The cells were treated with peptides at their respective MIC and incubated for 24 h. The image panel shows the effects of peptides in reducing biofilm formation. On the right, there are bar graphs representing the quantitative analysis of the number of cells under each treatment condition for each species. The quantitative cell number of the control is provided in the text but is not displayed in the plot because of variation in the Y-axis scale. Peptide treatment has invariably decreased the <span class="html-italic">Candida</span> spp. Cell numbers compared to the control. Statistical significance representations are * <span class="html-italic">p</span>  &lt;  0.5, ** <span class="html-italic">p</span>  &lt;  0.1, and *** <span class="html-italic">p</span>  &lt;  0.01.</p>
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<p>Scanning electron microscopic (SEM) image of <span class="html-italic">C. albicans</span> (MTCC 227) (<b>a</b>) Control, and (<b>b</b>–<b>d</b>) peptide-treated cells. The cells were treated with the peptides at their respective MIC and incubated for 6 h. The reduction in the number of cells noted as empty spaces between the cells and the disruption of the cell membrane demonstrates the membrane disruption activity of the peptides. The peptide-treated cells show an aberrant surface with internally collapsed morphology due to cytoplasm leakage and appear rugged. Bottom panel shows an enlarged view of the red inset box marked in the upper panel.</p>
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<p>Docking interactions of epinecidin-1 and its variants with <span class="html-italic">Candida</span> Exo-B-(1,3)-Glucanase: PDB ID 1CZ1 (<b>a</b>–<b>c</b>), Secreted aspartic proteinase (Sap) 1: PDB ID 2QZW (<b>d</b>–<b>f</b>) and N-terminal domain of Als 9-2: PDB ID 2Y7L (<b>g</b>–<b>i</b>) biofilm forming membrane receptors. The <span class="html-italic">Candida</span> cell wall proteins are represented green in colour and the peptides are yellow in colour. The amino acids originating from the <span class="html-italic">Candida</span> cell wall proteins interacting with the peptides are represented in white with their three-letter code and their corresponding position. The amino acid side chain of the <span class="html-italic">Candida</span> protein (TYR, VAL, GLY, TYR, CYS, GLU, CYS, GLY, ASN, ASP, THR) acting as a binding pocket for the peptides is shown red in colour.</p>
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<p>Effect of epinecidin-1 and its variants on the production of ROS by in <span class="html-italic">C. albicans</span>. Column (<b>a</b>) control cells, (<b>b</b>) epi-1, (<b>c</b>) var-1, and (<b>d</b>) var-2 peptide-treated cells and (<b>e</b>) Quantitation of number of cells in the field. Logarithmically growing <span class="html-italic">C. albicans</span> cells (1 × 10<sup>5</sup>) were pre-incubated with 20 μM DCFHDA and grown for 24 h at 37 °C. Cells were treated with and without peptides. After 24 h, the culture dish was washed with phosphate-buffered saline pH 7.4 and observed under a fluorescent microscope. Hoechst staining was performed alongside to determine the number of live cells. The images were captured at an excitation wavelength of 488 nm and an emission wavelength of 525 nm. The bar graphs on the right show the quantitation of cells in blue and green channels. The bar on the extreme right shows the percentage of cells with ROS calculated from the number of cells in the blue and green channels. The control does not have ROS-positive cells. The variants have more than 90% ROS production.</p>
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17 pages, 3630 KiB  
Article
Porcine Epidemic Diarrhea Virus Infection of Porcine Intestinal Epithelial Cells Causes Mitochondrial DNA Release and the Activation of the NLRP3 Inflammasome to Mediate Interleukin-1β Secretion
by Di Bao, Shushuai Yi, Luobing Zhao, Han Zhao, Jiuyuan Liu, Yiming Wei, Guixue Hu and Xinxin Liu
Vet. Sci. 2024, 11(12), 643; https://doi.org/10.3390/vetsci11120643 (registering DOI) - 12 Dec 2024
Abstract
Porcine epidemic diarrhea virus (PEDV) induces enteritis and diarrhea in piglets. Mitochondrial DNA (mtDNA) contributes to virus-induced inflammatory responses; however, the involvement of inflammasomes in PEDV infection responses remains unclear. We investigated the mechanism underlying inflammasome-mediated interleukin (IL)-1β secretion during the PEDV infection [...] Read more.
Porcine epidemic diarrhea virus (PEDV) induces enteritis and diarrhea in piglets. Mitochondrial DNA (mtDNA) contributes to virus-induced inflammatory responses; however, the involvement of inflammasomes in PEDV infection responses remains unclear. We investigated the mechanism underlying inflammasome-mediated interleukin (IL)-1β secretion during the PEDV infection of porcine intestinal epithelial (IPEC-J2) cells. IL-1β production and caspase-1 activity were assessed by quantitative PCR and enzyme-linked immunosorbent assay. NLRP3 inflammasome activation was assessed using immunoprecipitation experiments. Mitochondrial damage was evaluated by analyzing the mitochondrial membrane potential and ATP levels and by the flow cytometry examination of mitochondrial reactive oxygen species (mtROS). Mitochondria and mtDNA localization were observed using immunofluorescence. The inhibition of mtROS and mtDNA production allowed NLRP3 inflammasome and IL-1β expression detection and the evaluation of the pathway underlying NLRP3 inflammasome activation in PEDV-infected IPEC-J2 cells. IPEC-J2 cells upregulated IL-1β upon PEDV infection, where mature IL-1β secretion depended on caspase-1 activity, triggered NLRP3 inflammasome expression and assembly, and caused mitochondrial dysfunction, leading to mtDNA release and NLRP3 inflammasome activation, while mtROS contributed to NF-κB pathway activation, enhancing IL-1β secretion. This is the first demonstration of the mechanism underlying mtDNA release and NLRP3 inflammasome activation facilitating IL-1β secretion from PEDV-infected IPEC-J2 cells. These data enhance our understanding of the inflammatory mechanisms triggered by PEDV. Full article
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<p>Porcine epidemic diarrhea virus (PEDV) infects IPEC-J2 cells to promote the secretion of mature interleukin (IL)-1β. PEDV (multiplicity of infection (MOI) = 1) was used to infect IPEC-J2 cells for the specified periods of time. Detection of PEDV N protein expression via Western blot following PEDV infection in IPEC-J2 cells (<b>a</b>). mRNA expression (<b>b</b>) and Western blot (<b>f</b>) analyses of IL-1β levels, as well as the results of an enzyme-linked immunosorbent assay (ELISA) to assess IL-1β secretion in the cell supernatants (<b>c</b>). IPEC-J2 cells were infected with PEDV at the specified dose for 24 h, followed by fluorescence quantification (<b>d</b>), Western blotting (<b>g</b>), and the ELISA detection of IL-1β secretion in the cell supernatant (<b>e</b>). Data represent the mean ± SD (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>PEDV promotes caspase-1 enzymatic activity in IPEC-J2 cells. Caspase-1 enzyme activity (<b>a</b>) and Western blot results (<b>b</b>) at the specified time points after the PEDV infection of IPEC-J2 cells (MOI = 1). Caspase-1 enzyme activity (<b>c</b>) and Western blot results (<b>d</b>) of IPEC-J2 cells infected with PEDV at the specified doses for 24 h. IPEC-J2 cells were treated with the caspase-1 inhibitor, Ac-YVAD-cmk, at the specified concentration for 1 h and then inoculated with PEDV (MOI = 1). ELISA was used to detect IL-1β secretion in the cell supernatant 24 h after infection (<b>e</b>). Data represent the mean ± SD (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>PEDV infection of IPEC-J2 cells activates the NLRP3 inflammasome. PEDV was used to infect IPEC-J2 cells at the specified doses for 24 h and then the <span class="html-italic">NLRP3</span> inflammasome (<b>a</b>) and <span class="html-italic">ASC</span> (<b>c</b>) mRNA expression were analyzed. <span class="html-italic">NLRP3</span> (<b>b</b>) and <span class="html-italic">ASC</span> (<b>d</b>) mRNA expression in IPEC-J2 cells infected with PEDV (MOI = 1) for the specified times. NLRP3 (<b>e</b>) and ASC (<b>f</b>) rabbit-derived primary antibodies were used as bait antibodies and rabbit serum was used as the negative control antibody in a co-immunoprecipitation experiment. Data represent the mean ± SD (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>PEDV-infected IPEC-J2 cells secrete IL-1β through NLRP3 inflammasome activity. IPEC-J2 cells were treated with 10 µM MCC950 for 1 h, and negative control cells were treated with DMSO for the same time. After 24 h of cell infection with PEDV (MOI = 1), IL-1β secretion in the cell supernatants was detected by ELISA (<b>a</b>) and <span class="html-italic">IL-1β</span> mRNA quantified by fluorescence (<b>c</b>). siNLRP3 and siCtrl (control) were transfected into IPEC-J2 cells. PEDV was used to infect cells (MOI = 1) for 24 h, IL-1β secretion in the cell supernatants was detected by ELISA (<b>b</b>), and <span class="html-italic">IL-1β</span> mRNA expression was quantified by fluorescence (<b>d</b>). Western blot of IPEC-J2 cells treated with 10 µM MCC950 (<b>e</b>) and siNLRP3 transfection of IPEC-J2 cells to detect the expression of inflammasome proteins (<b>f</b>). Data represent the mean ± SD (<span class="html-italic">n</span> = 3),** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>PEDV infection causes mitochondrial dysfunction and results in mitochondrial reactive oxygen species (mtROS) production and mitochondrial DNA (mtDNA) release. Mitochondrial membrane potential was decreased (<b>a</b>). Reduced ATP production after PEDV infection (<b>b</b>). Flow cytometry fluorescence intensity analysis of mtROS production after PEDV infection (<b>c</b>). Immunofluorescence showing mtDNA release after PEDV infection (<b>d</b>). Data represent mean ± SD (<span class="html-italic">n</span> = 3), ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>mtROS are involved in NF-κB activation in PEDV-infected IPEC-J2 cells. IPEC-J2 cells were infected with PEDV at the specified doses for 24 h and the expression levels of pp65 and p65 detected by Western blot (<b>a</b>). IPEC-J2 cells were treated with the NF-κB inhibitor BAY11-7082 (10 μM). <span class="html-italic">IL-1β</span> mRNA expression was detected 24 h after PEDV infection (MOI = 1) (<b>b</b>) and IL-1β secretion in the cell supernatant detected by ELISA (<b>c</b>). Western blot to assess the expression of pp65, p65, and IL-1β proteins (<b>d</b>). After IPEC-J2 cells were treated with 10 µM Mito-TEMPO for 1 h, <span class="html-italic">IL-1β</span> mRNA expression was detected following PEDV infection (MOI = 1) for 24 h (<b>e</b>) and IL-1β secretion in the cell supernatant detected by ELISA (<b>f</b>). Data represent mean ± SD (<span class="html-italic">n</span> = 3), ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>mtDNA participates in NLRP3 inflammasome activation in PEDV-infected IPEC-J2 cells. After transfection with DNase I, IPEC-J2 cells were infected with PEDV (MOI = 1). IL-1β mRNA expression (<b>a</b>) and secretion in cell supernatants detected by ELISA (<b>b</b>). <span class="html-italic">NLRP3</span> inflammasome mRNA expression (<b>c</b>) and caspase-1 enzyme activity (<b>d</b>). Western blot detection of NLRP3 inflammasome and downstream protein expression after transfection with DNase I protein (<b>e</b>). Data represent mean ± SD (<span class="html-italic">n</span> = 3). ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Schematic showing that PEDV infection leads to cytoplasmic mitochondrial DNA release and the activation of the NLPR3 inflammasome.</p>
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24 pages, 4342 KiB  
Review
Lipid Nanovesicles for Antioxidant Delivery in Skin: Liposomes, Ufasomes, Ethosomes, and Niosomes
by Agnese Ricci, Luca Stefanuto, Tecla Gasperi, Fabio Bruni and Daniela Tofani
Antioxidants 2024, 13(12), 1516; https://doi.org/10.3390/antiox13121516 (registering DOI) - 12 Dec 2024
Abstract
The skin, being the largest organ of the human body, serves as the primary barrier against external insults, including UV radiation, pollutants, and microbial pathogens. However, prolonged exposure to these environmental stressors can lead to the generation of reactive oxygen species (ROS), causing [...] Read more.
The skin, being the largest organ of the human body, serves as the primary barrier against external insults, including UV radiation, pollutants, and microbial pathogens. However, prolonged exposure to these environmental stressors can lead to the generation of reactive oxygen species (ROS), causing oxidative stress, inflammation, and ultimately, skin aging and diseases. Antioxidants play a crucial role in neutralizing ROS and preserving skin health by preventing oxidative damage. In recent years, nanotechnology has emerged as a powerful tool for enhancing the delivery of antioxidants onto the skin. In particular, liposomal formulations have offered unique advantages such as improved stability, controlled release, and enhanced penetration through the skin barrier. This has led to a surge in research focused on developing liposomal-based antioxidant delivery systems tailored for skin health applications. Through a comprehensive analysis of the literature from the 2019–2024 period, this review provides an overview of emerging trends in the use of liposomal delivery systems developed for antioxidants aimed at improving skin health. It explores the latest advancements in liposomal formulation strategies, vesicle characterization, and their applications in delivering antioxidants to combat oxidative stress-induced skin damage and other associated skin pathologies. A comparison of various delivery systems is conducted for the most common antioxidants. Finally, a brief analysis of lipid nanovesicles used in the cosmeceutical industry is provided. Full article
(This article belongs to the Special Issue Methodologies for Improving Antioxidant Properties and Absorption)
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Figure 1
<p>Structure of layers of epidermis (<a href="https://commons.wikimedia.org/wiki/File:502_Layers_of_epidermis.jpg" target="_blank">https://commons.wikimedia.org/wiki/File:502_Layers_of_epidermis.jpg</a> (accessed on 16 October 2024)).</p>
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<p>Principal techniques for producing liposomes [<a href="#B42-antioxidants-13-01516" class="html-bibr">42</a>].</p>
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<p>Schematic representation of the different types of liposomal drug delivery systems (<a href="https://commons.wikimedia.org/wiki/File:Liposomas_articulo_liberaci%C3%B3n_activos.jpg" target="_blank">https://commons.wikimedia.org/wiki/File:Liposomas_articulo_liberaci%C3%B3n_activos.jpg</a> (accessed on 16 October 2024)).</p>
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<p>Different sizes and lamellarity of liposomes. V: vesicles, U: unilamellar; G: giant, L: large; M: medium; S: small; OLV: oligolamellar vesicles; MLV: multilamellar vesicles; MVV: multi vesicular vesicles (<a href="https://commons.wikimedia.org/wiki/File:1-s2.0-S0168365921005034-gr6_lrg.jpg#filelinks" target="_blank">https://commons.wikimedia.org/wiki/File:1-s2.0-S0168365921005034-gr6_lrg.jpg#filelinks</a> (accessed on 16 October 2024)).</p>
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<p>(<b>a</b>) HPLC–DAD chromatogram of myrtle berry extract at λ = 280 nm. Chromatographic conditions are described in the reference article. 1: gallic acid; 2: gallic acid derivative; 3: Delphinidin-3-O-glucoside; 4: Cyanidin-3-O-glucoside; 5: Petunidin-3-O-glucoside; 6: Peonidin-3-O-glucoside; 7: Malvidin-3-O-glucoside; 8: Myricetin-3-O-galactoside; 9: Myricetin-3-O-rhamnoside; and 10: ellagic acid. (<b>b</b>) Myrtle liposomes through cryo-TEM observation [<a href="#B24-antioxidants-13-01516" class="html-bibr">24</a>].</p>
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<p>(<b>a</b>) Release of antioxidant activity (DPPH) and total phenolic compounds (TPC) of propolis extract (PE) loaded in LIP during 48 h. (<b>b</b>) Cytotoxicity profile of PE loaded in human immortalized keratinocyte (HaCaT) cells. HaCaT cells were incubated for 72 h with increasing concentrations (0–5%) of PE loaded in LIP, and their cell viability was estimated via sulforhodamine B (SRB) assay. The EC50 and EC10 values (efficient concentration that causes 50% and 10% decrease in cell viability, respectively) of PE loaded in LIP were determined from the dose–response curves. The results are shown as the mean ± SD of three independent experiments [<a href="#B26-antioxidants-13-01516" class="html-bibr">26</a>].</p>
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<p>TEM images of fibroblasts in cells treated with ETH (0.9 concentration of PC in ethanol) (<b>a</b>,<b>b</b>). Panel (<b>a</b>), 31.5K magnification; panel (<b>b</b>), 50K magnification. ETH are indicated by arrows. Lower bar corresponds to 500 nm [<a href="#B30-antioxidants-13-01516" class="html-bibr">30</a>].</p>
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<p>Quercetin (QT) release kinetics from ETH<sub>0.9</sub>-QT (blue circles), TETH<sub>0.9</sub>-QT (red squares), TETH<sub>2.7</sub>-QT (green triangles), and SOL-QT (black crosses), as determined by Franz cell associated with NY (<b>a</b>) and PTFE (<b>b</b>). Data are the mean of 6 independent experiments ± s.d.</p>
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<p>Curcumin lipid nanoparticle delivery systems analyzed.</p>
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19 pages, 8094 KiB  
Article
Improving the Quality of the Water Flowing over a Stepped Spillway in Open Canals by Increasing Its Degree of Aeration
by Mohamed A. Ashour, Mohamed Khairy Ali and Tawab E. Aly
Limnol. Rev. 2024, 24(4), 653-671; https://doi.org/10.3390/limnolrev24040038 (registering DOI) - 12 Dec 2024
Abstract
As spillways are hydraulic structures constructed for the safe release of floodwater from the upstream (US) side of a dam to the downstream side, or from the end of canals and drains to a lower stream, the upstream water flow of such structures [...] Read more.
As spillways are hydraulic structures constructed for the safe release of floodwater from the upstream (US) side of a dam to the downstream side, or from the end of canals and drains to a lower stream, the upstream water flow of such structures gains significant amounts of potential energy. As this water flows over a spillway or escapes, the gained potential energy is converted into kinetic energy, resulting in the water gaining an increasing velocity, thereby enhancing the flow’s destructive potential. This can have a harmful impact on the hydraulic performance and the structural stability of the spillway itself. To avoid such harmful effects, engineers and designers of such structures usually provide the spillways and water escapes with some tools for dissipating that kinetic energy and decreasing the flowing water’s velocity. The present study aims to enhance the performance efficiency of such dissipating tools, as well as to improve the quality of the flowing water by leveraging the significant turbulence generated by the existing energy dissipators on the back of the spillway body. The aeration process enabled by this turbulence increases the dissolved oxygen contents, thereby enhancing the water quality, which is one of the main objectives of this work. On the back surface of the spillway, various dissipater shapes with different geometrical configurations, dimensions, and combinations were tested, in order to determine the most suitable engineering treatments for maximizing the dissolved oxygen content and improving the water quality for various uses, as the study’s main goal. By testing 21 different model configurations with the available laboratory discharges, the study successfully identified the most effective shape and properties of the desired dissipator, which increased the dissolved oxygen content by an average of 21.70% and dissipated water energy by about 69%. Full article
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<p>Definition sketch of the experimental model.</p>
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<p>Experimental set-up.</p>
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<p>Models used for estimating the best effective ratio of step height to step length (<span class="html-italic">h</span>/<span class="html-italic">l</span>) on the back surface of the spillway.</p>
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<p>Models used for estimating the best effective ratio of step height to spillway height (<span class="html-italic">h</span>/<span class="html-italic">H</span>) on the back surface of the spillway.</p>
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<p>Models used for estimating the most effective ratio of length of the reverse inclined step to step length (<span class="html-italic">lr</span>/<span class="html-italic">l</span>) on the back surface of the spillway.</p>
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<p>Models used for estimating the best effective ratio of end sill radius to step height (r/h) on the back surface of the spillway.</p>
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<p>Models used in comparison between different spillway geometrical arrangements.</p>
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<p>Relationship between relative energy loss (<span class="html-italic">∆E</span>/<span class="html-italic">E<sub>o</sub></span>) and step height to step length ratio (<span class="html-italic">h</span>/<span class="html-italic">l</span>).</p>
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<p>Relationship between relative dissolved oxygen concentration (<span class="html-italic">∆DO</span>/<span class="html-italic">Doo</span>) and step height to step length ratio (<span class="html-italic">h</span>/<span class="html-italic">l</span>).</p>
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<p>Relationship between relative residual energy at the toe of the spillway (E1/E1(<span class="html-italic">h</span>/<span class="html-italic">l</span> = 2.0)) and the cost of the spillway at discharge <span class="html-italic">Q</span> = 8.17 L/s.</p>
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<p>Relationship between relative energy loss (<span class="html-italic">∆E</span>/<span class="html-italic">E<sub>o</sub></span>) and number of steps (<span class="html-italic">Ns</span>).</p>
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<p>Relationship between relative dissolved oxygen concentration (<span class="html-italic">∆DO</span>/<span class="html-italic">DO<sub>o</sub></span>) and number of steps (<span class="html-italic">Ns</span>).</p>
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<p>Relationship between relative energy loss (<span class="html-italic">∆E</span>/<span class="html-italic">Eo</span>) and inclined step length to total step length ratio (<span class="html-italic">lr</span>/<span class="html-italic">l</span>).</p>
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<p>Relationship between relative dissolved oxygen concentration (<span class="html-italic">∆DO</span>/<span class="html-italic">DO<sub>o</sub></span>) and inclined step length to total step length ratio (<span class="html-italic">lr</span>/<span class="html-italic">l</span>).</p>
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<p>Relationship between relative energy loss (<span class="html-italic">∆E</span>/<math display="inline"><semantics> <mrow> <msub> <mrow> <mi>E</mi> </mrow> <mrow> <mi>o</mi> </mrow> </msub> </mrow> </semantics></math>) and end sill radius to step height ratio (<span class="html-italic">r</span>/<span class="html-italic">h</span>).</p>
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<p>Relationship between relative dissolved oxygen concentration (<span class="html-italic">∆DO</span>/<math display="inline"><semantics> <mrow> <msub> <mrow> <mi>D</mi> <mi>O</mi> </mrow> <mrow> <mn>0</mn> </mrow> </msub> </mrow> </semantics></math>) and end sill radius to step height ratio (<span class="html-italic">r</span>/<span class="html-italic">h</span>).</p>
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<p>(<b>a</b>,<b>b</b>). Relationship between the relative energy loss (∆E/Eo) and the flow rate (Q) (L/s) with variable back surface arrangements.</p>
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<p>(<b>a</b>,<b>b</b>). Relationship between dissolved oxygen content (<span class="html-italic">∆DO</span>/<span class="html-italic">DOo</span>) and the flow rate (<span class="html-italic">Q</span>)(L/s) with variable back surface arrangements.</p>
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17 pages, 6521 KiB  
Article
Rational Fabrication of Ag2S/g-C3N4 Heterojunction for Photocatalytic Degradation of Rhodamine B Dye Under Natural Solar Radiation
by Ali Alsalme, Ahmed Najm, Nagy N. Mohammed, M. F. Abdel Messih, Ayman Sultan and Mohamed Abdelhay Ahmed
Catalysts 2024, 14(12), 914; https://doi.org/10.3390/catal14120914 (registering DOI) - 11 Dec 2024
Abstract
Near-infrared light-triggered photocatalytic water treatment has attracted significant attention in recent years. In this novel research, rational sonochemical fabrication of Ag2S/g-C3N4 nanocomposites with various compositions of Ag2S (0–25) wt% was carried out to eliminate hazardous rhodamine [...] Read more.
Near-infrared light-triggered photocatalytic water treatment has attracted significant attention in recent years. In this novel research, rational sonochemical fabrication of Ag2S/g-C3N4 nanocomposites with various compositions of Ag2S (0–25) wt% was carried out to eliminate hazardous rhodamine B dye in a cationic organic pollutant model. g-C3N4 sheets were synthesized via controlled thermal annealing of microcrystalline urea. However, black Ag2S nanoparticles were synthesized through a precipitation-assisted sonochemical route. The chemical interactions between various compositions of Ag2S and g-C3N4 were carried out in an ultrasonic bath with a power of 300 W. XRD, PL, DRS, SEM, HRTEM, mapping, BET, and SAED analysis were used to estimate the crystalline, optical, nanostructure, and textural properties of the solid specimens. The coexistence of the diffraction peaks of g-C3N4 and Ag2S implied the successful production of Ag2S/g-C3N4 heterojunctions. The band gap energy of g-C3N4 was exceptionally reduced from 2.81 to 1.5 eV with the introduction of 25 wt% of Ag2S nanoparticles, implying the strong absorbability of the nanocomposites to natural solar radiation. The PL signal intensity of Ag2S/g-C3N4 was reduced by 40% compared with pristine g-C3N4, implying that Ag2S enhanced the electron–hole transportation and separation. The rate of the photocatalytic degradation of rhodamine B molecules was gradually increased with the introduction of Ag2S on the g-C3N4 surface and reached a maximum for nanocomposites containing 25 wt% Ag2S. The radical trapping experiments demonstrated the principal importance of reactive oxygen species and hot holes in destroying rhodamine B under natural solar radiation. The charge transportation between Ag2S and g-C3N4 semiconductors proceeded through the type I straddling scheme. The enriched photocatalytic activity of Ag2S/g-C3N4 nanocomposites resulted from an exceptional reduction in band gap energy and controlling the electron–hole separation rate with the introduction of Ag2S as an efficient photothermal photocatalyst. The novel as-synthesized nanocomposites are considered a promising photocatalyst for destroying various types of organic pollutants under low-cost sunlight radiation. Full article
(This article belongs to the Section Photocatalysis)
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<p>XRD of g-C<sub>3</sub>N<sub>4</sub>, Ag<sub>2</sub>S, and CNAgS25 nanocomposites.</p>
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<p>N<sub>2</sub>-adsorption isotherm of (<b>a</b>) g-C<sub>3</sub>N<sub>4</sub> and (<b>b</b>) CNAgS25.</p>
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<p>(<b>a</b>) SEM of CNAgS25, (<b>b</b>) mapping of CNAgS25, (<b>c</b>) mapping of C, (<b>d</b>) mapping of (N), (<b>e</b>) mapping of Ag, (<b>f</b>) mapping of S, (<b>g</b>) EDX of CNAgS25.</p>
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<p>(<b>a</b>) TEM of CNAgS25, (<b>b</b>) HRTEM of CNAgS25 and (<b>c</b>) SAED of CNAgS25.</p>
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<p>(<b>a</b>) TEM of CNAgS25, (<b>b</b>) HRTEM of CNAgS25 and (<b>c</b>) SAED of CNAgS25.</p>
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<p>(<b>a</b>) DRS of g-C<sub>3</sub>N<sub>4</sub>, Ag<sub>2</sub>S, CNAgS15, and CNAgS25. (<b>b</b>) Tauc plot of g-C<sub>3</sub>N<sub>4</sub>, Ag2S, CNAgS15, and CNAgS25.</p>
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<p>PL analysis of g-C<sub>3</sub>N<sub>4</sub>, NAgS15, and CNAgS25.</p>
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<p>The absorption spectrum for photocatalytic degradation of rhodamine B over the surfaces of g-C3N4, CNAg10, CNAg15, and CNAg25.</p>
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<p>(<b>a</b>) The variations in the amount of RhB removed (%) under dark and light reactions with the illumination time over the surfaces of g-C<sub>3</sub>N<sub>4</sub>, CNAg10, CNAg15, and CNAg25. (<b>b</b>) The kinetic first-order plot for photocatalytic degradation of RhB dye over the surfaces of g-C<sub>3</sub>N<sub>4</sub>, CNAg10, CNAg15, and CNAg25.</p>
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<p>Photocatalytic degradation of rhodamine B (2 × 10<sup>−5</sup> M) over CNAgS25 nanocomposite in the presence of 2 × 10<sup>−5</sup> M of the following scavengers: (<b>a</b>) benzoquinone, (<b>b</b>) ammonium oxalate, and (<b>c</b>) isopropanol. (<b>d</b>) PL spectrum of terephthalic acid 2 × 10<sup>−4</sup> M over CNAgS25 nanocomposite at 325 nm excitation wavelength.</p>
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<p>Photocatalytic degradation of rhodamine B (2 × 10<sup>−5</sup> M) over CNAgS25 nanocomposite in the presence of 2 × 10<sup>−5</sup> M of the following scavengers: (<b>a</b>) benzoquinone, (<b>b</b>) ammonium oxalate, and (<b>c</b>) isopropanol. (<b>d</b>) PL spectrum of terephthalic acid 2 × 10<sup>−4</sup> M over CNAgS25 nanocomposite at 325 nm excitation wavelength.</p>
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<p>Regeneration of CNAgS25 for five consecutive cycles for removal of RhB dye over CNAgS25 nanocomposite.</p>
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<p>A scheme for electron transportation between g-C<sub>3</sub>N<sub>4</sub> and Ag<sub>2</sub>S semiconductors.</p>
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<p>Scheme for synthesis of (<b>a</b>) g-C<sub>3</sub>N<sub>4</sub>, (<b>b</b>) Ag<sub>2</sub>S and (<b>c</b>) Ag<sub>2</sub>S/g-C<sub>3</sub>N<sub>4</sub> heterojunction.</p>
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9 pages, 1392 KiB  
Review
The Evolution of Kidney Graft Preservation Through the Years
by Andres Calva Lopez, Jose Enrique Robles Garcia, Carlos Andres Yanez Ruiz, Mario Daniel Tapia Tapia, Vanessa Talavera Cobo, Carmina Alejandra Muñoz Bastidas, Daniel Sanchez Zalabardo and Bernardino Miñana Lopez
Life 2024, 14(12), 1647; https://doi.org/10.3390/life14121647 - 11 Dec 2024
Abstract
Chronic kidney disease (CKD) is a prevalent disease affecting almost 10% of the world’s population, with many cases progressing to end-stage kidney disease (ESKD). Kidney transplantation (KT) is the gold-standard treatment for ESKD. Due to growing KT waitlists, the deceased kidney donor (DKDs) [...] Read more.
Chronic kidney disease (CKD) is a prevalent disease affecting almost 10% of the world’s population, with many cases progressing to end-stage kidney disease (ESKD). Kidney transplantation (KT) is the gold-standard treatment for ESKD. Due to growing KT waitlists, the deceased kidney donor (DKDs) criteria have expanded to increase the number of available kidney grafts. Kidney graft preservation ensures optimal graft function after KT. Static cold storage (SCS) as a preservation method is still widely used. Hypothermic machine perfusion (HMP) has proven to decrease delayed graft function (DGF) and increase graft survival. Most recent studies advocate for the use of HMP regardless of donor type. However, emerging technologies, such as hypothermic oxygenated machine perfusion (HOPE) and normothermic machine perfusion (NMP), have shown promising results in specific scenarios. This review aims to provide a summary of the well-established kidney graft preservation methods and their outcomes, as well as novel technological advances that allow for newer preservation strategies. Full article
(This article belongs to the Special Issue Kidney Transplantation: What’s Hot and What’s New)
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<p>Steps for static cold storage (SCS) organ preservation.</p>
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<p>Steps for hypothermic machine perfusion (HMP) organ preservation.</p>
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<p>(<b>a</b>) LifePort<sup>®</sup> Kidney Transporter; (<b>b</b>) Kidney Assist<sup>TM</sup>; (<b>c</b>) RM3<sup>®</sup>.</p>
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20 pages, 8137 KiB  
Article
A Simple and Rapid “Turn-On” Fluorescent Probe Based on Binuclear Schiff Base for Zn2+ and Its Application in Cell Imaging and Test Strips
by Jinghui Cheng, Yi Li, Zhiye Zhu, Huijuan Guan, Jinsong Zhai, Yibing Xiang and Man Wang
Molecules 2024, 29(24), 5850; https://doi.org/10.3390/molecules29245850 - 11 Dec 2024
Abstract
A series of colorful binuclear Schiff bases derived from the different diamine bridges including 1,2- ethylenediamine (bis-Et-SA, bis-Et-4-NEt2, bis-Et-5-NO2, bis-Et-Naph), 1,2-phenylenediamine (bis-Ph-SA, bis-Ph-4-NEt2, bis-Ph-5-NO2, bis-Ph-Naph), dicyano-1,2-ethenediamine (bis-CN-SA, bis-CN-4-NEt2, bis-CN-5-NO2, bis-CN-Naph) have [...] Read more.
A series of colorful binuclear Schiff bases derived from the different diamine bridges including 1,2- ethylenediamine (bis-Et-SA, bis-Et-4-NEt2, bis-Et-5-NO2, bis-Et-Naph), 1,2-phenylenediamine (bis-Ph-SA, bis-Ph-4-NEt2, bis-Ph-5-NO2, bis-Ph-Naph), dicyano-1,2-ethenediamine (bis-CN-SA, bis-CN-4-NEt2, bis-CN-5-NO2, bis-CN-Naph) have been designed and prepared. The optical properties of these binuclear Schiff base ligands were fully determined by UV–Vis absorption spectroscopy, fluorescence emission spectroscopy, and time-dependent-density functional theory (TD-DFT) calculations. The inclusion of D-A systems and/or π-extended systems in these binuclear Schiff base ligands not only enables adjustable RGB light absorption and emission spectra (300~700 nm) but also yields high fluorescence quantum efficiencies of up to 0.84 in MeCN solution. Then, with the ESIPT (excited-state intramolecular proton transfer) property, fluorescence analysis showed that the probe bis-Et-SA and bis-Ph-SA could recognize Zn2+ via the “turn on” mode in the MeCN solution. During the detection process, bis-Et-SA and bis-Ph-SA demonstrate rapid response and high selectivity upon the addition of Zn2+. The coordination of Zn2+ with the oxygen atom and Schiff base nitrogen atom in a tetrahedral geometry is confirmed by Job’s plot, FT-IR, and 1H NMR spectroscopy. In addition, the paper test and Hela cells were successfully carried out to detect Zn2+. Moreover, the sensitivity of bis-Et-SA and bis-Ph-SA is much better than that of those Schiff base ligands containing only one chelating unit [O^N^N^O]. Full article
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Figure 1
<p>Normalized absorption spectra of the bis-Schiff ligands with the same bridges and different diethylamino-phenol in MeCN.</p>
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<p>Normalized UV–Vis absorption spectra of the bis-Schiff ligands with the same diethylamino-phenol different bridges in MeCN.</p>
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<p>Images captured of the chosen binuclear Schiff base ligands in MeCN at ambient temperature. (Top: under natural sunlight; bottom: under 360 nm UV light).</p>
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<p>Normalized emission spectra were obtained for bis-Schiff ligands featuring the same bridges but varying diethylamino-phenol substituents in MeCN solvent.</p>
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<p>Normalized emission spectra were obtained for bis-Schiff ligands featuring various bridges and a consistent di-ethylamino-phenol in MeCN.</p>
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<p>Frontier molecular orbitals for the bis-Schiff ligands with the different diethylamino-phenol and same bridges in MeCN at B3LYP 6-31G(d,p) level of theory.</p>
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<p>Frontier molecular orbitals for the bis-Schiff ligands with different bridges and the same diethylamino-phenol in MeCN at B3LYP 6-31G(d,p) level of theory.</p>
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<p>The UV–Vis absorption spectra of bis-Et-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN) were observed upon the addition of 2 equiv. of different metal ions.</p>
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<p>The UV–Vis absorption spectra of bis-Ph-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN) were observed upon the addition of 2 equiv. of different metal ions.</p>
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<p>The emission spectra of bis-Et-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN, excited at 360 nm) upon the addition of 2 equiv. of different metal ions.</p>
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<p>The emission spectra of bis-Ph-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN, excited at 340 nm) were observed upon the addition of 2 equiv of various metal ions.</p>
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<p>The selectivity of bis-Et-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN, with emission measured at 450 nm and excited at 360 nm) towards 2.0 equiv. of other metal ions, and Zn<sup>2+</sup> was investigated.</p>
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<p>Selectivity of bis-Ph-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN, emission measured at 405 nm and excited at 340 nm) toward 2.0 equiv. of other metal ions and Zn<sup>2+</sup>.</p>
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<p>Plot of (I − I0)/I (emission at 450 nm) as a function of Zn<sup>2+</sup> concentration based on bis-Et-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN).</p>
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<p>Plot of emission intensity at 405 nm as a function of Zn<sup>2+</sup> concentration based on bis-Ph-SA (1.0 × 10<sup>−5</sup> mol dm<sup>−3</sup> in MeCN).</p>
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<p>Job’s plot of bis-Et-SA and Zn<sup>2+</sup> in CH<sub>3</sub>CN solution. The total concentration of bis-Et-SA and Zn<sup>2+</sup> was 10 μM. The emission intensity at 450 nm.</p>
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<p>Job’s plot of bis-Ph-SA and Zn<sup>2+</sup> in CH<sub>3</sub>CN solution. The total concentration of bis-Ph-SA and Zn<sup>2+</sup> was 10 μM. The emission intensity at 405 nm.</p>
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<p>The fraction <sup>1</sup>HNMR spectroscopy of bis-Et-SA and bis-Et-SA+Zn<sup>2+</sup> in DMSO-d<sub>6</sub>.</p>
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<p>The IR spectra of bis-Et-SA and bis-Et-SA+Zn<sup>2+</sup>.</p>
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<p>Energy level diagram for the frontier molecular orbitals of bis-Et-SA (<b>left</b>) and its zinc complex (<b>right</b>) calculated by using the B3LYP/6–31G basis set.</p>
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<p>The proposed binding mechanism of bis-Et-SA with Zn<sup>2+</sup>.</p>
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<p>Fluorescent images were captured of Hela cells. (1) Bis-Ph-SA (30 μM) was added to the culture medium and incubated with Hela cells for 40 min. (2) Prior to treatment, Hela cells were exposed to bis-Ph-SA (30 μM) for 40 min, followed by an additional 40 min incubation with Zn<sup>2+</sup> ions (60 μM).</p>
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<p>The cytotoxicity test of bis-Ph-SA.</p>
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<p>Pictures of bis-Et-SA and bis-Ph-SA test strips treated with Zn<sup>2+</sup>. The photographs were captured upon exposure to 360 nm UV light.</p>
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<p>The synthetic route and chemical structures of binuclear Schiff base ligands in this work.</p>
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26 pages, 2294 KiB  
Protocol
The Multifunctional Catalytic Hemoglobin from Amphitrite ornata: Protocols on Isolation, Taxonomic Identification, Protein Extraction, Purification, and Characterization
by Anna L. Husted, Victoria R. Sutton, Lauren A. Presnar, R. Kevin Blackburn, Joseph L. Staton, Stephen A. Borgianini and Edward L. D’Antonio
Methods Protoc. 2024, 7(6), 100; https://doi.org/10.3390/mps7060100 - 11 Dec 2024
Abstract
The multifunctional catalytic hemoglobin from the terebellid polychaete Amphitrite ornata, also named dehaloperoxidase (AoDHP), utilizes the typical oxygen transport function in addition to four observed activities involved in substrate oxidation. The multifunctional ability of AoDHP is presently a rare [...] Read more.
The multifunctional catalytic hemoglobin from the terebellid polychaete Amphitrite ornata, also named dehaloperoxidase (AoDHP), utilizes the typical oxygen transport function in addition to four observed activities involved in substrate oxidation. The multifunctional ability of AoDHP is presently a rare observation, and there exists a limitation for how novel dehaloperoxidases can be identified from macrobenthic infauna. In order to discover more infaunal DHP-bearing candidates, we have devised a facilitated method for an accurate taxonomic identification that places visual and molecular taxonomic approaches in parallel. Traditional visual taxonomic species identification by the non-specialist, at least for A. ornata or even for other marine worms, is a very difficult and time-consuming task since a large diversity is present and the method is restricted to adult worm specimens. The work herein aimed to describe a method that simplifies the taxonomic identification of A. ornata in particular through the assessment of its mitochondrial cytochrome c oxidase subunit I gene by employing the DNA barcoding technique. Furthermore, whole-worm specimens of A. ornata were used to extract and purify AoDHP followed by an H2O2-dependent peroxidase activity assay evaluation against substrate 2,4,6-trichlorophenol. AoDHP isoenzyme A was also overexpressed as the recombinant protein in Escherichia coli, and its peroxidase activity parameters were compared to AoDHP from the natural source. The activity assay assessment indicated a tight correlation for all Michaelis–Menten parameters evaluated. We conclude that the method described herein exhibits a streamlined approach to identify the polychaete A. ornata, which can be adopted by the non-specialist, and the full procedure is predicted to facilitate the discovery of novel dehaloperoxidases from other marine invertebrates. Full article
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<p>Methodological flowchart for the isolation and taxonomic identification of <span class="html-italic">A. ornata</span> marine worms, followed by the extraction, purification, and characterization of <span class="html-italic">Ao</span>DHP from the natural source and through recombinant overexpression.</p>
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<p>Field collection site for <span class="html-italic">A. ornata</span> marine worms at Hickory Forest Beach in Hilton Head Island, South Carolina, USA; coordinates: 32.2509536, −80.6974940. This private beach includes a variety of healthy oyster reefs in soft plough mud and high salinity, which are the ideal areas to find <span class="html-italic">A. ornata</span> in addition to other polychaetes. The image was taken from one of the oyster reefs in the intertidal zone during low tide.</p>
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<p>Field collection and cleaning of marine polychaete worms. (<b>a</b>) Excavated polychaetes from plough mud, including the host burrower, (1) the ornate terebellid worm (<span class="html-italic">Amphitrite ornata</span>) (length: 7.5 cm × width: 0.6 cm), in the presence of one of its symbionts, (2) <span class="html-italic">Lepidametria commensalis</span> (length: 1.9 cm × width: 0.4 cm). (<b>b</b>) The <span class="html-italic">A. ornata</span> specimen was cleaned from mud debris in a neutral saline buffer, 100 mM potassium phosphate (pH 7.0).</p>
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<p>Purification of a truncated <span class="html-italic">Ao</span>COI gene amplicon (578 bp) by agarose gel electrophoresis. The <span class="html-italic">Ao</span>COI amplification by PCR involved <span class="html-italic">A. ornata</span> genomic DNA from one worm specimen and <span class="html-italic">Ao</span>COI-specific primers. Lane 1 is a 1-kb DNA ladder and lane 2 is a single sample of PCR amplicon product. Electrophoresis was performed using a 1.2% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) agarose gel containing ethidium bromide with a 40 μL sample loading volume.</p>
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<p>Extraction of <span class="html-italic">Ao</span>DHP<sub>NS</sub> from <span class="html-italic">A. ornata</span> whole-worm tissue. (<b>a</b>) <span class="html-italic">A. ornata</span> marine worms undergoing a thawing procedure from −80 °C to room temperature in a weighing-boat. (<b>b</b>) Ground worm tissue. (<b>c</b>) Impure <span class="html-italic">Ao</span>DHP<sub>NS</sub> recovered in a 50 mL conical tube after being thoroughly sonicated, passed through four layers of cheesecloth, and centrifuged to remove cellular debris.</p>
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<p>SDS-PAGE of sample 1 representing 6×-His-tagged <span class="html-italic">Ao</span>DHP<sub>REC</sub>-A (control-1; MW of 16,540 Da); sample 2 representing non-His-tagged <span class="html-italic">Ao</span>DHP<sub>REC, NHT</sub>-A (control-2; MW of 15,660 Da); sample 3: <span class="html-italic">Ao</span>DHP<sub>NS</sub> (approx. 15.7 kDa). Sample L represents the molecular weight marker ranging from 10–250 kDa (Precision Plus Protein Dual Color Standards; Bio-Rad). The 15 kDa standard (within L) exhibited slightly different electrophoretic mobility compared to samples 2 and 3, which have similar molecular masses. For trypsin digestion, four bands of sample 2 and four bands of sample 3 were cut from the gel using a cleaned razor blade. Electrophoresis was performed using a 4–15% polyacrylamide gel that was stained with Coomassie Brilliant Blue R-250 with a 12 μL sample loading volume per well.</p>
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<p>Analysis of trypsin-digested samples for (<b>a</b>) <span class="html-italic">Ao</span>DHP<sub>NS</sub> and (<b>b</b>) <span class="html-italic">Ao</span>DHP<sub>REC, NHT</sub>-A by LC-MS/MS. With respect to <span class="html-italic">A. ornata</span> dehaloperoxidase A (138 residues) (GenBank accession no. AAF97245.1), blue color-coding represents the matched peptides. Additionally, <span class="html-italic">Ao</span>DHP<sub>NS</sub> and <span class="html-italic">Ao</span>DHP<sub>REC, NHT</sub>-A revealed protein sequence coverages of 86.2% and 96.4%, respectively. The proteolytic enzyme trypsin cleaves peptide bonds from the C-terminal direction of its target protein on lysine (K) or arginine (R) residues, unless they are next to a proline (P) residue.</p>
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<p>Peroxidase reaction for the oxidative dechlorination of 2,4,6-trichlorophenol into 2,6-dichloro-1,4-benzoquinone catalyzed by <span class="html-italic">Ao</span>DHP and hydrogen peroxide.</p>
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<p>Enzyme kinetics of <span class="html-italic">Ao</span>DHP (natural source vs. recombinant) as represented by Michaelis–Menten plots for the peroxidase reaction. Panels (<b>a</b>,<b>b</b>) show rates of 2,6-dichloro-1,4-benzoquinone (DCQ) formation (90 s time course) as a function of (<b>a</b>) TCP and (<b>b</b>) H<sub>2</sub>O<sub>2</sub> in the presence of <span class="html-italic">Ao</span>DHP<sub>NS</sub>. Panels (<b>c</b>,<b>d</b>) show rates of DCQ formation (90 s time course) as a function of (<b>c</b>) TCP and (<b>d</b>) H<sub>2</sub>O<sub>2</sub> in the presence of <span class="html-italic">Ao</span>DHP<sub>REC</sub>-A. In these representative single experiments, the following Michaelis–Menten parameters were determined: in panel (<b>a</b>) <span class="html-italic">Ao</span>DHP<sub>NS</sub> [K<sub>M</sub> (TCP) = 69.2 μM, V<sub>MAX</sub> (TCP) = 0.1519 μM s<sup>−1</sup> μg prot<sup>−1</sup>, R<sup>2</sup> = 0.9644] and in panel (<b>b</b>) <span class="html-italic">Ao</span>DHP<sub>NS</sub> [K<sub>M</sub> (H<sub>2</sub>O<sub>2</sub>) = 2.3 μM, V<sub>MAX</sub> (H<sub>2</sub>O<sub>2</sub>) = 0.8198 μM s<sup>−1</sup> μg prot<sup>−1</sup>, R<sup>2</sup> = 0.9773]. In panel (<b>c</b>) <span class="html-italic">Ao</span>DHP<sub>REC</sub>-A [K<sub>M</sub> (TCP) = 5.9 μM, V<sub>MAX</sub> (TCP) = 0.04786 μM s<sup>−1</sup> μg prot<sup>−1</sup>, R<sup>2</sup> = 0.9262] and in panel (<b>d</b>) <span class="html-italic">Ao</span>DHP<sub>REC</sub>-A [K<sub>M</sub> (H<sub>2</sub>O<sub>2</sub>) = 12.1 μM, V<sub>MAX</sub> (H<sub>2</sub>O<sub>2</sub>) = 0.6057 μM s<sup>−1</sup> μg prot<sup>−1</sup>, R<sup>2</sup> = 0.9617].</p>
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23 pages, 3021 KiB  
Article
Increased Levels of hsa-miR-199a-3p and hsa-miR-382-5p in Maternal and Neonatal Blood Plasma in the Case of Placenta Accreta Spectrum
by Angelika V. Timofeeva, Ivan S. Fedorov, Anastasia D. Nikonets, Alla M. Tarasova, Ekaterina N. Balashova, Dmitry N. Degtyarev and Gennady T. Sukhikh
Int. J. Mol. Sci. 2024, 25(24), 13309; https://doi.org/10.3390/ijms252413309 - 11 Dec 2024
Abstract
Despite the increasing number of placenta accreta spectrum (PAS) cases in recent years, its impact on neonatal outcomes and respiratory morbidity, as well as the underlying pathogenetic mechanism, has not yet been extensively studied. Moreover, no study has yet demonstrated the effectiveness of [...] Read more.
Despite the increasing number of placenta accreta spectrum (PAS) cases in recent years, its impact on neonatal outcomes and respiratory morbidity, as well as the underlying pathogenetic mechanism, has not yet been extensively studied. Moreover, no study has yet demonstrated the effectiveness of antenatal corticosteroid therapy (CT) for the prevention of respiratory distress syndrome (RDS) in newborns of mothers with PAS at the molecular level. In this regard, microRNA (miRNA) profiling by small RNA deep sequencing and quantitative real-time PCR was performed on 160 blood plasma samples from preterm infants (gestational age: 33–36 weeks) and their mothers who had been diagnosed with or without PAS depending on the timing of the antenatal RDS prophylaxis. A significant increase in hsa-miR-199a-3p and hsa-miR-382-5p levels was observed in the blood plasma of the newborns from mothers with PAS compared to the control group. A clear trend toward the normalization of hsa-miR-199a-3p and hsa-miR-382-5p levels in the neonatal blood plasma of the PAS groups was observed when CT was administered within 14 days before delivery, but not beyond 14 days. Direct correlations were found among the hsa-miR-382-5p level in neonatal blood plasma and the hsa-miR-199a-3p level in the same sample (r = 0.49; p < 0.001), the oxygen requirements in the NICU (r = 0.41; p = 0.001), the duration of the NICU stay (r = 0.31; p = 0.019), and the severity of the newborn’s condition based on the NEOMOD scale (r = 0.36; p = 0.005). Logistic regression models based on the maternal plasma levels of hsa-miR-199a-3p and hsa-miR-382-5p predicted the need for cardiotonic therapy, invasive mechanical ventilation, or high-frequency oscillatory ventilation in newborns during the early neonatal period, with a sensitivity of 95–100%. According to the literary data, these miRNAs regulate fetal organogenesis via IGF-1, the formation of proper lung tissue architecture, surfactant synthesis in alveolar cells, and vascular tone. Full article
(This article belongs to the Special Issue The Role of miRNA in Human Diseases)
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<p>PLS-A analysis of deep sequencing data of miRNA in the peripheral blood plasma of day-old newborns from mothers with PAS and without PAS (control).</p>
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<p>The dependence of hsa-miR-382-5p and hsa-miR-199a-3p content in the blood plasma of newborns and their mothers on the severity of placenta accreta spectrum (PAS) and the timing of antenatal corticosteroid therapy (CT). Levels of miR-382-5p (−∆Ct, PCR data) in the blood plasma of newborns from mothers with placenta accreta or placenta increta or placenta percreta without CT or with CT 2–14 days before delivery in comparison with control group—without PAS and without CT (<b>A</b>). Levels of miR-382-5p (−∆Ct, PCR data) in the blood plasma of pregnant women with placenta accreta or placenta increta or placenta percreta without CT or with CT 2–14 days before delivery in comparison with control group—without PAS and without CT (<b>B</b>). Levels of miR-199a-3p (−∆Ct, PCR data) in the blood plasma of newborns from mothers with placenta accreta or placenta increta or placenta percreta without CT or with CT 2–14 days before delivery in comparison with control group—without PAS and without CT (<b>C</b>). Levels of miR-199a-3p (−∆Ct, PCR data) in the blood plasma of pregnant women with placenta accreta or placenta increta or placenta percreta without CT or with CT 2–14 days before delivery in comparison with control group—without PAS and without CT (<b>D</b>). “Wo” means “without”.</p>
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<p>Dynamics of changes in hsa-miR-199a-3p levels in the blood plasma of newborns relative to their mothers’ blood plasma, with and without PAS, depending on the antenatal corticosteroid therapy (CT). “Wo” means “without”.</p>
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<p>Levels of hsa-miR-199a-3p and hsa-miR-382-5p in the blood plasma of newborns with PAS, categorized by their severity score according to the Neomod scale.</p>
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<p>Levels of miR-181a-5p, miR-199a-3p and miR-382-5p in blood plasma of pregnant women with/without PAS and with/without antenatal corticosteroid therapy. “Wo” means “without”.</p>
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<p>Logistic regression models for predicting neonatal complications by plasma miR-199a-3p and/or miR-382-5p levels in pregnant women with PAS using miR-181a-5p as a reference endogenous RNA. (<b>A</b>) Respiratory complications probability models. (<b>B</b>) Cardiovascular complications probability models. Se—sensitivity, Sp—specificity.</p>
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<p>Enrichment analysis of gene targets of hsa-miR-382-5p and hsa-miR-199a-3p using FunRich software tool.</p>
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25 pages, 873 KiB  
Review
Extraction Methods, Encapsulation Techniques, and Health Benefits of Astaxanthin
by Ioannis Panagiotakopoulos and Constantina Nasopoulou
Sustainability 2024, 16(24), 10859; https://doi.org/10.3390/su162410859 - 11 Dec 2024
Abstract
Astaxanthin, a red carotenoid pigment found in marine species like microalgae, shrimp, and salmon, is a powerful bioactive molecule with several health effects. Astaxanthin, despite its potential, is highly vulnerable to degradation from external elements, including light, oxygen, and temperature, requiring meticulous extraction [...] Read more.
Astaxanthin, a red carotenoid pigment found in marine species like microalgae, shrimp, and salmon, is a powerful bioactive molecule with several health effects. Astaxanthin, despite its potential, is highly vulnerable to degradation from external elements, including light, oxygen, and temperature, requiring meticulous extraction and stabilization methods. Astaxanthin can be extracted using solvent extraction, ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), supercritical fluid extraction (SFE), and enzymatic extraction. Additionally, encapsulation methods that improve the stability and bioavailability of astaxanthin are examined, highlighting their efficacy in maintaining the chemical in unfavorable conditions. This review discusses the extensive range of astaxanthin’s medicinal capabilities, encompassing its antioxidant, anticancer, antidiabetic, neuroprotective, and skin-protective activities. This research seeks to emphasize the significance of astaxanthin as a functional bioactive component with substantial potential in nutraceutical and pharmaceutical applications by describing the extraction and encapsulation techniques and their health-promoting attributes. Full article
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<p>Chemical structure of astaxanthin.</p>
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<p>Cavitation principle.</p>
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15 pages, 1569 KiB  
Article
Electrochemical Oxidation of Selected Micropollutants from Environment Matrices Using Boron-Doped Diamond Electrodes: Process Efficiency and Transformation Product Detection
by Filip Gamoń, Sebastian Żabczyński, Małgorzata Szopińska, Mattia Pierpaoli, Dawid Zych, Robert Bogdanowicz, Wojciech Artichowicz, Aneta Łuczkiewicz and Sylwia Fudala-Książek
Water 2024, 16(24), 3567; https://doi.org/10.3390/w16243567 - 11 Dec 2024
Abstract
Bisphenol A (BPA) and diclofenac (DCF) are among the most prevalent micropollutants in aquatic environments, with concentrations reaching up to several hundred µg/L. These compounds pose significant risks to biodiversity and environmental health, necessitating the development of effective removal methods. However, both BPA [...] Read more.
Bisphenol A (BPA) and diclofenac (DCF) are among the most prevalent micropollutants in aquatic environments, with concentrations reaching up to several hundred µg/L. These compounds pose significant risks to biodiversity and environmental health, necessitating the development of effective removal methods. However, both BPA and DCF can be resistant to conventional treatment technologies, highlighting the need for innovative approaches. Electrochemical oxidation (EO) has emerged as a promising solution. In this study, we assessed the effectiveness of EO using boron-doped diamond (BDD) anodes to remove BPA and DCF from two types of treated wastewater (TWW-W and TWW-D) and landfill leachate (LL). The evaluation included an analysis of the removal efficiency of BPA and DCF and the identification of transformation products generated during the process. Additionally, the feasibility of the EO-BDD process to remove ammonium nitrogen (N-NH4+) and organic compounds present in these environmental matrices was investigated. The EO-BDD treatment achieved remarkable removal efficiencies, reducing BPA and DCF concentrations by over 96% in LL and TWW-W. Transformation product analyses identified four intermediates formed from parent compounds during the oxidation process. Furthermore, the EO-BDD process effectively removed both chemical oxygen demand (COD) and ammonium nitrogen from LL, although weaker results were observed for TWWs. These findings underscore the potential of the EO-BDD process as an effective method for the removal of BPA and DCF from challenging matrices, such as wastewater containing micropollutants. It also shows promise as a complementary technology for enhancing current conventional wastewater treatment methods, especially biological degradation. Full article
(This article belongs to the Section Wastewater Treatment and Reuse)
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<p>Set for electrolytic oxidation of micropollutants. (<b>A</b>): electrolyser with thermostatic bath (1—electrochemical reactor, 2—power supply, and 3—electromagnetic stirrer) and (<b>B</b>): holder including BDD anode.</p>
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<p>Reactions showing direct and indirect electrochemical oxidation of ammonium compounds in the presence of chloride ions [<a href="#B34-water-16-03567" class="html-bibr">34</a>,<a href="#B35-water-16-03567" class="html-bibr">35</a>].</p>
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16 pages, 2182 KiB  
Article
Enhancements of Triethanolamine CO2 Absorption Rate and Degradation in the Presence of Nickel Nanoparticles Catalysts
by Harold W. Orendi, Kevin Joby and Lidija Šiller
Atmosphere 2024, 15(12), 1479; https://doi.org/10.3390/atmos15121479 (registering DOI) - 11 Dec 2024
Abstract
Here, the catalytic and degradation effect of nickel nanoparticles (NiNPs) on triethanolamine (TEA) with CO2 at 20 °C and 50 °C and a range of TEA concentrations (3–30 wt%) was studied. We show that TEA absorption rate of CO2 can be [...] Read more.
Here, the catalytic and degradation effect of nickel nanoparticles (NiNPs) on triethanolamine (TEA) with CO2 at 20 °C and 50 °C and a range of TEA concentrations (3–30 wt%) was studied. We show that TEA absorption rate of CO2 can be enhanced with NiNPs, the maximum enhancement was 8.3% when compared to a control solution found at 50 °C with 30 wt% TEA alone. Additionally, the time for TEA to be fully loaded with CO2 is reduced; compared to the control, NiNPs enhanced solutions were up to 26.3% faster. Also, to the best of our knowledge, this is the first time the degradation of TEA with NiNPs has been studied. TEA was subject to both oxygen (30 wt%, 55 °C, 0.35 L/min of air, 0.4 molCO2/molTEA, 7.5 mL/min of CO2) and thermal degradation with and without NiNPs (30 wt%, 0.5 molCO2/molTEA, 135 °C). In both degradation experiments, surprisingly, there was no significant difference in TEA degradation in the presence of NiNPs. At high temperature (135 °C), the solution lost 19.2% and 20.3% of the original TEA, with and without NiNPs, respectively. In the presence of oxygen, the solution lost 30.5% and 33.6% of the original TEA, with and without NiNPs, respectively. This indicates that TEA or its mixture with other amines and NiNPs could improve post-combustion CO2 capture. Full article
(This article belongs to the Special Issue Advances in CO2 Capture and Absorption)
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Graphical abstract
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<p>Schematic of the reaction mechanism of hydration of CO<sub>2</sub> by NiNPs, adopted from [<a href="#B17-atmosphere-15-01479" class="html-bibr">17</a>,<a href="#B29-atmosphere-15-01479" class="html-bibr">29</a>].</p>
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<p>Diagram of CO<sub>2</sub> loading apparatus used for the CO<sub>2</sub> capture and oxygen degradation experiments. The thermal degradation experiment uses this apparatus to load the TEA solution before it is placed into an autoclave reactor. See <a href="#sec2dot1-atmosphere-15-01479" class="html-sec">Section 2.1</a>, <a href="#sec2dot2-atmosphere-15-01479" class="html-sec">Section 2.2</a> and <a href="#sec2dot3-atmosphere-15-01479" class="html-sec">Section 2.3</a> for more information.</p>
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<p>Mean percentage CO<sub>2</sub> loading over time at 20 °C. CO<sub>2</sub> is sparged into TEA solution with/without 30 ppm NiNPs at concentrations of TEA (<b>a</b>) 3 wt%, (<b>b</b>) 5 wt%, (<b>c</b>) 7 wt%, (<b>d</b>) 10 wt%, (<b>e</b>), 15 wt%, (<b>f</b>), 20 wt%, and (<b>g</b>) 30 wt%. The horizontal dotted lines show the point at which there is the largest difference in CO<sub>2</sub> loading between the NiNPs and control samples; the time of this is shown with the right vertical dotted line and below in <a href="#atmosphere-15-01479-t001" class="html-table">Table 1</a>. The left vertical dotted line shows the time of the intercept with the NiNPs results line.</p>
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<p>Mean percentage CO<sub>2</sub> loading over time at 50 °C. CO<sub>2</sub> is sparged into TEA solution with/without 30 ppm NiNPs at concentrations of TEA (<b>a</b>) 3 wt%, (<b>b</b>) 5 wt%, (<b>c</b>) 7 wt%, (<b>d</b>) 10 wt%, (<b>e</b>), 15 wt%, (<b>f</b>), 20 wt%, and (<b>g</b>) 30 wt%. The horizontal dotted lines show the point at which there is the largest difference in CO<sub>2</sub> loading between the NiNPs and control samples; the time of this is shown with the right vertical dotted line and below in <a href="#atmosphere-15-01479-t003" class="html-table">Table 3</a>. The left vertical dotted line shows the time of the intercept with the NiNPs results line.</p>
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<p>Comparison of percentage enhancements of CO<sub>2</sub> absorption with NiNPs in TEA at 3–30 wt% concentrations and at 20 °C and 50 °C. See <a href="#app1-atmosphere-15-01479" class="html-app">Supplementary Materials Section S1</a> for the description of how enhancement is calculated.</p>
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<p>Photo of TEA (30 wt%) without and with NiNPs samples that are subjected to oxidative degradation.</p>
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<p>TEA % loss from day 0 due to oxidative degradation of TEA (30 wt%, 0.4 mole CO<sub>2</sub>/mole TEA, 55 °C, 0.35 L/min air, 7.5 mL/min CO<sub>2</sub>) without and with NiNPs over 7 days. The loss of TEA was calculated by the reduction in the size of the GC-MS TEA peak area compared to the TEA peak on day 0. Error bars show mean percentage error.</p>
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<p>Photo of TEA samples without and with NiNPs subjected to thermal degradation (the darker the color, the greater the degradation in the sample).</p>
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<p>TEA % loss from week 0 due to thermal degradation products in TEA (30 wt%, 0.5 mole CO<sub>2</sub>/mole TEA, 135 °C) without and with NiNPs over 5 weeks. The loss of TEA was calculated by the reduction in the size of the GC-MS TEA peak area compared to the TEA peak of day 0. Error bars show mean percentage error.</p>
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14 pages, 5130 KiB  
Article
Sodium Phenylbutyrate Attenuates Cisplatin-Induced Acute Kidney Injury Through Inhibition of Pyruvate Dehydrogenase Kinase 4
by Chang Joo Oh, Wooyoung Choi, Ha Young Lee, In-Kyu Lee, Min-Ji Kim and Jae-Han Jeon
Biomedicines 2024, 12(12), 2815; https://doi.org/10.3390/biomedicines12122815 - 11 Dec 2024
Abstract
Background/Objectives: Cisplatin nephrotoxicity is a significant clinical issue, and currently, no approved drug exists to prevent cisplatin-induced acute kidney injury (AKI). This study investigated whether sodium phenylbutyrate (4-PBA), a chemical chaperone, can prevent cisplatin-induced AKI. Methods: Six consecutive days of intraperitoneal injections of [...] Read more.
Background/Objectives: Cisplatin nephrotoxicity is a significant clinical issue, and currently, no approved drug exists to prevent cisplatin-induced acute kidney injury (AKI). This study investigated whether sodium phenylbutyrate (4-PBA), a chemical chaperone, can prevent cisplatin-induced AKI. Methods: Six consecutive days of intraperitoneal injections of 4-PBA were administered in a murine model before and after the cisplatin challenge. This study evaluated tubular injury, serum blood urea nitrogen (BUN) and creatinine levels, and inflammatory markers such as tumor necrosis factor-alpha (TNF-α) and intercellular adhesion molecule 1 (ICAM-1). Additionally, apoptosis, mitochondrial membrane potential, oxygen consumption ratio, and reactive oxygen species (ROS) were assessed in renal tubular cells. The expression levels of pyruvate dehydrogenase kinase 4 (Pdk4) were also analyzed. Results: 4-PBA prevented tubular injury and normalized serum BUN and creatinine levels. Inflammatory markers TNF-α and ICAM-1 were suppressed. In renal tubular cells, 4-PBA reduced apoptosis, restored mitochondrial membrane potential and oxygen consumption ratio, and reduced ROS production. Mechanistically, 4-PBA suppressed the expression of Pdk4, which is known to be induced during cisplatin-induced renal injury. The protective effect of 4-PBA was abolished in Pdk4-overexpressing renal tubular cells, indicating that the efficacy of 4-PBA partially depends on the suppression of Pdk4 expression. In cancer cells, 4-PBA did not interfere with the anti-cancer efficacy of cisplatin. Conclusions: These findings suggest that 4-PBA effectively prevents cisplatin-induced acute kidney injury by suppressing Pdk4. Full article
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<p>4-PBA attenuates cisplatin-induced acute kidney injury in mice. (<b>A</b>) Hematoxylin and eosin (H&amp;E) and periodic acid–Schiff (PAS) staining of mice kidney sections (original magnification 200×, scale bar = 50 μm). (<b>B</b>) Immunohistochemical staining for NGAL in mice kidney sections (original magnification 200×, scale bar = 100 μm). (<b>C</b>) Tubular injury score based on histopathological analysis of mice kidney sections. (<b>D</b>) Percentage of NGAL-positive area in kidney sections. (<b>E</b>,<b>F</b>) Serum creatinine (<b>E</b>) and blood urea nitrogen (BUN) levels (<b>F</b>) in mice. Data are presented as mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 versus Control (Saline); # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 versus Cisplatin (Saline).</p>
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<p>4-PBA attenuates cisplatin-induced apoptosis in mice and NRK-52E cells. (<b>A</b>,<b>B</b>) TUNEL staining and quantification of TUNEL-positive cells in mice kidney sections (original magnification 200×, scale bar = 50 μm). Arrows indicate TUNEL-positive areas. (<b>C</b>,<b>D</b>) Protein expression levels of cleaved caspase-3 in mice kidneys and the corresponding quantitative analysis. (<b>E</b>) Morphological changes in NRK-52E cells with or without cisplatin and 4-PBA treatment (original magnification 40×, scale bar = 500 μm). (<b>F</b>) Time-dependent and (<b>G</b>) 4-PBA dose-dependent protein expression of cleaved caspase-3 in NRK-52E cells. (<b>H</b>) Quantification of cleaved caspase-3 levels in NRK-52E cells. Data are presented as mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 versus Control (Saline); # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 versus Cisplatin (Saline).</p>
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<p>4-PBA attenuates cisplatin-induced inflammation in mice. (<b>A</b>) Immunohistochemical staining for TNF-α, p65, and ICAM-1 in mice kidney sections (original magnification 200×, scale bar = 100 μm). (<b>B</b>–<b>D</b>) Quantification of TNF-α, p65, and ICAM-1 positive areas in kidney sections. (<b>E</b>,<b>F</b>) Relative mRNA expression levels of TNF-α and ICAM-1 in kidney tissues. Data are presented as mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.01 versus Control (Saline); # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 versus Cisplatin (Saline).</p>
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<p>4-PBA attenuates cisplatin-induced mitochondrial damage in mice and NRK-52E cells. (<b>A</b>) Nitrotyrosine (NT) staining in mice kidney sections (original magnification 200×, scale bar = 100 μm). (<b>B</b>) Quantification of nitrotyrosine-positive areas in kidney sections. (<b>C</b>–<b>E</b>) Mitochondrial ROS (MitoSOX) and membrane potential (TMRE) in NRK-52E cells (original magnification 200×, scale bar = 250 μm). (<b>F</b>) Mitochondrial oxygen consumption rate (OCR) and (<b>G</b>) ATP production in NRK-52E cells. (<b>H</b>) Protein expression levels of mitochondrial complexes IV and V, and PGC-1α in kidney tissues. (<b>I</b>) Quantification of Complex IV, V, and PGC-1α in kidney tissues. Data are presented as mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 versus Control (Saline); # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 versus Cisplatin (Saline).</p>
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<p>The renoprotective effect of 4-PBA is mediated by the inhibition of Pdk4 in mice and NRK-52E cells. (<b>A</b>) Immunohistochemical staining for Pdk4 in mice kidney sections (original magnification 200×, scale bar = 30 μm). (<b>B</b>) Quantification of Pdk4-positive areas in kidney sections. (<b>C</b>) Gene expression was analyzed by measuring variations in <span class="html-italic">Pdk</span> mRNA levels. (<b>D</b>) Quantification of relative mRNA expressions in <span class="html-italic">Pdk1</span>, <span class="html-italic">Pdk2</span>, <span class="html-italic">Pdk3</span>, and <span class="html-italic">Pdk4</span>. (<b>E</b>) Protein expression levels of Pdk subtypes in mice kidneys. (<b>F</b>) Quantitative analysis of protein expression levels of Pdk subtypes in mice kidneys. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 versus Control (Saline); # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.01 versus Cisplatin (Saline). (<b>G</b>) Protein expression levels of Pdk4 and β-tubulin in Ad-Pdk4 and Ad-GFP. (<b>H</b>) Protein expression levels of cleaved caspase-3 and β-tubulin in NRK-52E cells. (<b>I</b>) Mitochondrial oxygen consumption rate (OCR) and (<b>J</b>) ATP production in NRK-52E cells. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 versus Control; ### <span class="html-italic">p</span> &lt; 0.001 versus Cisplatin.</p>
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<p>4-PBA does not inhibit the anti-cancer effect in MBT-2 and DU145 cells. (<b>A</b>,<b>B</b>) Cell survival ratio of MBT-2 and DU145 cells with 4-PBA and cisplatin (8 μM) treatment. * <span class="html-italic">p</span> &lt; 0.05 versus control (0). (<b>C</b>) Diagram of the proposed mechanism.</p>
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18 pages, 2114 KiB  
Article
Synthesis of Oxazoles Containing CF3-Substituted Alcohol Unit via Tandem Cycloisomerization/Hydroxyalkylation from N-Propargylamides with Trifluoropyruvates
by Juan-Juan Gao, Long-Hui Wu, Shu-Qin Yu, Xue Zhu, Yu Zeng, Kai Yang and Zhao-Yang Wang
Molecules 2024, 29(24), 5848; https://doi.org/10.3390/molecules29245848 - 11 Dec 2024
Abstract
Oxazoles are important five-membered heterocycles that contain both nitrogen and oxygen atoms. Due to their wide range of biological activities, many oxazoles demonstrate potential for extensive application in various fields, including medicinal chemistry. Trifluoromethyl carbinol, an important pharmacophore, contains both trifluoromethyl and hydroxyl [...] Read more.
Oxazoles are important five-membered heterocycles that contain both nitrogen and oxygen atoms. Due to their wide range of biological activities, many oxazoles demonstrate potential for extensive application in various fields, including medicinal chemistry. Trifluoromethyl carbinol, an important pharmacophore, contains both trifluoromethyl and hydroxyl groups and is common in molecules with important biological activities. Constructing oxazoles that contain a trifluoromethyl carbinol unit is undoubtedly important and valuable for expanding the chemical space in drug discovery. In this study, a simple and efficient method was developed for the synthesis of oxazoles containing a CF3-substituted alcohol unit via the tandem cycloisomerization/hydroxyalkylation of N-propargylamides with trifluoropyruvates through a rational Lewis acid catalytic mechanism. This Zn(OTf)2-catalyzed synthetic protocol is operationally simple and provides a series of oxazoles in moderate to good yields. The protocol demonstrates broad substrate scope, high functional group tolerance, and high atom economy and can achieve gram-level reactions, indicating the strong possibility of its practical application. Full article
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Figure 1
<p>Examples of bioactive molecules containing trifluoromethyl carbinol.</p>
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<p>Examples of natural products and biologically active molecules containing an oxazole unit.</p>
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<p>Typical examples of synthesis methods using tandem reactions to obtain fluorinated oxazoles.</p>
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<p>Control experiments: (<b>a</b>) Explore whether there is a free radical pathway. (<b>b</b>) Synthesis of <b>4a</b> catalyzed by FeCl<sub>3</sub>. (<b>c</b>) Synthesis of <b>5a</b> catalyzed by ZnI<sub>2</sub>. (<b>d</b>) Explore whether <b>4a</b> is an intermediate in the reaction. (<b>e</b>) Explore whether <b>5a</b> is an intermediate in the reaction.</p>
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<p>Proposed mechanisms.</p>
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