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14 pages, 3332 KiB  
Article
Myrosin Cells and Myrosinase Expression Pattern in Nasturtium (Tropaeolum majus L.)
by Ivana Restović, Nives Kević, Laura Kurić, Ivana Bočina, Elma Vuko and Ivana Vrca
Agronomy 2024, 14(9), 2108; https://doi.org/10.3390/agronomy14092108 - 16 Sep 2024
Abstract
Plants from the Brassicales order are known for the presence of a glucosinolate–myrosinase link, which is an important protection strategy against multiple stressors. The main goal of this study was to investigate the presence of the myrosinase enzyme and reveal the myrosin cell [...] Read more.
Plants from the Brassicales order are known for the presence of a glucosinolate–myrosinase link, which is an important protection strategy against multiple stressors. The main goal of this study was to investigate the presence of the myrosinase enzyme and reveal the myrosin cell ultrastructure in the vegetative organs of nasturtium. The presence, localisation and expression of the enzyme myrosinase type 1 (TGG1) at different developmental stages of Tropaeolum majus L. (nasturtium) were investigated using immunohistochemical and immunofluorescent techniques. The expression of myrosinase was detected in the vegetative organs of T. majus. During plant development, within four consecutive weeks, a decrease in myrosinase expression was noticed in all studied plant organs. The location of greater myrosinase accumulation and activity is shown to be the root, contrary to the nasturtium stem and leaf, where we found the lowest myrosinase expression. Transmission electron microscopy was used to reveal the ultrastructural features of the myrosin cells of nasturtium. Myrosin cells are usually scattered between parenchyma cells and S-cells. Mostly, they are rectangular or slightly elongated in shape and can be recognised by an electron-dense large central vacuole and an expanded rough endoplasmic reticulum. The results of this study provide new data on myrosin cell morphology and the expression pattern of myrosinase in T. majus. Full article
(This article belongs to the Section Plant-Crop Biology and Biochemistry)
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Figure 1

Figure 1
<p><span class="html-italic">Tropaeolum majus</span> L. sample growth in four consecutive weeks. (<b>a</b>) The samples measured in the first phase (P1) of the study: 19-day-old plants; (<b>b</b>) The samples measured in the second phase (P2) of the study: 26-day-old plants; (<b>c</b>) The samples measured in the third phase (P3): 33-day-old plants; (<b>d</b>) Morphological difference in the leaves, with older jagged-shaped leaves and younger round-shaped leaves; (<b>e</b>) The samples measured in the fourth phase (P4): 40-day-old plants; the morphological difference in the stem with some of the stems having turned brown, while most of the stems are still the usual light green (arrow).</p>
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<p>Results of sample measurements for four consecutive experimental weeks of sampling. A one-way ANOVA test was used for statistical data processing, after which Tukey’s multiple comparisons test was used to examine the difference in the (<b>a</b>) root height, (<b>b</b>) stem height, (<b>c</b>) leaf length and (<b>d</b>) total plant height in the four developmental stages. The results are presented as mean (n = 5) ± standard deviation (SD). Significance was set at * <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, and **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Immunofluorescence staining of myrosinase in sections of the vegetative nasturtium (<span class="html-italic">Tropaeolum majus</span> L.) organs using TGG1 myrosinase 1 rabbit primary antibody. Expression of the enzyme myrosinase type 1 (arrows) in the parenchyma cells of the root (<b>a</b>–<b>d</b>), stem (<b>e</b>–<b>h</b>) and leaf (<b>i</b>–<b>l</b>) is shown during four different developmental phases (P1–P4). Localisation of myrosinase type 1 in the meristematic zone of a young root (panel (<b>a</b>)) and the cells of the vascular tissue of the root (panels b, d) and the stem (panel (<b>e</b>–<b>h</b>)). Cells of the ground tissue in the vascular part of the leaf (panels (<b>i</b>–<b>l</b>)). Transverse (<b>a</b>,<b>c</b>,<b>e</b>–<b>g</b>,<b>i</b>–<b>l</b>) and longitudinal sections (<b>b</b>,<b>d</b>,<b>h</b>). pd—protoderm; gm—ground meristem; xyl—xylem; phl—phloem; arrows—myrosinase expression.</p>
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<p>Statistical analysis of the expression pattern of myrosinase type 1 in the root, stem and leaf at different developmental stages after plants sprout. Myrosinase expression was quantified by measuring the percentage of immunoreactive cell areas (% area). Two-way ANOVA and Tukey’s multiple comparison test were used for statistical analyses. Statistically significant differences were set as * <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; and **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Immunohistochemical staining of myrosinase in nasturtium tissue. Presence of myrosinase type 1 (brown colour) was observed in the root area (<b>a</b>–<b>c</b>), the vascular elements of the stem (<b>d</b>), the leaf parenchyma tissue (<b>e</b>) and the xylem vessels (<b>f</b>). P1—Phase 1; P2—Phase 2; P3—Phase 3; P4—Phase 4; arrows—myrosin cells; p phl—phloem parenchyma cells; S—S cell; arrowhead—parenchyma cells; xyl—xylem; phl—phloem.</p>
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<p>Ultrastructure of myrosin cells in the root (<b>a</b>,<b>b</b>), the stem (<b>c</b>,<b>d</b>) and the leaf (<b>e</b>,<b>f</b>) in the first phase of development. Myrosin cells (M) differ from the surrounding S-cells by their higher electron density and the presence of one large and several smaller vacuoles. S-cells (S) surround the myrosin cells and can be recognised by the very thin layer of cytoplasm and the large volume of the central vacuole. M—myrosin cell; S—S-cell; V—vacuole; ve—vessel elements; mg—myrosin granule; rER—rough endoplasmic reticulum; G—Golgi apparatus; m—mitochondrion; cw—cell wall; n—nucleus; ch—chloroplast; PPC—phloem parenchyma cell; *—protein storage vesicle; arrow—plasmodesmal opening; CC—companion cell; ST—sieve tube element.</p>
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15 pages, 7531 KiB  
Article
Synthesis, Characterization, and Evaluation of the Antimicrobial and Anticancer Activities of Zinc Oxide and Aluminum-Doped Zinc Oxide Nanocomposites
by Muhammad Asif, Muhammad Fakhar-e-Alam, Muhammad Tahir, Farah Jamil, Hassan Sardar, Javed Rehman and Kholood A. Dahlous
Pharmaceuticals 2024, 17(9), 1216; https://doi.org/10.3390/ph17091216 - 16 Sep 2024
Viewed by 139
Abstract
In this research, we developed undoped and aluminum-doped zinc oxide for antimicrobial and anticancer activities. This study focuses on the synthesis, characterization, and biological activities of zinc oxide nanoparticles (ZnO NPs) and aluminum-doped zinc oxide nanocomposites (Zn1−xAlxO NCs) at [...] Read more.
In this research, we developed undoped and aluminum-doped zinc oxide for antimicrobial and anticancer activities. This study focuses on the synthesis, characterization, and biological activities of zinc oxide nanoparticles (ZnO NPs) and aluminum-doped zinc oxide nanocomposites (Zn1−xAlxO NCs) at varying concentrations (x = 0, 0.25, 0.5, and 0.75 wt%) using the coprecipitation method. Various characterization techniques such as XRD, UV-Vis, FTIR, EDX, and SEM were performed to analyze the crystal structure, optical properties, functional group identification, elemental composition, and surface morphology. The antimicrobial activity test showed that Zn0.75Al0.25O NCs exhibited the strongest inhibition zone against Bacillus cereus compared to Staphylococcus aureus > Pasteurella multocida > Escherichia coli. Moreover, the cytotoxicity and cell viability of liver cancer (HepG-2), breast cancer (MCF-7), ovarian cancer (SKOV3), and normal liver cell lines) were evaluated using the MTT assay, demonstrating that Zn0.75Al0.25O NCs not only enhance cell destruction but also show low cytotoxicity and high biocompatibility at low concentrations. These results suggest that Zn0.75Al0.25O NCs could be a promising candidate for in vivo anticancer applications and should be further investigated. Full article
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Figure 1

Figure 1
<p>XRD spectrum of ZnO NPs and Al-doped ZnO NCs at varying concentrations 0.25%, 0.5%, and 0.75%.</p>
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<p>FTIR spectrum of ZnO NPs and Al-doped ZnO NCs at varying concentrations 0.25%, 0.5%, and 0.75%.</p>
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<p>UV-visible spectrums of ZnO NPs and Al-doped ZnO NCs at varying concentrations 0.25%, 0.5%, and 0.75%.</p>
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<p>SEM results of (<b>a</b>) ZnO NPs and Al-doped ZnO NCs at varying concentrations (<b>b</b>) 0.25%, (<b>c</b>) 0.5%, and (<b>d</b>) 0.75%.</p>
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<p>Zeta potential of ZnO NPs and Al-doped ZnO NCs at varying concentrations 0.25%, 0.5%, and 0.75%.</p>
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<p>EDX spectrum of (<b>a</b>) ZnO NPs and Al-doped ZnO NCs at varying concentrations (<b>b</b>) 0.25%, (<b>c</b>) 0.5%, and (<b>d</b>) 0.75%.</p>
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<p>EDX spectrum of (<b>a</b>) ZnO NPs and Al-doped ZnO NCs at varying concentrations (<b>b</b>) 0.25%, (<b>c</b>) 0.5%, and (<b>d</b>) 0.75%.</p>
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<p>The antimicrobial activity of samples DMSO (control), standard antibiotic (cephalosporin), M1 (ZnO NPs), M2 (Zn<sub>0.75</sub>Al<sub>0.25</sub>O), M3 (Zn<sub>0.5</sub>Al<sub>0.5</sub>O), and M4 (Zn<sub>0.25</sub>Al<sub>0.75</sub>O NCs) against <span class="html-italic">S. aureus</span>, <span class="html-italic">B. cereus</span>, <span class="html-italic">P. multocida</span>, and <span class="html-italic">E. coli</span>.</p>
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<p>The antimicrobial activity of DMSO (control), standard antibiotic (cephalosporin), ZnO NPs, Zn<sub>0.75</sub>Al<sub>0.25</sub>O NCs, Zn<sub>0.5</sub>Al<sub>0.5</sub>O NCs, and Zn<sub>0.25</sub>Al<sub>0.75</sub>O NCs at 40 mg/mL, where * is significance for <span class="html-italic">p</span> ≤ 0.05 and ** is highly significance for <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>The antimicrobial activity of Zn<sub>0.75</sub>Al<sub>0.25</sub>O NCs at 150 mg/mL, where * is significance for <span class="html-italic">p</span> ≤ 0.05 and ** is highly significance for <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>The anticancer activity of samples M1 (ZnO NPs), M2 (Zn<sub>0.75</sub>Al<sub>0.25</sub>O), M3 (Zn<sub>0.5</sub>Al<sub>0.5</sub>O), and M4 (Zn<sub>0.25</sub>Al<sub>0.75</sub>O NCs) against HepG-2, MCF-7, SKOV3 cancer cell lines, and normal liver cell line, where * is significance for <span class="html-italic">p</span> ≤ 0.05 and ** is highly significance for <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>% cell viability of Zn<sub>0.75</sub>Al<sub>0.25</sub>O NCs at varying concentration (μg/mL) against HepG-2, MCF-7, SKOV3, and normal liver cell line, where * is significance for <span class="html-italic">p</span> ≤ 0.05 and ** is highly significance for <span class="html-italic">p</span> ≤ 0.01.</p>
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<p>Synthesis process of ZnO NPs and Al-doped ZnO NCs.</p>
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16 pages, 6432 KiB  
Article
Enhanced Antibacterial Activity of Carbon Dots: A Hybrid Approach with Levofloxacin, Curcumin, and Tea Polyphenols
by Khurram Abbas, Haimei Zhu, Weixia Qin, Meiyan Wang, Zijian Li and Hong Bi
C 2024, 10(3), 84; https://doi.org/10.3390/c10030084 (registering DOI) - 15 Sep 2024
Viewed by 250
Abstract
Bacterial infections and their increasing resistance to antibiotics pose a significant challenge in medical treatment. This study presents the synthesis and characterization of novel carbon dots (CDs) using levofloxacin (Lf), curcumin (Cur), and tea polyphenols (TP) through a facile hydrothermal method. The synthesized [...] Read more.
Bacterial infections and their increasing resistance to antibiotics pose a significant challenge in medical treatment. This study presents the synthesis and characterization of novel carbon dots (CDs) using levofloxacin (Lf), curcumin (Cur), and tea polyphenols (TP) through a facile hydrothermal method. The synthesized curcumin-tea polyphenol@carbon dots (Cur-TP@CDs) and levofloxacin-tea polyphenol@carbon dots (Lf-TP@CDs) were characterized using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy, confirming their unique structural and chemical properties. Cur-TP@CDs exhibited an average particle size of 1.32 nanometers (nm), while Lf-TP@CDs averaged 1.58 nm. Both types demonstrated significant antibacterial activity, with Lf-TP@CDs showing superior effectiveness against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in broth dilution and disc diffusion assays. Biofilm inhibition assays revealed a significant reduction in biofilm formation at higher concentrations. The ultraviolet-visible (UV-vis) and photoluminescence (PL) spectral analyses indicated efficient photon emission, and electron paramagnetic resonance (EPR) analysis showed increased singlet oxygen generation, enhancing bactericidal effects. Live and dead bacterial staining followed by scanning electron microscopy (SEM) analysis confirmed dose-dependent bacterial cell damage and morphological deformities. These findings suggest that Cur-TP@CDs and Lf-TP@CDs are promising antibacterial agents, potentially offering a novel approach to combat antibiotic-resistant bacterial infections. Full article
(This article belongs to the Special Issue Carbon Nanohybrids for Biomedical Applications)
16 pages, 8326 KiB  
Article
Cytoarchitecture of Breast Cancer Cells under Diabetic Conditions: Role of Regulatory Kinases—Rho Kinase and Focal Adhesion Kinase
by Diganta Dutta, Matthew Ziemke, Payton Sindelar, Hernan Vargas, Jung Yul Lim and Surabhi Chandra
Cancers 2024, 16(18), 3166; https://doi.org/10.3390/cancers16183166 - 15 Sep 2024
Viewed by 224
Abstract
Diabetes greatly reduces the survival rates in breast cancer patients due to chemoresistance and metastasis. Reorganization of the cytoskeleton is crucial to cell migration and metastasis. Regulatory cytoskeletal protein kinases such as the Rho kinase (ROCK) and focal adhesion kinase (FAK) play a [...] Read more.
Diabetes greatly reduces the survival rates in breast cancer patients due to chemoresistance and metastasis. Reorganization of the cytoskeleton is crucial to cell migration and metastasis. Regulatory cytoskeletal protein kinases such as the Rho kinase (ROCK) and focal adhesion kinase (FAK) play a key role in cell mobility and have been shown to be affected in cancer. It is hypothesized that diabetes/high-glucose conditions alter the cytoskeletal structure and, thus, the elasticity of breast cancer cells through the ROCK and FAK pathway, which can cause rapid metastasis of cancer. The aim of the study was to investigate the role of potential mediators that affect the morphology of cancer cells in diabetes, thus leading to aggressive cancer. Breast cancer cells (MDA-MB-231 and MCF-7) were treated with 5 mM glucose (low glucose) or 25 mM glucose (high glucose) in the presence of Rho kinase inhibitor (Y-27632, 10 mM) or FAK inhibitor (10 mM). Cell morphology and elasticity were monitored using atomic force microscopy (AFM), and actin staining was performed by fluorescence microscopy. For comparative study, normal mammary breast epithelial cells (MCF-10A) were used. It was observed that high-glucose treatments modified the cytoskeleton of the cells, as observed through AFM and fluorescence microscopy, and significantly reduced the elasticity of the cells. Blocking the ROCK or FAK pathway diminished the high-glucose effects. These changes were more evident in the breast cancer cells as compared to the normal cells. This study improves the knowledge on the cytoarchitecture properties of diabetic breast cancer cells and provides potential pathways that can be targeted to prevent such effects. Full article
(This article belongs to the Special Issue Application of Imaging in Breast Cancer)
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Figure 1
<p>Atomic force microscopy of triple negative breast cancer cells (MDA-MB-231) treated with varying concentrations of glucose. Cells treated with (<b>a</b>) low glucose 5 mM and (<b>b</b>) high glucose 25 mM.</p>
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<p>Atomic force microscopy of triple negative breast cancer cells (MDA-MB-231) treated with varying concentrations of glucose in presence of Rho kinase inhibitor Y-27,632 (Y, 10 mM) for 24 h. Cells treated with (<b>a</b>) low glucose (5 mM) in combination with Y (10 mM) and (<b>b</b>) high glucose (25 mM) in combination with Y (10 mM).</p>
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<p>Atomic force microscopy of triple negative breast cancer cells (MDA-MB-231) treated with varying glucose in presence of FAK inhibitor (F, 10 mM) for 24 h. Cells treated with low glucose (5 mM) in combination with F (10 mM). The red dotted square highlights a region further magnified to the right.</p>
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<p>Atomic force microscopy of breast cancer cells (MCF-7) treated with varying concentrations of glucose for 24 h. Cells treated with (<b>a</b>) low glucose 5 mM and (<b>b</b>) high glucose 25 mM.</p>
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<p>Atomic force microscopy of breast cancer cells (MCF-7) treated with varying concentrations of glucose in presence of Rho kinase inhibitor Y-27632 (Y, 10 mM) for 24 h. Cells treated with (<b>a</b>) low glucose (5 mM) in combination with Y (10 mM) and (<b>b</b>) high glucose (25 mM) in combination with Y (10 mM).</p>
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<p>Atomic force microscopy of normal mammary epithelial cells (MCF-10A) treated with varying concentrations of glucose for 24 h. Cells treated with (<b>a</b>) low glucose 5 mM and (<b>b</b>) high glucose 25 mM.</p>
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<p>Modulus of elasticity of MDA-MB-231 cells treated with varying concentrations of glucose in the presence of the Y compound. Cells were treated for 24 h with low glucose (5 mM, 5G), high glucose (25 mM, 25G), low glucose with inhibitor (5G + Y), and high glucose with inhibitor (25G + Y). N = 28~75.</p>
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<p>Modulus of elasticity of MDA-MB-231 cells treated with varying concentrations of glucose in the presence of the F compound. Cells were treated for 24 h with low glucose (5 mM, 5G), high glucose (25 mM, 25G), low glucose with inhibitor (5G + F), and high glucose with inhibitor (25G + F). N = 50~83.</p>
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<p>Modulus of elasticity of MCF-7 cells treated with varying concentrations of glucose in the presence of the F compound. Cells were treated for 24 h with low glucose (5 mM, 5G), high glucose (25 mM, 25G), low glucose with inhibitor (5G + F), and high glucose with inhibitor (25G + F). N = 40~50.</p>
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<p>Modulus of elasticity of MCF-10A cells treated with varying concentrations of glucose in the presence of the Y compound. Cells were treated for 24 h with low glucose (5 mM, 5G), high glucose (25 mM, 25G), low glucose with inhibitor (5G + Y), and high glucose with inhibitor (25G + Y). N = 78~38.</p>
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<p>Actin staining of MDA-MB-231 cells treated with varying concentrations of glucose in the presence of the F compound. Cells were treated for 24 h with (<b>a</b>) low glucose (5 mM, 5G), (<b>b</b>) high glucose (25 mM, 25G), (<b>c</b>) low glucose with inhibitor (5G + F), and (<b>d</b>) high glucose with inhibitor (25G + F).</p>
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17 pages, 2502 KiB  
Article
Impact of Uniaxial Static Strain on Myoblast Differentiation in Collagen-Coated PCL Microfilament Scaffolds: Role of Onset Time of Mechanical Stimulation
by María Laura Espinoza-Álvarez, Laura Rojas-Rojas, Johan Morales-Sánchez and Teodolito Guillén-Girón
Bioengineering 2024, 11(9), 919; https://doi.org/10.3390/bioengineering11090919 - 13 Sep 2024
Viewed by 367
Abstract
Tissue engineering endeavors to create in vitro constructs that replicate the properties of native tissue, such as skeletal muscle. This study investigated the use of mechanical stimulation to promote myogenic differentiation and enhance the functionality of bioengineered tissues. Specifically, it aimed to facilitate [...] Read more.
Tissue engineering endeavors to create in vitro constructs that replicate the properties of native tissue, such as skeletal muscle. This study investigated the use of mechanical stimulation to promote myogenic differentiation and enhance the functionality of bioengineered tissues. Specifically, it aimed to facilitate the differentiation of myoblasts within a three-dimensional scaffold using a defined pattern of mechanical stimulation. C2C12 cells were cultured on a collagen-coated PCL microfilament scaffold and subjected to 24 h of uniaxial static strain using a biomechanical stimulation system. Two onset times of stimulation, 72 h and 120 h post-seeding, were evaluated. Cell proliferation, myogenic marker expression, and alterations in cell morphology and orientation were assessed. Results indicate that static strain on the scaffold promoted myoblast differentiation, evidenced by morphological and molecular changes. Notably, strain initiated at 72 h induced an early differentiation stage marked by MyoD expression, whereas stimulation beginning at 120 h led to a mid-stage differentiation characterized by the co-expression of MyoD and Myogenin, culminating in myotube formation. These results highlight the critical influence of myoblast maturity at the time of strain application on the differentiation outcome. This study provides insights that could guide the optimization of mechanical stimulation protocols in tissue engineering applications. Full article
(This article belongs to the Special Issue Mechanobiology in Biomedical Engineering)
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Graphical abstract

Graphical abstract
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<p>(<b>a</b>) Diagram of the horizontal microfilament grip. (<b>b</b>) Diagram of the bioreactor. (<b>c</b>) Diagram of the biomechanical stimulation system.</p>
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<p>Loading pattern used on the cell-seeded scaffold.</p>
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<p>Stress–strain curve of microfilaments during mechanical stimulation. Onset time of mechanical stimulation was 72 h. Colored lines represent different replicas of the experiment.</p>
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<p>Proliferation and immunofluorescence of mechanically stimulated cells after 72 h of culture. (<b>a</b>) Cell proliferation percentage before and after mechanical stimulation. Error bars are standard deviation. * <span class="html-italic">p</span> ≤ 0.05. (<b>b</b>) Immunofluorescent staining after mechanical stimulation. Merge shows the overlay of DAPI (blue), Phalloidin (green), and MyoD/Myogenin (red) 20×. (<b>c</b>) Nuclei elongation and alignment after mechanical stimulation. DAPI staining (blue). Electronically zoomed-in micrograph, original magnification in microscope 20×. (<b>d</b>) Expression of MyoD myogenic marker after mechanical stimulation (red). Electronically zoomed-in micrograph, original magnification in microscope 20×.</p>
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<p>Stress–strain curve of microfilaments during mechanical stimulation. Onset time of mechanical stimulation was 120 h. Colored lines represent different replicas of the experiment.</p>
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<p>Proliferation and immunofluorescence of mechanically stimulated cells after 120 h of culture. (<b>a</b>) Cell proliferation percentage before and after mechanical stimulation. Error bars are standard deviation. * <span class="html-italic">p</span> ≤ 0.05. (<b>b</b>) Immunofluorescent staining after mechanical stimulation. Merge shows the overlay of DAPI (blue), Phalloidin (green), and MyoD/Myogenin (red). 20×. (<b>c</b>) Expression of MyoD myogenic marker after mechanical stimulation (red). Electronically zoomed-in micrograph, original magnification in microscope 20×. (<b>d</b>) Expression of Myogenin myogenic marker after mechanical stimulation (red). Electronically zoomed-in micrograph, original magnification in microscope 20×.</p>
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<p>Multinucleated cells on the strained scaffold. Onset time of mechanical stimulation was 120 h. DAPI (blue), Phalloidin (green), Myogenin (red). Electronically zoomed-in micrograph, original magnification in microscope is 20×.</p>
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17 pages, 16291 KiB  
Article
Tumor Heterogeneity in Gastrointestinal Cancer Based on Multimodal Data Analysis
by Dongmei Ai, Yang Du, Hongyu Duan, Juan Qi and Yuduo Wang
Genes 2024, 15(9), 1207; https://doi.org/10.3390/genes15091207 - 13 Sep 2024
Viewed by 279
Abstract
Background: Gastrointestinal cancer cells display both morphology and physiology diversity, thus posing a significant challenge for precise representation by a single data model. We conducted an in-depth study of gastrointestinal cancer heterogeneity by integrating and analyzing data from multiple modalities. Methods: We used [...] Read more.
Background: Gastrointestinal cancer cells display both morphology and physiology diversity, thus posing a significant challenge for precise representation by a single data model. We conducted an in-depth study of gastrointestinal cancer heterogeneity by integrating and analyzing data from multiple modalities. Methods: We used a modified Canny algorithm to identify edges from tumor images, capturing intricate nonlinear interactions between pixels. These edge features were then combined with differentially expressed mRNA, miRNA, and immune cell data. Before data integration, we used the K-medoids algorithm to pre-cluster individual data types. The results of pre-clustering were used to construct the kernel matrix. Finally, we applied spectral clustering to the fusion matrix to identify different tumor subtypes. Furthermore, we identified hub genes linked to these subtypes and their biological roles through the application of Weighted Gene Co-expression Network Analysis (WGCNA) and Gene Ontology (GO) enrichment analysis. Results: Our investigation categorized patients into three distinct tumor subtypes and pinpointed hub genes associated with each. Genes MAGI2-AS3, MALAT1, and SPARC were identified as having a differential impact on the metastatic and invasive capabilities of cancer cells. Conclusion: By harnessing multimodal features, our study enhances the understanding of gastrointestinal tumor heterogeneity and identifies biomarkers for personalized medicine and targeted treatments. Full article
(This article belongs to the Section Bioinformatics)
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Figure 1
<p>Significantly different characteristics between cancer and normal samples: (<b>A</b>) top 10 most significant mRNAs; (<b>B</b>) most significant differential miRNAs; (<b>C</b>) differential boxplots of 22 immune cells.</p>
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<p>Heat maps of the top 20 significantly differentially expressed genes (DEGs): (<b>A</b>) mRNA expression heat map; (<b>B</b>) miRNA expression heat map; (<b>C</b>) mRNA expression heat map after taking the mean for the corresponding feature of the same subtype sample; (<b>D</b>) miRNA expression heat map after taking the mean for the corresponding feature of the same subtype sample.</p>
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<p>Six features with the most significant differences among the three subtypes: (<b>A</b>) Top six most significantly differentially expressed mRNAs; (<b>B</b>) Top six most significantly differentially expressed miRNAs.</p>
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<p>Immune cell characteristics of different subtypes: (<b>A</b>) Heat map of immune cell characteristics of samples of the same subtype after taking the mean value; (<b>B</b>) Difference in the percentage of immune cells in samples of different subtypes.</p>
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<p>Results of Weighted Gene Co-expression Network Analysis (WGCNA): (<b>A</b>) Scale-Free Topology Analysis, frequency distribution of the number of connections (i.e., node degree, k) in the network (<b>left</b>), and a test of the scale-independent nature of the network (<b>right</b>); (<b>B</b>) Clustering of Module Eigengenes; (<b>C</b>) Gene Dendrogram and Module Colors, different colors represent different modules; (<b>D</b>) Module Eigengene Correlation Heatmap.</p>
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<p>Correlation of different modules with different subtypes.</p>
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<p>GO analysis results: (<b>A</b>) subtype I (<b>B</b>) subtype III.</p>
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13 pages, 12746 KiB  
Article
Characterization and Pathogenicity of Colletotrichum truncatum Causing Hylocereus undatus Anthracnose through the Changes of Cell Wall-Degrading Enzymes and Components in Fruits
by Shuwu Zhang, Yun Liu, Jia Liu, Enchen Li and Bingliang Xu
J. Fungi 2024, 10(9), 652; https://doi.org/10.3390/jof10090652 - 13 Sep 2024
Viewed by 311
Abstract
Anthracnose is one of the destructive diseases of pitaya that seriously affects the plant growth and fruit quality and causes significant yield and economic losses worldwide. However, information regarding the species of pathogens that cause anthracnose in pitaya (Hylocereus undatus) fruits [...] Read more.
Anthracnose is one of the destructive diseases of pitaya that seriously affects the plant growth and fruit quality and causes significant yield and economic losses worldwide. However, information regarding the species of pathogens that cause anthracnose in pitaya (Hylocereus undatus) fruits in Gansu Province, China, and its pathogenic mechanism is unknown. Thus, the purposes of our present study were to identify the species of pathogens causing H. undatus fruits anthracnose based on the morphological and molecular characteristics and determine its pathogenic mechanism by physiological and biochemical methods. In our present study, forty-six isolates were isolated from the collected samples of diseased H. undatus fruits and classified as three types (named as H-1, H-2, and H-3), according to the colony and conidium morphological characteristics. The isolation frequencies of H-1, H-2, and H-3 types were 63.04%, 21.74%, and 15.22%, respectively. The representative single-spore isolate of HLGTJ-1 in H-1 type has significant pathogenicity, and finally we identified Colletotrichum truncatum as the pathogen based on the morphological characteristics as well as multi-locus sequence analysis. Moreover, the H. undatus fruits inoculated with C. truncatum had a significantly increased activity of cell wall-degrading enzymes (CWDEs) cellulase (Cx), β-glucosidase (β-Glu), polygalacturonase (PG), and pectin methylgalacturonase (PMG), while having a decreased level of cell wall components of original pectin and cellulose in comparison to control. The average increased activities of Cx, β-Glu, PG, and PMG were 30.73%, 40.40%, 51.55%, and 32.23% from day 0 to 6 after inoculation, respectively. In contrast, the average decreased contents of original pectin and cellulose were 1.82% and 16.47%, respectively, whereas the average increased soluble pectin content was 38.31% in comparison to control. Our results indicate that C. truncatum infection increased the activities of CWDEs in H. undatus fruits to disassemble their cell wall components, finally leading to the fruits’ decay and deterioration. Thus, our findings will provide significant evidence in the controlling of pitaya anthracnose in the future. Full article
(This article belongs to the Special Issue Control of Postharvest Fungal Diseases)
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<p>Symptoms of <span class="html-italic">Hylocereus undatus</span> fruit anthracnose at different time periods in Wuwei city, China. (<b>A</b>) and (<b>B</b>): the symptoms at the initial stage; (<b>C</b>–<b>F</b>): the symptoms at a later stage.</p>
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<p>The pathogenicity test of the representative isolate of HLGTJ-1 on <span class="html-italic">Hylocereus undatus</span> fruit after inoculation. (<b>A</b>) Fruit inoculation with the PDA discs without the HLGTJ-1 isolate (control); (<b>B</b>,<b>C</b>) fruits inoculation with the mycelial discs of the HLGTJ-1 isolate at 3 and 7 days after inoculation, respectively.</p>
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<p>Morphological characteristics of the representative isolate of HLGTJ-1. (<b>A</b>,<b>B</b>) The front and reverse views of the colony, respectively; (<b>C</b>) the conidial mass produced on the front of the colony; (<b>D</b>) the conidial mass observed under a stereoscope; (<b>E</b>) the acervuli and setae observed under a stereoscope; (<b>F</b>) the acervulus and setae observed under a microscope using the hand-sliced method; (<b>G</b>,<b>H</b>) conidia; (<b>I</b>,<b>J</b>) appressorium.</p>
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<p>Multi-locus phylogenetic tree of the single-spore isolate of HLGTJ-1 based on the combined sequences (HJ-ITS region and <span class="html-italic">HJ-GAPDH</span> and <span class="html-italic">HJ-HIS3</span> genes) by the maximum likelihood (ML) method. Bootstraps supporting values higher than 50% from the 1000 replicates are presented at the nodes.</p>
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<p>Changes in activities of Cx (<b>A</b>), β-Glu (<b>B</b>), PG (<b>C</b>), and PMG (<b>D</b>) in <span class="html-italic">Hylocereus undatus</span> fruits at different days after inoculation with the isolate. Different letters in Figure are significantly different at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Changes in original pectin (<b>A</b>), soluble pectin (<b>B</b>), and cellulose (<b>C</b>) contents in <span class="html-italic">Hylocereus undatus</span> fruits at different days after inoculation with the isolate. Different letters in Figure are significantly different at <span class="html-italic">p</span> &lt; 0.05.</p>
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26 pages, 11072 KiB  
Article
Exploring the Potential Effects of Cryopreservation on the Biological Characteristics and Cardiomyogenic Differentiation of Rat Adipose-Derived Mesenchymal Stem Cells
by Ahmed Farag, Sai Koung Ngeun, Masahiro Kaneda, Mohamed Aboubakr, Asmaa Elhaieg, Hanan Hendawy and Ryou Tanaka
Int. J. Mol. Sci. 2024, 25(18), 9908; https://doi.org/10.3390/ijms25189908 (registering DOI) - 13 Sep 2024
Viewed by 400
Abstract
Cryopreservation is essential for the broad clinical application of mesenchymal stem cells (MSCs), yet its impact on their cellular characteristics and cardiomyogenic differentiation potential remains a critical concern in translational medicine. This study aimed to evaluate the effects of cryopreservation on the biological [...] Read more.
Cryopreservation is essential for the broad clinical application of mesenchymal stem cells (MSCs), yet its impact on their cellular characteristics and cardiomyogenic differentiation potential remains a critical concern in translational medicine. This study aimed to evaluate the effects of cryopreservation on the biological properties and cardiomyogenic capacity of rat adipose-derived MSCs (AD-MSCs). We examined their cellular morphology, surface marker expression (CD29, CD90, CD45), trilineage differentiation potential (adipogenic, osteogenic, chondrogenic), and gene expression profiles for the pluripotency marker REX1 and immunomodulatory markers TGFβ1 and IL-6. After inducing cardiomyocyte differentiation, we assessed cardiac-specific gene expressions (Troponin I, MEF2c, GSK-3β) using quantitative RT-qPCR, along with live/dead cell staining and immunofluorescence for cardiac-specific proteins (Troponin T, α-actinin, Myosin Heavy Chain). Cryopreserved AD-MSCs preserved their morphology, surface markers, and differentiation potential, but exhibited a reduced expression of REX1, TGFβ1, and IL-6. Additionally, cryopreservation diminished cardiomyogenic differentiation, as indicated by the lower levels of Troponin I, MEF2c, and GSK-3β seen compared to non-cryopreserved cells. Despite this, high cell viability (>90%) and maintained cardiac protein expression were observed post-cryopreservation. These findings highlight the necessity of optimizing cryopreservation protocols to ensure the full therapeutic potential of AD-MSCs, particularly in applications related to cardiac regenerative medicine. Full article
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<p>Morphological characteristics of plastic-adherent rat adipose-derived mesenchymal stem cells (AD-MSCs) at their fourth passage, both prior to and following cryopreservation. Abbreviations: AD-MSCs, adipose-derived mesenchymal stem cells; BF, before cryopreservation; AF, after cryopreservation; P4, passage 4. The scale bar represents 100 μm.</p>
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<p>Representative histograms illustrating the expression of MSC surface markers (CD29 and CD90) and a hematopoietic marker (CD45). (<b>A</b>) Samples prior to cryopreservation. (<b>B</b>) Samples following cryopreservation.</p>
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<p>Adipogenic differentiation of AD-MSCs. (<b>A</b>) Representative histological images of both undifferentiated and Oil Red O-stained differentiated cells from rat AD-MSCs, before and after cryopreservation. (<b>B</b>) The percentage of Oil Red O-stained fat vacuoles was quantified using ImageJ (1.8.0-345). The scale bar represents 100 µm.</p>
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<p>Osteogenic differentiation of AD-MSCs. (<b>A</b>) Representative histological images of both undifferentiated and Alizarin Red-stained osteogenic differentiated cells from rat AD-MSCs, before and after cryopreservation. (<b>B</b>) The percentage of Alizarin Red-stained mineral matrix deposition was quantified using ImageJ. The scale bar represents 100 µm.</p>
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<p>Chondrogenic differentiation of AD-MSCs. (<b>A</b>) Representative histological images showing both undifferentiated and Alcian Blue-stained chondrogenic differentiated cells from rat AD-MSCs, before and after cryopreservation. (<b>B</b>) The percentage of Alcian Blue-stained cartilage matrix was quantified using ImageJ. The scale bar represents 100 µm.</p>
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<p>Bar graph representing the fold change expression levels of pluripotency and immunomodulatory markers in AD-MSCs after their cryopreservation, normalized with pre-cryopreservation AD-MSCs as the reference. All genes showed a decrease in their fold change, with values below 1. All experiments were conducted in triplicate to ensure the reliability of the results.</p>
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<p>Morphological changes following cardiomyocyte differentiation in plastic-adherent rat AD-MSCs before and after cryopreservation. The scale bar represents 200 μm. Abbreviations: AD-MSCs, adipose-derived mesenchymal stem cells; BF, before cryopreservation; AF, after cryopreservation; AF. Cardio. Diff., after cardiomyocyte differentiation.</p>
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<p>Viability of AD-MSCs assessed by live/dead (green/red) cell staining before and after cryopreservation. The scale bar represents 100 μm.</p>
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<p>Immunofluorescence staining of cardiac-specific proteins in rat AD-MSCs before and after cryopreservation: (<b>A</b>) Troponin T, (<b>B</b>) α-Actinin, and (<b>C</b>) Myosin Heavy Chain 1. All experiments were conducted in triplicate to ensure the reliability of the results. The scale bar represents 100 μm.</p>
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<p>Immunofluorescence staining of cardiac-specific proteins in rat AD-MSCs before and after cryopreservation: (<b>A</b>) Troponin T, (<b>B</b>) α-Actinin, and (<b>C</b>) Myosin Heavy Chain 1. All experiments were conducted in triplicate to ensure the reliability of the results. The scale bar represents 100 μm.</p>
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<p>Immunofluorescence staining of cardiac-specific proteins in rat AD-MSCs before and after cryopreservation: (<b>A</b>) Troponin T, (<b>B</b>) α-Actinin, and (<b>C</b>) Myosin Heavy Chain 1. All experiments were conducted in triplicate to ensure the reliability of the results. The scale bar represents 100 μm.</p>
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<p>Quantitative analysis of cardiac-specific gene expression in rat AD-MSCs before (BF) and after (AF) cryopreservation using RT-qPCR. The expression levels of <span class="html-italic">Troponin I</span>, <span class="html-italic">MEF2c</span>, and <span class="html-italic">GSK-3β</span> are presented for the various study groups. All experiments were conducted in triplicate to ensure the reliability of the results. Statistical significance is indicated as follows: *** <span class="html-italic">p</span>  &lt;  0.001 and **** <span class="html-italic">p</span>  &lt;  0.0001.</p>
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<p>Overview of the study’s experimental design.</p>
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16 pages, 1308 KiB  
Review
Correlating Ultrastructural Changes in the Invasion Area of Colorectal Cancer with CT and MRI Imaging
by Joanna Urbaniec-Stompór, Maciej Michalak and Janusz Godlewski
Int. J. Mol. Sci. 2024, 25(18), 9905; https://doi.org/10.3390/ijms25189905 - 13 Sep 2024
Viewed by 206
Abstract
The cancer invasion of the large intestine, a destructive process that begins within the mucous membrane, causes cancer cells to gradually erode specific layers of the intestinal wall. The normal tissues of the intestine are progressively replaced by a tumour mass, leading to [...] Read more.
The cancer invasion of the large intestine, a destructive process that begins within the mucous membrane, causes cancer cells to gradually erode specific layers of the intestinal wall. The normal tissues of the intestine are progressively replaced by a tumour mass, leading to the impairment of the large intestine’s proper morphology and function. At the ultrastructural level, the disintegration of the extracellular matrix (ECM) by cancer cells triggers the activation of inflammatory cells (macrophages) and connective tissue cells (myofibroblasts) in this area. This accumulation and the functional interactions between these cells form the tumour microenvironment (TM). The constant modulation of cancer cells and cancer-associated fibroblasts (CAFs) creates a specific milieu akin to non-healing wounds, which induces colon cancer cell proliferation and promotes their survival. This review focuses on the processes occurring at the “front of cancer invasion”, with a particular focus on the role of the desmoplastic reaction in neoplasm development. It then correlates the findings from the microscopic observation of the cancer’s ultrastructure with the potential of modern radiological imaging, such as computer tomography (CT) and magnetic resonance imaging (MRI), which visualizes the tumour, its boundaries, and the tissue reactions in the large intestine. Full article
(This article belongs to the Special Issue Molecular Mechanisms and Therapies of Colorectal Cancer 3.0)
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<p>The role of enzymes and signalling molecules involved in promotion and progression of CRC.</p>
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<p>Comparison of two different growth patterns of large intestine cancer. The first one is expansive, with a well-demarcated tumour mass boundary, which pushes away the healthy tissue. The second one is infiltrative, without a demarcated mass boundary, with cords of cancer cells invading the healthy tissue.</p>
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<p>Illustration of different T-stages of CRC and their relation to layers of rectum.</p>
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25 pages, 6783 KiB  
Article
Metabolomic and Transcriptomic Analyses Reveal the Potential Mechanisms of Dynamic Ovarian Development in Goats during Sexual Maturation
by Yanyan Wang, Tianle Chao, Qing Li, Peipei He, Lu Zhang and Jianmin Wang
Int. J. Mol. Sci. 2024, 25(18), 9898; https://doi.org/10.3390/ijms25189898 (registering DOI) - 13 Sep 2024
Viewed by 314
Abstract
The ovary is a crucial reproductive organ in mammals, and its development directly influences an individual’s sexual maturity and reproductive capacity. To comprehensively describe ovarian sexual maturation in goats, we integrated phenotypic, hormonal, metabolomic, and transcriptomic data from four specific time points: after [...] Read more.
The ovary is a crucial reproductive organ in mammals, and its development directly influences an individual’s sexual maturity and reproductive capacity. To comprehensively describe ovarian sexual maturation in goats, we integrated phenotypic, hormonal, metabolomic, and transcriptomic data from four specific time points: after birth (D1), at 2 months old (M2), at 4 months old (M4), and at 6 month old (M6). The study showed that during the early stage (D1–M2), ovarian growth was the most rapid, with weight and morphology increasing by 284% and 65%, respectively, and hormone levels rose significantly, with estradiol increasing by 57%. Metabolomic analysis identified 1231 metabolites, primarily lipids, lipid molecules, and organic acids, which can support hormone balance and follicle development by providing energy and participating in signaling transduction. Transcriptomic analysis identified 543 stage-specific differentially expressed genes, mainly enriched in steroid biosynthesis, amino acid metabolism, and the PI3K/AKT pathway, which are key factors influencing ovarian cell proliferation, apoptosis, hormone secretion, and metabolism. The integrated analysis revealed the key processes in the ovarian steroid hormone biosynthesis pathway and gene/metabolite networks associated with ovarian phenotypes and hormone levels, ultimately highlighting scavenger receptor class B type 1 (SCARB1), Cytochrome P450 Family 1 Subfamily A Member 1 (CYP11A1), 3beta-hydroxysteroid dehydrogenase (3BHSD), progesterone, estradiol, and L-phenylalanine as key regulators of ovarian morphological and functional changes at different developmental stages. This study is the first to reveal the metabolic changes and molecular regulatory mechanisms during ovarian sexual maturation in goats, providing valuable insights for understanding reproductive system development and optimizing reproductive performance and breeding efficiency. Full article
(This article belongs to the Section Molecular Endocrinology and Metabolism)
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<p>Analysis of ovarian phenotypic characteristics and serum hormone levels at four developmental stages. (<b>A</b>) Analysis of the ovarian weight (OW), ovarian length (OL), ovarian width (OD), and ovarian thickness (OT). (<b>B</b>) Levels of gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (PROG) and estradiol (E2) in serum. D1: after birth, M2: at 2 months old, M4: at 4 months old, M6: at 6 months old. Values represent mean ± standard error. Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). * <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. ns = not statistically significant.</p>
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<p>Basic metabolomics analysis of ovarian tissues at different developmental stages. (<b>A</b>) Principal component analysis of metabolites at four developmental stages. (<b>B</b>) Top 10 classes of metabolites identified during ovarian development. (<b>C</b>) The number of DAMs in pairwise comparisons. (<b>D</b>) The Venn diagram of DAMs in pairwise comparisons. (<b>E</b>) The heatmap of DAMs during ovarian development.</p>
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<p>The expression patterns and functional enrichment analysis of DAMs. (<b>A</b>) Expression pattern analysis of DAMs. (<b>B</b>) KEGG pathway enrichment analysis of all DAMs.</p>
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<p>Basic transcriptome analysis of ovarian tissues at different developmental stages. (<b>A</b>) Venn diagram of the number of mRNA in ovary tissues at different developmental stages. (<b>B</b>) Validation of transcriptome data by qRT-PCR. (<b>C</b>) The number of DAMs in pairwise comparisons. (<b>D</b>) The Upset Plot for DEGs in different comparison groups.</p>
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<p>The expression patterns and functional enrichment analysis of DEGs. (<b>A</b>) The heatmap of DEGs during ovarian development. (<b>B</b>) KEGG pathway enrichment analysis of all DEGs. (<b>C</b>) Expression pattern analysis of DEGs.</p>
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<p>The integrated metabolomic and transcriptomic analysis of ovarian tissues at different developmental stages. (<b>A</b>) The O2PLS analysis of DAMs and DEGs. (<b>B</b>) Bar plot of the top 20 transcriptomics and metabolites pq1.</p>
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<p>The pathway related to steroid hormone synthesis and the heatmap of changes in metabolites and genes. (<b>A</b>) The steroid biosynthesis pathway. (<b>B</b>) The ovarian steroid synthesis pathway. (<b>C</b>) Correlation analysis of DAMs and DEGs involved in steroid synthesis. Blue indicates a negative correlation and red indicates a positive correlation. The “*” and “**” represent significant differences at levels of <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.01 according to Pearson’s correlation coefficient. (<b>D</b>) The estrogen signaling pathway during ovarian development.</p>
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<p>Analysis of the WGCNA of the ovarian metabolome at different developmental stages. (<b>A</b>) The clustering dendrogram of all metabolites. The color rows below the dendrogram show module assignments based on the dynamic hybrid branch cutting method. Each color represents a different module of co-expressed metabolites, while the “Merged dynamic” row indicates merged modules, with gray representing unassigned metabolites. The y-axis (Height) represents the dissimilarity between metabolites, with higher values indicating less similarity. (<b>B</b>) Heatmap of the correlations between modules and phenotypic traits. Modules are color-coded on the left, and phenotypic traits are listed below. The numbers in the cells represent correlation coefficients, with red for positive correlations and green for negative correlations. Color intensity indicates correlation strength. (<b>C</b>,<b>D</b>) Visualization of connections of metabolites in pink (<b>C</b>) and brown (<b>D</b>) modules. According to the connection weight value, only the first 100 functional relationships are shown in the figures. Yellow nodes represent hub metabolites with high connectivity, while blue nodes represent others. The thickness of the edges between nodes reflects the strength of their connections, with thicker edges indicating stronger associations, highlights key metabolites in the modules. OW: ovarian weight. OL: ovarian length. OD: ovarian width. OT: ovarian thickness. E2: estradiol. PROG: progesterone.</p>
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<p>Analysis of the WGCNA of ovarian transcriptome at different developmental stages. (<b>A</b>) The clustering dendrogram of all genes. The color rows below the dendrogram show module assignments based on the dynamic hybrid branch cutting method. Each color represents a different module of co-expressed genes, while the “Merged dynamic” row indicates merged modules, with gray representing unassigned metabolites. The y-axis (Height) represents the dissimilarity between genes, with higher values indicating less similarity. (<b>B</b>) Heatmap of the correlations between modules and phenotypic traits. The modules are color-coded on the left, and phenotypic traits are listed below. The numbers in the cells represent correlation coefficients, with red for positive correlations and green for negative correlations. Color intensity indicates correlation strength. (<b>C</b>) KEGG enrichment analysis of genes in the yellow module. The y-axis shows enriched biological pathways, and the x-axis (Count) indicates the number of genes involved. (<b>D</b>) Network visualization of the top 50 genes in the yellow module, based on connection weights. Yellow nodes represent hub genes with high connectivity, while blue nodes represent other genes. Edge thickness reflects connection strength, with thicker edges indicating stronger associations. OL: ovarian length. OD: ovarian width. OT: ovarian thickness. E2: estradiol. PROG: progesterone.</p>
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10 pages, 1309 KiB  
Review
Iatrogenic Sigma Perforation in a Patient with Localized Rectal and Sigma AL Amyloidosis: A Forensic Case and a Literature Review
by Giuseppe Davide Albano, Stefania Zerbo, Simone Di Franco, Elisabetta Orlando, Eleonora Formisano, Antonina Argo and Beatrice Belmonte
Forensic Sci. 2024, 4(3), 453-462; https://doi.org/10.3390/forensicsci4030029 - 13 Sep 2024
Viewed by 219
Abstract
Amyloidosis is defined as a rare group of 30 protein-folding diseases characterized by the extracellular deposition of a specific soluble precursor protein that aggregates in the form of insoluble fibrils. The gastrointestinal tract (GI) is a common site for amyloid deposits: Among patients [...] Read more.
Amyloidosis is defined as a rare group of 30 protein-folding diseases characterized by the extracellular deposition of a specific soluble precursor protein that aggregates in the form of insoluble fibrils. The gastrointestinal tract (GI) is a common site for amyloid deposits: Among patients with systemic amyloidosis, at least 70% present with gastrointestinal deposition. Rarely, the deposition is exclusively localized in this area, leading to various gastrointestinal symptoms (bleeding, weight loss, etc.). In this case report, we present a rare and unusual form of localized gastrointestinal amyloidosis, diagnosed after a post-mortem examination of an 83-year-old woman who died due to septic shock resulting from post-colonoscopy iatrogenic perforation of the sigma, in a suspected medical liability case. Morphological examination revealed AL amyloid deposits within the muscular wall of the submucosal vessels of the rectum, which caused increased friability of the vessels and ischemic changes in the intestinal mucosa. A renal cell carcinoma (RCC) was found, which might be related to amyloid deposits, as reported by the literature. Amyloid deposits are an unknown and unpredictable pathological substrate that increase the risk of iatrogenic perforation. Analysis of the medical documentation did not reveal any censurable conduct in terms of prescribing the procedure, technical execution, or subsequent management of the patient following the perforation. GI amyloidosis should be part of the risk stratification of patients with rectal bleeding and gastrointestinal symptoms, and awareness is essential to guide subsequent diagnostic and therapeutic approaches and investigate underlying causes. Full article
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<p>(<b>A</b>,<b>B</b>) Representative microphotographs showing the amyloid deposits (black arrows) within the muscular wall of submucosal vessels in rectum. Sampled using hematoxylin and eosin staining. Scale bars: 50 µm. (<b>C</b>–<b>H</b>) Presence of amyloid deposited within the muscular wall vessels of the rectal mucosa (black arrows in the top images under bright field; white arrows in the bottom images under dark field). Scale bars: 50 µm.</p>
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<p>(<b>A</b>–<b>D</b>) Presence of amyloid deposits, lambda light chains-positive, within the muscular wall vessels of the rectal mucosa (black arrows). Sampled using DAB chromogen. Scale bars: 50 µm.</p>
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<p>Representative pictures of renal carcinoma. (<b>A</b>) Macroscopic appearance, (<b>B</b>,<b>C</b>) Hematoxylin- and eosin-stained sections, and (<b>D</b>,<b>E</b>) CD10 immunostaining of clear cell renal carcinoma. Scale bars: 50 µm.</p>
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15 pages, 707 KiB  
Review
Biomarkers Involved in the Pathogenesis of Hemophilic Arthropathy
by Oana Viola Badulescu, Dragos-Viorel Scripcariu, Minerva Codruta Badescu, Manuela Ciocoiu, Maria Cristina Vladeanu, Carmen Elena Plesoianu, Andrei Bojan, Dan Iliescu-Halitchi, Razvan Tudor, Bogdan Huzum, Otilia Elena Frasinariu and Iris Bararu-Bojan
Int. J. Mol. Sci. 2024, 25(18), 9897; https://doi.org/10.3390/ijms25189897 (registering DOI) - 13 Sep 2024
Viewed by 228
Abstract
Hemophilia, which is a rare disease, results from congenital deficiencies of coagulation factors VIII and IX, respectively, leading to spontaneous bleeding into joints, resulting in hemophilic arthropathy (HA). HA involves complex processes, including synovial proliferation, angiogenesis, and tissue remodeling. Despite ongoing research, factors [...] Read more.
Hemophilia, which is a rare disease, results from congenital deficiencies of coagulation factors VIII and IX, respectively, leading to spontaneous bleeding into joints, resulting in hemophilic arthropathy (HA). HA involves complex processes, including synovial proliferation, angiogenesis, and tissue remodeling. Despite ongoing research, factors contributing to HA progression, especially in adults with severe HA experiencing joint pain, remain unclear. Blood markers, particularly collagen-related ones, have been explored to assess joint health in hemophilia. For example, markers like CTX-I and CTX-II reflect bone and cartilage turnover, respectively. Studies indicate elevated levels of certain markers post-bleeding episodes, suggesting joint health changes. However, longitudinal studies on collagen turnover and basement membrane or endothelial cell markers in relation to joint outcomes, particularly during painful episodes, are scarce. Given the role of the CX3CL1/CX3XR1 axis in arthritis, other studies investigate its involvement in HA. The importance of different inflammatory and bone damage biomarkers should be assessed, alongside articular cartilage and synovial membrane morphology, aiming to enhance understanding of hemophilic arthropathy progression. Full article
(This article belongs to the Special Issue Advances in Rare Diseases Biomarkers)
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<p>Biomarkers in HA [<a href="#B38-ijms-25-09897" class="html-bibr">38</a>].</p>
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26 pages, 3627 KiB  
Article
Unveiling the Performance of Co-Assembled Hybrid Nanocarriers: Moving towards the Formation of a Multifunctional Lipid/Random Copolymer Nanoplatform
by Efstathia Triantafyllopoulou, Diego Romano Perinelli, Aleksander Forys, Pavlos Pantelis, Vassilis G. Gorgoulis, Nefeli Lagopati, Barbara Trzebicka, Giulia Bonacucina, Georgia Valsami, Natassa Pippa and Stergios Pispas
Pharmaceutics 2024, 16(9), 1204; https://doi.org/10.3390/pharmaceutics16091204 - 13 Sep 2024
Viewed by 252
Abstract
Despite the appealing properties of random copolymers, the use of these biomaterials in association with phospholipids is still limited, as several aspects of their performance have not been investigated. The aim of this work is the formulation of lipid/random copolymer platforms and the [...] Read more.
Despite the appealing properties of random copolymers, the use of these biomaterials in association with phospholipids is still limited, as several aspects of their performance have not been investigated. The aim of this work is the formulation of lipid/random copolymer platforms and the comprehensive study of their features by multiple advanced characterization techniques. Both biomaterials are amphiphilic, including two phospholipids (1,2-dioctadecanoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)) and a statistical copolymer of oligo (ethylene glycol) methyl ether methacrylate (OEGMA) and 2-(diisopropylamino) ethyl methacrylate (DIPAEMA). We examined the design parameters, including the lipid composition, the % comonomer ratio, and the lipid-to-polymer ratio that could be critical for their behavior. The structures were also probed in different conditions. To the best of the authors’ knowledge, this is the first time that P(OEGMA-co-DIPAEMA)/lipid hybrid colloidal dispersions have been investigated from a membrane mechanics, biophysical, and morphological perspective. Among other parameters, the copolymer architecture and the hydrophilic to hydrophobic balance are deemed fundamental parameters for the biomaterial co-assembly, having an impact on the membrane’s fluidity, morphology, and thermodynamics. Exploiting their unique characteristics, the most promising candidates were utilized for methotrexate (MTX) loading to explore their encapsulation capability and potential antitumor efficacy in vitro in various cell lines. Full article
(This article belongs to the Special Issue Polymer-Based Delivery System)
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<p>(<b>a</b>) The chemical structure of the random copolymer P(OEGMA950-co-DIPAEMA) synthesized by RAFT polymerization; (<b>b</b>) Graphic illustration of P(OEGMA-co-DIPAEMA)-1 or copolymer 1 and P(OEGMA-co-DIPAEMA)-2 or copolymer 2, respectively, with a different % comonomer ratio.</p>
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<p>Charts derived from DLS measurements at 25 °C: (<b>a</b>) The hydrodynamic radius (R<sub>h</sub>, nm); (<b>b</b>) the scattered intensity (I, kilocounts per second or kcps) of hybrid colloidal dispersions the day of their preparation, utilizing water for injection as the dispersion medium. The standard deviation (SD) is less than 10% in both diagrams. * Hybrid systems with more than one population; the predominant (higher intensity) one is presented in the graph.</p>
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<p>The hydrodynamic radius (R<sub>h</sub>, nm) of the hybrid colloidal dispersions in: (<b>a</b>) different dispersion media at body temperature (37 °C); (<b>b</b>) FBS:PBS biorelevant medium at different temperatures. The standard deviation (SD) is less than 10% in both diagrams. The DSPC:DOPC:1 9:1 hybrid system at 37 °C in both diagrams refers to a very high R<sub>h</sub> compared with the rest of the systems exceeding the scale of the graph.</p>
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<p>GP parameter vs. lipid composition of P(OEGMA<sub>950</sub>-co-DIPAEMA) hybrid systems at a steady lipid to polymer weight ratio (9:1).</p>
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<p>Cryo-TEM images of (<b>a</b>) P(OEGMA-co-DIPAEMA)-1. (<b>b</b>) P(OEGMA-co-DIPAEMA)-2 hybrid systems with different lipid compositions: (<b>i</b>) DSPC; (<b>ii</b>) DSPC:DOPC (9:1 weight ratio) and constant lipid to polymer ratio (9:1) or a constant lipid composition (DSPC) with different lipid to copolymer weight ratios: (<b>iii</b>) 7:3 and (<b>iv</b>) 5:5. The arrows represent the following: green color: spherical or irregularly shaped particles with distinct membrane; red color: “patchy” spherical- or pentagon-shaped vesicles; black color: rods; yellow color: small spherical particles.</p>
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<p>mDSC traces of (<b>a</b>) DSPC; (<b>b</b>) DSPC: DOPC (9:1 weight ratio) hybrid systems integrating P(OEGMA-co-DIPAEMA)-1 or -2 at different lipid to polymer weight ratios into aqueous medium.</p>
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<p>Thermodynamic evaluation of DSPC:P(OEGMA-co-DIPAEMA)-1 and -2 in an acidic environment (pH 4.5): (<b>a</b>) mDSC profiles; (<b>b</b>) (<b>i</b>) sound speed or (<b>ii</b>) attenuation vs. temperature from HR-US.</p>
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<p>(<b>a</b>) Size distributions from the DLS of DSPC:P(OEGMA-co-DIPAEMA)-2 hybrid systems incorporating MTX at two different lipid to polymer ratios, 9:1 (black line) and 5:5 (red line), on the day of their preparation; (<b>b</b>) the systems’ stability assessment (R<sub>h</sub> vs. time) under storage conditions (4 °C) for 21 days.</p>
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<p>Cell viability vs. different concentrations of MTX-DSPC:2 9:1 (blue line) and MTX-DSPC:2 5:5 (orange line) on (<b>a</b>) HEK293 and (<b>b</b>) HeLa cells. The concentration levels refer to MTX concentration, and the obtained data represent the means ± standard deviation from three experiments conducted in triplicates. The asterisks (*) in (<b>b</b>) correspond to p values of less than 0.05 (<span class="html-italic">p</span> &lt; 0.05) that are considered as statistically significant.</p>
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12 pages, 274 KiB  
Review
Atypical Spindle Cell/Pleomorphic Lipomatous Tumor: A Review and Update
by Jun Nishio, Shizuhide Nakayama, Yoshiro Chijiiwa, Mikiro Koga and Mikiko Aoki
Cancers 2024, 16(18), 3146; https://doi.org/10.3390/cancers16183146 - 13 Sep 2024
Viewed by 195
Abstract
Atypical spindle cell/pleomorphic lipomatous tumor (ASCPLT) is a rare and recently described adipocytic neoplasm that primarily occurs in the subcutis of the limbs and limb girdles, particularly of middle-aged adults. It has locally recurrent potential if incompletely excised but no risk for distant [...] Read more.
Atypical spindle cell/pleomorphic lipomatous tumor (ASCPLT) is a rare and recently described adipocytic neoplasm that primarily occurs in the subcutis of the limbs and limb girdles, particularly of middle-aged adults. It has locally recurrent potential if incompletely excised but no risk for distant metastasis. ASCPLT is histologically similar to spindle cell/pleomorphic lipoma and atypical lipomatous tumor and shows a mixture of atypical spindle cells, adipocytes, lipoblasts, floret-like multinucleated giant cells, and/or pleomorphic cells. It has been recently recognized that ASCPLT can undergo sarcomatous transformation. However, the biological significance of morphological sarcomatous transformation in ASCPLT remains uncertain. Immunohistochemically, the tumor cells show variable expression of CD34, S-100 protein, and desmin. Loss of nuclear Rb expression is observed in the majority of cases. ASCPLT lacks MDM2 gene amplification but can show RB1 gene deletion in a significant subset of cases. Complete surgical excision is the treatment of choice. This review provides an overview of the current knowledge on the clinicoradiological features, pathogenesis, histopathology, and treatment of ASCPLT. In addition, we will discuss the differential diagnosis of this new entity. Full article
(This article belongs to the Special Issue Recent Advances in Orthopaedic Oncology: 2nd Edition)
17 pages, 6378 KiB  
Article
Bioprospecting of Mangrove Filamentous Fungi for the Biodegradation of Polyethylene Microplastics
by Arthur Aguiar, Letícia Gama, Milene Fornari, Almir Neto, Rodrigo de Souza, Rafael Perna, Laura Castro, Stella Kovacs, Marta Filipa Simões, Nelson Ferreira, Yoannis Domínguez, Leandro de Castro and Cristiane Ottoni
J. Mar. Sci. Eng. 2024, 12(9), 1629; https://doi.org/10.3390/jmse12091629 - 12 Sep 2024
Viewed by 405
Abstract
The accumulation of microplastics (MPs) in the environment has been a bottleneck for scientific society. Several approaches have been described as possibilities for reducing MPs in aquatic and terrestrial ecosystems; however, most of them are not environmentally friendly. Filamentous fungi (Ff) cells are [...] Read more.
The accumulation of microplastics (MPs) in the environment has been a bottleneck for scientific society. Several approaches have been described as possibilities for reducing MPs in aquatic and terrestrial ecosystems; however, most of them are not environmentally friendly. Filamentous fungi (Ff) cells are currently considered a promising solution as a treatment for MPs. Therefore, the present study reports the potential ability of Ff isolated from mangrove sediments to biodegrade low-density polyethylene MPs (LDPEMPs). Six Ff strains were grown in batch cultures for 28 days, and one of them, Aspergillus sp. (AQ3A), showed the most prominent profile to biodegrade polymeric compounds. After morphological and molecular analysis, all strains were identified as belonging to the genera Aspergillus (MQ1C, AQ2A and AQ3A), Penicillium (MQ1A), and Trichoderma (MQ1B and MQ2A). The strain Aspergillus sp. (AQ3A) showed the most promising results with a LDPEMPs reduction rate of 47% and biomass formation of 0.0890 g·mL−1. Complementary studies with Aspergillus sp. (AQ3A) using Fourier-transform infrared spectroscopy (FTIR) highlighted changes in the molecular structure of LDPEMPs. These results indicate that Ff can contribute to the biodegradation of LDPEMPs. However, other parameters, mainly associated with the enzymes that are involved in this biodegradation process, need to be explored. Full article
(This article belongs to the Special Issue Effects of Ocean Plastic Pollution on Aquatic Life)
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Figure 1
<p>Location of the sampling site at the Ecological Station of Juréia-Itatins, Peruíbe, Brazil.</p>
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<p>Morphology of low density polyethylene microplastics: (<b>A</b>) microfiber, (<b>B</b>) microfragments, and (<b>C</b>) microsphere.</p>
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<p>Bayesian tree of internal transcribed spacers (ITS) sequences, obtained from fungal isolates (AQ2A, AQ3A, MQ1A, MQ1B, MQ1C, and MQ2A) and related species previously recorded in Brazil. Numbers above the branches indicate posterior probabilities, and the bar indicates the number of nucleotide substitutions per site.</p>
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<p>Fourier-transform infrared spectroscopy (FTIR) analysis of low-density polyethylene microplastics (LDPE<sub>MPs</sub>), before and after biodegradation test, using AQ3A strain. (<b>A</b>) Treatment after seven days, (<b>B</b>) treatment after 14 days, (<b>C</b>) treatment after 21 days, (<b>D</b>) treatment after 28 days, (<b>E</b>) control containing only LDPE<sub>MPs</sub> after 7 days, and (<b>F</b>) control containing only LDPE<sub>MPs</sub> after 28 days.</p>
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