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Search Results (1,988)

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13 pages, 2648 KiB  
Article
Comparative Study of Deep Transfer Learning Models for Semantic Segmentation of Human Mesenchymal Stem Cell Micrographs
by Maksim Solopov, Elizaveta Chechekhina, Anna Kavelina, Gulnara Akopian, Viktor Turchin, Andrey Popandopulo, Dmitry Filimonov and Roman Ishchenko
Int. J. Mol. Sci. 2025, 26(5), 2338; https://doi.org/10.3390/ijms26052338 - 6 Mar 2025
Viewed by 151
Abstract
The aim of this study is to conduct a comparative assessment of the effectiveness of neural network models—U-Net, DeepLabV3+, SegNet and Mask R-CNN—for the semantic segmentation of micrographs of human mesenchymal stem cells (MSCs). A dataset of 320 cell micrographs annotated by cell [...] Read more.
The aim of this study is to conduct a comparative assessment of the effectiveness of neural network models—U-Net, DeepLabV3+, SegNet and Mask R-CNN—for the semantic segmentation of micrographs of human mesenchymal stem cells (MSCs). A dataset of 320 cell micrographs annotated by cell biology experts was created. The models were trained using a transfer learning method based on ImageNet pre-trained weights. As a result, the U-Net model demonstrated the best segmentation accuracy according to the metrics of the Dice coefficient (0.876) and the Jaccard index (0.781). The DeepLabV3+ and Mask R-CNN models also showed high performance, although slightly lower than U-Net, while SegNet exhibited the least accurate results. The obtained data indicate that the U-Net model is the most suitable for automating the segmentation of MSC micrographs and can be recommended for use in biomedical laboratories to streamline the routine analysis of cell cultures. Full article
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Figure 1
<p>Training graphs of neural network models for segmenting micrographs of mesenchymal stem cells (MSCs). Dynamics of changes in pixel accuracy (PA) and loss function for the investigated models on training and validation samples of micrographs during training: (<b>a</b>) U-Net, (<b>b</b>) DeepLabV3+, (<b>c</b>) SegNet, and (<b>d</b>) Mask R-CNN.</p>
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<p>Optimal prediction thresholds for U-Net, DeepLabV3+, SegNet, and Mask R-CNN segmentation models (from top to bottom) according to the Dice coefficient (DC), Jaccard index (JI) and PA metrics (from left to right). The optimal thresholds are defined as the maximum values of the functional dependencies of the metric on the threshold value. To plot the dependencies, the average value of each metric was calculated over 64 images from the validation sample at a given value of the varying threshold. The graphs show the mean values (blue line) with standard deviations (highlighted in gray).</p>
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<p>Comparison of the performance of segmentation models based on DC (<b>a</b>), JI (<b>b</b>), and PA (<b>c</b>) metrics. The charts show the distribution of metric values for each model. * <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; 0.0001; ns—differences are not significant.</p>
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<p>Examples of segmentation of MSC micrographs by neural network models: original images, ground truth masks, and masks predicted by U-Net, DeepLabV3+, SegNet, and Mask R-CNN models. The micrographs were captured at a magnification of 40×.</p>
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17 pages, 9975 KiB  
Article
Oropouche Virus: Isolation and Ultrastructural Characterization from a Human Case Sample from Rio de Janeiro, Brazil, Using an In Vitro System
by Ana Luisa Teixeira de Almeida, Igor Pinto Silva da Costa, Maycon Douglas do Nascimento Garcia, Marcos Alexandre Nunes da Silva, Yasmim Gonçalves Lazzaro, Ana Maria Bispo de Filippis, Fernanda de Bruycker Nogueira and Debora Ferreira Barreto-Vieira
Viruses 2025, 17(3), 373; https://doi.org/10.3390/v17030373 - 5 Mar 2025
Viewed by 85
Abstract
The Oropouche virus (OROV) is a segmented negative-sense RNA arbovirus member of the Peribunyaviridae family, associated with recurring epidemics of Oropouche fever in Central and South America. Since its identification in 1955, OROV has been responsible for outbreaks in both rural and urban [...] Read more.
The Oropouche virus (OROV) is a segmented negative-sense RNA arbovirus member of the Peribunyaviridae family, associated with recurring epidemics of Oropouche fever in Central and South America. Since its identification in 1955, OROV has been responsible for outbreaks in both rural and urban areas, with transmission involving sylvatic and urban cycles. This study focuses on the characterization of an OROV isolate from a human clinical sample collected in the state of Rio de Janeiro, a non-endemic region in Brazil, highlighting ultrastructural and morphological aspects of the viral replicative cycle in Vero cells. OROV was isolated in Vero cell monolayers which, following viral inoculation, exhibited marked cytopathic effects (CPEs), mainly represented by changes in cell morphology, including membrane protrusions and vacuolization, as well as cell death. Studies by transmission electron microscopy (TEM) revealed significant ultrastructural changes, such as apoptosis, intense remodeling of membrane-bound organelles and signs of rough endoplasmic reticulum and mitochondrial stress. Additionally, the formation of specialized cytoplasmic vacuoles and intra- and extracellular vesicles emphasized trafficking and intercellular communication as essential mechanisms in OROV infection. RT-qPCR studies confirmed the production of viral progeny in high titers, corroborating the efficiency of this experimental model. These findings contribute to a better understanding of the cytopathogenic mechanisms of OROV infection and the contribution of cellular alterations in OROV morphogenesis. Full article
(This article belongs to the Special Issue Oropouche Virus (OROV): An Emerging Peribunyavirus (Bunyavirus))
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<p>Monolayers of Vero cells in control conditions and infected with Oropouche virus (OROV) at 48, 72, and 96 h post-infection (h p.i.). Bright-field microscopy analysis revealed conformational changes in the membrane as the infection progresses, including the formation of projections and membrane partitioning (insets), which are indicative of apoptosis.</p>
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<p>Cytopathic effects in Vero cells infected with Oropouche virus—cellular response and damage. (<b>A</b>) Uninfected Vero cell. Membrane blebbing (MB). (<b>B</b>–<b>F</b>) OROV-infected cells. (<b>B</b>) Apoptotic bodies (AB), cytoplasmic vacuoles (CV), mitochondrial clustering (MC). (<b>C</b>) Filopodia formation (FF), mitochondrial clustering (MC). (<b>D</b>) Apoptotic bodies (AB), pyknotic nucleus (PN). (<b>E</b>) Mitochondrial cristae disorganization (CD), hypercondensation of chromatin (HC). (<b>F</b>) Lipid droplets (LD), mitochondrial clustering (MC), myelin figure (MF), multilamellar bodies (ML).</p>
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<p>Cytopathic effects in Vero cells infected with Oropouche virus—membrane remodeling. (<b>A</b>) Cytoplasmic vacuoles (CV), plasma membrane blebbing (MB), mitochondrial clustering (MC), nuclear membrane invagination (NI). (<b>B</b>) Multilamellar body (ML), multivesicular body (MVB), rough endoplasmic reticulum (RER) thickening (RT). (<b>C</b>) Autophagy components (AC), budding vesicles (BV). (<b>D</b>) Budding vesicles (BV), RER cisterns (RC). (<b>E</b>) Budding vesicles (BV), filopodia formation (FF), RER cisterns (RC). (<b>F</b>) Lipid droplet (LD), multivesicular cargo (MV), RER cisterns (RC). (<b>G</b>) Multivesicular cargo (MV), RER thickening (RT).</p>
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<p>Cytopathic effects in Vero cells infected with Oropouche virus—rough endoplasmic reticulum (RER) and mitochondrial stress. (<b>A</b>) Mitophagy (MA), mitochondrial swelling (MS), RER thickening (RT). (<b>B</b>) Cytoplasmic vacuole (CV), mitochondrial swelling (MS), pyknotic nucleus (PN), RER cisterns (RC), RER thickening (RT). (<b>C</b>) Myelin figures (MF), mitochondrial swelling (MS), vacuolar degeneration of mitochondria (VD). (<b>D</b>) Mitochondrial swelling (MS), vacuolar degeneration of mitochondria (VD). (<b>E</b>) Cytoplasmic vacuole (CV), giant mitochondria (GM), lipid droplets (LD), multilamellar bodies (ML).</p>
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<p>Cytopathic effects in Vero cells infected with Oropouche virus—viral replication insights. (<b>A</b>,<b>B</b>) Multivesicular body (MVB), viral particles (VP). (<b>A</b>) Immature viral particles (IP), intraluminal vesicle (IV). (<b>B</b>–<b>D</b>) Clathrin-coated vesicles (CCV). (<b>B</b>) Apoptotic body (AB). (<b>C</b>) Intraluminal vesicle (IV), multivesicular body (MVB). (<b>D</b>) Filopodia formation (FF). (<b>E</b>,<b>F</b>) Viral particles (VP). (<b>E</b>) Multivesicular body (MVB), myelin figures (MF). (<b>F</b>) Mitochondrial swelling (MS).</p>
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<p>Morphology of Oropouche viral particles. (<b>A</b>) Viral particle (arrowhead) in extracellular space, adsorbed to the membrane surface. (<b>B</b>–<b>F</b>) Viral particles (arrowhead) into cytoplasm. (<b>C</b>–<b>E</b>) Morphological diversity noted within multivesicular bodies, showcasing particles at different stages of morphogenesis (arrowhead), leading to variations in size and electron density. (<b>C</b>) Reduced diameter observed in indicated particles (arrowhead) reflects incomplete particles. (<b>F</b>) Additional location of viral particles (arrowhead) within cytoplasm, associated with Golgi apparatus stacks. Inset: (<b>C</b>,<b>D</b>) <a href="#viruses-17-00373-f005" class="html-fig">Figure 5</a>A. (<b>E</b>) <a href="#viruses-17-00373-f005" class="html-fig">Figure 5</a>E. (<b>F</b>) <a href="#viruses-17-00373-f005" class="html-fig">Figure 5</a>F. All particles exhibited a spherical shape, with variation in diameter indirectly reflecting maturation status of these particles.</p>
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<p>Morphometric analysis of the mitochondria. (<b>A</b>) Micrograph of the uninfected cell. (<b>B</b>) Micrograph of the OROV-infected cell culture to illustrate morphometric procedure. All mitochondria were counted, excluding those that were not fully within the field (black arrow). For the mitochondrial density of each micrography, the evaluated areas (yellow rectangle) were defined as the boundary surrounding the outermost mitochondria (yellow crosses). The cross-sectional area (black dashed line) and minor and major axes (continuous and dashed white line, respectively) were measured in all mitochondria (<b>C</b>) Mitochondrial density of 20 micrographs per group. Median: 0.29 mitochondria/μm<sup>2</sup> (95% CI, 0.25–0.31)—control group), and 1.78 mitochondria/μm<sup>2</sup> (95% CI, 1.27–2.3)—OROV-infected group. The data presented in panels (<b>C</b>,<b>D</b>) correspond to the micrographs totaling 500 mitochondria per group (control and OROV-infected). (<b>D</b>) Mitochondrial cross-sectional area. Median: 0.37 μm<sup>2</sup> (95% CI, 0.32–0.36)—control group, and 0.08 μm<sup>2</sup> (95% CI, 0.07–0.08)—OROV-infected group. **** <span class="html-italic">p</span> &lt; 0.0001—Mann–Whitney test. (<b>E</b>) Distribution of the frequencies of mitochondrial circularity index. The y-axis indicates the number of mitochondria located within the circularity index range specified by the center bins on the x-axis. Median: 0.72—control group, and 0.68—OROV-infected group.</p>
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15 pages, 2042 KiB  
Review
Insights into CYP1B1-Related Ocular Diseases Through Genetics and Animal Studies
by Elizabeth M. Bolton, Andy Drackley, Antionette L. Williams and Brenda L. Bohnsack
Life 2025, 15(3), 395; https://doi.org/10.3390/life15030395 - 3 Mar 2025
Viewed by 201
Abstract
The CYP1B1 gene encodes a cytochrome p450 monooxygenase enzyme, and over 150 variants have been associated with a spectrum of eye diseases, including primary congenital glaucoma, anterior segment dysgenesis, juvenile open-angle glaucoma, and primary open-angle glaucoma. Clinical genetics has yielded insights into the [...] Read more.
The CYP1B1 gene encodes a cytochrome p450 monooxygenase enzyme, and over 150 variants have been associated with a spectrum of eye diseases, including primary congenital glaucoma, anterior segment dysgenesis, juvenile open-angle glaucoma, and primary open-angle glaucoma. Clinical genetics has yielded insights into the functions of the various CYP1B1 gene domains; however, animal studies are required to investigate the molecular role of CYP1B1 in the eye. While both zebrafish and mice express CYP1B1 in the developing eye, embryonic studies have shown disparate species-specific functions. In zebrafish, CYP1B1 regulates ocular fissure closure such that overexpression causes a remarkable phenotype consisting of the absence of the posterior eye wall. Adult CYP1B1 null zebrafish lack an ocular phenotype but show mild craniofacial abnormalities. In contrast, CYP1B1−/− mice display post-natal mild to severe trabecular meshwork degeneration due to increased oxidative stress damage. Interestingly, the retinal ganglion cells in CYP1B1 null mice may be more susceptible to damage secondary to increased intraocular pressure. Future studies, including detailed genotype–phenotype information and animal work elucidating the regulation, substrates, and downstream effects of CYP1B1, will yield important insights for developing molecularly targeted therapies that will aim to prevent vision loss in CYP1B1-related eye diseases. Full article
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<p>Clinical phenotypes of <span class="html-italic">CYP1B1</span>-related congenital eye diseases. (<b>A</b>,<b>B</b>) Primary congenital glaucoma is diagnosed between birth and 3 years of age and is characterized by elevated IOP typically due to trabeculodysgenesis. Importantly, there is an absence of other types of anterior segment dysgenesis. Typical clinical signs include buphthalmos with increased corneal diameter and axial length, corneal edema (<b>A</b>), Haabs striae ((<b>B</b>) black arrow, breaks in Descemets membrane), and glaucomatous optic neuropathy. (<b>C</b>) Congenital ectropion uvea occurs due to the failure of remnant neural crest cells within the anterior segment to undergo apoptosis. This results in a membrane that pulls the pigmented iris epithelium through the pupil (ectropion uvea, white arrow) and covers the iris and angle. Glaucoma is common and may initially be due to trabeculodysgenesis, but eventually, the membrane causes angle closure. (<b>D</b>) Congenital corneal opacities are considered Peters Anomaly if there is an absence of Descemet’s membrane underlying the corneal defect. Peters Anomaly is due to abnormal separation of the lens vesicle from the overlying surface ectoderm and is divided into two types based on whether the lens is involved (Type 2) or not (Type 1). Other congenital corneal opacities not classified as Peters Anomaly are typically avascular, and the Descemet’s membrane is present under the corneal stromal haze. Glaucoma is diagnosed in more than 50% of affected individuals and occurs due to trabeculo-iridogoniodysgenesis and/or angle closure. (<b>E</b>) Axenfeld–Rieger Syndrome is characterized by Axenfeld Anomaly (posterior embryotoxon (black arrowhead) with iridocorneal touch (white arrowhead)) and Rieger anomaly (iris hypoplasia with pseudopolycoria and/or corectopia). More than 50% of affected individuals develop glaucoma due to iridogoniodysgenesis. (<b>F</b>) Sclerocornea is the absence of demarcation between the cornea and sclera resulting in diffuse corneal opacification often with neovascularization. Sclerocornea is often accompanied by congenital aphakia and glaucoma due to trabeculo-iridogoniodysgenesis. (<b>G</b>,<b>H</b>) Aniridia classically shows pan-ocular defects, including foveal hypoplasia, optic nerve dysplasia/hypoplasia, iris hypoplasia, cataract, and keratopathy, due to limbal stem cell deficiency. In partial aniridia, there is remnant iris (white open arrows) with varying degrees of other ocular findings. Both types are associated with open-angle glaucoma (iridogoniodysgenesis) or closed-angle glaucoma (anterior rotation of the remnant iris root).</p>
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<p><span class="html-italic">CYP1B1</span> gene structure. The <span class="html-italic">CYP1B1</span> gene consists of 3 exons with the start codon in the 2nd exon followed by the N-terminal transmembrane domain. The hinge region gives flexibility to the remaining cytosolic globular portion of the protein. The conserved core structures include the four helices (I-, J-, K-, and L-), between which the meander region and the heme binding sites are located. The location of variants discussed within this review are denoted. The p.M1T variant disrupts the start codon, thereby preventing translation. The p.W57* and G61E variants are both within the hinge region. The W57* is a nonsense variant, while the G61E variant leads to decreased enzymatic activity. The p.Y81N and p.E229K variants are between the hinge region and the I-helix and are predicted to decrease enzymatic activity and disrupt the helical structure, respectively. The p.R355Hfs*69, p.R368H, p.E387K, and p.R390H variants are within the J- and K-helices. The p.E387K and p.R390H variants decrease protein stability and enzymatic activity. The p.368H variant is a VUS but is predicted to have decreased enzymatic activity. The p.R444Q variant in the meander region and the p.R469W variant in the heme-binding region both decrease heme binding, thereby inhibiting protein activity.</p>
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<p><span class="html-italic">CYP1B1</span> protein sequence. The protein sequences for human <span class="html-italic">CYP1B1</span> show 81% and 56% homology with mouse and zebrafish <span class="html-italic">CYP1B1</span>, respectively. There is a high level of conservation between the 3 species in the hinge region, the four helices (I-, J-, K- and L-), the meander region, and the heme-binding domain, which are all denoted. Thus, clinically relevant variants tend to be clustered within these regions. Notably, the amino acids affected by clinically relevant variants discussed within this review, namely W57, G61, R355, R368, E387, R390, R444, and R469 (denoted by boxes), are also conserved between humans, mice, and zebrafish.</p>
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20 pages, 3591 KiB  
Article
Novel HSA-PMEMA Nanomicelles Prepared via Site-Specific In Situ Polymerization-Induced Self-Assembly for Improved Intracellular Delivery of Paclitaxel
by Yang Chen, Shuang Liang, Binglin Chen, Fei Jiao, Xuliang Deng and Xinyu Liu
Pharmaceutics 2025, 17(3), 316; https://doi.org/10.3390/pharmaceutics17030316 - 1 Mar 2025
Viewed by 253
Abstract
Background/Objectives: Paclitaxel (PTX) is a potent anticancer drug that is poorly soluble in water. To enhance its delivery efficiency in aqueous environments, amphiphilic polymer micelles are often used as nanocarriers for PTX in clinical settings. However, the hydrophilic polymer segments on the [...] Read more.
Background/Objectives: Paclitaxel (PTX) is a potent anticancer drug that is poorly soluble in water. To enhance its delivery efficiency in aqueous environments, amphiphilic polymer micelles are often used as nanocarriers for PTX in clinical settings. However, the hydrophilic polymer segments on the surface of these micelles may possess potential immunogenicity, posing risks in clinical applications. To address this issue, nanomicelles based on human serum albumin (HSA)–hydrophobic polymer conjugates constructed via site-specific in situ polymerization-induced self-assembly (SI-PISA) are considered a promising alternative. The HSA shell not only ensures good biocompatibility but also enhances cellular uptake because of endogenous albumin trafficking pathways. Moreover, compared to traditional methods of creating protein–hydrophobic polymer conjugates, SI-PISA demonstrates higher reaction efficiency and better preservation of protein functionality. Methods: We synthesized HSA-PMEMA nanomicelles via SI-PISA using HSA and methoxyethyl methacrylate (MEMA)—a novel hydrophobic monomer with a well-defined and stable chemical structure. The protein activity and the PTX intracellular delivery efficiency of HSA-PMEMA nanomicelles were evaluated. Results: The CD spectra of HSA and HSA-PMEMA exhibited similar shapes, and the relative esterase-like activity of HSA-PMEMA was 94% that of unmodified HSA. Flow cytometry results showed that Cy7 fluorescence intensity in cells treated with HSA-PMEMA-Cy7 was approximately 1.35 times that in cells treated with HSA-Cy7; meanwhile, HPLC results indicated that, under the same conditions, the PTX loading per unit protein mass on HSA-PMEMA was approximately 1.43 times that of HSA. These collectively contributed to a 1.78-fold overall PTX intracellular delivery efficiency of HSA-PMEMA compared to that of HSA. Conclusions: In comparison with HSA, HSA-PMEMA nanomicelles exhibit improved cellular uptake and higher loading efficiency for PTX, effectively promoting the intracellular delivery of PTX. Tremendous potential lies in these micelles for developing safer and more efficient next-generation PTX formulations for tumor treatment. Full article
(This article belongs to the Special Issue Advanced Materials Science and Technology in Drug Delivery)
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Graphical abstract

Graphical abstract
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<p>Scheme of novel HSA-PMEMA nanomicelles prepared via site-specific in situ polymerization-induced self-assembly (SI-PISA) for the improved intracellular delivery of paclitaxel. The blue spheres represent HSA, the yellow lines represent PMEMA, and the red explosion-shaped icons represent PTX. The purple cells are indicative of tumor cells. The solid lines illustrate the synthetic pathway of HSA-PMEMA@PTX and its interactions with the cells, while the dashed lines depict that of HSA@PTX.</p>
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<p>Synthesis and characterization of HSA-Br. (<b>a</b>) Scheme of the synthesis of HSA-Br. (<b>b</b>) <sup>1</sup>H NMR spectrum of DEEBMP in CDCl<sub>3</sub>. Via the identification of peaks in the spectrum, we confirmed that the synthesized DEEBMP has a chemical structure consistent with our expectations. (<b>c</b>) ESI-MS spectrum of DEEBMP. The signals at <span class="html-italic">m</span>/<span class="html-italic">z</span> 334 and 336 correspond to the M + H<sup>+</sup> signals of DEEBMP containing <sup>79</sup>Br and <sup>81</sup>Br, respectively, confirming the presence of DEEBMP (theoretical molecular weight: 334) in the synthesized product. (<b>d</b>) MALDI-TOF MS characterization of HSA and HSA-Br. Given that the theoretical molecular weight of HSA is approximately 66.5 kDa, the signal peaks in the figure correspond to z = 1. The <span class="html-italic">m</span>/<span class="html-italic">z</span> signal peak of HSA-Br is shifted to the right compared to that of HSA, indicating an increase in molecular weight, which confirms the successful conjugation of DEEBMP to HSA.</p>
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<p>Physicochemical characterization of HSA-PMEMA. (<b>a</b>) <sup>1</sup>H NMR spectrum of HSA-PMEMA in deuterated dimethyl sulfoxide. The figure shows the chemical structure of HSA-PMEMA. All proton signals corresponding to the PMEMA chain are observed in the spectrum, confirming the successful in situ growth of PMEMA on HSA. X (Water) represents the water peak, and X (DMSO) represents the solvent peak of DMSO. (<b>b</b>) GPC traces of HSA and HSA-PMEMA. The table shows the number-averaged molecular weight (<span class="html-italic">M</span><sub>n</sub>), weight-averaged molecular weight (<span class="html-italic">M</span><sub>w</sub>), and dispersity (<span class="html-italic">Đ</span>) of the samples.</p>
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<p>Physicochemical characterization of HSA-PMEMA. (<b>a</b>) SDS-PAGE results before and after the in situ growth of PMEMA on HSA. Lane 1: Protein marker; Lane 2: HSA; Lane 3: HSA-Br; Lane 4: HSA-PMEMA. (<b>b</b>) DLS results before and after the in situ growth of PMEMA on HSA. <span class="html-italic">D</span><sub>h</sub> represents the hydrodynamic diameter. The illustrations show the difference between the appearance of HSA and HSA-PMEMA in PBS buffer. (<b>c</b>) Representative TEM images of HSA-PMEMA micelles.</p>
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<p>Evaluation of the protein activity of HSA-PMEMA. (<b>a</b>) CD spectra of HSA, HSA-Br, and HSA-PMEMA. The similar shapes of the CD spectra indicate that the in situ growth of PMEMA on HSA does not affect its secondary structure. (<b>b</b>) Normalized lipase-like activity of HSA, HSA-Br, and HSA-PMEMA.</p>
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<p>Evaluation of the PTX intracellular delivery efficiency of HSA-PMEMA. (<b>a</b>) CLSM images of Cal27 cells co-cultured with HSA-Cy7 or HSA-PMEMA-Cy7 for 2 h. In the CLSM images, green, blue, and red fluorescence correspond to FITC (cytoskeleton), DAPI (nucleus), and Cy7 (HSA or HSA-PMEMA), respectively. (<b>b</b>) Flow cytometry analysis of Cal27 cells co-cultured with HSA-Cy7 or HSA-PMEMA-Cy7. (<b>c</b>) Relative fluorescence intensity of Cal27 cells co-cultured with HSA-Cy7 or HSA-PMEMA-Cy7 compared to the control group (n = 3). The result was derived from flow cytometry analysis. (<b>d</b>) Normalized PTX loading content of HSA@PTX and HSA-PMEMA@PTX. (<b>e</b>) Normalized intracellular PTX delivery efficiency of HSA@PTX and HSA-PMEMA@PTX. *** represents <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Anticancer activity of HSA@PTX and HSA-PMEMA@PTX against 4T1 cells (measurement was based on PTX concentration). The half-maximal inhibitory concentration (IC<sub>50</sub>) was calculated from the inhibition curves fitted.</p>
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<p>In vivo delivery behavior of HSA-PMEMA. (<b>a</b>) Time-dependent changes in plasma HSA concentration in Balb/c mice following the intravenous injection of HSA-Cy7 and HSA-PMEMA-Cy7. (<b>b</b>) Heatmap of Cy7 fluorescence intensity at the tumor site of C3H mice at 12 and 24 h after the intravenous injection of HSA-Cy7 and HSA-PMEMA-Cy7. The blue box indicates the tumor region, with the color scale on the right. (<b>c</b>) Normalized fluorescence intensity derived from panel (<b>b</b>), with the average fluorescence intensity at the tumor site of mice injected with HSA-PMEMA-Cy7 at 12 h set to 100%.</p>
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9 pages, 1058 KiB  
Article
Simultaneous Packaging of Two Different RNA Segments into an Influenza C Virus-like Particle Occurs Inefficiently
by Yasushi Muraki
Viruses 2025, 17(3), 350; https://doi.org/10.3390/v17030350 - 28 Feb 2025
Viewed by 207
Abstract
Reverse genetics systems for influenza C virus encounter challenges due to the inefficient production of infectious virus particles. In the present study, we explored the underlying cause by assessing the efficiency of generating influenza C virus-like particles (C-VLPs) containing specific virus RNA (vRNA) [...] Read more.
Reverse genetics systems for influenza C virus encounter challenges due to the inefficient production of infectious virus particles. In the present study, we explored the underlying cause by assessing the efficiency of generating influenza C virus-like particles (C-VLPs) containing specific virus RNA (vRNA) segments. Using 293T cells transfected with plasmids encoding GFP- and DsRed2-vRNAs (each flanked by the non-coding regions of Segments 5 and 6, respectively), along with plasmids expressing virus proteins, we observed that C-VLPs containing a single vRNA segment were generated efficiently. However, the simultaneous packaging of two vRNA segments into a single C-VLP was less frequent, as demonstrated by flow cytometry and reverse-transcription PCR analyses. Statistical evaluations revealed a decreased efficiency of incorporating multiple vRNA segments into single particles, which likely contributes to the reduced production of infectious virus particles in reverse genetics systems. These findings highlight a critical limitation in the vRNA incorporation mechanism of influenza C virus, contrasting with that of influenza A virus. Hence, further studies are required to elucidate specific vRNA packaging signals and optimize vRNA expression levels to improve the production of infectious influenza C virus particles. Full article
(This article belongs to the Section General Virology)
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<p>A flow chart of the experimental design. Eleven plasmids expressing reporter gene RNA genomes (GFP-vRNA and DsRed2-vRNA) and virus proteins (PB2, PB1, P3, HEF, NP, M1, CM2, NS1, and NS2) were transfected into 293T cells. In these cells, GFP-vRNA and DsRed2-vRNA s (each flanked by the non-coding regions of Segments 5 and 6, respectively) were replicated and transcribed by the RNA polymerase complex comprising the PB2, PB1, P3, and NP proteins, resulting in the expression of GFP and DsRed2 proteins. At 48 h post transfection, the cells were subjected to flow cytometry, and cells expressing both GFP and DsRed2 were sorted. The sorted cells were then incubated for 48 h, and the influenza C virus-like particles (C-VLPs) generated from the cells were collected. HMV-II cells were infected with these C-VLPs, followed by superinfection with a helper virus (C/Ann Arbor/1/50), and the fluorescence-positive cells were examined.</p>
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<p>Gene expressions of plasmid-transfected 293T cells. (<b>A</b>) The transfected 293T cells were subjected to flow cytometry 48 h post transfection. The x- and y-axes in the graph indicate the intensities of GFP and DsRed2, respectively. Each 293T cell analyzed is expressed as a dot. The graph area is divided into four regions according to the proteins expressed: DsRed2, cells expressing DsRed2 alone; EGFP, cells expressing GFP alone; Q2, cells expressing both GFP and DsRed2; and Q3, cells expressing neither GFP nor DsRed2. (<b>B</b>) The sorted 293T cells were observed under a fluorescence microscope to detect GFP and DsRed2 expression. Two independent fields are shown (fields 1 and 2). Scale bar, 20 µm. (<b>C</b>) Total RNA extracted from the sorted 293T cells was treated with DNase I, reverse-transcribed, and PCR-amplified with a primer set specific to <span class="html-italic">GFP</span> or <span class="html-italic">DsRed2</span>. The cDNA preparation setup without reverse transcription was PCR-amplified using primers specific to <span class="html-italic">GFP</span> and electrophoresed in the lane RT(−). The main products in the corresponding lanes are shown (DsRed2, 537 bp; GFP, 504 bp). M: DNA size marker (bp).</p>
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<p>HMV-II cells infected with C-VLPs. The culture supernatant of the sorted 293T cells was added to the HMV-II monolayers, followed by superinfection with the helper virus. At 48 h post infection, the cells were observed under a fluorescence microscope (<b>A</b>). Cells expressing GFP alone, DsRed2 alone, and both GFP and DsRed2 are shown in the upper, middle, and lower panels, respectively. Merged images are shown on the right side. Scale bar, 20 µm. (<b>B</b>) The proportion of fluorescence-positive HMV-II cells expressing GFP or DsRed2 (left bar), or GFP and DsRed2 (right bar). Data obtained from three independent experiments are shown. Comparisons between groups were statistically evaluated using paired <span class="html-italic">t</span>-tests.</p>
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20 pages, 8319 KiB  
Article
Shortening the Saturation Time of PBAT Sheet Foaming via the Pre-Introducing of Microporous Structures
by Fangwei Tian, Junjie Jiang, Yaozong Li, Hanyi Huang, Yushu Wang, Ziwei Qin and Wentao Zhai
Materials 2025, 18(5), 1044; https://doi.org/10.3390/ma18051044 - 26 Feb 2025
Viewed by 236
Abstract
Poly (butylene adipate-co-terephthalate) (PBAT) foam sheets prepared by foaming supercritical fluids are characterized by high resilience, homogeneous cellular structure, and well-defined biodegradability. However, the inert chemical structure and the rigid hard segments restrict the diffusion of CO2 within the PBAT matrix, resulting [...] Read more.
Poly (butylene adipate-co-terephthalate) (PBAT) foam sheets prepared by foaming supercritical fluids are characterized by high resilience, homogeneous cellular structure, and well-defined biodegradability. However, the inert chemical structure and the rigid hard segments restrict the diffusion of CO2 within the PBAT matrix, resulting in extremely long gas saturation times as long as 9 h at a thickness of 12 mm. In this study, microporous structures were pre-introduced into the PBAT matrix to provide a fast gas diffusion pathway during the saturation process. After 2 h of saturation, PBAT foam sheets with expansion ratio of 10 to 13.8 times were prepared. The interaction of CO2 with PBAT was systematically investigated, and the CO2 sorption process was evaluated kinetically and thermodynamically using the Fickian diffusion theory. The solubility and diffusion rate of CO2 in pretreated PBAT sheets with different microporous sizes and densities were investigated, and the effects of pretreatment strategies on the foaming behavior and cell structure of PBAT foam sheets were discussed. The introduction of a microporous structure not only reduces saturation time but also enhances solubility, enabling the successful preparation of soft foams with high expansion ratios and resilience. After undergoing foaming treatment, the PBAT pretreated sheets with a 10 μm microporous structure and a density of 0.45 g/cm3 demonstrated improved mechanical properties: their hardness decreased to 35 C while resilience increased to 58%, reflecting enhanced elastic recovery capabilities. The pretreatment method, which increases the diffusion rate of CO2 in PBAT sheets, offers a straightforward approach that provides valuable insights into achieving rapid and efficient foaming of thick PBAT sheets in industrial applications. Full article
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<p>Schematic of PBAT foam preparation. (<b>a</b>) One-step foaming of PBAT sheets; (<b>b</b>) foaming of PBAT pretreated sheets; (<b>c</b>) schematic diagram of the PBAT sheet pretreatment–short-duration foaming process; (<b>d</b>) parameter distributions during pretreatment–short-duration saturated foaming process.</p>
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<p>CO<sub>2</sub> sorption and diffusion in untreated PBAT sheets. (<b>a</b>) Schematic diagram of CO<sub>2</sub> diffusion in PBAT matrix; (<b>b</b>) CO<sub>2</sub> sorption in PBAT sheets of different thicknesses at 100 °C—18 MPa; (<b>c</b>) fitting of Fick’s diffusion model; (<b>d</b>) isothermal sorption of CO<sub>2</sub> in PBAT; (<b>e</b>) kinetic linear fitting of diffusion coefficients at different temperatures; and (<b>f</b>) kinetic linear fitting of diffusion coefficients at different pressures.</p>
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<p>PBAT foam thermal behavior. (<b>a</b>) DSC curves of the PBAT samples treated under various pressures and temperatures; (<b>b</b>) degree of crystallinity based on DSC pattern.</p>
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<p>SEM image of the evolution of the cell structure of untreated PBAT sheet (6 mm thickness) with saturation time at 100 °C—18 MPa. (<b>a<sub>1</sub></b>–<b>e<sub>1</sub></b>) evolution of the cell structure in the edge region with saturation time (10–150 min); (<b>a<sub>2</sub></b>–<b>e<sub>2</sub></b>) evolution of the cell structure in the middle region with saturation time (10–150 min); and (<b>a<sub>3</sub></b>–<b>e<sub>3</sub></b>) evolution of the cell structure in the core region with saturation time (10–150 min).</p>
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<p>Dynamic information on the cell structure of PBAT foams obtained at 100 °C—18 MPa. (<b>a</b>) Cell size across different positions at different times; (<b>b</b>) cell density across different positions at different times.</p>
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<p>SEM image of microporous structures of typical PBAT pretreated sheets. (<b>a<sub>1</sub></b>–<b>c<sub>1</sub></b>) samples with different microporous sizes at a density of 0.35 g/cm<sup>3</sup>; (<b>a<sub>2</sub></b>–<b>c<sub>2</sub></b>) samples with different microporous sizes at a density of 0.45 g/cm<sup>3</sup>; (<b>a<sub>3</sub></b>–<b>c<sub>3</sub></b>) samples with different microporous sizes at a density of 0.65 g/cm<sup>3</sup>.</p>
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<p>Diffusive behavior of CO<sub>2</sub> in pretreated PBAT sheets. (<b>a</b>) Effect of microporous structure on cell wall; (<b>b</b>) CO<sub>2</sub> sorption process under different microporous structures; (<b>c</b>) linear fitting of diffusion coefficients; (<b>d</b>) schematic diagram of rapid CO<sub>2</sub> diffusion in pretreated PBAT sheets.</p>
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<p>SEM images of foamed PBAT sheets with different microporous structures (red circles show graded cell structures). (<b>a<sub>1</sub></b>–<b>c<sub>1</sub></b>) samples with different micropore sizes of 10 μm, 75 μm and 140 μm at a density of 0.35 g/cm<sup>3</sup>; (<b>a<sub>2</sub></b>–<b>c<sub>2</sub></b>) samples with different micropore sizes of 10 μm, 75 μm and 140 μm at a density of 0.35 g/cm<sup>3</sup>; (<b>a<sub>3</sub></b>–<b>c<sub>3</sub></b>) samples with different micropore sizes of 10 μm, 75 μm and 140 μm at a density of 0.35 g/cm<sup>3</sup>.</p>
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<p>Cell morphology of PBAT foamed sheets with different microporous structures. (<b>a</b>) Cell size; (<b>b</b>) cell density; (<b>c</b>) expansion ratio.</p>
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<p>Mechanical properties of PBAT foams. (<b>a</b>,<b>b</b>) Compression properties, (<b>c</b>) hardness, and (<b>d</b>) resilience.</p>
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15 pages, 4678 KiB  
Article
Genetic Characterization of SWEET Genes in Coconut Palm
by Jiepeng Chen, Weiming Zeng, Jiali Mao, Runan Chen, Ran Xu, Ying Wang, Ruibo Song, Zifen Lao, Zhuang Yang, Zhihua Mu, Ruohan Li, Hongyan Yin, Yong Xiao, Jie Luo and Wei Xia
Plants 2025, 14(5), 686; https://doi.org/10.3390/plants14050686 - 23 Feb 2025
Viewed by 250
Abstract
Sugar-Will-Eventually-be-Exported Transporters (SWEETs) play a crucial role in sugar transport in plants, mediating both plant development and stress responses. Despite their importance, there has been limited research characterizing the functional characteristics of CnSWEET genes in coconut (Cocos nucifera). In this study, [...] Read more.
Sugar-Will-Eventually-be-Exported Transporters (SWEETs) play a crucial role in sugar transport in plants, mediating both plant development and stress responses. Despite their importance, there has been limited research characterizing the functional characteristics of CnSWEET genes in coconut (Cocos nucifera). In this study, we conducted a systematic analysis of SWEET genes in coconut using bioinformatics, subcellular localization studies, in silico promoter analysis, and functional assays with yeast mutants. A total of 16 CnSWEET genes were identified and grouped into four clades. Clade I contained the highest number of genes (eight), derived from four pairs of duplicated genomic segments. In contrast, the other clades had fewer genes (one to four) compared to those in Arabidopsis and other species in the Arecaceae family. An extensive analysis of gene expansion using MSCanX indicated significant divergence in gene expansion patterns, both between and within monocots and dicots, as well as among closely related species within the same family. Notable variations in conserved protein motifs and the number of transmembrane helices (TMHs) were detected within Clade I compared to other clades, affecting the subcellular localization of CnSWEET proteins. Specifically, seven TMHs were associated with proteins located in the cell membrane, while CnSWEET2A, which had five TMHs, was found in both the cell membrane and cytosol. Promoter analysis revealed that some CnSWEET genes contained endosperm or seed specific motifs associated with specific endosperm expression, consistent with expression patterns observed in transcriptome data. Functional analysis of five CnSWEET genes, with transcript sequences supported by transcriptome data, was conducted using yeast mutant complementation assays. This analysis demonstrated diverse transport activities for sucrose, fructose, glucose, galactose, and mannose. Our findings provide valuable insights into the functional diversity of SWEET genes in coconuts and their potential roles in stress responses and plant development. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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<p>The gene structures and protein features of <span class="html-italic">CnSWEETs</span>. (<b>A</b>) The gene structures of <span class="html-italic">CnSWEETs</span> displayed according to gene model information and ISOseq data modification. The gene structures were displayed using TBtools. Transposon and repeat sequences were analyzed by RepeatMasker (v4.1.1). (<b>B</b>) The distribution of conserved protein motifs and transmembrane regions. The former were deduced from the MEME online software. The TMHMM online software was used to analyze the transmembrane regions.</p>
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<p><span class="html-italic">CnSWEET</span> gene members in coconut palm and compared with <span class="html-italic">AtSWEETs</span>. (<b>A</b>) The phylogenetic tree of <span class="html-italic">CnSWEETs</span> and <span class="html-italic">AtSWEETs</span> was constructed using the neighbor-joining method in MEGA 7.0. (<b>B</b>) The genomic locations of <span class="html-italic">AtSWEET</span> genes situated within the duplicated genomic segments. (<b>C</b>) The genomic locations of <span class="html-italic">CnSWEET</span> genes situated within the duplicated genomic segments. (<b>D</b>) Homologous genomic segments between coconut and Arabidopsis contain <span class="html-italic">CnSWEET</span> and <span class="html-italic">AtSWEET</span> genes, respectively. The genomic locations of <span class="html-italic">CnSWEETs</span> and <span class="html-italic">AtSWEETs</span>, along with duplicated genomic segments containing <span class="html-italic">SWEETs</span> deduced from MCScanX analysis, were visualized by TBtools.</p>
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<p>The <span class="html-italic">SWEET</span> gene expansion of coconut and fourteen other species in the context of angiosperms. The ML phylogenetic tree of these 15 species was constructed using 140 single-copy genes detected by orthfinder, which was the same method used in our previous research [<a href="#B33-plants-14-00686" class="html-bibr">33</a>]. The method used in this study determined the number, subclade, and duplicated events of <span class="html-italic">SWEETs</span> in each species, following the approach used for coconut and Arabidopsis. The length of the colored bar with numbers represents the number of <span class="html-italic">SWEETs</span> detected in each clade. The genomic segment duplication events were identified by MCScanX analysis. “Dup”, “Tri”, and “Tetra” represent that the numbers of homologous segments in each duplication event were two, three, and four, respectively. The number before these letters represents the frequency of this type of duplication detected. The synteny regions in chromosomes were displayed in circles using TBtools. The length of chromosomes is measured in megabases. The Materials and Methods Section lists the information about the three characters representing the species. Three-letter abbreviations are used to represent each species: <span class="html-italic">Amborella trichopoda</span> (Atr), <span class="html-italic">Daucus carota</span> (Dca), <span class="html-italic">Solanum tuberosum</span> (Stu), <span class="html-italic">Vitis vinifera</span> (Vvi), <span class="html-italic">Malus domestica</span> (Mdo), <span class="html-italic">Citrus sinensis</span> (Csi), <span class="html-italic">Arabidopsis thaliana</span> (Ath), <span class="html-italic">Dioscorea alata</span> (Dal), <span class="html-italic">Phoenix dactylifera</span> (Pda), <span class="html-italic">Elaeis guineensis</span> (Egu), <span class="html-italic">Musa acuminata</span> (Mac), <span class="html-italic">Ananas comosus</span> (Aco), <span class="html-italic">Brachypodium distachyon</span> (Bdi), and <span class="html-italic">Oryza sativa</span> (Osa).</p>
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<p>The distribution of the promoter conserved motifs and expression patterns of <span class="html-italic">CnSWEETs</span>. (<b>A</b>) Motifs in all <span class="html-italic">CnSWEET</span> promoters. The motifs in the putative promoters of <span class="html-italic">CnSWEETs</span>, located 2000 bp upstream from the ATG start codon, were analyzed using the Plantcare and TSSP software. (<b>B</b>) The heatmap of <span class="html-italic">CnSWEET</span> expression based on log2-transformed mean FPKM for coconut leaf, shoot, endosperm, and mesocarp tissues was generated using the transcriptomes used in this study. (<b>C</b>) The Pearson correlation coefficients of FPKM values for gene pairs were derived from segmental duplication. The Pearson correlation coefficient (P.C.C.) between duplicated <span class="html-italic">CnSWEET</span> gene pairs was calculated using the coconut transcriptomes, and significance testing was performed using a <span class="html-italic">t</span>-test (* <span class="html-italic">p</span> &lt; 0.05) in R (cor.test).</p>
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<p>The subcellular location and 3D protein structures of CnSWEET proteins. (<b>A</b>) CnSWEET proteins were localized to the membranes. (<b>B</b>) The 3D protein structures of CnSWEET protein were deduced and displayed using the online SWISS-MODEL. The 35S::CnSWEET: eGFP fusion protein and the cell membrane marker 35S::OsCBL1:RFP fusion protein were transiently expressed in tobacco epidermal cells. GFP and RFP signals were detected with time intervals between 48 and 72 h post-infiltration using a confocal microscope (SESIS, LMS980). GFP: green fluorescence; RFP: red fluorescence; Bright field: visible light; Merge: visible light merged with fluorescence. Scalebars: 50 μm or 100 μm.</p>
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<p>Substrate specificity analysis of five selected CnSWEET proteins in yeast mutant strains EBY.VW4000 (<b>A</b>) and SUSY7/ura3 (<b>B</b>). Cells were serially diluted 10-fold (10-, 100-, and 1000-fold) and spotted on solid SD media supplemented with 2% concentration of different sugar substrates. Maltose and glucose were only carbon sources used in positive controls for EBY, VW4000 and SUSY7/ura3 cells, respectively.</p>
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5 pages, 146 KiB  
Editorial
Nanomaterials Applied to Fuel Cells and Catalysts
by András Tompos
Nanomaterials 2025, 15(5), 347; https://doi.org/10.3390/nano15050347 - 23 Feb 2025
Viewed by 153
Abstract
Hydrogen and fuel cell technologies are accepted by consensus as being part of the future energy system, especially in hard-to-abate segments where electrification is not an efficient solution [...] Full article
(This article belongs to the Special Issue Nanomaterials Applied to Fuel Cells and Catalysts)
23 pages, 6468 KiB  
Article
Urban Signalized Intersection Traffic State Prediction: A Spatial–Temporal Graph Model Integrating the Cell Transmission Model and Transformer
by Anran Li, Zhenlin Xu, Wenhao Li, Yanyan Chen and Yuyan Pan
Appl. Sci. 2025, 15(5), 2377; https://doi.org/10.3390/app15052377 - 23 Feb 2025
Viewed by 242
Abstract
This paper presents the Cell Transformer (CeT), which utilizes high-definition (HD) map data to predict future traffic states at signalized intersections, thereby aiding trajectory planning for autonomous vehicles. CeT employs discretized lane segments to emulate the cell transmission model, creating a cell space [...] Read more.
This paper presents the Cell Transformer (CeT), which utilizes high-definition (HD) map data to predict future traffic states at signalized intersections, thereby aiding trajectory planning for autonomous vehicles. CeT employs discretized lane segments to emulate the cell transmission model, creating a cell space to forecast vehicle counts across all segments based on historical traffic data. CeT enhances prediction accuracy by distinguishing between different vehicle types by incorporating vehicle-type attributes into vehicle-state representations through multi-head attention. In this framework, cells are modeled as nodes in a directed graph, with dynamic connections representing variations in signal phases, thereby embedding spatial relationships and signal information within dynamic graphs. Temporal embeddings derived from time attributes are integrated with these graphs to generate comprehensive spatial–temporal representations. Utilizing an encoder–decoder architecture, CeT captures dependencies and correlations from past cell states to predict future traffic conditions. Validation using real traffic data from pNEUMA demonstrates that CeT significantly outperforms baseline models in two-phase signalized intersection scenarios, achieving reductions of 11.47% in Mean Absolute Error (MAE), 13.48% in Root Mean Square Error (RMSE), and an increase of 4.36% in Accuracy (ACC). In four-phase signalized intersection scenarios, CeT shows even greater effectiveness, with improvements of 13.36% in MAE, 12.93% in RMSE, and 4.78% in ACC. These results underscore CeT’s superior predictive capabilities and highlight the contributions of its core components. Full article
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<p>A three-dimensional representation in which the horizontal axis denotes lanes, the vertical axis indicates time, and the depth axis represents the road. The ego vehicle and background vehicles are situated along lane segments. The orange space–time corridor delineates the unoccupied future lane segments available for AV, while the gray curve illustrates the optimized planning trajectory within this corridor. This visualization emphasizes the consideration of future traffic states, which is essential for ensuring safe and efficient trajectory planning.</p>
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<p>The illustration of the cell space in conventional CTM at a signalized intersection.</p>
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<p>The architecture of CeT and its key components.</p>
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<p>The data collection area in pNEUMA and the zone for each drone in the swarms.</p>
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<p>The establishment of the experimental scenarios. (<b>a</b>) The OSM map for the data collection area in pNEUMA. (<b>b</b>) The densely distributed zones of signalized intersections in CommonRoad. (<b>c</b>) The illustration of the two adjacent signalized intersections in Scenario 1. (<b>d</b>) The illustration of the constructed Scenario 1. The color ring denotes the azimuths of vehicles.</p>
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<p>The comparison of predictive performance between CeT and baseline models in Scenario 1 and Scenario 2.</p>
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<p>Ablation study for vehicle-state embeddings in Scenario 1 and Scenario 2.</p>
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<p>Ablation study for spatial-temporal embeddings in Scenario 1 and Scenario 2.</p>
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<p>Ablation study for spatial and temporal attention in Scenario 1 and Scenario 2.</p>
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<p>Analysis of the impact of hyperparameters on predictive performance in Scenario 2.</p>
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<p>Analysis of learning rates and batch sizes on model convergence in Scenario 2.</p>
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17 pages, 4488 KiB  
Article
Early-Life Stress Caused by Maternal Deprivation Impacts Dendritic Morphology of Adult Male Mouse Neocortical Interneurons
by Mohammed M. Nakhal, Lidya K. Yassin, Shaikha Al Houqani, Ayishal B. Mydeen, Marwa F. Ibrahim, Safa Shehab, Mohammed Z. Allouh and Mohammad I. K. Hamad
Int. J. Mol. Sci. 2025, 26(5), 1909; https://doi.org/10.3390/ijms26051909 - 23 Feb 2025
Viewed by 266
Abstract
A substantial body of research suggests that early-life stress (ELS) is associated with neuropathology in adulthood. Maternal deprivation (MD) is a commonly utilised model in mice for the study of specific neurological diseases. The appropriate growth of dendrites is essential for the optimal [...] Read more.
A substantial body of research suggests that early-life stress (ELS) is associated with neuropathology in adulthood. Maternal deprivation (MD) is a commonly utilised model in mice for the study of specific neurological diseases. The appropriate growth of dendrites is essential for the optimal functioning of the nervous system. However, the impact of ELS on interneuron dendritic morphology remains unclear. To ascertain whether ELS induces alterations in the morphology of GABAergic inhibitory interneurons in layers II/III of the medial entorhinal cortex (mEC), the somatosensory cortex (SSC), the motor cortex (MC), and the CA1 region of the hippocampus (Hp), 9-day-old male GAD-67-EGFP transgenic mice were subjected to a 24 h MD. At postnatal day 60 (P60), the animals were sacrificed, and their brains were subjected to morphological analyses. The results indicated that MD affected the dendritic morphology of GABAergic interneurons. The mean dendritic length and mean dendritic segments of the examined cortical areas, except for the MC, were significantly decreased, whereas the number of primary dendrites was unaffected. Furthermore, the density of GAD67-EGFP-positive interneurons was decreased in the mEC and Hp, but not in the somatosensory and MC. The induction of ELS through MD in a developmental time window when significant morphological changes occur rendered the developing cells particularly susceptible to stress, resulting in a significant reduction in the number of surviving interneurons at the adult stage. Full article
(This article belongs to the Special Issue Current Insights on Neuroprotection)
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<p>Experimental procedure. The day of delivery was P0. On P9, the litters were subjected to the MD procedure. The dams were removed from the litter, and the pups remained in their home cage at room temperature. A day later, the dams were returned to their cages. As a control experiment, the dams were removed from their home cages for 5 min. The animals were sacrificed at P60, perfused transcardially, immunostained against EGFP, and the dendritic morphology was quantified.</p>
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<p>MD effect on the morphology of the interneuron of the mEC. Parasagittal brain slices from the mEC region were stained with EGFP to visualise morphology by light microscopy. GAD67-positive interneurons were reconstructed using Neurolucida. (<b>A</b>–<b>C</b>,<b>G</b>) The median value is represented by the horizontal lines within the box plots, while the variabilities outside the upper and lower quartiles are indicated with whiskers. The middle half of the sample is represented with the box. (<b>A</b>) Mean dendritic length. (<b>B</b>) Mean dendritic segments. (<b>C</b>) Mean of the number of primary dendrites. The number of reconstructed cells is shown in (<b>A</b>) for the control and MD groups. The number of cells analysed was obtained from 8 control mice (2–3 slices corresponding only to the mEC area) and 8 MD mice (2–3 slices). Example images at 40× magnification from a control (<b>D</b>) and an MD interneuron are shown (<b>E</b>). The traces are shown next to the images. Scale bars = 50 μm. (<b>F</b>) Sholl analysis of the control and MD groups. The error bars in (<b>F</b>) represent the standard error mean. MD-induced decrease in the number of intersections was observed between 60 and 170 μm from the soma using <span class="html-italic">t</span>-test post hoc Bonferroni corrections. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001. (<b>G</b>) The mean of the total number of dendritic intersections. In the statistical analyses conducted for the experiment in (<b>F</b>), the number of dendritic intersections was compared between the control and MD groups at each distance point. For the graphs in (<b>A</b>–<b>C</b>,<b>G</b>), the <span class="html-italic">p</span> values were calculated using the Mann–Whitney U test and reported only if they were statistically significant. ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>MD effect on the morphology of the interneuron of the SSC. Parasagittal brain slices from the SSC region were stained with EGFP to visualise morphology by light microscopy. GAD67-positive interneurons were reconstructed using Neurolucida. (<b>A</b>–<b>C</b>,<b>G</b>) The median value is represented by the horizontal lines within the box plots, while the variabilities outside the upper and lower quartiles are indicated with whiskers. The middle half of the sample is represented with the box. (<b>A</b>) Mean dendritic length. (<b>B</b>) Mean dendritic segments. (<b>C</b>) Mean of the number of primary dendrites. The number of reconstructed cells is shown in (<b>A</b>) for the control and MD groups. The number of cells analysed were obtained from 8 control mice (2–3 slices corresponding only to the SSC area) and 8 MD mice (2–3 slices). Example images at 40× magnification from a control (<b>D</b>) and an MD interneuron are shown (<b>E</b>). The traces are shown next to the images. Scale bars = 50 μm. (<b>F</b>) Sholl analysis of the control and MD groups. The error bars in (<b>F</b>) represent the standard error mean. MD-induced decrease in the number of intersections was observed between 70 and 110 μm from the soma using <span class="html-italic">t</span>-test post hoc Bonferroni corrections. * <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. (<b>G</b>) The mean of the total number of dendritic intersections. In the statistical analyses conducted for the experiment in (<b>F</b>), the number of dendritic intersections was compared between the control and MD groups at each distance point. For the graphs in (<b>A</b>–<b>C</b>,<b>G</b>), the <span class="html-italic">p</span> values were calculated using the Mann–Whitney U test and reported only if they were statistically significant. ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>MD does not affect MC interneuron morphology. Parasagittal brain slices from the MC region were stained with EGFP to visualise morphology by light microscopy. GAD67-positive interneurons were reconstructed using Neurolucida. (<b>A</b>–<b>C</b>,<b>G</b>) The median value is represented by the horizontal lines within the box plots, while the variabilities outside the upper and lower quartiles are indicated with whiskers. The middle half of the sample is represented with the box. (<b>A</b>) Mean dendritic length. (<b>B</b>) Mean dendritic segments. (<b>C</b>) Mean of the number of primary dendrites. The number of reconstructed cells is shown in (<b>A</b>) for the control and MD groups. The number of cells analysed was obtained from 8 control mice (2–3 slices corresponding only to the MC area) and 8 MD mice (2–3 slices). Example images at 40× magnification from a control (<b>D</b>) and an MD interneuron are shown (<b>E</b>). The traces are shown next to the images. Scale bars = 50 μm. (<b>F</b>) Sholl analysis of the control and MD groups. The error bars in (<b>F</b>) represent the standard error mean. MD did not affect the number of intersections using 2-way repeated measures ANOVA. (<b>G</b>) The mean of total number of dendritic intersections. In the statistical analyses conducted for the experiment in (<b>F</b>), the number of dendritic intersections was compared between the control and MD groups at each distance point.</p>
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<p>MD effect on the morphology of the interneuron of the Hp. Parasagittal brain slices from the Hp region were stained with EGFP to visualise morphology by light microscopy. GAD67-positive interneurons were reconstructed using Neurolucida. (<b>A</b>–<b>C</b>,<b>G</b>) The median value is represented by the horizontal lines within the box plots, while the variabilities outside the upper and lower quartiles are indicated with whiskers. The middle half of the sample is represented with the box. (<b>A</b>) Mean dendritic length. (<b>B</b>) Mean dendritic segments. (<b>C</b>) Mean of the number of primary dendrites. The number of reconstructed cells is shown in (<b>A</b>) for the control and MD groups. The number of cells analysed was obtained from 8 control mice (2–3 slices corresponding only to the Hp area) and 8 MD mice (2–3 slices). Example images at 40× magnification from a control (<b>D</b>) and an MD interneuron are shown (<b>E</b>). The traces are shown next to the images. Scale bars = 50 μm. (<b>F</b>) Sholl analysis of the control and MD groups. The error bars in (<b>F</b>) represent the standard error mean. MD-induced decrease in the number of intersections was observed between 70 and 200 μm from the soma using <span class="html-italic">t</span>-test post hoc Bonferroni corrections. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001 (<b>G</b>) The mean of the total number of dendritic intersections. In the statistical analyses conducted for the experiment in (<b>F</b>), the number of dendritic intersections was compared between the control and MD groups at each distance point. For the graphs in (<b>A</b>–<b>C</b>,<b>G</b>), the <span class="html-italic">p</span> values were calculated using the Mann–Whitney U test and reported only if they were statistically significant. ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effect of MD on GAD67-positive interneuron density. (<b>A</b>) Virtual parasagittal brain slice from the Allen Brain Atlas reference showing the 4 main regions examined. (<b>B</b>) An example of a parasagittal brain slice from a GAD67-GFP-stained brain slice. (<b>C</b>–<b>F</b>) The median value is represented by the horizontal lines within the box plots, while the variabilities outside the upper and lower quartiles are indicated with whiskers. The middle half of the sample is represented with the box. (<b>C</b>) Mean values of the number of GAD67-positive interneurons per 1 mm<sup>2</sup> in the mEC region. The accompanying illustration depicts a photomicrograph captured at 4× magnification (scale bar = 200 μm) and a zoomed area captured at 40× magnification (scale bar = 20 μm) from the control and MD mice from the mEC region. It is presented alongside the corresponding graph. (<b>D</b>) The box plot in the graph represents the mean values of the number of GAD67-positive interneurons per 1 mm<sup>2</sup> in the SSC region. (<b>E</b>) Mean values of the number of GAD67-positive interneurons per 1 mm<sup>2</sup> in the MC region. (<b>F</b>) Mean values of the number of GAD67-positive interneurons per 1 mm<sup>2</sup> in the Hp region. The total number of the analysed regions of interest (ROIs) is indicated above the box plot for each condition. 5–6 ROI were analysed per animal from control and MD groups. The number of analysed ROIs is shown in the graphs for the control and MD groups. The analysed ROIs were obtained from 7 control mice (2–3 slices corresponding to each area) and 8 MD mice (2–3 slices corresponding to each area). Mann–Whitney U test, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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20 pages, 3715 KiB  
Article
L-GABS: Parametric Modeling of a Generic Active Lumbar Exoskeleton for Ergonomic Impact Assessment
by Manuel Pérez-Soto, Javier Marín and José J. Marín
Sensors 2025, 25(5), 1340; https://doi.org/10.3390/s25051340 - 22 Feb 2025
Viewed by 284
Abstract
Companies increasingly implement exoskeletons in their production lines to reduce musculoskeletal disorders. Studies have been conducted on the general ergonomic effects of exoskeletons in production environments; however, it remains challenging to predict the biomechanical effects these devices may have in specific jobs. This [...] Read more.
Companies increasingly implement exoskeletons in their production lines to reduce musculoskeletal disorders. Studies have been conducted on the general ergonomic effects of exoskeletons in production environments; however, it remains challenging to predict the biomechanical effects these devices may have in specific jobs. This article proposes the parametric modeling of an active lumbar exoskeleton using the Forces ergonomic method, which calculates the ergonomic risk using motion capture in the workplace, considering the internal joint forces. The exoskeleton was studied to model it in the Forces method using a four-phase approach based on experimental observations (Phase 1) and objective data collection via motion capture with inertial sensors and load cells for lifting load movements. From the experimentation the angles of each body segment, the effort perceived by the user, and the activation conditions were obtained (Phase 2). After modeling development (Phase 3), the experimental results regarding the force and risk were evaluated obtaining differences between model and experimental data of 0.971 ± 0.171 kg in chest force and 1.983 ± 0.678% in lumbar risk (Phase 4). This approach provides a tool to evaluate the biomechanical effects of this device in a work task, offering a parametric and direct approximation of the effects prior to implementation. Full article
(This article belongs to the Special Issue Wearable Robotics and Assistive Devices)
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<p>Apogee active exoskeleton diagram (images from Apogee User Manual).</p>
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<p>Apogee exoskeleton configuration examples and legend.</p>
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<p>Model diagram forces (right: R, left: L) from Delgado et al. [<a href="#B15-sensors-25-01340" class="html-bibr">15</a>] modified for Apogee Exoskeleton.</p>
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<p>Load cell assembly for force measurement. Experimentation with motion capture and synchronized force measurement. CAD drawing of load cell assembly (1: Load cell, 2: Ad hoc 3D printed part with a threaded insert, 3: Adjustable back support, 4: Casing with electric motors at the side).</p>
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<p>Experimental diagram.</p>
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<p>(<b>a</b>) Averaged chest force curves experimentally obtained concerning lumbar flexion. (<b>b</b>) Modeled passive torque curves on each side.</p>
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<p>(<b>a</b>) Averaged chest force curves obtained experimentally concerning lumbar angular velocity. (<b>b</b>) Modeled active torque curves on each side.</p>
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<p>Force measured on the chest compared to the force simulated by the model. (<b>a</b>) Passive assistance comparison. (<b>b</b>) Active assistance comparison. (<b>c</b>) Active Mod. assistance comparison.</p>
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<p>Force measured on the chest compared to the force simulated by the model. (<b>a</b>) Passive assistance comparison. (<b>b</b>) Active assistance comparison. (<b>c</b>) Active Mod. assistance comparison.</p>
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<p>Comparison of the hybrid assistance. Force measured on the chest compared to force simulated by the model.</p>
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13 pages, 4993 KiB  
Article
Smooth Muscle Silent Information Regulator 1 Contributes to Colitis in Mice
by Xiaoqin Liu, Yu Song, Mengmeng Shen, Xinlong Liu, Wendi Zhang, Haibin Jiang and Mei Han
Int. J. Mol. Sci. 2025, 26(5), 1807; https://doi.org/10.3390/ijms26051807 - 20 Feb 2025
Viewed by 208
Abstract
Smooth muscle cells (SMCs) are an essential component of the intestine, play an important role to maintain intestine structure, and produce peristaltic and segmentation movements. The silent information regulator 1 (SIRT1) has a dual role along with possible mechanisms in the different experimental [...] Read more.
Smooth muscle cells (SMCs) are an essential component of the intestine, play an important role to maintain intestine structure, and produce peristaltic and segmentation movements. The silent information regulator 1 (SIRT1) has a dual role along with possible mechanisms in the different experimental models of inflammatory bowel disease (IBD). However, very little is known about other putative roles that overexpression of SIRT1 in SMCs may have. Here, we explored the role of SMC SIRT1 in colonic mucosa regeneration and recovery after DSS-induced colitis. We showed that smooth-muscle-specific SIRT1 transgene (Sirt1-Tg) mice have abnormal baseline intestinal architecture. The overexpression of SIRT1 impaired the recovery after DSS-induced injury. Furthermore, we showed that smooth-muscle SIRT1 affected the intestinal epithelial regeneration after damage by releasing cZFP609, which inhibited the hypoxia-inducible factor (HIF)-1α nuclear translocation. Together, we identify an important signaling axis cZFP609-HIF-1α linking SMCs and intestinal epithelium, which is involved in colitis development. Full article
(This article belongs to the Section Molecular Biology)
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<p>Abnormal baseline intestinal architecture in mice <span class="html-italic">Sirt1</span>-Tg mice compared to WT mice. (<b>A</b>) qRT-PCR for the expression of SIRT1 in mouse colon smooth muscle tissue. (<b>B</b>) Representative images showing colon length of WT and <span class="html-italic">Sirt1</span>-Tg mice. (<b>C</b>) Statistics of colon length of WT and <span class="html-italic">Sirt1</span>-Tg mice. (<b>D</b>) Representative H&amp;E-stained images, out of three independently acquired, of colon sections from the indicated groups of mice (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>E</b>) Representative Alcian-Blue-stained images, out of three independently acquired, of colon sections from the indicated groups (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>F</b>) Representative photomicrographs of colonic PCNA IHC staining in each group (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>G</b>) Quantified colonic crypt depth. (<b>H</b>) Quantification of the average number of goblet cells per crypt. (<b>I</b>) AOD analysis of IHC results of CLAUDIN2, <span class="html-italic">n</span> = 6. (<b>J</b>) Western blot detection of ZO1 and CLAUDIN1. (<b>K</b>) qRT-PCR for the expression of inflammation-related cytokines. All quantifications are represented as mean ± SD and statistical significance was assessed by two-tailed unpaired Student’s <span class="html-italic">t</span>-test. Actual <span class="html-italic">p</span>-values are indicated in each graph. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; ns indicates no significant change. AOD: average optical density; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.</p>
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<p><span class="html-italic">Sirt1</span>-Tg mice exhibit impaired intestinal regeneration in DSS-induced colitis. (<b>A</b>) Timeline of DSS treatment. Mice were exposed to 2.5% DSS for 5 days followed by 6 days of regular water to allow for epithelial restoration. (<b>B</b>) Representative images showing colon length at end point. (<b>C</b>) Statistics of colon length of WT and <span class="html-italic">Sirt1</span>-Tg mice with DSS. (<b>D</b>) Weight loss relative to % to initial weight, <span class="html-italic">n</span> = 6 mice per genotype, determined daily and compared to the weights at the start of DSS treatment for each mouse. (<b>E</b>) Disease activity index measured daily in mice described in a during DSS treatment schedule. (<b>F</b>) Representative H&amp;E-stained images, out of three independently acquired, of colon sections from the indicated groups of mice treated at the end of DSS administration (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>G</b>) Histopathological changes in colon tissue of WT and <span class="html-italic">Sirt1</span>-Tg mice with DSS. (<b>H</b>) Representative Alcian-Blue-stained images, out of three independently acquired, of colon sections from the indicated groups of mice treated at the end of DSS administration (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>I</b>) Representative immunohistochemistry images of the PCNA in the colon, out of three independently acquired, of colon sections from the indicated groups of mice treated at the end of DSS administration (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>J</b>) AOD analysis of IHC results of PCNA, <span class="html-italic">n</span> = 6. (<b>K</b>) qPCR analysis of cZFP609 levels in colon tissue of mice at the end of DSS administration. Data are presented as mean ± SD of <span class="html-italic">n</span> = 6. All quantifications are represented as mean ± SD and statistical significance was assessed by two-tailed unpaired Student’s <span class="html-italic">t</span>-test. Actual <span class="html-italic">p</span>-values are indicated in each graph. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 (two-tailed Student’s <span class="html-italic">t</span>-test).</p>
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<p>The conditioned medium (CM) of <span class="html-italic">Sirt1</span>-Tg CSMCs inhibits the proliferation of Caco-2 cells in vitro. (<b>A</b>) qRT-PCR of cZFP609 expression in CSMCs from WT or <span class="html-italic">Sirt1</span>-Tg mice treated with TNF-α for 24 h. (<b>B</b>) qRT-PCR of cZFP609 expression in Caco-2 cells incubated with the TNF-α-induced CSMC conditioned medium (CM) for 24 h and exposed to hypoxia. (<b>C</b>) Ratio of cell counting of Caco-2 cells incubated with the TNF-α-induced CSMC (CM) for 24 h and exposed to hypoxia. (<b>D</b>,<b>E</b>) Migration of Caco-2 cells was assessed using a scratch wound assay. Caco-2 cells were incubated with the TNF-α-induced CSMC exosomes (CM) for 24 h and exposed to hypoxia treated with TNF-α. (Scale bar: 200 µm.) Data are presented as mean ± SEM. (<b>F</b>) Immunofluorescent confocal microscopy of HIF1α nuclear translocation in the Caco-2 cells, Scale bars: 25 μm. Bar graphs show mean ± SEM. Student’s <span class="html-italic">t</span>-test or one-way ANOVA was used. ** <span class="html-italic">p</span> &lt; 0.01 versus the corresponding control.</p>
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<p>cZFP609 inhibits proliferation in Caco-2 cells via inhibiting HIF1α nuclear translocation. (<b>A</b>) qRT-PCR of cZFP609 expression in Caco-2 cells treated with vector or cZFP609. (<b>B</b>) Ratio of cell counting of Caco-2 cells treated with vector or cZFP609. (<b>C</b>,<b>D</b>) Migration of Caco-2 cells was assessed using a scratch wound assay. Caco-2 cells treated with vector of cZFP609. (Scale bar: 200 µm). Data are presented as mean ± SEM. (<b>E</b>) Immunofluorescent confocal microscopy of HIF1α nuclear translocation in the Caco-2 cells. (Scale bars: 25 μm). Bar graphs show mean ± SEM. Student’s <span class="html-italic">t</span>-test or one-way ANOVA was used. ** <span class="html-italic">p</span> &lt; 0.01 versus the corresponding control.</p>
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<p>cZFP609 inhibits endothelial regeneration after DSS-induced colitis. (<b>A</b>) Macroscopic colon appearance of tail-injected WT mice treated with DSS-induced colitis. (<b>B</b>) The colon length of vector and cZFP609 tail-injection mice. (<b>C</b>) Weight loss relative to % to initial weight, <span class="html-italic">n</span> = 6 mice per group, determined daily and compared to the weights at the start of DSS treatment for each mouse. (<b>D</b>) Disease activity index measured daily in tail-injection mice described during the DSS treatment schedule. (<b>E</b>) Representative H&amp;E-stained images, out of three independently acquired, of colon sections from the indicated groups of mice treated at the end of DSS administration (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>F</b>) Histopathological changes in colon tissue of WT and <span class="html-italic">Sirt1</span>-Tg mice with DSS. (<b>G</b>) Representative Alcian-Blue-stained images, out of three independently acquired, of colon sections from the indicated groups of mice treated at the end of DSS administration (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>H</b>) Representative immunohistochemistry images of the PCNA in the colon, out of three independently acquired, of colon sections from the indicated groups of mice treated at the end of DSS administration (scale bar: 200 µm; scale bar in the enlarged image is 100 µm). (<b>I</b>) AOD analysis of IHC results of PCNA, <span class="html-italic">n</span> = 6. (<b>J</b>) qPCR analysis of cZFP609 levels in colon tissue of tail-injection treated mice at the end of DSS administration. All quantifications are represented as mean ± SD and statistical significance was assessed by two-tailed unpaired Student’s <span class="html-italic">t</span>-test. Actual <span class="html-italic">p</span>-values are indicated in each graph. * <span class="html-italic">p</span> &lt; 0.05.</p>
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24 pages, 5275 KiB  
Article
Force Map-Enhanced Segmentation of a Lightweight Model for the Early Detection of Cervical Cancer
by Sabina Umirzakova, Shakhnoza Muksimova, Jushkin Baltayev and Young Im Cho
Diagnostics 2025, 15(5), 513; https://doi.org/10.3390/diagnostics15050513 - 20 Feb 2025
Viewed by 171
Abstract
Background/Objectives: Accurate and efficient segmentation of cervical cells is crucial for the early detection of cervical cancer, enabling timely intervention and treatment. Existing segmentation models face challenges with complex cellular arrangements, such as overlapping cells and indistinct boundaries, and are often computationally intensive, [...] Read more.
Background/Objectives: Accurate and efficient segmentation of cervical cells is crucial for the early detection of cervical cancer, enabling timely intervention and treatment. Existing segmentation models face challenges with complex cellular arrangements, such as overlapping cells and indistinct boundaries, and are often computationally intensive, which limits their deployment in resource-constrained settings. Methods: In this study, we introduce a lightweight and efficient segmentation model specifically designed for cervical cell analysis. The model employs a MobileNetV2 architecture for feature extraction, ensuring a minimal parameter count conducive to real-time processing. To enhance boundary delineation, we propose a novel force map approach that drives pixel adjustments inward toward the centers of cells, thus improving cell separation in densely packed areas. Additionally, we integrate extreme point supervision to refine segmentation outcomes using minimal boundary annotations, rather than full pixel-wise labels. Results: Our model was rigorously trained and evaluated on a comprehensive dataset of cervical cell images. It achieved a Dice Coefficient of 0.87 and a Boundary F1 Score of 0.84, performances that are comparable to those of advanced models but with considerably lower inference times. The optimized model operates at approximately 50 frames per second on standard low-power hardware. Conclusions: By effectively balancing segmentation accuracy with computational efficiency, our model addresses critical barriers to the widespread adoption of automated cervical cell segmentation tools. Its ability to perform in real time on low-cost devices makes it an ideal candidate for clinical applications and deployment in low-resource environments. This advancement holds significant potential for enhancing access to cervical cancer screening and diagnostics worldwide, thereby supporting broader healthcare initiatives. Full article
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<p>Improved contextual integration and feature extraction with MobileNetV2 for accurate cell segmentation.</p>
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<p>Description of the SipakMed dataset.</p>
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<p>Results of image segmentation using the proposed model.</p>
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<p>Performance curves for model robustness and overfitting prevention.</p>
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25 pages, 5090 KiB  
Article
Research on Intelligent Verification of Equipment Information in Engineering Drawings Based on Deep Learning
by Zicheng Zhang and Yurou He
Electronics 2025, 14(4), 814; https://doi.org/10.3390/electronics14040814 - 19 Feb 2025
Viewed by 229
Abstract
This paper focuses on the crucial task of automatic recognition and understanding of table structures in engineering drawings and document processing. Given the importance of tables in information display and the urgent need for automated processing of tables in the digitalization process, an [...] Read more.
This paper focuses on the crucial task of automatic recognition and understanding of table structures in engineering drawings and document processing. Given the importance of tables in information display and the urgent need for automated processing of tables in the digitalization process, an intelligent verification method is proposed. This method integrates multiple key techniques: YOLOv10 is used for table object recognition, achieving a precision of 0.891, a recall rate of 0.899, mAP50 of 0.922, and mAP50-95 of 0.677 in table recognition, demonstrating strong target detection capabilities; the improved LORE algorithm is adopted to extract table structures, breaking through the limitations of the original algorithm by segmenting large-sized images, with a table extraction accuracy rate reaching 91.61% and significantly improving the accuracy of handling complex tables; RapidOCR is utilized to achieve text recognition and cell correspondence, solving the problem of text-cell matching; for equipment name semantic matching, a method based on BERT is introduced and calculated using a comprehensive scoring method. Meanwhile, an improved cuckoo search algorithm is proposed to optimize the adjustment factors, avoiding local optima through sine optimization and the catfish effect. Experiments show the accuracy of equipment name matching in semantic similarity calculation approaches 100%. Finally, the paper provides a concrete system practice to prove the effectiveness of the algorithm. In conclusion, through experimental comparisons, this method exhibits excellent performance in table area location, structure recognition, and semantic matching and is of great significance and practical value in advancing table data processing technology in engineering drawings. Full article
(This article belongs to the Section Artificial Intelligence)
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<p>The framework of intelligent verification methods.</p>
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<p>The framework of YOLOv10.</p>
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<p>Illustration of the improved LORE algorithm.</p>
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<p>First-last layer average pooling.</p>
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<p>Improved cuckoo search algorithm flowchart.</p>
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<p>Iteration curve of algorithm training effectiveness.</p>
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<p>Display of recognition results.</p>
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<p>Comparison of the recognition process of this paper’s algorithm with the original LORE algorithm.</p>
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<p>Iteration curves of the three functions for CS and ICS. (<b>a</b>) Iteration Curves of Function F1 (<b>b</b>) Iteration Curves of Function F2 (<b>c</b>) Iteration Curves of Function F3.</p>
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<p>Iteration curves of the three algorithms.</p>
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<p>Schematic diagram of system process, model and components.</p>
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<p>Matching result system screenshot.</p>
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11 pages, 1276 KiB  
Article
Prognostic Role of Pan-Immune-Inflammatory Value in Patients with Non-ST-Segment Elevation Acute Coronary Syndrome
by Jeong Tae Byoun, Kyeong Ho Yun, Sungho Jo, Donghyeon Joo and Jae Young Cho
J. Cardiovasc. Dev. Dis. 2025, 12(2), 79; https://doi.org/10.3390/jcdd12020079 - 18 Feb 2025
Viewed by 283
Abstract
Blood cell-derived indices are potential predictors of clinical outcomes in coronary artery disease. This study assessed the prognostic value of the pan-immune-inflammatory value (PIV) for predicting 1-year major adverse cardiovascular events (MACEs) in patients with non-ST-segment elevation acute coronary syndrome (ACS). A retrospective [...] Read more.
Blood cell-derived indices are potential predictors of clinical outcomes in coronary artery disease. This study assessed the prognostic value of the pan-immune-inflammatory value (PIV) for predicting 1-year major adverse cardiovascular events (MACEs) in patients with non-ST-segment elevation acute coronary syndrome (ACS). A retrospective cohort of 1651 patients receiving percutaneous coronary intervention was analyzed. PIV, calculated from blood cell counts, was categorized with a cut-off value of 256.3 (sensitivity 60.7%, specificity 59.3%) based on receiver operating characteristic curve analysis. MACEs were operationalized as a composite of all-cause mortality, myocardial infarction (MI), stroke, any revascularization, and rehospitalization for heart failure. The incidence of MACEs was 5.0% in patients with low PIV and 9.7% in those with high PIV (log-rank p < 0.001). Multivariate analysis identified age 65 > years, renal dysfunction (eGFR < 60 mL/min/1.73 m2), and high PIV (>256.3) (HR 1.49, 95% CI 1.01–2.22, p = 0.048) as independent predictors of MACEs. Subgroup analyses revealed no statistically significant interaction between MI status or C-reactive protein levels and PIV. PIV was an independent predictor of 1-year MACEs in patients with non-ST-segment elevation ACS. It may serve as a reliable prognostic marker independently of MI or C-reactive protein levels. Full article
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<p>Participant flow in the present study.</p>
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<p>Event-free survival curve based on the pan-immune-inflammatory value (PIV).</p>
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<p>Prognostic impact of pan-immune-inflammatory value as a continuous variable on the risk of 1-year major adverse cardiovascular events. A restricted cubic spline curve was drawn to demonstrate the continuous prognostic effect of PIV on the risk of 1-year MACEs in patients with non-ST-segment elevation acute coronary syndrome. Adjusted variables were age, gender, hypertension, diabetes, hemoglobin, eGFR, C-reactive protein, ejection fraction, and troponin T.</p>
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