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28 pages, 10930 KiB  
Article
Comprehensive Physiology, Cytology, and Transcriptomics Studies Reveal the Regulatory Mechanisms Behind the High Calyx Abscission Rate in the Bud Variety of Korla Pear (Pyrus sinkiangensis ‘Xinnonglinxiang’)
by Xian’an Yang, Shiwei Wang, Zhenbin Jiang, Cuifang Zhang, Long Zhao and Yutong Cui
Plants 2024, 13(24), 3504; https://doi.org/10.3390/plants13243504 (registering DOI) - 15 Dec 2024
Abstract
Whether the calyx tube of the Korla fragrant pear falls off seriously affects the fruit quality. ‘Xinnonglinxiang’ is a mutant variety of the Korla fragrant pear, which has a high calyx removal rate under natural conditions, and calyx tube fall seriously affects the [...] Read more.
Whether the calyx tube of the Korla fragrant pear falls off seriously affects the fruit quality. ‘Xinnonglinxiang’ is a mutant variety of the Korla fragrant pear, which has a high calyx removal rate under natural conditions, and calyx tube fall seriously affects the fruit quality. The mechanism behind the high calyx removal rate of ‘Xinnonglinxiang’ remains unclear; thus, Korla fragrant pear (PT) and ‘Xinnonglinxiang’ (YB) with different degrees of calyx abscission were used as examples and the abscission areas of calyx tubes were collected in the early (21 April), middle (23 April), and late (25 April) shedding stages to explore the regulatory mechanism behind the abscission. The combination of the results of physiological, cytological, and transcriptomic methods indicated the highest number of differentially expressed genes (DEGs) in the middle of shedding. GO (Gene Ontology) enrichment analysis showed that the expression levels of genes related to the CEL (cellulase) and PG (polygalacturonase) activity functional pathways differed significantly in the two varieties during the three periods, whereas Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the DEGs were significantly concentrated in the plant hormone signal transduction pathway in all three periods. The expression levels of genes related to the plant hormone signal transduction pathway differed significantly for the two varieties during calyx shedding. Five gene modules were obtained using Weighted Gene Co-Expression Network Analysis (WGCNA), and transcriptome data were correlated with five physiological index values. Two key modules that highly correlated with the Eth (ethylene) response were then screened, and 20 core genes were identified, with IRX10, IRX9, and OXI1 likely the hub genes that are involved in the regulation of calyx shedding in the YB variety. The obtained results provide reliable data for the screening of candidate genes for calyx shedding and analysis of the regulatory mechanism behind a high calyx shedding rate, providing a theoretical basis upon which the calyx shedding rate of fruits can be improved through genetic improvement. Full article
(This article belongs to the Section Horticultural Science and Ornamental Plants)
18 pages, 10223 KiB  
Article
Integrating Single-Cell RNA-Seq and ATAC-Seq Analysis Reveals Uterine Cell Heterogeneity and Regulatory Networks Linked to Pimpled Eggs in Chickens
by Wenqiang Li, Xueying Ma, Xiaomin Li, Xuguang Zhang, Yifei Sun, Chao Ning, Qin Zhang, Dan Wang and Hui Tang
Int. J. Mol. Sci. 2024, 25(24), 13431; https://doi.org/10.3390/ijms252413431 (registering DOI) - 15 Dec 2024
Viewed by 227
Abstract
Pimpled eggs have defective shells, which severely impacts hatching rates and transportation safety. In this study, we constructed single-cell resolution transcriptomic and chromatin accessibility maps from uterine tissues of chickens using single-cell RNA sequencing (scRNA-seq) and single-cell ATAC sequencing (scATAC-seq). We identified 11 [...] Read more.
Pimpled eggs have defective shells, which severely impacts hatching rates and transportation safety. In this study, we constructed single-cell resolution transcriptomic and chromatin accessibility maps from uterine tissues of chickens using single-cell RNA sequencing (scRNA-seq) and single-cell ATAC sequencing (scATAC-seq). We identified 11 major cell types and characterized their marker genes, along with specific transcription factors (TFs) that determine cell fate. CellChat analysis showed that fibroblasts had the most extensive intercellular communication network and that the chickens laying pimpled eggs had amplified immune-related signaling pathways. Differential expression and enrichment analyses indicated that inflammation in pimpled egg-laying chickens may lead to disruptions in their circadian rhythm and changes in the expression of ion transport-related genes, which negatively impacts eggshell quality. We then integrated TF analysis to construct a regulatory network involving TF–target gene–Gene Ontology associations related to pimpled eggs. We found that the transcription factors ATF3, ATF4, JUN, and FOS regulate uterine activities upstream, while the downregulation of ion pumps and genes associated with metal ion binding directly promotes the formation of pimpled eggs. Finally, by integrating the results of scRNA-seq and scATAC-seq, we identified a rare cell type—ionocytes. Our study constructed single-cell resolution transcriptomic and chromatin accessibility maps of chicken uterine tissue and explored the molecular regulatory mechanisms underlying pimpled egg formation. Our findings provide deeper insights into the structure and function of the chicken uterus, as well as the molecular mechanisms of eggshell formation. Full article
(This article belongs to the Special Issue Big Data in Multi-Omics)
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<p>Single-cell transcriptome analysis and clustering identification of chicken uterine cells. (<b>A</b>) Unsupervised clustering revealed 21 distinct transcriptional cell clusters, which were visualized using a UMAP plot. Each point represents an individual cell, with colors indicating cluster assignments. (<b>B</b>) A UMAP plot was utilized to visualize 11 uterine cell types. Each point represents an individual cell and is color-coded according to its cell type. (<b>C</b>) The dot plot illustrates the distinct expression patterns of canonical marker genes across 11 cell populations. (<b>D</b>) Bar plot showing the percentage of different cell types within each of the 6 samples.</p>
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<p>Regulatory network of DE transcription factors (TFs) and their DE target genes in chicken uterine tissue, along with GO terms. In the diagram, inverted triangles represent transcription factors (TFs), circles denote target genes, and rectangles indicate GO terms. Red highlights TFs and target genes that are upregulated in the NE group, while green highlights those upregulated in the PE group. Blue represents GO terms. The size of the shapes corresponds to the degree of involvement, with larger shapes indicating greater participation in regulatory networks.</p>
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<p>scRNA-seq reveals significant changes in cell communication between the NE and PE groups. (<b>A</b>) Number of interactions between all cells in the NE group. Thicker links indicate a higher number of interactions. (<b>B</b>) Number of interactions between all cells in the PE group. Thicker links indicate a higher number of interactions. (<b>C</b>) Cell–cell communication chord diagram for the COLLAGEN signaling pathway in the NE group, with thicker links indicating stronger interactions between cells. (<b>D</b>) Cell–cell communication chord diagram for the COLLAGEN signaling pathway in the PE group, with thicker links indicating stronger interactions between cells. (<b>E</b>) Relative signaling pathway diagram showing the pathways identified in the NE and PE groups. Larger pathways in the NE group are depicted in cyan, while those in the PE group are depicted in red. Black indicates pathways with no significant difference.</p>
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<p>Single-Cell Chromatin Accessibility Analysis of Chicken Uterine Tissue. (<b>A</b>) Annotation and statistics of the distribution of peaks in different genomic functional regions (such as promoters, 5′UTR, 3′UTR, exons, introns, downstream regions, and intergenic regions) for the N1 sample (<b>top</b>) and P1 sample (<b>bottom</b>). (<b>B</b>) UMAP plot of the scATAC-seq dataset for sample P1, with cell type assignments based on scRNA-seq data. (<b>C</b>) Histogram of cell frequency distributions for scRNA and scATAC data in the P1 sample. (<b>D</b>) Feature plot of inferred marker gene activities, including epithelial cells (<span class="html-italic">KRT7</span>), ionocytes (<span class="html-italic">PDE1C</span>), luminal epithelial cells (<span class="html-italic">ATP8A2</span>), macrophages (<span class="html-italic">ENSGALG00010005330</span>), T cells (<span class="html-italic">ENSGALG00010005352</span>), fibroblasts (<span class="html-italic">COL5A1</span>), B cells (<span class="html-italic">ENSGALG00010003777</span>), and endothelial cells (<span class="html-italic">APOLD1</span>).</p>
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<p>Identification of Ionocyte through marker genes and enrichment analysis. (<b>A</b>) UMAP plot showing the expression patterns of ionocyte marker genes (from the literature) in scRNA-seq data. (<b>B</b>) Enriched GO terms for genes upregulated in Ionocytes, as identified in scRNA-seq data. (<b>C</b>) UMAP visualization of PDE1C, a marker gene for ionocytes identified in the literature, showing inferred gene activity within ionocyte clusters from scATAC-seq data.</p>
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26 pages, 7457 KiB  
Article
Digitalizing Material Knowledge: A Practical Framework for Ontology-Driven Knowledge Graphs in Process Chains
by Elena Garcia Trelles, Christoph Schweizer, Akhil Thomas, Philipp von Hartrott and Marina Janka-Ramm
Appl. Sci. 2024, 14(24), 11683; https://doi.org/10.3390/app142411683 (registering DOI) - 14 Dec 2024
Viewed by 293
Abstract
This paper proposes a robust methodology for integrating process-specific data and domain expert knowledge into linked knowledge graphs. These graphs utilize an ontology that provides a standardized vocabulary for material science and facilitates the creation of semantic models for various processes along the [...] Read more.
This paper proposes a robust methodology for integrating process-specific data and domain expert knowledge into linked knowledge graphs. These graphs utilize an ontology that provides a standardized vocabulary for material science and facilitates the creation of semantic models for various processes along the digital process chain. A generic template for structuring processes is proposed, simplifying subsequent data retrieval. The templates of specific processes are designed collaboratively by domain and ontology experts, aided by a proposed interview template that bridges the knowledge gap. Following the digitalization of material data through semantic modeling, machine-readable data with contextual metadata is stored in a graph database, which can be efficiently queried using the SPARQL language, enabling seamless integration into data pipelines. To demonstrate this approach, a knowledge graph is developed to represent the process chain of AlSi10Mg objects manufactured via permanent mold casting, capturing their complete history from the initial manufacturing step to final non-destructive testing and mechanical characterization. This methodology enhances data interoperability and accessibility while providing context-rich data for training AI models, potentially accelerating new knowledge discovery in material science. Full article
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<p>The BWMD process instance modeling philosophy: BFO classes depicted in black, BWMD classes in green, and BWMD object properties in red.</p>
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<p>Schematic representation of the permanent mold casting process including the ingot as input and the cast part as output.</p>
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<p>The Graph Designer Workflow. Extended documentation regarding its application and tools can be found in [<a href="#B36-applsci-14-11683" class="html-bibr">36</a>].</p>
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<p>Detail of the interview Excel sheet regulating the information exchange between the domain expert and the ontology developer: one section of the interview is reserved for collecting the list of parameters that determine a specific outcome of the process to be modeled, in the BWMD Ontology referred to through the class <span class="html-italic">ProcessControlParameters</span>.</p>
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<p>The generic Process Graph template: a common pattern to describe a process.</p>
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<p>Details of the Excel template for metadata acquisition: (1.) <span class="html-italic">user_variables</span>: user view including the node names of the literals from the process graphs; (2.) <span class="html-italic">gdtriples</span>: logic of the process graph; (3.) <span class="html-italic">namespaces</span>: ontology IRI and namespace.</p>
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<p>Exemplary use of the {1..n} construct with the example of user-specific duplication of chemical elements within a chemical composition.</p>
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<p>Description of the assignment of an ID and chemical composition to a cast part.</p>
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<p>Description of process parameters and handling of physical quantities and unit symbols.</p>
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<p>Visualization of the complete knowledge graph.</p>
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<p>Visualization of Brinell hardness SPARQL query algorithm.</p>
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<p>Visualization of the SPARQL query algorithm retrieving the process control parameters corresponding to one specific object identifier.</p>
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<p>Visualization of the process chain corresponding to the object <tt>ARI_Al6</tt> with the help of the Visual Graph tool of GraphDB database engine [<a href="#B39-applsci-14-11683" class="html-bibr">39</a>].</p>
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<p>Decision Tree Regression model to predict the tensile strength of the cast AlSi10Mg based on process chain graph data.</p>
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<p>Validation of the trained Decision Tree Regression model with a correlation coefficient (<span class="html-italic">R</span>) of 95%, a coefficient of determination (<math display="inline"><semantics> <msup> <mi>R</mi> <mn>2</mn> </msup> </semantics></math>) of 89%, and a relative standard error (RSE) of the slope equal to 15.36%.</p>
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20 pages, 596 KiB  
Article
Predictors of Adaptive Behaviors in Individuals on the Autism Spectrum as Assessed by Teachers and Parents: An Analysis Based on ABAS-3
by Janusz Kirenko, Anna Prokopiak and Maciej Wodziński
J. Clin. Med. 2024, 13(24), 7607; https://doi.org/10.3390/jcm13247607 - 13 Dec 2024
Viewed by 233
Abstract
Objectives: This present study focuses on analyzing the adaptive behaviors of individuals on the autism spectrum as perceived by parents and teachers of these individuals. Methods: This study was conducted in Poland with the use of the ABAS-3 (Adaptive Behavior Assessment [...] Read more.
Objectives: This present study focuses on analyzing the adaptive behaviors of individuals on the autism spectrum as perceived by parents and teachers of these individuals. Methods: This study was conducted in Poland with the use of the ABAS-3 (Adaptive Behavior Assessment System). The ABAS-3 tool involves both parents (or primary caregivers) and teachers in the diagnostic process and monitoring of adaptive development. The study included 99 individuals (29 girls, 70 boys) aged between 5 and 21 years. Results: The analysis of the results showed statistically significant discrepancies in the perception of adaptive skills diagnosed as assessed by parents and teachers. Furthermore, differences were found in the predictors of the General Adaptive Composite and adaptive domains. Conclusions: The results indicate the complexity of the assessment of adaptive skills by a parent of a child with autism spectrum disorder, as well as a teacher, and the need to include different perspectives in the process of diagnosing and supporting individuals with ASD. Full article
(This article belongs to the Section Mental Health)
7 pages, 479 KiB  
Brief Report
Dynamic Dysregulation of Ribosomal Protein Genes in Mouse Brain Stress Models
by Vandana Sharma and Rammohan Shukla
Stresses 2024, 4(4), 916-922; https://doi.org/10.3390/stresses4040061 - 12 Dec 2024
Viewed by 251
Abstract
Emphasizing their evolutionarily conserved role in stress adaptation mechanisms, ribosomal protein genes (RPGs) are observed to be downregulated in various stressors and across phyla. However, this evolutionarily conserved stress response is not well explored in mouse models of neurobiological stress. This study investigates [...] Read more.
Emphasizing their evolutionarily conserved role in stress adaptation mechanisms, ribosomal protein genes (RPGs) are observed to be downregulated in various stressors and across phyla. However, this evolutionarily conserved stress response is not well explored in mouse models of neurobiological stress. This study investigates the dysregulation patterns of RPGs in various murine preclinical stress paradigms across different brain regions using available transcriptomic data and identifies the non-canonical ribosomal functions using synaptic gene-ontology terms. Without a discernible structure across different brain areas, we observed heterogeneous dysregulation, encompassing either up or downregulation in both cytoplasmic and mitochondrial RPGs. However, downregulation was more prominent than upregulation, and the overall dysregulation seems more prevalent in the chronic stress paradigm compared to stress paradigms involving acute and early-life stress. Enrichment analysis significantly associates dysregulated RPGs with post-synaptic gene ontology terms, emphasizing their involvement in synaptic modulation. Overall, the study demonstrates ribosomal dysregulation as an evolutionarily conserved stress response mechanism during different mouse stress paradigms. We discuss the possibility that the variability in the directionality of dysregulation may emerge as a potential marker of neuronal activity in response to diverse stress paradigms and the involvement of paradigm-specific RPG dysregulation either in the process of global downscaling of ribosome biogenesis or in the process of ribosomal heterogeneity, each leading to a different effect. Full article
(This article belongs to the Collection Feature Papers in Human and Animal Stresses)
20 pages, 1956 KiB  
Article
Enhancing Ontological Metamodel Creation Through Knowledge Extraction from Multidisciplinary Design and Optimization Frameworks
by Esma Karagoz, Olivia J. Pinon Fischer and Dimitri N. Mavris
Systems 2024, 12(12), 555; https://doi.org/10.3390/systems12120555 - 12 Dec 2024
Viewed by 297
Abstract
The design of complex aerospace systems requires a broad multidisciplinary knowledge base and an iterative approach to accommodate changes effectively. Engineering knowledge is commonly represented through engineering analyses and descriptive models with underlying semantics. While guidelines from systems engineering methodologies exist to guide [...] Read more.
The design of complex aerospace systems requires a broad multidisciplinary knowledge base and an iterative approach to accommodate changes effectively. Engineering knowledge is commonly represented through engineering analyses and descriptive models with underlying semantics. While guidelines from systems engineering methodologies exist to guide the development of system models, creating a system model from scratch with every new application/system requires research into more adaptable and reusable modeling frameworks. In this context, this research demonstrates how a physics-based multidisciplinary analysis and optimization tool, SUAVE, can be leveraged to develop a system model. By leveraging the existing physics-based knowledge captured within SUAVE, the process benefits from the expertise embedded in the tool. To facilitate the systematic creation of the system model, an ontological metamodel is created in SysML. This metamodel is designed to capture the inner workings of the SUAVE tool, representing its concepts, relationships, and behaviors. By using this ontological metamodel as a modeling template, the process of creating the system model becomes more structured and organized. Overall, this research aims to streamline the process of building system models from scratch by leveraging existing knowledge and utilizing an ontological metamodel as a modeling template. This approach enhances formal knowledge representation and its consistency, and promotes reusability in multidisciplinary design problems. Full article
(This article belongs to the Section Systems Engineering)
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<p>Knowledge types in engineering design [<a href="#B14-systems-12-00555" class="html-bibr">14</a>,<a href="#B16-systems-12-00555" class="html-bibr">16</a>].</p>
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<p>Projections of knowledge on to the system model [<a href="#B23-systems-12-00555" class="html-bibr">23</a>].</p>
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<p>Research methodology.</p>
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<p>Structure of SUAVE.</p>
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<p>Logical decomposition: In the system model, the logical components are represented by disciplinary analyses within SUAVE.</p>
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<p>SUAVE—aerodynamics analyses: The decomposition of aerodynamics analyses involves different levels of fidelity.</p>
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<p>Fidelity zero aerodynamics method.</p>
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<p>Data flow between the analysis method functions.</p>
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<p>Parametric diagram showing the relation between the components and inputs/outputs.</p>
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<p>Physical decomposition: This decomposition reflects the structure of SUAVE. It is important to note that this figure provides a high-level view of the physical decomposition to avoid overcrowded representations.</p>
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<p>Ontological metamodel.</p>
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<p>A high-level representation of the mission analysis for a Boeing 737-800, including the required analyses and the physical components whose values serve as input for the analysis.</p>
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22 pages, 8896 KiB  
Review
Framing Concepts of Agriculture 5.0 via Bipartite Analysis
by Ivan Bergier, Jayme G. A. Barbedo, Édson L. Bolfe, Luciana A. S. Romani, Ricardo Y. Inamasu and Silvia M. F. S. Massruhá
Sustainability 2024, 16(24), 10851; https://doi.org/10.3390/su162410851 - 11 Dec 2024
Viewed by 395
Abstract
Cultural diversity often complicates the understanding of sustainability, sometimes making its concepts seem vague. This issue is particularly evident in food systems, which rely on both renewable and nonrenewable resources and drive significant environmental changes. The widespread impacts of climate change, aggravated by [...] Read more.
Cultural diversity often complicates the understanding of sustainability, sometimes making its concepts seem vague. This issue is particularly evident in food systems, which rely on both renewable and nonrenewable resources and drive significant environmental changes. The widespread impacts of climate change, aggravated by the overuse of natural resources, have highlighted the urgency of balancing food production with environmental preservation. Society faces a pivotal challenge: ensuring that food systems produce ample, accessible, and nutritious food while also reducing their carbon footprint and protecting ecosystems. Agriculture 5.0, an innovative approach, combines digital advancements with sustainability principles. This study reviews current knowledge on digital agriculture, analyzing scientific data through an undirected bipartite network that links journals and author keywords from articles retrieved from Clarivate Web of Science. The main goal is to outline a framework that integrates various sustainability concepts, emphasizing both well-studied (economic) and underexplored (socioenvironmental) aspects of Agriculture 5.0. This framework categorizes sustainability concepts into material (tangible) and immaterial (intangible) values based on their supporting or influencing roles within the agriculture domain, as documented in the scientific literature. Full article
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<p>PRISMA methodology for extracting relevant articles in Web of Science. The symbol * stands for any additional character.</p>
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<p>Schematic representation of a bipartite analysis of two sets of nodes, <span class="html-italic">D</span> (purple) and <span class="html-italic">J</span> (green).</p>
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<p>Exponential growth rate (~30%.y<sup>−1</sup>) of scientific interest in included articles.</p>
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<p>Two representations of the same undirected bipartite graph with 943 nodes and 1129 links between journals (in blue, 120 nodes) and keywords (in red, 823 nodes). The size of the nodes is proportional to the weighted degree centrality.</p>
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<p>Log-binned (2<span class="html-italic"><sup>n</sup></span> for <span class="html-italic">n</span> = 0, 1, …, 7) node degree distribution of the keyword–journal network extracted from the 210 selected publications. Dark circles were disregarded in the statistical regression.</p>
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<p>Keywords semantics from the bipartite analysis. The size of the nodes (labels) is proportional to the weighted degree (betweeness) centrality.</p>
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<p>Details of subsets of underexplored keywords among journals.</p>
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<p>Network of conceptual assets of the Economic (technological application) dimension of Sustainability obtained from the bipartite analysis between “economic keywords” and the nine conceptual assets of the economic dimension of sustainability. The bipartite network is shown in <a href="#sustainability-16-10851-f0A2" class="html-fig">Figure A2</a>. The size of the nodes (labels) is proportional to the weighted degree (betweeness) centrality.</p>
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<p>Framework of material (red) and immaterial (blue) conceptual assets in Agriculture 5.0 as a directed network graph of weighted support (larger labels) and influence (larger nodes).</p>
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<p><span class="html-italic">J</span> (journals) set from the bipartite analysis of the keyword–journal network.</p>
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<p>Bipartite undirected network between superhub keywords (blue) and application categories (red). The size of nodes (labels) is proportional to weighted degree (betweeness) centrality, while the thickness of the edges is related to ties strength.</p>
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15 pages, 5486 KiB  
Article
Genome-Wide Identification and Analysis of Glycosyltransferases in Colletotrichum graminicola
by Yafei Wang, Honglian Li, Jiaxin Chang, Yu Zhang, Jinyao Li, Shaofeng Jia and Yan Shi
Microorganisms 2024, 12(12), 2551; https://doi.org/10.3390/microorganisms12122551 - 11 Dec 2024
Viewed by 332
Abstract
Corn leaf blight and stem rot caused by Colletotrichum graminicola are significant diseases that severely affect corn crops. Glycosyltransferases (GTs) catalyze the transfer of sugar residues to diverse receptor molecules, participating in numerous biological processes and facilitating functions ranging from structural support to [...] Read more.
Corn leaf blight and stem rot caused by Colletotrichum graminicola are significant diseases that severely affect corn crops. Glycosyltransferases (GTs) catalyze the transfer of sugar residues to diverse receptor molecules, participating in numerous biological processes and facilitating functions ranging from structural support to signal transduction. This study identified 101 GT genes through functional annotation of the C. graminicola TZ–3 genome. Subsequent analyses revealed differences among the C. graminicola GT (CgGT) genes. Investigation into subcellular localization indicated diverse locations of CgGTs within subcellular structures, while the presence of multiple domains in CgGTs suggests their involvement in diverse fungal biological processes through versatile functions. The promoter regions of CgGT genes are enriched with diverse cis-acting regulatory elements linked to responses to biotic and abiotic stresses, suggesting a key involvement of CgGT genes in the organism’s multi-faceted stress responses. Expression pattern analysis reveals that most CgGT genes were differentially expressed during the interaction between C. graminicola and corn. Integrating gene ontology functional analysis revealed that CgGTs play important roles in the interaction between C. graminicola and corn. Our research contributes to understanding the functions of CgGT genes and investigating their involvement in fungal pathogenesis. At the same time, our research has laid a solid foundation for the development of sustainable agriculture and the utilization of GT genes to develop stress-resistant and high-yield crop varieties. Full article
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<p>Phylogenetic tree of glycosyltransferases from <span class="html-italic">Colletotrichum graminicola</span>. Note: CgGTs are divided into nine groups (I–IX) and each color represents a group.</p>
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<p>The motifs of CgGTs. Note: Boxes of different colors represent different conserved motifs.</p>
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<p>Cis-acting regulatory elements in the promoter regions of <span class="html-italic">CgGT</span> genes. Note: Different colored boxes represent different cis-acting regulatory elements.</p>
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<p>The numbers of predicted cis-acting regulatory elements in the promoter regions of <span class="html-italic">CgGT</span> genes.</p>
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<p>Gene ontology enrichment analysis of <span class="html-italic">CgGT</span> genes.</p>
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<p>The expression level of <span class="html-italic">CgGTs</span> based on RNA-seq data. Note: <span class="html-italic">CgGT</span> genes were categorized into seven distinct classes (I–VII) based on expression profiles. Red and green indicate high and low expression levels of <span class="html-italic">CgGTs</span>, respectively.</p>
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<p>RT-qPCR verification of <span class="html-italic">CgGT</span> expression pattern. Note: * <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.</p>
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21 pages, 3589 KiB  
Article
Transcriptome Analysis Unveils Molecular Mechanisms of Acaricide Resistance in Two-Spotted Spider Mite Populations on Hops
by Sonu Koirala B K, Gaurab Bhattarai, Adekunle W. Adesanya, Timothy W. Moural, Laura C. Lavine, Douglas B. Walsh and Fang Zhu
Int. J. Mol. Sci. 2024, 25(24), 13298; https://doi.org/10.3390/ijms252413298 - 11 Dec 2024
Viewed by 284
Abstract
Broad-spectrum crop protection technologies, such as abamectin and bifenthrin, are globally relied upon to curb the existential threats from economic crop pests such as the generalist herbivore Tetranychus urticae Koch (TSSM). However, the rising cost of discovering and registering new acaricides, particularly for [...] Read more.
Broad-spectrum crop protection technologies, such as abamectin and bifenthrin, are globally relied upon to curb the existential threats from economic crop pests such as the generalist herbivore Tetranychus urticae Koch (TSSM). However, the rising cost of discovering and registering new acaricides, particularly for specialty crops, along with the increasing risk of pesticide resistance development, underscores the urgent need to preserve the efficacy of currently registered acaricides. This study examined the overall genetic mechanism underlying adaptation to abamectin and bifenthrin in T. urticae populations from commercial hop fields in the Pacific Northwestern region of the USA. A transcriptomic study was conducted using four populations (susceptible, abamectin-resistant, and two bifenthrin-resistant populations). Differential gene expression analysis revealed a notable disparity, with significantly more downregulated genes than upregulated genes in both resistant populations. Gene ontology enrichment analysis revealed a striking consistency among all three resistant populations, with downregulated genes predominately associated with chitin metabolism. In contrast, upregulated genes in the resistant populations were linked to biological processes, such as peptidase activity and oxidoreductase activity. Proteolytic activity by peptidase enzymes in abamectin- and bifenthrin-resistant TSSM populations may suggest their involvement in acaricide metabolism. These findings provide valuable insights into the molecular mechanisms underlying acaricide resistance in the TSSM. This knowledge can be utilized to develop innovative pesticides and molecular diagnostic tools for effectively monitoring and managing resistant TSSM populations. Full article
(This article belongs to the Section Molecular Toxicology)
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<p>Principal component analysis (PCA) of variance-stabilization-transformed (vst) normalized gene expression values in all three biological replicates of acaricide-resistant and -susceptible TSSM populations. The first (PC1) and second (PC2) principal components explain 69% and 14% of the total variance observed for gene expression, respectively. Each colored dot represents a biological replicate. SUS: susceptible; ABA_1X: abamectin-resistant; BIF_1X and BIF_100X: bifenthrin-resistant.</p>
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<p>Volcano plots showing differentially expressed genes (|log<sub>2</sub>fold change| ≥ 1.5) in (<b>A</b>) abamectin- and (<b>B</b>,<b>C</b>) bifenthrin-resistant two-spotted spider mite (TSSM) populations. Orange and blue dots represent upregulated (log<sub>2</sub>fold change ≥ 1.5) and downregulated (log<sub>2</sub>fold change ≤ −1.5) genes (FDR-adjusted <span class="html-italic">p</span>-value &lt; 0.05), respectively. SUS: susceptible; ABA_1X: abamectin-resistant; BIF_1X and BIF_100X: bifenthrin-resistant.</p>
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<p>Venn diagrams showing (<b>A</b>) all differentially expressed (both up- and downregulated), (<b>B</b>) only upregulated, and (<b>C</b>) only downregulated common and unique genes among three acaricide-resistant two-spotted spider mite (TSSM) populations: ABA_1X, BIF_1X, and BIF_100X, respectively. ABA_1X: abamectin-resistant; BIF_1X and BIF_100X: bifenthrin-resistant.</p>
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<p>The ontological relationship of up- and downregulated genes in three acaricide-resistant populations. The gene ontology is presented as biological processes (BPs) (<b>top</b>), molecular functions (MFs) (<b>middle</b>), and cellular components (<b>bottom</b>), and the color scales represent the number of differentially expressed genes in the corresponding gene ontology. Up- and downregulated genes were selected based on the following criteria: |log<sub>2</sub>fold change| &gt; 1.5 and Benjamini–Hochberg (BH)-adjusted <span class="html-italic">p</span>-values &lt; 0.05.</p>
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<p>log<sub>2</sub>fold change in genes belonging to various detoxification gene classes in abamectin (ABA_1X)- and bifenthrin (BIF_1X and BIF_100X)-resistant TSSM populations. The value “-” indicates that the gene does not meet the statistical criteria to be called differentially expressed in this study (|log<sub>2</sub>fold| ≥ 1.5 and Benjamin–Hochberg-adjusted contrast <span class="html-italic">p</span>-value ≤ 0.05). The color intensity in each gene category indicates the level of gene expression, with darker shades representing more |log<sub>2</sub>fold| changes. Dark blue indicates greater downregulation, while dark orange denotes higher upregulation. ABA_1X: abamectin-resistant; BIF_1X and BIF_100X: bifenthrin-resistant.</p>
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<p>Percentage of (<b>A</b>,<b>C</b>,<b>E</b>) single-nucleotide polymorphisms (SNPs) with all consequences and (<b>B</b>,<b>D</b>,<b>F</b>) consequences in coding regions in the ABA_1X, BIF_1X, and BIF_100X TSSM populations, respectively. SNPs were identified in resistant populations in reference to the homozygous genotype of the susceptible TSSM population. N: number of variants. ABA_1X: abamectin-resistant; BIF_1X and BIF_100X: bifenthrin-resistant.</p>
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17 pages, 4703 KiB  
Article
Robotics Classification of Domain Knowledge Based on a Knowledge Graph for Home Service Robot Applications
by Yiqun Wang, Rihui Yao, Keqing Zhao, Peiliang Wu and Wenbai Chen
Appl. Sci. 2024, 14(24), 11553; https://doi.org/10.3390/app142411553 - 11 Dec 2024
Viewed by 304
Abstract
The representation and utilization of environmental information by service robots has become increasingly challenging. In order to solve the problems that the service robot platform has, such as high timeliness requirements for indoor environment recognition tasks and the small scale of indoor scene [...] Read more.
The representation and utilization of environmental information by service robots has become increasingly challenging. In order to solve the problems that the service robot platform has, such as high timeliness requirements for indoor environment recognition tasks and the small scale of indoor scene data, a method and model for rapid classification of household environment domain knowledge is proposed, which can achieve high recognition accuracy by using a small-scale indoor scene and tool dataset. This paper uses a knowledge graph to associate data for home service robots. The application requirements of knowledge graphs for home service robots are analyzed to establish a rule base for the system. A domain ontology of the home environment is constructed for use in the knowledge graph system, and the interior functional areas and functional tools are classified. This designed knowledge graph contributes to the state of the art by improving the accuracy and efficiency of service decision making. The lightweight network MobileNetV3 is used to pre-train the model, and a lightweight convolution method with good feature extraction performance is selected. This proposal adopts a combination of MobileNetV3 and transfer learning, integrating large-scale pre-training with fine-tuning for the home environment to address the challenge of limited data for home robots. The results show that the proposed model achieves higher recognition accuracy and recognition speed than other common methods, meeting the work requirements of service robots. With the Scene15 dataset, the proposed scheme has the highest recognition accuracy of 0.8815 and the fastest recognition speed of 63.11 microseconds per sheet. Full article
(This article belongs to the Special Issue Artificial Intelligence in Complex Networks (2nd Edition))
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<p>Home service robot service system.</p>
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<p>Domain ontology of the home environment.</p>
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<p>Template for developing the service inference SWRL rule base.</p>
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<p>The acquisition mechanism of missing attributes of objects.</p>
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<p>The acquisition mechanism of missing attributes category, physical, and visual attributes of objects.</p>
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<p>Network structure of MobileNetV3.</p>
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<p>The proposed transfer learning strategy.</p>
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<p>The structure of the semantic cognitive framework.</p>
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<p>Examples of the CIFAR-100 dataset.</p>
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<p>Examples of the Scene15 dataset.</p>
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<p>Examples of the UMD part affordance dataset.</p>
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<p>Loss values and accuracy during training for the classification of indoor functional areas.</p>
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<p>Loss values and accuracy.</p>
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23 pages, 3173 KiB  
Article
The Cellular and Transcriptomic Early Innate Immune Response to BCG Vaccination in Mice
by Liya G. Kondratyeva, Olga A. Rakitina, Victor V. Pleshkan, Alexey I. Kuzmich, Irina A. Linge, Sofia A. Kondratieva, Eugene V. Snezhkov, Irina V. Alekseenko and Eugene D. Sverdlov
Cells 2024, 13(24), 2043; https://doi.org/10.3390/cells13242043 - 11 Dec 2024
Viewed by 546
Abstract
It is established that BCG vaccination results in the development of both a specific immune response to mycobacterial infections and a nonspecific (heterologous) immune response, designated as trained immunity (TRIM), to other pathogens. We hypothesized that local BCG immunization may induce an early [...] Read more.
It is established that BCG vaccination results in the development of both a specific immune response to mycobacterial infections and a nonspecific (heterologous) immune response, designated as trained immunity (TRIM), to other pathogens. We hypothesized that local BCG immunization may induce an early immune response in bone marrow and spleen innate immunity cells. The early transcriptomic response of various populations of innate immune cells, including monocytes, neutrophils, and natural killer (NK) cells, to BCG vaccination was examined. To this end, C57Bl/6J mice were subcutaneously immunized with 106 CFU of BCG. Three days following BCG administration, the three cell populations were collected from the control and BCG-vaccinated groups using FACS. All cell populations obtained were utilized for the preparation and sequencing of RNA-seq libraries. The analysis of FACS data revealed an increase in the proportion of splenic NK cells and monocytes 3 days post-vaccination. Transcriptomic analysis revealed the deregulation of genes associated with the regulation of immune response (according to Gene Ontology terms) in NK cells, monocytes, and unsorted bone marrow cells. Two NK cell-specific immune ligands (Tnfsf14 and S100a8) and two bone marrow-specific immune receptors (C5ar1 and Csf2rb) were identified among differentially expressed genes. No alterations were identified in neutrophils in either their percentage or at the transcriptomic level. Thus, in this study, we demonstrated that BCG vaccination provides an early increase in the proportion of murine bone marrow and spleen immune cell populations, as well as transcriptomic alterations in monocytes, NK cells, and non-sorted bone marrow cells. This early innate immune response may be beneficial for enhancing TRIM. Full article
(This article belongs to the Special Issue Innate Immunity in Health and Disease)
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<p>RNA-Seq experiment and sequencing analysis workflow.</p>
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<p>The analysis of murine bone marrow and spleen immune cell populations following BCG vaccination. (<b>A</b>) An example of the gating strategy for the bone marrow sample: First, a debris-free and DAPI-negative (live cells) cell population was isolated. Then, among them, a population of leukocytes (CD45+) was distinguished, which was subdivided into two subpopulations: CD45+ CD11b+ (myeloid cells) and CD45+ NK1.1+ (NK cells) immune cells. The population of myeloid cells (CD45+ and CD11b+) was further divided into subpopulations of monocytes (CD45+, CD11b+, and Ly6C+) and neutrophils (CD45+, CD11b+, and Ly6G+). (<b>B</b>,<b>C</b>) Changes in the proportions of innate immune cell populations in the spleen (<b>B</b>) and bone marrow (<b>C</b>) 3 days following subcutaneous BCG vaccination. The percentage of neutrophils, monocytes, and NK cells was calculated relative to the total number of leukocytes in the samples. Bars represent the mean among 5 mice per group ± S.D. * <span class="html-italic">p</span> &lt; 0.05 (Mann–Whitney test).</p>
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<p>The analysis of transcriptomic data using DESeq2. (<b>A</b>) Principal component analysis (PCA) of the normalized RNAseq data of innate immune cells in response to subcutaneous administration of BCG. (<b>B</b>) Volcano plots of DESeq2 results based on RNA-seq analysis of BCG-induced innate immune cells over control. Changes in transcript levels are represented by log2fc, the log2 fold change in normalized read counts between the immune populations derived from BCG-vaccinated and control mice, as obtained from the DESeq2 analysis [<a href="#B32-cells-13-02043" class="html-bibr">32</a>] (<span class="html-italic">x</span>-axis). The statistical significance of the change is represented as −log10 <span class="html-italic">p</span>-adj (<span class="html-italic">y</span>-axis). Differentially expressed genes (DEGs) are highlighted in red (upregulated) and blue (downregulated) with significant adjusted <span class="html-italic">p</span>-values (<span class="html-italic">p</span>-adj &lt; 0.05).</p>
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<p>The functional annotation of the revealed DEGs. (<b>A</b>) The functional annotation of 162 DEGs from splenic NK cells from mice after BCG vaccination was conducted using Metascape web-platform, resulting in the representation of the top 25 terms in a bar plot based on their <span class="html-italic">p</span>-value (log10 scale). (<b>B</b>) The functional annotation of 30 DEGs from Bone Marrow Monocytes from mice after BCG vaccination was conducted using Metascape web-platform, resulting in the representation of overall 20 terms in a bar plot based on their <span class="html-italic">p</span>-value (log10 scale). (<b>C</b>) The functional annotation of 184 DEGs from bulk Bone Marrow cells from mice after BCG vaccination was conducted using Metascape web-platform, resulting in the representation of top 25 terms in a bar plot based on their <span class="html-italic">p</span>-value (log10 scale).</p>
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<p>The analysis of common altered processes. (<b>A</b>) Venn diagram represents common processes found for DEGs of NK cells, monocytes, and bone marrow cells after BCG treatment. The percentage in the parenthesis reflects the percentage of processes of the overall number of unique processes among 3 populations. (<b>B</b>) TreeMap visualization of clustered common deregulated processes in NK cells, monocytes, and bone marrow cells after BCG treatment.</p>
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<p>Analysis of immune ligand and receptor gene expression. The expression levels were extracted from the RNA-seq data and presented as the mean TPM (transcripts per million) values with standard deviation bars (<span class="html-italic">y</span> axis) calculated from 3 independent samples. Gene abbreviations: C5ar1, complement C5a Receptor 1; Csf2rb, cytokine receptor common subunit beta; Tnfsf14, tumor necrosis factor superfamily member 14; S100a8, S100 calcium-binding protein A8. * <span class="html-italic">p</span>-adj &lt; 0.05 according to DeSeq2 data.</p>
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20 pages, 6078 KiB  
Article
A Smart Motor Rehabilitation System Based on the Internet of Things and Humanoid Robotics
by Yasamin Moghbelan, Alfonso Esposito, Ivan Zyrianoff, Giulia Spaletta, Stefano Borgo, Claudio Masolo, Fabiana Ballarin, Valeria Seidita, Roberto Toni, Fulvio Barbaro, Giusy Di Conza, Francesca Pia Quartulli and Marco Di Felice
Appl. Sci. 2024, 14(24), 11489; https://doi.org/10.3390/app142411489 - 10 Dec 2024
Viewed by 455
Abstract
The Internet of Things (IoT) is gaining increasing attention in healthcare due to its potential to enable continuous monitoring of patients, both at home and in controlled medical environments. In this paper, we explore the integration of IoT with human-robotics in the context [...] Read more.
The Internet of Things (IoT) is gaining increasing attention in healthcare due to its potential to enable continuous monitoring of patients, both at home and in controlled medical environments. In this paper, we explore the integration of IoT with human-robotics in the context of motor rehabilitation for groups of patients performing moderate physical routines, focused on balance, stretching, and posture. Specifically, we propose the I-TROPHYTS framework, which introduces a step-change in motor rehabilitation by advancing towards more sustainable medical services and personalized diagnostics. Our framework leverages wearable sensors to monitor patients’ vital signs and edge computing to detect and estimate motor routines. In addition, it incorporates a humanoid robot that mimics the actions of a physiotherapist, adapting motor routines in real-time based on the patient’s condition. All data from physiotherapy sessions are modeled using an ontology, enabling automatic reasoning and planning of robot actions. In this paper, we present the architecture of the proposed framework, which spans four layers, and discuss its enabling components. Furthermore, we detail the current deployment of the IoT system for patient monitoring and automatic identification of motor routines via Machine Learning techniques. Our experimental results, collected from a group of volunteers performing balance and stretching exercises, demonstrate that we can achieve nearly 100% accuracy in distinguishing between shoulder abduction and shoulder flexion, using Inertial Measurement Unit data from wearable IoT devices placed on the wrist and elbow of the test subjects. Full article
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<p>Framework and architecture of I-TROPHYTS.</p>
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<p>Implementation of the first two layers of I-TROPHYTS.</p>
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<p>Illustration of two exercises performed during the experiments.</p>
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<p>Accelerometer and gyroscope raw data—AR exercise.</p>
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<p>Accelerometer and gyroscope raw data—BL exercise.</p>
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<p>Comparison of accuracy and F1-Score metrics in evaluating different learning algorithms.</p>
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<p>Comparison of Accuracy for evaluating FFNN performance using different signals on a variable number of devices.</p>
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<p>Comparison of F1-Score for evaluating FFNN performance using different signals on a variable number of devices.</p>
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<p>Accuracy and F1-Score for predicting motion using FFNN across different time windows.</p>
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<p>Heart rate comparison of two subjects—AL exercise.</p>
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<p>Peak detection—AR exercise.</p>
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<p>Predicted versus actual repetitions of exercises.</p>
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<p>Agent-based cognitive architecture for structuring robotic systems that can monitor, suggest, explain in complex scenarios.</p>
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12 pages, 1330 KiB  
Article
Magnesium Supplementation Modifies Arthritis Synovial and Splenic Transcriptomic Signatures Including Ferroptosis and Cell Senescence Biological Pathways
by Teresina Laragione, Carolyn Harris and Pércio S. Gulko
Nutrients 2024, 16(23), 4247; https://doi.org/10.3390/nu16234247 - 9 Dec 2024
Viewed by 546
Abstract
Background: Rheumatoid arthritis (RA) is a common systemic autoimmune inflammatory disease that can cause joint damage. We have recently reported that oral magnesium supplementation significantly reduces disease severity and joint damage in models of RA. Methods: In the present study, we analyzed the [...] Read more.
Background: Rheumatoid arthritis (RA) is a common systemic autoimmune inflammatory disease that can cause joint damage. We have recently reported that oral magnesium supplementation significantly reduces disease severity and joint damage in models of RA. Methods: In the present study, we analyzed the transcriptome of spleens and synovial tissues obtained from mice with KRN serum-induced arthritis (KSIA) consuming either a high Mg supplemented diet (Mg2800; n = 7) or a normal diet (Mg500; n = 7). Tissues were collected at the end of a 15-day KSIA experiment. RNA was extracted and used for sequencing and analyses. Results: There was an enrichment of differentially expressed genes (DEGs) belonging to Reactome and Gene Ontology (GO) pathways implicated in RA pathogenesis such as RHO GTPases, the RUNX1 pathway, oxidative stress-induced senescence, and the senescence-associated secretory phenotype. Actc1 and Nr4a3 were among the genes with the highest expression, while Krt79 and Ffar2 were among the genes with the lowest expression in synovial tissues of the Mg2800 group compared with the Mg500 group. Spleens had an enrichment for the metabolism of folate and pterines and the HSP90 chaperone cycle for the steroid hormone receptor. Conclusions: We describe the tissue transcriptomic consequences of arthritis-protecting Mg supplementation in KSIA mice. These results show that oral Mg supplementation may interfere with the response to oxidative stress and senescence and other processes known to participate in RA pathogenesis. We provide new evidence supporting the disease-suppressing effect of increased Mg intake in arthritis and its potential to become a new addition to the therapeutic options for RA and other autoimmune and inflammatory diseases. Full article
(This article belongs to the Special Issue Magnesium Homeostasis and Magnesium Transporters in Human Health)
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<p>Arthritis severity scores of mice with KRN serum-induced arthritis (KSIA). (<b>A</b>) Mice were placed on either a normal Mg500 (n = 7) or a high Mg2800 (n = 7) diet 14 days prior to the induction of KSIA and kept on the same diet for an additional 15 days and scored for disease severity (** <span class="html-italic">p</span> = 0.004993 and <span class="html-italic">p</span> = 0.001462, respectively; non-paired <span class="html-italic">t</span>-test). Representative histology sections of KSIA mice on (<b>B</b>) the normal Mg500 diet, showing pronounced synovial hyperplasia and joint damage, and (<b>C</b>) the high Mg2800 diet, showing a protected and normal-looking joint without synovial hyperplasia or damage (H&amp;E staining, 200× magnification).</p>
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<p>Biological pathways enriched in the DEGs between KSIA arthritic mice on Mg2800 and Mg500 diets. (<b>A</b>) Selected Reactome biological pathways and cellular processes enriched in synovial tissues (top section) and spleens (bottom section). (<b>B</b>) Selected Gene Ontology (GO) pathways enriched in the DEGs between mice on Mg2800 and Mg500 diets in the synovial tissues (top section), and spleens (bottom section). (See <a href="#app1-nutrients-16-04247" class="html-app">Supplemental Tables S1 and S2</a> for additional details).</p>
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<p>Volcano plots of the DEGs between KSIA arthritic mice on the Mg2800 diet and those on the Mg500 diet and selected genes’ qPCR confirmation. (<b>A</b>) Volcano plot of DEGs in synovial tissues. (<b>B</b>) Volcano plot of DEGs in spleens. (<b>C</b>) Quantitative PCR (qPCR) confirmation of selected genes expressed in increased and decreased levels in the Mg2700 diet synovial tissues showing a trend in the same direction as seen in the RNA sequencing analyses (<span class="html-italic">p</span> &gt; 0.05).</p>
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12 pages, 1707 KiB  
Article
Pemafibrate Induces a Low Level of PPARα Agonist-Stimulated mRNA Expression of ANGPTL4 in ARPE19 Cell
by Hiroshi Ohguro, Nami Nishikiori, Tatsuya Sato, Megumi Watanabe, Megumi Higashide and Masato Furuhashi
Bioengineering 2024, 11(12), 1247; https://doi.org/10.3390/bioengineering11121247 - 9 Dec 2024
Viewed by 427
Abstract
To elucidate the unidentified roles of a selective peroxisome proliferator-activated receptor α (PPARα) agonist, pemafibrate (Pema), on the pathogenesis of retinal ischemic diseases (RID)s, the pharmacological effects of Pema on the retinal pigment epithelium (RPE), which is involved in the pathogenesis of RID, [...] Read more.
To elucidate the unidentified roles of a selective peroxisome proliferator-activated receptor α (PPARα) agonist, pemafibrate (Pema), on the pathogenesis of retinal ischemic diseases (RID)s, the pharmacological effects of Pema on the retinal pigment epithelium (RPE), which is involved in the pathogenesis of RID, were compared with the pharmacological effects of the non-fibrate PPARα agonist GW7647 (GW). For this purpose, the human RPE cell line ARPE19 that was untreated (NT) or treated with Pema or GW was subjected to Seahorse cellular metabolic analysis and RNA sequencing analysis. Real-time cellular metabolic function analysis revealed that pharmacological effects of the PPARα agonist actions on essential metabolic functions in RPE cells were substantially different between Pema-treated cells and GW-treated cells. RNA sequencing analysis revealed the following differentially expressed genes (DEGs): (1) NT vs. Pema-treated cells, 37 substantially upregulated and 72 substantially downregulated DEGs; (2) NT vs. GW-treated cells, 32 substantially upregulated and 54 substantially downregulated DEGs; and (3) Pema vs. GW, 67 substantially upregulated and 51 markedly downregulated DEGs. Gene ontology (GO) analysis and ingenuity pathway analysis (IPA) showed several overlaps or differences in biological functions and pathways estimated by the DEGs between NT and Pema-treated cells and between NT and GW-treated cells, presumably due to common PPARα agonist actions or unspecific off-target effects to each. For further estimation, overlaps of DEGs among different pairs of comparisons (NT vs. Pema, NT vs. GW, and Pema vs. GW) were listed up. Angiopoietin-like 4 (ANGPTL4), which has been shown to cause deterioration of RID, was the only DEG identified as a common significantly upregulated DEG in all three pairs of comparisons, suggesting that ANGPTL4 was upregulated by the PPARα agonist action but that its levels were substantially lower in Pema-treated cells than in GW-treated cells. In qPCR analysis, such lower efficacy for upregulation of the mRNA expression of ANGPTL4 by Pema than by GW was confirmed, in addition to substantial upregulation of the mRNA expression of HIF1α by both agonists. However, different Pema and GW-induced effects on mRNA expression of HIF1α (Pema, no change; GW, significantly downregulated) and mRNA expression of ANGPTL4 (Pema, significantly upregulated; GW, significantly downregulated) were observed in HepG2 cells, a human hepatocyte cell line. The results of this study suggest that actions of the PPARα agonists Pema and GW are significantly organ-specific and that lower upregulation of mRNA expression of the DR-worsening factor ANGPTL4 by Pema than by GW in ARPE19 cells may minimize the risk for development of RID. Full article
(This article belongs to the Special Issue Pathophysiology and Translational Research of Retinal Diseases)
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<p>Effects of the PPARα agonists pemafibrate (Pema) and GW7647 (GW) on metabolism in ARPE19 cells. ARPE19 cells that were not treated with (Ctrl) and those that were treated with Pema or GW were subjected to a Seahorse real-time metabolic function analysis. Measurement of oxygen consumption rate (OCR, panel (<b>A</b>)). Measurement of extracellular acidification rate (ECAR, panel (<b>B</b>)). Indices of mitochondrial function (panel (<b>C</b>)). Indices of glycolytic function (panel (<b>D</b>)). Baseline OCR/ECAR ratio (panel (<b>E</b>)). Freshly prepared specimens were used in all experiments (n = 6). Data are shown as means ± standard error of the mean (SEM). * <span class="html-italic">p</span> &lt; 0.05. Oligo, oligomycin; FCCP, carbonyl cyanide p-trifluoromethoxyphenylhydrazone; R/A, rotenone/antimycin A; 2-DG, 2-deoxyglucose.</p>
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<p>Heatmaps for DEGs in ARPE19 cells in three comparison conditions: not treated with a PPARα agonist (NT 1-3) vs. treated with Pema 1-3 ((<b>upper</b>) panel), not treated with a PPARα agonist (NT 1-3) vs. treated with GW 1-3 ((<b>middle</b>) panel), and treated with Pema 1-3 vs. treated with GW 1-3 ((<b>lower</b>) panel). Two-dimensionally cultured ARPE19 cells not treated with a PPARα agonist (NT, n = 3) and those treated with 10 μM of Pema (n = 3) or 20 μM of GW (n = 3) were subjected to RNA sequencing analysis. A hierarchical clustering heatmap of differentially expressed genes (DEGs) is shown. Either overexpressed (red) or underexpressed (blue) DEGs in NT cells compared with those in Pema cells are shown in the (<b>upper</b>) panel. Either overexpressed (red) or underexpressed (blue) DEGs in NT cells compared with those in GW cells are shown in the (<b>middle</b>) panel. Either overexpressed (red) or underexpressed (blue) DEGs in Pema cells compared with those in GW cells are shown in the (<b>lower</b>) panel.</p>
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<p>Venn diagram to represent DEGs detected in different categories. DEGs obtained from three different comparisons, namely NT vs. Pema, NT vs. GW, and Pema vs. GW, are shown in a Venn diagram. Gene names are listed in the overlapping area of each circle. ANGPTL4 was commonly observed as an upregulated DEG in these three comparisons. H1-5, histone H1.5; ACTL10, Actin-Like 10; EFHC2, EF-hand domain containing 2; LINC00910, Long Intergenic Non-Protein Coding RNA 910; PDE7B, phosphodiesterase 7B; SNORA66, small nucleolar RNA H/ACA box 66; SMIM11, small integral membrane protein 11; HCLS1, hematopoietic cell-specific Lyn substrate 1; MIR193BHG; MIR193b-365a host gene; ANGPTL4, angiopoietin-like 4; IKBKGP1, Inhibitor of nuclear factor kappa B kinase subunit gamma pseudogene 1; PKD1P1, polycystic kidney disease 1 pseudogene 1; SEPTIN7P3, septin 7 pseudogene 3; FRMPD2B, farnesyl diphosphate synthase pseudogene 2; EXOC5P1, exocyst complex component 5 pseudogene 1; SLC4A1APP1, Solute Carrier Family 4 Member 1 Adaptor Protein Pseudogene 1; TCAF2P1, TRPM8 channel-associated factor 2 pseudogene 1; ZNF890P, zinc finger protein 890 pseudogene.</p>
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<p>qPCR analysis for ANGPTL4 and HIF1α in PPARα agonist-treated ARPE19 cells and HepG2 cells. Two-dimensionally cultured ARPE19 cells and HepG2 cells not treated or treated with a PPARα agonist, GW or Pema, were subjected to qPCR analysis, and the mRNA expression of <span class="html-italic">ANGPTL4</span> and <span class="html-italic">HIF1α</span> was estimated. Experiments were repeated three times, using freshly prepared cells (n = 3 each), in each experiment. * <span class="html-italic">p</span> &lt; 0.05, **** <span class="html-italic">p</span> &lt; 0.001 (Pema vs. GW), <span style="color:red">* <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 </span>(among different concentrations of GW), <span style="color:#0070C0">*** <span class="html-italic">p</span> &lt; 0.005, and **** <span class="html-italic">p</span> &lt; 0.001</span> (among different concentrations of Pema).</p>
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Article
Transcriptome Analysis Reveals Sertoli Cells Adapting Through Redox and Metabolic Pathways Under Heat Stress in Goats
by Guang Yang, Yiwei Wang, Pengyun Ji, Bingyuan Wang and Guoshi Liu
Genes 2024, 15(12), 1582; https://doi.org/10.3390/genes15121582 - 9 Dec 2024
Viewed by 396
Abstract
Background/Objectives: Climate change-induced temperature elevations pose significant challenges to livestock reproduction, particularly affecting testicular function in small ruminants. This study investigates the acute heat-stress response in goat Sertoli cells (SCs), aiming to elucidate the molecular mechanisms underlying heat-induced damage to male reproductive tissues. [...] Read more.
Background/Objectives: Climate change-induced temperature elevations pose significant challenges to livestock reproduction, particularly affecting testicular function in small ruminants. This study investigates the acute heat-stress response in goat Sertoli cells (SCs), aiming to elucidate the molecular mechanisms underlying heat-induced damage to male reproductive tissues. Methods: SCs were isolated from testes of 4-month-old black goats and exposed to heat stress (44 °C for 2.5 h). We employed transcriptome sequencing, CCK-8 assay, electron microscopy, ROS measurement, autophagy detection, Western blot analysis, and lactate concentration measurement. Bioinformatics analyses including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and protein–protein interaction network analyses were performed on the transcriptome data. Results: Heat stress significantly reduced SC viability, induced oxidative stress and autophagy, and altered gene expression profiles. We identified 1231 significantly differentially expressed genes, with significant enrichment in membrane-related processes and metabolic pathways. Metabolism-related genes, including PKLR, ACOT11, and LPCT12, were significantly downregulated. Protein–protein interaction network analysis revealed ten hub genes potentially crucial in the heat-stress response: HSP90AA1, HSPA5, BAG3, IGF1, HSPH1, IL1A, CCL2, CXCL10, ALB, and CALML4. Conclusions: This study provides comprehensive insights into the molecular mechanisms underlying goat SC response to heat stress. The identified genes and pathways, particularly those related to metabolism and stress response, offer potential targets for developing strategies to mitigate heat-stress effects on livestock reproduction. These findings contribute to our understanding of climate change impacts on animal husbandry and may inform the development of heat-stress resistant livestock lines. Full article
(This article belongs to the Special Issue Genetics and Genomics of Sheep and Goat)
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<p>The isolation and identification of primary goat SCs. (<b>A</b>) Bright-field microscopy image of primary SCs. (<b>B</b>) H&amp;E staining of primary SCs. (<b>C</b>) Oil Red O staining of primary SCs; black arrows indicate red lipid droplets. (<b>D</b>,<b>E</b>) Immunofluorescence staining of SC-specific proteins. (<b>D</b>) Vimentin (green fluorescence); (<b>E</b>) WT1 (red fluorescence). Cell nuclei were counterstained with DAPI (blue fluorescence). Scale bars are shown in the bottom right corner of each image.</p>
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<p>The effects of heat stress on goat testicular SC viability and ultrastructure. (<b>A</b>) CCK-8 analysis of SC viability at different temperatures. (<b>B</b>) The impact of different heat-stress durations on SC viability. (<b>C</b>) The effect of various recovery times on SC viability after heat stress. Data are presented as mean ± SEM (n = 6). ** <span class="html-italic">p</span> &lt; 0.01 compared to control. (<b>D</b>) Transmission electron microscopy images of SCs before and after heat stress. Left: control group at 32 °C; right: heat-stress group at 44 °C. Scale bars are shown in the bottom right corner of each image. N: Nucleus; M: Mitochondria; RER: rough endoplasmic reticulum; LD: lipid droplets; ASS: autophagic lysosome; AP: autophagosome.</p>
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<p>Heat stress induces ROS production and autophagy in goat testicular SCs. (<b>A</b>) Representative fluorescence images of ROS levels in SCs at 32 °C and after heat-stress treatment. Green fluorescence indicates DCFH-DA probe staining. Scale bars are shown in the bottom right corner of each image. (<b>B</b>) The quantification of relative fluorescence intensity (OD488/525 nm) in the control and heat-stress groups. ** <span class="html-italic">p</span> &lt; 0.01. (<b>C</b>) Representative fluorescence images of autophagy levels in SCs at 32 °C and after heat-stress treatment. Green fluorescence indicates MDA probe staining. Scale bars are shown in the bottom right corner of each image. (<b>D</b>) The quantification of relative fluorescence intensity (OD335/512 nm) in the control and heat-stress groups. ** <span class="html-italic">p</span> &lt; 0.01. (<b>E</b>) Western blot analysis of autophagy-related proteins p62 and LC3-I/LC3-II. β-actin was used as an internal reference.</p>
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<p>Heat stress induces gene expression changes and pathway enrichment in SCs. (<b>A</b>) A volcano plot of DEGs. Red dots represent upregulated genes in the heat−stress group, while blue dots represent downregulated genes. (<b>B</b>) RT−qPCR validation of DEGs. The bar graph shows the expression levels of 5 upregulated and 5 downregulated genes, randomly selected. *** <span class="html-italic">p</span> &lt; 0.001. (<b>C</b>) A heatmap of the top 40 upregulated and 57 downregulated genes with the most significant expression changes. Color intensity indicates gene expression levels in the control and heat-stress groups. (<b>D</b>) GO enrichment analysis of DEGs. Shows the most significantly enriched biological processes, cellular components, and molecular functions. (<b>E</b>) KEGG pathway enrichment analysis of DEGs. Displays the most significantly enriched signaling pathways.</p>
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<p>Heat stress activates multiple metabolic pathways in goat testicular SCs and affects lactate production. (<b>A</b>) A metabolic pathway map of DEGs based on iPath3 website analysis. Red lines indicate metabolic pathways activated by heat stress. (<b>B</b>) A bar graph of DEGs related to various metabolic processes in the heat-stress group. Shows expression changes in genes associated with glucose metabolism, lipid metabolism, amino acid metabolism, energy metabolism, and nucleotide metabolism. (<b>C</b>) The effect of heat stress on lactate content in SCs’ culture supernatant. The bar graph compares lactate concentrations between the control and heat-stress groups. *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>STRING interaction analysis of all differentially expressed genes DEGs between 32 °C and heat-stressed SCs. Protein–protein interaction network visualized using STRING, with interactions shown at a confidence level of 0.4, while edges between nodes indicate various types of interactions, color-coded and defined in the figure legend. Isolated nodes without edges were removed from the visualization.</p>
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<p>Core genes related to heat-stress injury associated with SCs. Node scores were calculated using Cytoscape software, and the top 10 nodes ranked by node scores were selected.</p>
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