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12 pages, 1959 KB  
Article
UVB Light as a Source of Vitamin D for Indoor-Housed Gestating Sows
by Sine Stricker Jakobsen, Jette Jakobsen, Sheeva Bhattarai and Jens Peter Nielsen
Animals 2025, 15(21), 3185; https://doi.org/10.3390/ani15213185 - 1 Nov 2025
Viewed by 287
Abstract
Vitamin D has been shown to influence growth in both fetuses and piglets. Pigs housed indoors rely solely on their feed as a source of vitamin D since they are never exposed to natural sunlight. UVB light is effective in improving the vitamin [...] Read more.
Vitamin D has been shown to influence growth in both fetuses and piglets. Pigs housed indoors rely solely on their feed as a source of vitamin D since they are never exposed to natural sunlight. UVB light is effective in improving the vitamin D status in sows and piglets housed indoors. This study aimed to investigate an innovative method for UVB lighting to enhance the vitamin D status of gestating sows to a level that could positively impact the growth of both fetuses and suckling piglets. A total of 386 sows were included at the time of insemination. Of these, 197 were exposed to a daily dose of UVB light, and 189 sows served as a control group. Both sows and piglets in the UVB group had significantly higher vitamin D levels in serum samples at the time of farrowing than sows and piglets from the control group. No statistically significant differences in litter weight or the number of liveborn or stillborn piglets were observed between groups. UVB light installed in an electronic sow feeder was successful, but, unfortunately, the lamp used in the study was unable to induce the production of vitamin D to the desired level in the sows. Further studies are needed to investigate the effect of higher doses of UVB light. Full article
(This article belongs to the Section Animal Nutrition)
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16 pages, 943 KB  
Article
Harmonic Mitigation and Energy Savings in 13.2 kV Distribution Feeders via P–Q-Based Shunt Active Filters and Luminaire Retrofit
by Brandon Condemaita and Milton Ruiz
Energies 2025, 18(21), 5582; https://doi.org/10.3390/en18215582 - 23 Oct 2025
Viewed by 415
Abstract
This article designs and validates a P-Q-based shunt active power filter (SAPF) to mitigate voltage harmonics in EERSA’s 13.2 kV feeder 1500080T03. A CYMDIST feeder model, calibrated with field measurements, reveals worst-case voltage THD up to 9.48% due to legacy high-pressure sodium (HPS) [...] Read more.
This article designs and validates a P-Q-based shunt active power filter (SAPF) to mitigate voltage harmonics in EERSA’s 13.2 kV feeder 1500080T03. A CYMDIST feeder model, calibrated with field measurements, reveals worst-case voltage THD up to 9.48% due to legacy high-pressure sodium (HPS) street lighting. Co-simulation with a MATLAB/Simulink R2024b, controller guides the sizing of a 150 kVA SAPF at Substation 8. Simulations reduce peak THD at a representative node from 9.48% to 1.51%; replacing HPS with LEDs further improves efficiency while lowering distortion. The retrofit complies with IEEE Std 519-2022, enhances supply reliability, and yields an internal rate of return above 17%, indicating a technically and financially attractive solution for Latin American distribution networks. Full article
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15 pages, 14339 KB  
Article
Origami Fresnel Zone Plate Lens Reflector Antennas for Satellite Applications
by Massimo Donelli, Sreedevi Menon, Viviana Mulloni, Giada Marchi and Irene Dal Chiele
Electronics 2025, 14(19), 3892; https://doi.org/10.3390/electronics14193892 - 30 Sep 2025
Viewed by 329
Abstract
This work presents a methodology for designing deployable reflector antennas that combine origami structures and the Fresnel zone plate lens to obtain a compact antenna structure. In particular, Miura and Yoshimura’s origami patterns have been considered for the design of the Fresnel reflector [...] Read more.
This work presents a methodology for designing deployable reflector antennas that combine origami structures and the Fresnel zone plate lens to obtain a compact antenna structure. In particular, Miura and Yoshimura’s origami patterns have been considered for the design of the Fresnel reflector mirror and the conical horn antenna feeder, respectively. A set of memory-form alloy (MFA) actuators have been used to deploy the antenna. The MFA actuators are activated by a direct current aimed at increasing the temperature and activating the memorized shape. The combination of these techniques provides light, inexpensive, and very compact antennas, particularly suitable for satellite applications. A numerical and experimental assessment campaign has been carried out on antenna prototypes operating in the Ku band at 15 GHz. The obtained experimental results are quite promising. Full article
(This article belongs to the Special Issue Theory, Methods and Applications of Electromagnetic Scattering)
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19 pages, 2947 KB  
Article
Mouthparts and Alimentary Tract of Flower-Visiting Monkey Beetles (Coleoptera: Scarabaeoidea: Hopliini): Insights into Feeding Preferences
by Michael Neulinger, Florian Karolyi, Jonathan F. Colville, Myriam E. Widmann, Jonas Kristl and Harald W. Krenn
Insects 2025, 16(9), 985; https://doi.org/10.3390/insects16090985 - 21 Sep 2025
Viewed by 699
Abstract
Monkey beetles (Hopliini, Scarabaeoidea) are a species-rich group of flower-visiting insects that are specialized to feed on floral tissue, pollen and/or nectar. We studied ten South African species, examining morphological features associated with their specific feeding preferences. This is the first attempt in [...] Read more.
Monkey beetles (Hopliini, Scarabaeoidea) are a species-rich group of flower-visiting insects that are specialized to feed on floral tissue, pollen and/or nectar. We studied ten South African species, examining morphological features associated with their specific feeding preferences. This is the first attempt in a limited number of beetle species to comparatively investigate both the mouthparts and the alimentary tract in relation to ingested food. Using light microscopy, we found cutting edges on the mandibles and galea teeth in flower tissue-feeding species. Pollen feeders have numerous bristles on the maxillae and a prominent mola on the mandibles that are likely used for gathering and grinding pollen. The elongate heads and mouthparts of the nectar feeders are considered an adaptation that enable these species to mop up nectar while probing flowers. Using µCT imaging and reconstructions of the entire alimentary tract, our morphometric results suggest that food preferences are not related to total relative gut length, although the ratio of foregut to body length was greater in pollen- and nectar-feeding monkey beetles than in floral tissue-feeders. The midgut of females tends to be longer relative to body size compared to males. Our work serves as a basis for generating hypotheses for future research that includes gut morphology in flower-visiting insects. Full article
(This article belongs to the Special Issue Beetles: Biology, Ecology, and Integrated Management)
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20 pages, 13462 KB  
Article
An AI-Based System for Monitoring Laying Hen Behavior Using Computer Vision for Small-Scale Poultry Farms
by Jill Italiya, Ahmed Abdelmoamen Ahmed, Ahmed A. A. Abdel-Wareth and Jayant Lohakare
Agriculture 2025, 15(18), 1963; https://doi.org/10.3390/agriculture15181963 - 17 Sep 2025
Viewed by 1126
Abstract
Small-scale poultry farms often lack access to advanced monitoring tools and rely heavily on manual observation, which is time-consuming, inconsistent, and insufficient for precise flock management. Feeding and drinking behaviors are critical, as they serve as early indicators of health and environmental issues. [...] Read more.
Small-scale poultry farms often lack access to advanced monitoring tools and rely heavily on manual observation, which is time-consuming, inconsistent, and insufficient for precise flock management. Feeding and drinking behaviors are critical, as they serve as early indicators of health and environmental issues. With global poultry production expanding, raising over 70 billion hens annually, there is an urgent need for intelligent, low-cost systems that can continuously and accurately monitor bird behavior in resource-limited farm settings. This paper presents the development of a computer vision-based chicken behavior monitoring system, specifically designed for small barn environments where at most 10–15 chickens are housed at any time. The developed system consists of an object detection model, created on top of the YOLOv8 model, trained with an imagery dataset of laying hen, feeder, and waterer objects. Although chickens are visually indistinguishable, the system processes each detection per frame using bounding boxes and movement-based approximation identification rather than continuous identity tracking. The approach simplifies the tracking process without losing valuable behavior insights. Over 700 frames were annotated manually for high-quality labeled data, with different lighting, hen positions, and interaction angles with dispensers. The images were annotated in YOLO format and used for training the detection model for 100 epochs, resulting in a model having an average mean average precision (mAP@0.5) metric value of 91.5% and a detection accuracy of over 92%. The proposed system offers an efficient, low-cost solution for monitoring chicken feeding and drinking behaviors in small-scale farms, supporting improved management and early health detection. Full article
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23 pages, 4418 KB  
Article
Optimization of Electric Transformer Operation Through Load Estimation Based on the K-Means Algorithm
by Pedro Torres-Bermeo, José Varela-Aldás, Kevin López-Eugenio, Nancy Velasco and Guillermo Palacios-Navarro
Energies 2025, 18(14), 3755; https://doi.org/10.3390/en18143755 - 15 Jul 2025
Viewed by 984
Abstract
This study presents an innovative methodology to optimize the operation of distribution transformers through the estimation of hourly load curves, aimed at minimizing technical losses due to oversizing, particularly in systems lacking advanced metering infrastructure. The proposed approach combines clustering techniques, K-Means with [...] Read more.
This study presents an innovative methodology to optimize the operation of distribution transformers through the estimation of hourly load curves, aimed at minimizing technical losses due to oversizing, particularly in systems lacking advanced metering infrastructure. The proposed approach combines clustering techniques, K-Means with DTW, to identify representative daily consumption patterns and a supervised model based on LightGBM to estimate hourly load curves for unmetered transformers, using customer characteristics as input. These estimated curves are integrated into a process that calculates technical losses, both no-load and load losses, for different transformer sizes, selecting the optimal rating that minimizes losses without compromising demand. Empirical validation showed accuracy levels of 95.6%, 95.29%, and 98.14% at an individual transformer, feeder, and a complete electrical system with 16,864 transformers, respectively. The application of the methodology to a real distribution system revealed a potential annual energy savings of 3004 MWh, equivalent to an estimated economic reduction of 150,238 USD. Full article
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25 pages, 11560 KB  
Article
Transcriptomic and Proteomic Analyses of the Liver and Ileum Identify Key Genes and Pathways Associated with Low and High Groups of Social Genetic Effect of Residual Feed Intake
by Patrick Kofi Makafui Tecku, Zhenjian Zhao, Kai Wang, Xiang Ji, Dong Chen, Qi Shen, Yang Yu, Shengdi Cui, Junge Wang, Ziyang Chen, Jia Xue and Guoqing Tang
Animals 2025, 15(9), 1345; https://doi.org/10.3390/ani15091345 - 7 May 2025
Viewed by 784
Abstract
Social genetic effects (SGEs) refer to how the genotypes of other individuals impact an individual’s phenotype within a population. These effects significantly influence the feeding behavior and production performance in pigs, though their mechanisms are not well understood. This study examined two pig [...] Read more.
Social genetic effects (SGEs) refer to how the genotypes of other individuals impact an individual’s phenotype within a population. These effects significantly influence the feeding behavior and production performance in pigs, though their mechanisms are not well understood. This study examined two pig groups with extreme SGE values for residual feed intake (RFI), analyzing their feeding behavior and the molecular mechanisms involved using transcriptomics and proteomics analysis of liver and ileum tissues. Pigs with higher SGE values exhibited distinct feeding patterns, spending more time at the feeder but making fewer visits. They consumed less overall feed but had a higher intake per visit. Differentially expressed genes and proteins were identified in the liver and ileum and were associated with processes such as mitochondrial functions, oxidative phosphorylation, and cholesterol metabolism. Integrated analysis supported these findings. Combined transcriptome and proteome analysis identified potential key genes that were associated with processes including mitochondrial processes, oxidative phosphorylation, fat digestion and absorption, and cholesterol metabolism. The results showed that pigs with differing SGE values display different feeding behaviors and utilize distinct molecular pathways affecting RFI. These findings offer valuable insights into how SGEs influence feed efficiency and shed light on the fundamental mechanisms underlying it. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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11 pages, 2089 KB  
Article
An Experimental Study of Radiated Energy from an Optical Fiber and the Potential for an Optical MIMO System
by Hasan Farahneh, Jamal S. Rahhal, Dia I. Abualnadi, Ibrahim Mansour, Ahmad K. Atieh and Xavier Fernando
Appl. Sci. 2025, 15(6), 2916; https://doi.org/10.3390/app15062916 - 7 Mar 2025
Cited by 1 | Viewed by 2719
Abstract
Leaky feeders provide seamless and uniform signal coverage in confined spaces like tunnels, mines, and buildings. Their easy scalability and integration with modern systems, like Multiple Input Multiple Output (MIMO), make them ideal for environments requiring reliable and consistent connectivity. However, using optical [...] Read more.
Leaky feeders provide seamless and uniform signal coverage in confined spaces like tunnels, mines, and buildings. Their easy scalability and integration with modern systems, like Multiple Input Multiple Output (MIMO), make them ideal for environments requiring reliable and consistent connectivity. However, using optical fiber as a radiating cable has never been investigated before. This may seem infeasible at first sight. However, our experimental study shows otherwise. We measured light leaking from a bent optical fiber transmitter. We also derived closed-form formulas to describe the amount of leakage energy and found that this energy exponentially varies with the square of the curvature radius. This allows us to design an Optical Leaky Feeder (OLF) transmission system for the first time. Then, we analytically show that a slotted optical fiber can be used as a MIMO receiver. The proposed system can ensure reliable, high-quality signal distribution even in challenging environments like tunnels, industrial settings, and dense urban areas. Full article
(This article belongs to the Special Issue Signal Processing and Communication for Wireless Sensor Network)
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17 pages, 1073 KB  
Article
Promoting the Use of Bicycles for Transportation and Access to Bus Stations in Riyadh, Saudi Arabia
by Muhammad Ahmad Al-Rashid and Sameeh Alarabi
Sustainability 2025, 17(5), 1921; https://doi.org/10.3390/su17051921 - 24 Feb 2025
Viewed by 2128
Abstract
Most research on bicycling and public transportation integration centers around the developed world and inter-modal bicycle–train travel. Similarly, existing research into the factors influencing bicycling to public transport focuses solely on one aspect of the issue, such as physical infrastructure, social environment, user [...] Read more.
Most research on bicycling and public transportation integration centers around the developed world and inter-modal bicycle–train travel. Similarly, existing research into the factors influencing bicycling to public transport focuses solely on one aspect of the issue, such as physical infrastructure, social environment, user or individual characteristics, etc. On the other hand, interventions to promote bicycling are less likely to be successful when carried out together since there can be multiple levels of barriers and enablers to bicycling. Hence, this study addresses this gap and utilizes a socio-ecological model as a theoretical foundation to understand the potential of using bicycles as a feeder for bus services by analyzing the barriers and enablers. Data were collected through a face-to-face questionnaire survey conducted in public spaces, including bus stops in Riyadh, Saudi Arabia. Two binary logistics models were developed using Statistical Package for the Social Sciences (SPSS) to analyze the key factors influencing bicycle use as a feeder mode to public buses. The findings reveal that individual socio-demographic factors, reported barriers, and enablers are essential in explaining the users’ likelihood of bicycle use in accessing public buses. The barriers model indicates that existing bicycle parking facilities, bicycle theft and safety issues, bus reliability, and distance to bus stops play an essential role. Effective shading, dedicated bicycle tracks, appropriate lighting, and vital public services are explanatory for the enablers model. The findings offer valuable insights and policy recommendations to improve bicycling infrastructure and promote comprehensive accessibility to public transportation. Full article
(This article belongs to the Special Issue New Trends in Sustainable Transportation)
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22 pages, 8046 KB  
Article
Advanced Deep Learning Methods for Multiple Behavior Classification of Cage-Free Laying Hens
by Sachin Subedi, Ramesh Bahadur Bist, Xiao Yang, Guoming Li and Lilong Chai
AgriEngineering 2025, 7(2), 24; https://doi.org/10.3390/agriengineering7020024 - 23 Jan 2025
Cited by 2 | Viewed by 1583
Abstract
The welfare of hens in cage-free systems is closely linked to their behaviors, such as feeding, drinking, pecking, perching, bathing, preening, and foraging. To monitor these behaviors, we developed and evaluated deep learning models based on YOLO (You Only Look Once), an advanced [...] Read more.
The welfare of hens in cage-free systems is closely linked to their behaviors, such as feeding, drinking, pecking, perching, bathing, preening, and foraging. To monitor these behaviors, we developed and evaluated deep learning models based on YOLO (You Only Look Once), an advanced object detection technology known for its high accuracy, speed, and compact size. Three YOLO-based models—YOLOv5s_BH, YOLOv5x_BH, and YOLOv7_BH—were created to track and classify the behaviors of laying hens in cage-free environments. A dataset comprising 1500 training images, 500 validation images, and 50 test images was used to train and validate the models. The models successfully detected poultry behaviors in test images with bounding boxes and objectness scores ranging from 0 to 1. Among the models, YOLOv5s_BH demonstrated superior performance, achieving a precision of 78.1%, surpassing YOLOv5x_BH and YOLOv7_BH by 1.9% and 2.2%, respectively. It also achieved a recall of 71.7%, outperforming YOLOv5x_BH and YOLOv7_BH by 1.9% and 2.8%, respectively. Additionally, YOLOv5s_BH recorded a mean average precision (mAP) of 74.6%, exceeding YOLOv5x_BH by 2.6% and YOLOv7_BH by 9%. While all models demonstrated high detection precision, their performance was influenced by factors such as stocking density, varying light conditions, and obstructions from equipment like drinking lines, perches, and feeders. This study highlights the potential for the automated monitoring of poultry behaviors in cage-free systems, offering valuable insights for producers. Full article
(This article belongs to the Section Livestock Farming Technology)
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17 pages, 1994 KB  
Article
Notes on Modified Planar Kelvin–Stuart Models: Simulations, Applications, Probabilistic Control on the Perturbations
by Nikolay Kyurkchiev, Tsvetelin Zaevski, Anton Iliev, Vesselin Kyurkchiev and Asen Rahnev
Axioms 2024, 13(10), 720; https://doi.org/10.3390/axioms13100720 - 17 Oct 2024
Cited by 3 | Viewed by 887
Abstract
In this paper, we propose a new modified planar Kelvin–Stuart model. We demonstrate some modules for investigating the dynamics of the proposed model. This will be included as an integral part of a planned, much more general Web-based application for scientific computing. Investigations [...] Read more.
In this paper, we propose a new modified planar Kelvin–Stuart model. We demonstrate some modules for investigating the dynamics of the proposed model. This will be included as an integral part of a planned, much more general Web-based application for scientific computing. Investigations in light of Melnikov’s approach are considered. Some simulations and applications are also presented. The proposed new modifications of planar Kelvin–Stuart models contain many free parameters (the coefficients gi,i=1,2,,N), which makes them attractive for use in engineering applications such as the antenna feeder technique (a possible generating and simulating of antenna factors) and the theory of approximations (a possible good approximation of a given electrical stage). The probabilistic control of the perturbations is discussed. Full article
(This article belongs to the Special Issue Differential Equations and Related Topics, 2nd Edition)
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22 pages, 3149 KB  
Article
Accurate Surge Arrester Modeling for Optimal Risk-Aware Lightning Protection Utilizing a Hybrid Monte Carlo–Particle Swarm Optimization Algorithm
by Amir Hossein Kimiai Asadi, Mohsen Eskandari and Hadi Delavari
Technologies 2024, 12(6), 88; https://doi.org/10.3390/technologies12060088 - 8 Jun 2024
Cited by 4 | Viewed by 3009
Abstract
The application of arresters is critical for the safe operation of electric grids against lightning. Arresters limit the consequences of lightning-induced over-voltages. However, surge arrester protection in electric grids is challenging due to the intrinsic complexities of distribution grids, including overhead lines and [...] Read more.
The application of arresters is critical for the safe operation of electric grids against lightning. Arresters limit the consequences of lightning-induced over-voltages. However, surge arrester protection in electric grids is challenging due to the intrinsic complexities of distribution grids, including overhead lines and power components such as transformers. In this paper, an optimal arrester placement technique is developed by proposing a multi-objective function that includes technical, safety and risk, and economic indices. However, an effective placement model demands a comprehensive and accurate modeling of an electric grid’s components. In this light, appropriate models of a grid’s components including an arrester, the earth, an oil-immersed transformer, overhead lines, and lightning-induced voltage are developed. To achieve accurate models, high-frequency transient mathematical models are developed for the grid’s components. Notably, to have an accurate model of the arrester, which critically impacts the performance of the arrester placement technique, a new arrester model is developed and evaluated based on real technical data from manufacturers such as Pars, Tridelta, and Siemens. Then, the proposed model is compared with the IEEE, Fernandez, and Pinceti models. The arrester model is incorporated in an optimization problem considering the performance of the over-voltage protection and the risk, technical, and economic indices, and it is solved using the particle swarm optimization (PSO) and Monte Carlo (MC) techniques. To validate the proposed arrester model and the placement technique, real data from the Chopoghloo feeder in Bahar, Hamedan, Iran, are simulated. The feeder is expanded over three different geographical areas, including rural, agricultural plain, and mountainous areas. Full article
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24 pages, 10911 KB  
Article
A Comprehensive Analysis of Structural Alternatives and Local Opening for a 2000 TEU Green Methanol-Powered Container Vessel
by Arturo Silva-Campillo and Francisco Pérez-Arribas
Appl. Sci. 2024, 14(7), 2810; https://doi.org/10.3390/app14072810 - 27 Mar 2024
Cited by 2 | Viewed by 1626
Abstract
In the context of global trade and environmental concerns, this research focuses on examining the influence of the principal dimensions of a container feeder vessel whose propulsion has been modified to operate with green methanol. Methanol emerges as a sustainable marine fuel, reducing [...] Read more.
In the context of global trade and environmental concerns, this research focuses on examining the influence of the principal dimensions of a container feeder vessel whose propulsion has been modified to operate with green methanol. Methanol emerges as a sustainable marine fuel, reducing emissions and dependence on fossil fuels. The study comprehensively examines the structural challenges posed by container ships, with a particular focus on torsional stresses and hydroelasticity effects. A significant novelty of this study is the inclusion of a comparative structural analysis evaluating how changes in the vessel’s main dimensions impact its structural response. This analysis sheds some light on crucial insights into the effects of structural modifications required for accommodating these changes, ensuring the structural strength of these vessels. The research also underscores the impact of permanent and transient springing and whipping phenomena on fatigue damage. The study’s significance lies in its role in the ongoing transition to sustainable maritime transportation, as it not only examines structural challenges but also provides solutions for achieving an optimal structural configuration in this new era of environmental responsibility. Full article
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13 pages, 958 KB  
Communication
On a Class of Orthogonal Polynomials as Corrections in Lienard Differential System: Applications
by Vesselin Kyurkchiev, Anton Iliev, Asen Rahnev and Nikolay Kyurkchiev
Algorithms 2023, 16(6), 297; https://doi.org/10.3390/a16060297 - 12 Jun 2023
Cited by 14 | Viewed by 2040
Abstract
In this paper we demonstrate some specialized modules for investigating the dynamics of differential models, an integral part of a planned much more general Web-based application for scientific computing. As “corrections” in the Lienard differential system is presented a class of orthogonal polynomials [...] Read more.
In this paper we demonstrate some specialized modules for investigating the dynamics of differential models, an integral part of a planned much more general Web-based application for scientific computing. As “corrections” in the Lienard differential system is presented a class of orthogonal polynomials (also known as “shell polynomials”). We will note that some specifics of the amplitudes of these polynomials open up the possibility of modeling signals from the field of antenna-feeder techniques. Algorithms and modules have been consistently used for: automatic generation of a theorem on the number and type of limit cycles (in the light of Melnikov’s considerations); study of the Hamiltonian of the system and “level curves”; for the study of catastrophic surfaces (in the light of Zeeman’s considerations), etc. Similar studies have been carried out for associated polynomials. Numerical examples, illustrating our results using CAS MATHEMATICA are given. Full article
(This article belongs to the Collection Feature Papers in Algorithms)
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13 pages, 8418 KB  
Article
Embryonic Stem Cells Can Generate Oral Epithelia under Matrix Instruction
by Ridhima Das, Lisa Harper, Kayoko Kitajima, Tarig Al-Hadi Osman, Mihaela Roxana Cimpan, Anne Chr. Johannssen, Salwa Suliman, Ian C. Mackenzie and Daniela-Elena Costea
Int. J. Mol. Sci. 2023, 24(9), 7694; https://doi.org/10.3390/ijms24097694 - 22 Apr 2023
Cited by 2 | Viewed by 2493
Abstract
We aimed to investigate whether molecular clues from the extracellular matrix (ECM) can induce oral epithelial differentiation of pluripotent stem cells. Mouse embryonic stem cells (ESC) of the feeder-independent cell line E14 were used as a model for pluripotent stem cells. They were [...] Read more.
We aimed to investigate whether molecular clues from the extracellular matrix (ECM) can induce oral epithelial differentiation of pluripotent stem cells. Mouse embryonic stem cells (ESC) of the feeder-independent cell line E14 were used as a model for pluripotent stem cells. They were first grown in 2D on various matrices in media containing vitamin C and without leukemia inhibitory factor (LIF). Matrices investigated were gelatin, laminin, and extracellular matrices (ECM) synthesized by primary normal oral fibroblasts and keratinocytes in culture. Differentiation into epithelial lineages was assessed by light microscopy, immunocytochemistry, and flow cytometry for cytokeratins and stem cell markers. ESC grown in 2D on various matrices were afterwards grown in 3D organotypic cultures with or without oral fibroblasts in the collagen matrix and examined histologically and by immunohistochemistry for epithelial (keratin pairs 1/10 and 4/13 to distinguish epidermal from oral epithelia and keratins 8,18,19 to phenotype simple epithelia) and mesenchymal (vimentin) phenotypes. ECM synthesized by either oral fibroblasts or keratinocytes was able to induce, in 2D cultures, the expression of cytokeratins of the stratified epithelial phenotype. When grown in 3D, all ESC developed into two morphologically distinct cell populations on collagen gels: (i) epithelial-like cells organized in islands with occasional cyst- or duct-like structures and (ii) spindle-shaped cells suggestive of mesenchymal differentiation. The 3D culture on oral fibroblast-populated collagen matrices was necessary for further differentiation into oral epithelia. Only ESC initially grown on 2D keratinocyte or fibroblast-synthesized matrices reached full epithelial maturation. In conclusion, ESC can generate oral epithelia under matrix instruction. Full article
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