22 results on '"Raška J"'
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2. Dynamic response of a material during a high-speed impact
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Raška, J., primary and Oberthor, M., additional
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- 2020
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3. Numerical evaluations of strain field modification induced by production flaws in loaded composite structures
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Raška Jan, Vlach Jarmil, and Horňas Jan
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Engineering (General). Civil engineering (General) ,TA1-2040 - Abstract
Structural health monitoring (SHM) involves observation and analysis of a system over time using periodically sampled response measurements to monitor the initiation and propagation of a flaw. The article is focused on finite element (FE) modelling of strain field modification induced by flaws in loaded composite structures. Shape of the current production flaw – delamination – was idealised as square with a side of 1 inch. One flaw per laminate in several positions in the layup was considered. The pristine structure was modelled with one element per thickness. For delaminated zone in the centre, the twin coincident elements per thickness were adopted: first element to simulate the lay-up from bottom to delamination, second from delamination to top. The loads and the boundary conditions of the FE model were issued from the tests, based on the ASTM standards: tension test – ASTM D3039, compression after impact test – ASTM D7137 and 4-point bending test – ASTM D7264. Generally, the strain modification is stronger for the delamination near the surface than for the delamination deep in the structure.
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- 2021
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4. Bird and hail stone impact resistance analysis on a jet engine composite air inlet
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Doubrava Radek, Oberthor Martin, Bělský Petr, and Raška Jan
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Engineering (General). Civil engineering (General) ,TA1-2040 - Abstract
Bird or hail stone impacts are an important phenomenon that must be taken into consideration when designing aircraft. As engines are the sole thrust-providing mechanisms of an aircraft, it is critical that the effects of bird or hail stone strikes on engine inlets and systems be investigated and mitigated to the greatest extent possible. A combination of experiments and numerical simulations is necessary to properly understand the behaviour of a bird or hail stone during impact and the reaction of the impacted material with the structure. A simulation methodology is developed and validated to certify the bird or hail stone strike resistance of composite air ducts designed for a new generation of jet training aircraft. Physical impact tests were performed on real composite parts. Numerical simulation results were compared with test results. Numerical simulation was also used for test preparation and optimization of the test rig design from the point of view of the influence of the stiffness of the surrounding aircraft structure. The validated modelling procedure allows the analysis of numerous impact scenarios, improving the optimization procedures for aircraft component design and reducing the cost of development by reducing the need to manufacture test prototypes.
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- 2018
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5. Mathematical Simulation of Drying Process of Fibrous Material
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Blejchař Tomáš, Raška Jiří, and Jablonská Jana
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Physics ,QC1-999 - Abstract
The article describes mathematical simulation of flowing air through porous zone and water vaporisation from mentioned porous area which actually represents dried fibrous material - cotton towel. Simulation is based on finite volume method. Wet towel is placed in pipe and hot air flow through the towel. Water from towel is evaporated. Simulation of airflow through porous element is described first. Eulerian multiphase model is then used for simulation of water vaporisation from porous medium. Results of simulation are compared with experiment. Ansys Fluent 13.0 was used for calculation.
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- 2018
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6. The evolution and ecology of multiple antipredator defences.
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Kikuchi DW, Allen WL, Arbuckle K, Aubier TG, Briolat ES, Burdfield-Steel ER, Cheney KL, Daňková K, Elias M, Hämäläinen L, Herberstein ME, Hossie TJ, Joron M, Kunte K, Leavell BC, Lindstedt C, Lorioux-Chevalier U, McClure M, McLellan CF, Medina I, Nawge V, Páez E, Pal A, Pekár S, Penacchio O, Raška J, Reader T, Rojas B, Rönkä KH, Rößler DC, Rowe C, Rowland HM, Roy A, Schaal KA, Sherratt TN, Skelhorn J, Smart HR, Stankowich T, Stefan AM, Summers K, Taylor CH, Thorogood R, Umbers K, Winters AE, Yeager J, and Exnerová A
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- Animals, Phenotype, Ecology, Predatory Behavior
- Abstract
Prey seldom rely on a single type of antipredator defence, often using multiple defences to avoid predation. In many cases, selection in different contexts may favour the evolution of multiple defences in a prey. However, a prey may use multiple defences to protect itself during a single predator encounter. Such "defence portfolios" that defend prey against a single instance of predation are distributed across and within successive stages of the predation sequence (encounter, detection, identification, approach (attack), subjugation and consumption). We contend that at present, our understanding of defence portfolio evolution is incomplete, and seen from the fragmentary perspective of specific sensory systems (e.g., visual) or specific types of defences (especially aposematism). In this review, we aim to build a comprehensive framework for conceptualizing the evolution of multiple prey defences, beginning with hypotheses for the evolution of multiple defences in general, and defence portfolios in particular. We then examine idealized models of resource trade-offs and functional interactions between traits, along with evidence supporting them. We find that defence portfolios are constrained by resource allocation to other aspects of life history, as well as functional incompatibilities between different defences. We also find that selection is likely to favour combinations of defences that have synergistic effects on predator behaviour and prey survival. Next, we examine specific aspects of prey ecology, genetics and development, and predator cognition that modify the predictions of current hypotheses or introduce competing hypotheses. We outline schema for gathering data on the distribution of prey defences across species and geography, determining how multiple defences are produced, and testing the proximate mechanisms by which multiple prey defences impact predator behaviour. Adopting these approaches will strengthen our understanding of multiple defensive strategies., (© 2023 The Authors. Journal of Evolutionary Biology published by John Wiley & Sons Ltd on behalf of European Society for Evolutionary Biology.)
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- 2023
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7. Ontogenetic change in effectiveness of chemical defence against different predators in Oxycarenus true bugs.
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Raška J, Chalušová K, Krajiček J, Čabala R, Bosáková Z, Štys P, and Exnerová A
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- Animals, Larva, Birds, Aldehydes, Predatory Behavior, Heteroptera physiology
- Abstract
Many prey species change their antipredator defence during ontogeny, which may be connected to different potential predators over the life cycle of the prey. To test this hypothesis, we compared reactions of two predator taxa - spiders and birds - to larvae and adults of two invasive true bug species, Oxycarenus hyalinipennis and Oxycarenus lavaterae (Heteroptera: Oxycarenidae) with life-stage-specific chemical defence mechanisms. The reactions to larvae and adults of both true bug species strikingly differed between the two predator taxa. The spiders were deterred by the defences of adult bugs, but the larval defences were ineffective against them. By contrast, birds attacked the larvae considerably less often than the adult bugs. The results indicate a predator-specific ontogenetic change in defence effectiveness of both Oxycarenus species. The change in defence is likely linked to the life-stage-specific composition of secretions in both species: whereas secretions of larvae are dominated by unsaturated aldehydes, secretions of adults are rich in terpenoids, which probably serve dual function of defensive chemicals and pheromones. Our results highlight the variation in defence between different life stages and the importance of testing responses of different types of predators., (© 2023 European Society for Evolutionary Biology.)
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- 2023
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8. Single Cerebral Organoid Mass Spectrometry of Cell-Specific Protein and Glycosphingolipid Traits.
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Nezvedová M, Jha D, Váňová T, Gadara D, Klímová H, Raška J, Opálka L, Bohačiaková D, and Spáčil Z
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- Humans, Reproducibility of Results, Neurons metabolism, Cell Differentiation, Mass Spectrometry, Organoids, Neural Stem Cells
- Abstract
Cerebral organoids are a prolific research topic and an emerging model system for neurological diseases in human neurobiology. However, the batch-to-batch reproducibility of current cultivation protocols is challenging and thus requires a high-throughput methodology to comprehensively characterize cerebral organoid cytoarchitecture and neural development. We report a mass spectrometry-based protocol to quantify neural tissue cell markers, cell surface lipids, and housekeeping proteins in a single organoid. Profiled traits probe the development of neural stem cells, radial glial cells, neurons, and astrocytes. We assessed the cell population heterogeneity in individually profiled organoids in the early and late neurogenesis stages. Here, we present a unifying view of cell-type specificity of profiled protein and lipid traits in neural tissue. Our workflow characterizes the cytoarchitecture, differentiation stage, and batch cultivation variation on an individual cerebral organoid level.
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- 2023
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9. Strain-Field Modifications in the Surroundings of Impact Damage of Carbon/Epoxy Laminate.
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Vlach J, Doubrava R, Růžek R, Raška J, Horňas J, and Kadlec M
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The relationship between deformation and stress is crucial for any elasto-plastic body. This paper deals with the experimental identification of the basic parameters of the composite laminate model in relation to the finite element model. Standardized tensile, impact, and post-impact tests on a carbon fiber-reinforced epoxy laminate were used. The method by which the elasticity and failure parameters were obtained from the initial components is described. In the article, the modes of initiation and complete failure of samples in tensile tests, which are compared with the simulation, are presented. Furthermore, the article deals with the issue of the generation and detection of damage by low-speed impact, which can be caused by contact with moving objects, due to improper handling or maintenance. The results of impact analysis simulations are shown in the context of strain-field distribution changes obtained with the help of digital image correlation. The results showed high agreement between the calculations and the experiments. Based on this agreement, simulations of impact damage for various energies were performed. These simulations were used to determine the approximate sizes of the affected zones in relation to the impact energy. The results are finally discussed in the context of the possible use of structural health monitoring based on strain modifications.
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- 2022
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10. Endocrine-disrupting chemicals affect Sertoli TM4 cell functionality through dysregulation of gap junctional intercellular communication in vitro.
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Yawer A, Sychrová E, Raška J, Babica P, and Sovadinová I
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- Cell Communication, Connexin 43 genetics, Connexin 43 metabolism, Gap Junctions metabolism, Humans, Male, Phosphorylation, Endocrine Disruptors metabolism, Testicular Neoplasms metabolism
- Abstract
The frequencies of adverse outcomes associated with male reproductive health, including infertility and testicular cancer, are increasing. These adverse trends are partially attributed to increased exposure to environmental agents such as endocrine-disrupting chemicals (EDCs). This study addresses effects on EDCs on adjacent prepubertal Sertoli TM4 cells, specifically on 1) testicular gap junctional intercellular communication (GJIC), one of the hallmarks of non-genotoxic carcinogenicity, 2) GJIC building blocks connexins (Cx), and 3) mitogen-activated protein kinases MAPKs. We selected eight representatives of EDCs: organochlorine chemicals such as pesticides dichlorodiphenyltrichloroethane, lindane, methoxychlor, and vinclozolin, industrial chemicals bisphenol A and 2,2',4,4',5,5'-hexachlorobiphenyl, and components of personal care products, triclocarban and triclosan. EDCs rapidly dysregulated GJIC in Sertoli TM4 cells mainly via MAPK p38 and/or Erk1/2 pathways by the intermediate hyper- or de-phosphorylation of Cx43 (Ser368, Ser282) and translocation of Cx43 from the plasma membrane, suggesting disturbed intracellular trafficking of Cx43 protein. Surprisingly, EDCs did not rapidly activate MAPK Erk1/2 or p38; on the contrary, TCC and TCS decreased their activity (phosphorylation). Our results indicate that EDCs might disrupt testicular homeostasis and development via testicular GJIC, junctional and non-junctional functions of Cx43 and MAPK-signaling pathways in Sertoli cells., (Copyright © 2022 Elsevier Ltd. All rights reserved.)
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- 2022
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11. Spatiotemporal variation in the role of floral traits in shaping tropical plant-pollinator interactions.
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Klomberg Y, Tropek R, Mertens JEJ, Kobe IN, Hodeček J, Raška J, Fominka NT, Souto-Vilarós D, Janečková P, and Janeček Š
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- Phenotype, Plants, Seasons, Flowers, Pollination
- Abstract
The pollination syndrome hypothesis predicts that plants pollinated by the same pollinator group bear convergent combinations of specific floral functional traits. Nevertheless, some studies have shown that these combinations predict pollinators with relatively low accuracy. This discrepancy may be caused by changes in the importance of specific floral traits for different pollinator groups and under different environmental conditions. To explore this, we studied pollination systems and floral traits along an elevational gradient on Mount Cameroon during wet and dry seasons. Using Random Forest (Machine Learning) models, allowing the ranking of traits by their relative importance, we demonstrated that some floral traits are more important than others for pollinators. However, the distribution and importance of traits vary under different environmental conditions. Our results imply the need to improve our trait-based understanding of plant-pollinator interactions to better inform the debate surrounding the pollination syndrome hypothesis., (© 2022 John Wiley & Sons Ltd.)
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- 2022
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12. An intergenerational approach to parasitoid fitness determined using clutch size.
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Samková A, Raška J, Hadrava J, and Skuhrovec J
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- Animals, Clutch Size, Ecosystem, Female, Host-Parasite Interactions, Reproduction, Wasps
- Abstract
Parasitoids, as important natural enemies, occur in high numbers and help maintain balance in natural ecosystems. Their fitness is traditionally studied as fertility based on the number of offspring in the F1 generation. Here, using gregarious parasitoids as models, we show that this traditional approach omits one important parameter: the clutch size-body size-fertility correlation among offspring. As a result of this correlation, when females adjust the number of offspring laid in a host, they determine not only the number of offspring produced but also the body size and reproductive potential of those offspring. Although parasitoid fertility has been determined several times from clutch size, here we use Anaphes flavipes to demonstrate the use of this relationship in an upgraded intergenerational approach to parasitoid fitness. We show that with a range of hosts simultaneously utilized by female parasitoids, identical fertility in the F1 generation can lead to distinctly different fertility values in the F2 generation. Even with the same number of hosts, lower fertility in the F1 generation can generate higher fertility in the F2 generation. Our approach provides an intergenerational perspective for determining individual fitness of gregarious parasitoids and new possibilities for the modelling of parasitoid population density., (© 2022. The Author(s).)
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- 2022
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13. Linearization of Composite Material Damage Model Results and Its Impact on the Subsequent Stress-Strain Analysis.
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Vlach J, Doubrava R, Růžek R, Raška J, Horňas J, and Kadlec M
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To solve problems in the field of mechanical engineering efficiently, individual numerical procedures must be developed, and solvers must be adapted. This study applies the results of a carbon-fibre reinforced polymer (CFRP) analysis along with the nonlinear finite element damage (FE) method to the translation of a linear solver. The analyzed tensile test sample is modelled using the ply-by-ply method. To describe the nonlinear post-damage behavior of the material, the Hashin model is used. To validate the transformation, an analysis and comparison of the damage results of the linearized and nonlinear model is carried out. Job linearization was performed by collecting elements into groups based on their level of damage and pairing them with unique material cards. Potentially suitable mathematical functions are tested for the grouping and consolidation of the elements. The results show that the agreement of some presented methods depends on the damage level. The influence of the selected statistical functions on the result is shown here. The optimal solution is demonstrated, and the most efficient method of linearization is presented. The main motivation behind this work is that the problem has not been discussed in the literature and that there is currently no commercial software translator that provides the transference of models between solvers.
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- 2022
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14. Effect of host switching simulation on the fitness of the gregarious parasitoid Anaphes flavipes from a novel two-generation approach.
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Samková A, Raška J, Hadrava J, and Skuhrovec J
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- Animals, Herbivory, Plants parasitology, Genetic Fitness, Host-Parasite Interactions, Insecta
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Herbivorous insects can escape the strong pressure of parasitoids by switching to feeding on new host plants. Parasitoids can adapt to this change but at the cost of changing their preferences and performance. For gregarious parasitoids, fitness changes are not always observable in the F1 generation but only in the F2 generation. Here, with the model species and gregarious parasitoid Anaphes flavipes, we examined fitness changes in the F1 generation under pressure from the simulation of host switching, and by a new two-generation approach, we determined the impact of these changes on fitness in the F2 generation. We showed that the parasitoid preference for host plants depends on hatched or oviposited learning in relation to the possibility of parasitoid decisions between different host plants. Interestingly, we showed that after simulation of parasitoids following host switching, in the new environment of a fictitious host plant, parasitoids reduced the fictitious host. At the same time, parasitoids also reduced fertility because in fictitious hosts, they are not able to complete larval development. However, from a two-generation approach, the distribution of parasitoid offspring into both native and fictitious hosts caused lower parasitoid clutch size in native hosts and higher individual offspring fertility in the F2 generation., (© 2021. The Author(s).)
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- 2021
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15. The Effects of Bilirubin and Lumirubin on the Differentiation of Human Pluripotent Cell-Derived Neural Stem Cells.
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Capková N, Pospíšilová V, Fedorová V, Raška J, Pospíšilová K, Dal Ben M, Dvořák A, Viktorová J, Bohačiaková D, and Vítek L
- Abstract
The 'gold standard' treatment of severe neonatal jaundice is phototherapy with blue-green light, which produces more polar photo-oxidation products that are easily excreted via the bile or urine. The aim of this study was to compare the effects of bilirubin (BR) and its major photo-oxidation product lumirubin (LR) on the proliferation, differentiation, morphology, and specific gene and protein expressions of self-renewing human pluripotent stem cell-derived neural stem cells (NSC). Neither BR nor LR in biologically relevant concentrations (12.5 and 25 µmol/L) affected cell proliferation or the cell cycle phases of NSC. Although none of these pigments affected terminal differentiation to neurons and astrocytes, when compared to LR, BR exerted a dose-dependent cytotoxicity on self-renewing NSC. In contrast, LR had a substantial effect on the morphology of the NSC, inducing them to form highly polar rosette-like structures associated with the redistribution of specific cellular proteins (β-catenin/N-cadherin) responsible for membrane polarity. This observation was accompanied by lower expressions of NSC-specific proteins (such as SOX1, NR2F2, or PAX6) together with the upregulation of phospho-ERK. Collectively, the data indicated that both BR and LR affect early human neurodevelopment in vitro, which may have clinical relevance in phototherapy-treated hyperbilirubinemic neonates.
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- 2021
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16. Investigation of Significant Parameters during Abrasive Waterjet Turning.
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Štefek A, Raška J, Hlaváč LM, and Spadło S
- Abstract
This paper presents an investigation of abrasive waterjet turning (AWJT). The purpose of the article was to investigate significant parameters of the turning process and to evaluate their impact on the turning product. The influence of the traverse speed, the rotational speed, and the relative position of the jet to the specimen (lateral jet shift) were investigated. Based on the previous research done in this field, the multi-pass tangential turning method was selected. Rotational speed does not seem to have a significant impact on the AWJ turning process. However, the relative position of the jet is a key parameter for improving the efficiency of the process. Increasing the lateral jet shift causes the volume of the material removed to increase until the optimal impact angle is reached. These findings need to be extended in order to adjust AWJT. Without these improvements, a comparison of jet to traditional technologies is inappropriate.
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- 2021
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17. Endocrine-disrupting chemicals rapidly affect intercellular signaling in Leydig cells.
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Yawer A, Sychrová E, Labohá P, Raška J, Jambor T, Babica P, and Sovadinová I
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- Animals, Cell Communication drug effects, Cell Survival drug effects, Connexin 43 genetics, Connexin 43 metabolism, Dose-Response Relationship, Drug, Male, Mice, Endocrine Disruptors toxicity, Leydig Cells drug effects, Phenols toxicity, Signal Transduction drug effects
- Abstract
A decline in male fertility possibly caused by environmental contaminants, namely endocrine-disrupting chemicals (EDCs), is a topic of public concern and scientific interest. This study addresses a specific role of testicular gap junctional intercellular communication (GJIC) between adjacent prepubertal Leydig cells in endocrine disruption and male reproductive toxicity. Organochlorine pesticides (lindane, methoxychlor, DDT), industrial chemicals (PCB153, bisphenol A, nonylphenol and octylphenol) as well as personal care product components (triclosan, triclocarban) rapidly dysregulated GJIC in murine Leydig TM3 cells. The selected GJIC-inhibiting EDCs (methoxychlor, triclosan, triclocarban, lindane, DDT) caused the immediate GJIC disruption by the relocation of gap junctional protein connexin 43 (Cx43) from the plasma membrane and the alternation of Cx43 phosphorylation pattern (Ser368, Ser279, Ser282) of its full-length and two N-truncated isoforms. After more prolonged exposure (24 h), EDCs decreased steady-state levels of full-length Cx43 protein and its two N-truncated isoforms, and eventually (triclosan, triclocarban) also tight junction protein Tjp-1. The disturbance of GJIC was accompanied by altered activity of mitogen-activated protein kinases MAPK-Erk1/2 and MAPK-p38, and a decrease in stimulated progesterone production. Our results indicate that EDCs might disrupt testicular homeostasis and development via disruption of testicular GJIC, a dysregulation of junctional and non-junctional functions of Cx43, activation of MAPKs, and disruption of an early stage of steroidogenesis in prepubertal Leydig cells. These critical disturbances of Leydig cell development and functions during a prepubertal period might be contributing to impaired male reproduction health later on., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)
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- 2020
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18. Structure-Dependent Effects of Phthalates on Intercellular and Intracellular Communication in Liver Oval Cells.
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Čtveráčková L, Jančula D, Raška J, Babica P, and Sovadinová I
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- Animals, Cell Communication drug effects, Cell Line, Cell Survival drug effects, Dose-Response Relationship, Drug, Gap Junctions drug effects, Liver metabolism, MAP Kinase Signaling System drug effects, Phthalic Acids administration & dosage, Rats, Structure-Activity Relationship, Liver cytology, Liver drug effects, Phthalic Acids chemistry, Phthalic Acids toxicity
- Abstract
Humans are exposed to phthalates released from plastics, cosmetics, or food on a daily basis. Phthalates have low acute liver toxicity, but their chronic exposures could induce molecular and cellular effects linked to adverse health outcomes, such as liver tumor promotion or chronic liver diseases. The alternation of gap junctional intercellular communication (GJIC) and MAPK-Erk1/2 pathways in liver progenitor or oval cells can disrupt liver tissue homeostatic mechanisms and affect the development and severity of these adverse outcomes. Our study with 20 different phthalates revealed their structurally dependent effects on liver GJIC and MAPK-Erk1/2 signaling in rat liver WB-F344 cell line with characteristics of liver oval cells. The phthalates with a medium-length side chain (3-6 C) were the most potent dysregulators of GJIC and activators of MAPK-Erk1/2. The effects occurred rapidly, suggesting the activation of non-genomic (non-transcriptional) mechanisms directly by the parental compounds. Short-chain phthalates (1-2 C) did not dysregulate GJIC even after longer exposures and did not activate MAPK-Erk1/2. Longer chain (≥7 C) phthalates, such as DEHP or DINP, moderately activated MAPK-Erk1/2, but inhibited GJIC only after prolonged exposures (>12 h), suggesting that GJIC dysregulation occurs via genomic mechanisms, or (bio)transformation. Overall, medium-chain phthalates rapidly affected the key tissue homeostatic mechanisms in the liver oval cell population via non-genomic pathways, which might contribute to the development of chronic liver toxicity and diseases.
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- 2020
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19. Cylindrospermopsin induces cellular stress and activation of ERK1/2 and p38 MAPK pathways in adult human liver stem cells.
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Raška J, Čtveráčková L, Dydowiczová A, Sovadinová I, Bláha L, and Babica P
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- Alkaloids, Cell Line, Cell Proliferation, Cell Survival drug effects, Cyanobacteria Toxins, DNA Damage, Hepatocytes drug effects, Humans, Liver metabolism, MAP Kinase Signaling System, Marine Toxins, Microcystins, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinase 3 metabolism, Oxidative Stress drug effects, Reactive Oxygen Species metabolism, Stem Cells, Toxicity Tests, Uracil toxicity, Bacterial Toxins toxicity, Uracil analogs & derivatives, p38 Mitogen-Activated Protein Kinases metabolism
- Abstract
Cyanobacterial toxin cylindrospermopsin (CYN) is an emerging freshwater contaminant, whose expanding environmental occurrence might result into increased human health risks. CYN is potent hepatotoxin, with cytotoxicity and genotoxicity documented in primary hepatocytes or hepatoma cell lines. However, there is only limited information about CYN effects on adult human liver stem cells (LSCs), which play an important role in liver tissue development, regeneration and repair. In our study with human liver cell line HL1-hT1 which expresses characteristics of LSCs, CYN was found to be cytotoxic and increasing cell death after 24-48 h exposure to concentrations >1 μM. Subcytotoxic 1 μM concentration did not induce cell death or membrane damage, but inhibited cellular processes related to energy production, leading to a growth stagnation after >72 h. Interestingly, these effects were not associated with increased DNA damage, reactive oxygen species production, or endoplasmic reticulum stress. However, CYN induced a sustained (24-48 h) activation of mitogen-activated protein kinases ERK1/2 and p38, and increased expression of stress-related transcription factor ATF3. Thus, LSCs were not primarily affected by CYN-induced genotoxicity and oxidative stress, but via activation of signaling and transcriptional pathways critical for regulation of cell proliferation, stress responses, cell survival and inflammation. Alterations of LSCs during CYN-induced liver injury, including the role of nongenotoxic mechanisms, should be therefore considered in mechanistic assessments of chronic CYN hepatotoxicity and hepatocarcinogenicity., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Published
- 2019
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20. Ligase 3-mediated end-joining maintains genome stability of human embryonic stem cells.
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Kohutova A, Raška J, Kruta M, Seneklova M, Barta T, Fojtik P, Jurakova T, Walter CA, Hampl A, Dvorak P, and Rotrekl V
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- Cells, Cultured, DNA End-Joining Repair radiation effects, DNA Ligase ATP genetics, DNA Repair radiation effects, Homologous Recombination, Human Embryonic Stem Cells cytology, Humans, Poly-ADP-Ribose Binding Proteins genetics, DNA Breaks, Double-Stranded radiation effects, DNA End-Joining Repair physiology, DNA Ligase ATP metabolism, DNA Repair physiology, Genomic Instability, Human Embryonic Stem Cells physiology, Poly-ADP-Ribose Binding Proteins metabolism
- Abstract
Maintenance of human embryonic stem cells (hESCs) with stable genome is important for their future use in cell replacement therapy and disease modeling. Our understanding of the mechanisms maintaining genomic stability of hESC and our ability to modulate them is essential in preventing unwanted mutation accumulation during their in vitro cultivation. In this study, we show the DNA damage response mechanism in hESCs is composed of known, yet unlikely components. Clustered oxidative base damage is converted into DNA double-strand breaks (DSBs) by base excision repair (BER) and then quickly repaired by ligase (Lig)3-mediated end-joining (EJ). If there is further induction of clustered oxidative base damage by irradiation, then BER-mediated DSBs become essential in triggering the checkpoint response in hESCs. hESCs limit the mutagenic potential of Lig3-mediated EJ by DNA break end protection involving p53 binding protein 1 (53BP1), which results in fast and error-free microhomology-mediated repair and a low mutant frequency in hESCs. DSBs in hESCs are also repaired via homologous recombination (HR); however, DSB overload, together with massive end protection by 53BP1, triggers competition between error-free HR and mutagenic nonhomologous EJ.-Kohutova, A., Raška, J., Kruta, M., Seneklova, M., Barta, T., Fojtik, P., Jurakova, T., Walter, C. A., Hampl, A., Dvorak, P., Rotrekl, V. Ligase 3-mediated end-joining maintains genome stability of human embryonic stem cells.
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- 2019
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21. Polycyclic Aromatic Hydrocarbons and Endocrine Disruption: Role of Testicular Gap Junctional Intercellular Communication and Connexins.
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Kubincová P, Sychrová E, Raška J, Basu A, Yawer A, Dydowiczová A, Babica P, and Sovadinová I
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- Animals, Bay-Region, Polycyclic Aromatic Hydrocarbon, Cell Line, Cell Survival drug effects, Connexin 43 genetics, Endocrine Disruptors chemistry, Gap Junctions metabolism, Gap Junctions pathology, Leydig Cells metabolism, Leydig Cells pathology, Male, Mice, Mitogen-Activated Protein Kinases metabolism, Phosphorylation, Polycyclic Aromatic Hydrocarbons chemistry, Sertoli Cells metabolism, Sertoli Cells pathology, Signal Transduction, Cell Communication drug effects, Connexin 43 metabolism, Endocrine Disruptors toxicity, Gap Junctions drug effects, Leydig Cells drug effects, Polycyclic Aromatic Hydrocarbons toxicity, Sertoli Cells drug effects
- Abstract
Ambient air pollution and smoking are well-documented risk factors for male infertility. Prevalent air pollutants and cigarette smoke components, polycyclic aromatic hydrocarbons (PAHs), are environmental and occupational toxicants that act as chemicals disrupting endocrine regulation and reproductive potential in males. Testicular gap junctional intercellular communication (GJIC) is critical for normal development and function of testicular tissue, thus we assessed GJIC as a process potentially targeted by PAHs in testes. Lower MW PAHs with a bay or bay-like region rapidly dysregulated GJIC in Leydig TM3 cells by relocalization of major testicular gap junctional protein connexin 43 (Cx43) from plasma membrane to cytoplasm. This was associated with colocalization between Cx43 and ubiquitin in intracellular compartments, but without any effect on Cx43 degradation rate or steady-state Cx43 mRNA levels. A longer exposure to active PAHs decreased steady-state levels of full-length Cx43 protein and its 2 N-truncated isoforms. Inhibition of GJIC by PAHs, similarly to a prototypic GJIC-inhibitor TPA, was mediated via the MAP kinase-Erk1/2 and PKC pathways. Polycyclic aromatic hydrocarbon-induced GJIC dysregulation in testes was cell-type-specific because neither PAH dysregulated GJIC in Sertoli TM4 cells, despite PAHs were rapidly taken up by both Leydig TM3 as well as Sertoli TM4 cells. Because TPA effectively dysregulated GJIC in both testicular cell types, a unique regulator of GJIC targeted by PAHs might exist in Leydig TM3 cells. Our results indicate that PAHs could be a potential etiological agent contributing to reproductive dysfunctions in males through an impairment of testicular GJIC and junctional and/or nonjunctional functions of Cx43., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2019
- Full Text
- View/download PDF
22. Mutation frequency dynamics in HPRT locus in culture-adapted human embryonic stem cells and induced pluripotent stem cells correspond to their differentiated counterparts.
- Author
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Krutá M, Šeneklová M, Raška J, Salykin A, Zerzánková L, Pešl M, Bártová E, Franek M, Baumeisterová A, Košková S, Neelsen KJ, Hampl A, Dvořák P, and Rotrekl V
- Subjects
- Cell Differentiation radiation effects, Cell Line, Gamma Rays, Humans, Hypoxanthine Phosphoribosyltransferase metabolism, Induced Pluripotent Stem Cells cytology, Genetic Loci, Hypoxanthine Phosphoribosyltransferase genetics, Induced Pluripotent Stem Cells metabolism, Mutation Rate
- Abstract
The genomic destabilization associated with the adaptation of human embryonic stem cells (hESCs) to culture conditions or the reprogramming of induced pluripotent stem cells (iPSCs) increases the risk of tumorigenesis upon the clinical use of these cells and decreases their value as a model for cell biology studies. Base excision repair (BER), a major genomic integrity maintenance mechanism, has been shown to fail during hESC adaptation. Here, we show that the increase in the mutation frequency (MF) caused by the inhibition of BER was similar to that caused by the hESC adaptation process. The increase in MF reflected the failure of DNA maintenance mechanisms and the subsequent increase in MF rather than being due solely to the accumulation of mutants over a prolonged period, as was previously suggested. The increase in the ionizing-radiation-induced MF in adapted hESCs exceeded the induced MF in nonadapted hESCs and differentiated cells. Unlike hESCs, the overall DNA maintenance in iPSCs, which was reflected by the MF, was similar to that in differentiated cells regardless of the time spent in culture and despite the upregulation of several genes responsible for genome maintenance during the reprogramming process. Taken together, our results suggest that the changes in BER activity during the long-term cultivation of hESCs increase the mutagenic burden, whereas neither reprogramming nor long-term propagation in culture changes the MF in iPSCs.
- Published
- 2014
- Full Text
- View/download PDF
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