Michaela Liegertová

Jan Evangelista Purkyně University in Ústí nad Labem

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Jan Malý

Jan Evangelista Purkyně University in Ústí nad Labem

MŠ

Marcel Štofik

Jan Evangelista Purkyně University in Ústí nad Labem

JŠ

Jiří Šmejkal

Jan Evangelista Purkyně University in Ústí nad Labem

Regina Herma

Jan Evangelista Purkyně University in Ústí nad Labem

Petr Aubrecht

Jan Evangelista Purkyně University in Ústí nad Labem

Alena Semerádtová

Jan Evangelista Purkyně University in Ústí nad Labem

Olga Janoušková

Jan Evangelista Purkyně University in Ústí nad Labem

Jiří Pergner

Czech Academy of Sciences, Institute of Molecular Genetics

Monika Müllerová

Czech Academy of Sciences, Institute of Experimental Medicine

Zbyněk Kozmík

Czech Academy of Sciences, Institute of Molecular Genetics

Cooperative Institutions

Jan Evangelista Purkyně University in Ústí nad Labem

Czech Academy of Sciences

University of Chemistry and Technology, Prague

Charles University

Czech Academy of Sciences, Institute of Molecular Genetics

Inje University

Czech Academy of Sciences, Institute of Chemical Process Fundamentals

Czech Academy of Sciences, Institute of Inorganic Chemistry

Nuclear Research Institute Rez (Czechia)

Research Centre Rez

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Cubozoan genome illuminates functional diversification of opsins and photoreceptor evolution

Scientific Reports (2015)

Michaela Liegertová Jiří Pergner Iryna Kozmiková Péter Fábián António R. Pombinho

Abstract Animals sense light primarily by an opsin-based photopigment present in a photoreceptor cell. Cnidaria are arguably the most basal phylum containing a well-developed visual system. The evolutionary history of opsins in the animal kingdom has not yet been resolved. Here, we study the evolution of animal opsins by genome-wide analysis of the cubozoan jellyfish Tripedalia cystophora , a cnidarian possessing complex lens-containing eyes and minor photoreceptors. A large number of opsin genes with distinct tissue- and stage-specific expression were identified. Our phylogenetic analysis unequivocally classifies cubozoan opsins as a sister group to c-opsins and documents lineage-specific expansion of the opsin gene repertoire in the cubozoan genome. Functional analyses provided evidence for the use of the Gs-cAMP signaling pathway in a small set of cubozoan opsins, indicating the possibility that the majority of other cubozoan opsins signal via distinct pathways. Additionally, these tests uncovered subtle differences among individual opsins, suggesting possible fine-tuning for specific photoreceptor tasks. Based on phylogenetic, expression and biochemical analysis we propose that rapid lineage- and species-specific duplications of the intron-less opsin genes and their subsequent functional diversification promoted evolution of a large repertoire of both visual and extraocular photoreceptors in cubozoans.

Opsin

Lineage (genetic)

10.1038/srep11885

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Citations (57)

Carbosilane dendrimers with phosphonium terminal groups are low toxic non-viral transfection vectors for siRNA cell delivery

International Journal of Pharmaceutics (2019)

Regina Herma Dominika Wróbel Michaela Liegertová Monika Müllerová Tomáš Strašák

Cationic polymerization

10.1016/j.ijpharm.2019.03.018

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Citations (24)

Conventional and Nonconventional Sources of Exosomes–Isolation Methods and Influence on Their Downstream Biomedical Application

Frontiers in Molecular Biosciences (2022)

Olga Janoušková Regina Herma Alena Semerádtová David Poustka Michaela Liegertová

Despite extensive study of extracellular vesicles (EVs), specifically exosomes (EXs) as biomarkers, important modulators of physiological or pathological processes, or therapeutic agents, relatively little is known about nonconventional sources of EXs, such as invertebrate or plant EXs, and their uses. Likewise, there is no clear information on the overview of storage conditions and currently used isolation methods, including new ones, such as microfluidics, which fundamentally affect the characterization of EXs and their other biomedical applications. The purpose of this review is to briefly summarize conventional and nonconventional sources of EXs, storage conditions and typical isolation methods, widely used kits and new "smart" technologies with emphasis on the influence of isolation techniques on EX content, protein detection, RNA, mRNA and others. At the same time, attention is paid to a brief overview of the direction of biomedical application of EXs, especially in diagnostics, therapy, senescence and aging and, with regard to the current situation, in issues related to Covid-19.

Isolation

Extracellular Vesicles

10.3389/fmolb.2022.846650

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Citations (24)

Class II biocompatible E-Shell 300 3D printing material causes severe developmental toxicity in Danio rerio embryos and reduced cell proliferation in vitro – implications for 3D printed microfluidics

RSC Advances (2021)

Zuzana Nejedlá David Poustka Regina Herma Michaela Liegertová Marcel Štofik

Additive manufacturing is a new technology that represents a highly promising, cheap, and efficient solution for the production of various tools in the biomedicine field. In our study, the toxicity of the commercially available E-Shell 300 series photopolymer, which is used in the manufacture of hearing aids and other implants and which could be potentially exploited in microfluidic device fabrication, was tested using in vivo and in vitro biological models. We examined B14 cell proliferation in direct contact with the three-dimensional (3D)-printed material as well as in water extracts to evaluate in vitro cytotoxicity. Similarly, in vivo tests were performed using an OECD-standardized fish embryo acute toxicity (FET) test on Danio rerio embryos in direct contact with the material and in extracts as well. Despite E-Shell 300 3D-printed material being declared as class-IIa biocompatible, in the case of direct contact with both biological models, the results demonstrated a considerable negative impact on cell proliferation and severe developmental toxicity. In this study, up to 84% reduced cell proliferation in vitro and 79% mortality of in vivo models were observed. In contrast, a negligible toxic influence of E-Shell 300 water extracts was present. Four different post-processing treatments to reduce the toxicity were also tested. We observed that post-printing treatment of 3D-printed material in 96% ethanol can reduce embryonic mortality in the FET test by 71% and also completely eliminate negative effects on cell proliferation. We analyzed leachates from the polymeric structures by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy, and we discovered the presence of surfactant residues. In summary, our results indicate the importance of biocompatibility testing of the 3D printing photopolymer material in direct contact with the given biological model. On the other hand, the possibility of eliminating toxic effects by an appropriate post-processing strategy opens the door for broader applications of E-Shell 300 photopolymers in the development of complex microfluidic devices for various biological applications.

Danio

Biocompatible material

3d printed

10.1039/d1ra00305d

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Citations (13)

Molecular analysis of the amphioxus frontal eye unravels the evolutionary origin of the retina and pigment cells of the vertebrate eye

Proceedings of the National Academy of Sciences (2012)

Pavel Vopálenský Jiří Pergner Michaela Liegertová Èlia Benito‐Gutiérrez Detlev Arendt

The origin of vertebrate eyes is still enigmatic. The “frontal eye” of amphioxus, our most primitive chordate relative, has long been recognized as a candidate precursor to the vertebrate eyes. However, the amphioxus frontal eye is composed of simple ciliated cells, unlike vertebrate rods and cones, which display more elaborate, surface-extended cilia. So far, the only evidence that the frontal eye indeed might be sensitive to light has been the presence of a ciliated putative sensory cell in the close vicinity of dark pigment cells. We set out to characterize the cell types of the amphioxus frontal eye molecularly, to test their possible relatedness to the cell types of vertebrate eyes. We show that the cells of the frontal eye specifically coexpress a combination of transcription factors and opsins typical of the vertebrate eye photoreceptors and an inhibitory Gi-type alpha subunit of the G protein, indicating an off-responding phototransductory cascade. Furthermore, the pigmented cells match the retinal pigmented epithelium in melanin content and regulatory signature. Finally, we reveal axonal projections of the frontal eye that resemble the basic photosensory-motor circuit of the vertebrate forebrain. These results support homology of the amphioxus frontal eye and the vertebrate eyes and yield insights into their evolutionary origin.

Chordate

Opsin

Eye Proteins

Forebrain

10.1073/pnas.1207580109

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Citations (124)

Correction: Corrigendum: Cubozoan genome illuminates functional diversification of opsins and photoreceptor evolution

Scientific Reports (2015)

Michaela Liegertová Jiří Pergner Iryna Kozmiková Péter Fábián António R. Pombinho

Animals sense light primarily by an opsin-based photopigment present in a photoreceptor cell. Cnidaria are arguably the most basal phylum containing a well-developed visual system. The evolutionary history of opsins in the animal kingdom has not yet been resolved. Here, we study the evolution of animal opsins by genome-wide analysis of the cubozoan jellyfish Tripedalia cystophora , a cnidarian possessing complex lens-containing eyes and minor photoreceptors. A large number of opsin genes with distinct tissue- and stage-specific expression were identified. Our phylogenetic analysis unequivocally classifies cubozoan opsins as a sister group to c-opsins and documents lineage-specific expansion of the opsin gene repertoire in the cubozoan genome. Functional analyses provided evidence for the use of the Gs-cAMP signaling pathway in a small set of cubozoan opsins, indicating the possibility that the majority of other cubozoan opsins signal via distinct pathways. Additionally, these tests uncovered subtle differences among individual opsins, suggesting possible fine-tuning for specific photoreceptor tasks. Based on phylogenetic, expression and biochemical analysis we propose that rapid lineage- and species-specific duplications of the intron-less opsin genes and their subsequent functional diversification promoted evolution of a large repertoire of both visual and extraocular photoreceptors in cubozoans.

Opsin

Lineage (genetic)

10.1038/srep14396

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Citations (2)

A millifluidic chip for cultivation of fish embryos and toxicity testing fabricated by 3D printing technology

RSC Advances (2021)

Petr Panuška Zuzana Nejedlá Jiří Šmejkal Petr Aubrecht Michaela Liegertová

Zebrafish (Danio rerio) serves as a popular animal model for in vivo acute toxicity evaluation with the Fish embryo test (FET). Over the last few years there has been an effort to develop various systems for a high-throughput zebrafish embryo cultivation and FET. In this paper, we present a novel design of a millifluidic system fabricated by 3D printing technology and we evaluate its functional properties on Danio rerio embryos cultivation and toxicity testing. The development and the optimization of the millifluidic chip was performed by experimental measurements supported by numerical simulations of mass and momentum transport. The cultivation chip with two inlets and one outlet consisted of two individual channels placed on top of each other and separated by a partition with cultivation chambers. An individual embryo removal functionality, which can be used during the cultivation experiments for selective unloading of any of the cultivated embryos out of the chip, was added to the chip design. This unique property raises the possibility of detailed studies of the selected embryos by additional methods. Long-term (96 hours) perfusion cultivation experiments showed a normal development of zebrafish embryos in the chip. Model toxicity tests were further performed with diluted ethanol as a teratogen. Compared to the FET assays, an increased toxic effect of the ethanol on the embryos cultivated in the chip was observed when the median lethal dose and the percentage of the morphological end-points were evaluated. We conclude that the presented 3D printed chip is suitable for long-term zebrafish embryo cultivations and toxicity testing and can be further developed for the automated assays.

10.1039/d1ra00846c

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Citations (7)

Cerium–Bismuth Oxides/Oxynitrates with Low Toxicity for the Removal and Degradation of Organophosphates and Bisphenols

ACS Applied Nano Materials (2022)

Jiří Henych Martin Šťastný Zuzana Němečková Martin Kormunda Zuzana Šanderová

Nanoscale cerium-bismuth oxides/oxynitrates were prepared by a scalable low-temperature method at ambient pressure using water as the sole solvent. Solid solutions were formed up to a 1:1 Ce/Bi molar ratio, while at higher doping levels, bismuth oxynitrate photocatalysts with a pronounced layered structure were formed. Bismuth caused significant changes in the structure and surface properties of nanoceria, such as the formation of defects, oxygen-containing surface groups, and Lewis and Brønsted acid sites. The prepared bifunctional adsorbents/photocatalysts were efficient in the removal of toxic organophosphate (methyl paraoxon) from water by reactive adsorption followed by photocatalytic decomposition of the parent compound and its degradation product (p-nitrophenol). Bi-doped ceria also effectively adsorbed and photodegraded the endocrine disruptors bisphenols A and S and outperformed pure ceria and the P25 photocatalyst in terms of efficiency, durability, and long-term stability. The very low toxicity of Bi-nanoceria to mammalian cells, aquatic organisms, and bacteria has been demonstrated by comprehensive in vivo/in vitro testing, which, in addition to its simple "green" synthesis, high activity, and durability, makes Bi-doped ceria promising for safe use in abatement of toxic chemicals.

Bismuth

4-Nitrophenol

Cerium oxide

10.1021/acsanm.2c03926

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Citations (10)

Mucus-derived exosome-like vesicles from the Spanish slug (Arion vulgaris): taking advantage of invasive pest species in biotechnology

Scientific Reports (2022)

Michaela Liegertová Alena Semerádtová Michaela Kocholatá Michaela Průšová Lenka Němcová

Abstract The slug Arion vulgaris has attracted major attention as one of the worst invasive herbivore pests in Europe and is renowned for the stiff mucus it secretes for locomotion. In this study we focused on the isolation and characterisation of extracellular vesicles, specifically exosomes and exosome-like vesicles, from Arion secretions. We developed a method for slug mucus collection and subsequent vesicle isolation by ultracentrifugation. The isolated vesicles with an average diameter of ~ 100 nm carry abundant proteins and short RNAs, as well as adhesion molecules similar to mammalian galectins. We demonstrated that the slug extracellular vesicles are internalised by plant cells and human cancer cells in in vitro assays and are loadable by bioactive compounds, which makes them an interesting tool for utilisation in biotechnology.

Slug

Exosome

10.1038/s41598-022-26335-3

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Citations (5)

Identification and characterization of main genetic components involved in phototransduction and vision of the cubozoan jellyfish Tripedalia cystophora

Michaela Liegertová

Visual phototransduction

Identification

Source

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Citations (0)