Mouse neuronal CAD 5 cell line effectively propagates various strains of prions. Previously, we have shown that it can also be differentiated into the cells morphologically resembling neurons. Here, we demonstrate that CAD 5 cells chronically infected with prions undergo differentiation under the same conditions. To make our model more realistic, we triggered the differentiation in the 3D culture created by gentle rocking of CAD 5 cell suspension. Spheroids formed within 1 week and were fully developed in less than 3 weeks of culture. The mature spheroids had a median size of ~300 μm and could be cultured for up to 12 weeks. Increased expression of differentiation markers GAP 43, tyrosine hydroxylase, β-III-tubulin and SNAP 25 supported the differentiated status of the spheroid cells. The majority of them were found in the G0/G1 phase of the cell cycle, which is typical for differentiated cells. Moreover, half of the PrPC on the cell membrane was N-terminally truncated, similarly as in differentiated CAD 5 adherent cells. Finally, we demonstrated that spheroids could be created from prion-infected CAD 5 cells. The presence of prions was verified by immunohistochemistry, western blot and seed amplification assay. We also confirmed that the spheroids can be infected with the prions de novo. Our 3D culture model of differentiated CAD 5 cells is low cost, easy to produce and cultivable for weeks. We foresee its possible use in the testing of anti-prion compounds and future studies of prion formation dynamics.

• MeSH
• buněčná diferenciace * fyziologie   MeSH
• buněčné kultury metody   MeSH
• buněčné linie MeSH
• buněčné sféroidy * metabolismus   MeSH
• myši MeSH
• neurony metabolismus   MeSH
• prionové nemoci * metabolismus   patologie   MeSH
• priony metabolismus   MeSH
• techniky 3D buněčné kultury metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Three-dimensional (3D) cell cultures are to date the gold standard in biomedical research fields due to their enhanced biological functions compared to conventional two-dimensional (2D) cultures. 3D cell spheroids, as well as organoids, are better suited to replicate tissue functions, which enables their use both as in vitro models for basic research and toxicology, as well as building blocks used in tissue/organ biofabrication approaches. Culturing 3D spheroids from bone-derived cells is an emerging technology for both disease modelling and drug screening applications. Bone tissue models are mainly limited by the implementation of sophisticated devices and procedures that can foster a tissue-specific 3D cell microenvironment along with a dynamic cultivation regime. In this study, we consequently developed, optimized and characterized an advanced perfused microfluidic platform to improve the reliability of 3D bone cell cultivation and to enhance aspects of bone tissue maturation in vitro. Moreover, biomechanical stimulation generated by fluid flow inside the arrayed chamber, was used to mimic a more dynamic cell environment emulating a highly vascularized bone we expected to improve the osteogenic 3D microenvironment in the developed multifunctional spheroid-array platform. The optimized 3D cell culture protocols in our murine bone-on-a-chip spheroid model exhibited increased mineralization and viability compared to static conditions. As a proof-of-concept, we successfully confirmed on the beneficial effects of a dynamic culture environment on osteogenesis and used our platform for analysis of bone-derived spheroids produced from primary human pre-osteoblasts. To conclude, the newly developed system represents a powerful tool for studying human bone patho/physiology in vitro under more relevant and dynamic culture conditions converging the advantages of microfluidic platforms with multi-spheroid array technologies.

• Publikační typ
• časopisecké články MeSH

Phytopathology is a highly complex scientific discipline. Initially, its focus was on the study of plant-pathogen interactions in agricultural and forestry production systems. Host-pathogen interactions in natural plant communities were generally overlooked until the 1970s when plant pathologists and evolutionary biologists started to take an interest in these interactions, and their dynamics in natural plant populations, communities, and ecosystems. This article introduces the general principles of plant pathosystems, provides a basic critical overview of current knowledge of host-pathogen interactions in natural plant pathosystems, and shows how this knowledge is important for future developments in plant pathology especially as it applies in cropping systems, ecology, and evolutionary biology. Plant pathosystems can be further divided according to the structure and origin of control, as autonomous (wild plant pathosystems, WPPs) or deterministic (crop plant pathosystems, CPPs). WPPs are characterized by the disease triangle and closed-loop (feedback) controls, and CPPs are characterized by the disease tetrahedron and open-loop (non-feedback) controls. Basic general, ecological, genetic, and population structural and functional differences between WPPs and CPPs are described. It is evident that we lack a focus on long-term observations and research of diseases and their dynamics in natural plant populations, metapopulations, communities, ecosystems, and biomes, as well as their direct or indirect relationships to CPPs. Differences and connections between WPPs and CPPs, and why, and how, these are important for agriculture varies. WPP and CPP may be linked by strong biological interactions, especially where the pathogen is in common. This is demonstrated through a case study of lettuce (Lactuca spp., L. serriola and L. sativa) and lettuce downy mildew (Bremia lactucae). In other cases where there is no such direct biological linkage, the study of WPPs can provide a deeper understanding of how ecology and genetics interacts to drive disease through time. These studies provide insights into ways in which farming practices may be changed to limit disease development. Research on interactions between pathosystems, the "cross-talk" of WPPs and CPPs, is still very limited and, as shown in interactions between wild and cultivated Lactuca spp.-B. lactucae associations, can be highly complex. The implications and applications of this knowledge in plant breeding, crop management, and disease control measures are considered. This review concludes with a discussion of theoretical, general and specific aspects, challenges and limits of future WPP research, and application of their results in agriculture.

• MeSH
• ekosystém * MeSH
• nemoci rostlin genetika   MeSH
• oomycety * genetika   MeSH
• rostliny MeSH
• salát (hlávkový) MeSH
• šlechtění rostlin MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The cyanobacterial genus Synechocystis is of particular interest to science and industry because of its efficient phototrophic metabolism, its accumulation of the polymer poly(3-hydroxybutyrate) (PHB) and its ability to withstand or adapt to adverse growing conditions. One such condition is the increased salinity that can be caused by recycled or brackish water used in cultivation. While overall reduced growth is expected in response to salt stress, other metabolic responses relevant to the efficiency of phototrophic production of biomass or PHB (or both) have been experimentally observed in three Synechocystis strains at stepwise increasing salt concentrations. In response to recent reports on metabolic strategies to increase stress tolerance of heterotrophic and phototrophic bacteria, we focused particularly on the stress-induced response of Synechocystis strains in terms of PHB, glycogen and photoactive pigment dynamics. Of the three strains studied, the strain Synechocystis cf. salina CCALA192 proved to be the most tolerant to salt stress. In addition, this strain showed the highest PHB accumulation. All the three strains accumulated more PHB with increasing salinity, to the point where their photosystems were strongly inhibited and they could no longer produce enough energy to synthesize more PHB.

• Publikační typ
• časopisecké články MeSH

Purpose: Nanomaterials for antimicrobial applications have gained interest in recent years due to the increasing bacteria resistance to conventional antibiotics. Wound sterilization, water treatment and surface decontamination all avail from multifunctional materials that also possess excellent antibacterial properties, eg zinc oxide (ZnO). Here, we assess and compare the effects of synthesized hedgehog-like ZnO structures and commercial ZnO particles with and without mixing on the inactivation of bacteria on surfaces and in liquid environments. Methods: Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria in microbial culture medium were added to reverse spin bioreactors that contained different concentrations of each ZnO type to enable dynamic mixing of the bacteria-ZnO suspensions. Optical density of the bacteria-ZnO suspensions was measured in real-time and the number of viable bacteria after 24 h exposure was determined using standard microbiological techniques. The concentration of zinc ion generated from ZnO dissolution in different liquid types was estimated from the dynamic interaction exposure. Static antibacterial tests without agitation in liquid media and on agar surface were performed for comparison. Results: A correlation between increasing ZnO particle concentration and reduction in viable bacteria was not monotonous. The lowest concentration tested (10 µg/mL) even stimulated bacteria growth. The hedgehog ZnO was significantly more antibacterial than commercial ZnO particles at higher concentrations (up to 1000 µg/mL tested), more against E. coli than S. aureus. Minimum inhibitory concentration in microwell plates was correlated with those results. No inhibition was detected for any ZnO type deposited on agar surface. Zinc ion release was greatly suppressed in cultivation media. Scanning electron microscopy images revealed that ZnO needles can pierce membrane of bacteria whereas the commercial ZnO nanoparticles rather agglomerate on the cell surface. Conclusion: The inhibition effects are thus mainly controlled by the interaction dynamics between bacteria and ZnO, where mixing greatly enhances antibacterial efficacy of all ZnO particles. The efficacy is modulated also by ZnO particle shapes, where hedgehog ZnO has superior effect, in particular at lower concentrations. However, at too low concentrations, ZnO can stimulate bacteria growth and must be thus used with caution.

• MeSH
• antibakteriální látky chemie   farmakologie   MeSH
• biomimetické materiály chemie   farmakologie   MeSH
• Escherichia coli účinky léků   růst a vývoj   MeSH
• ježkovití * MeSH
• mikrobiální testy citlivosti MeSH
• oxid zinečnatý chemie   farmakologie   MeSH
• Staphylococcus aureus účinky léků   růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Wild barley is abundant, occupying large diversity of sites, ranging from the northern mesic Mediterranean meadows to the southern xeric deserts in Israel. This is also reflected in its wide phenotypic heterogeneity. We investigated the dynamics of DNA content changes in seed tissues in ten wild barley accessions that originated from an environmental gradient in Israel. The flow cytometric measurements were done from the time shortly after pollination up to the dry seeds. We show variation in mitotic cell cycle and endoreduplication dynamics in both diploid seed tissues (represented by seed maternal tissues and embryo) and in the triploid endosperm. We found that wild barley accessions collected at harsher xeric environmental conditions produce higher proportion of endoreduplicated nuclei in endosperm tissues. Also, a comparison of wild and cultivated barley strains revealed a higher endopolyploidy level in the endosperm of wild barley, that is accompanied by temporal changes in the timing of the major developmental phases. In summary, we present a new direction of research focusing on connecting spatiotemporal patterns of endoreduplication in barley seeds and possibly buffering for stress conditions.

• MeSH
• DNA rostlinná genetika   MeSH
• endosperm genetika   MeSH
• genetická variace genetika   MeSH
• ječmen (rod) genetika   MeSH
• polyploidie MeSH
• populační genetika metody   MeSH
• semena rostlinná genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Izrael MeSH

Tento článek přináší zamyšlení nad možnostmi, které nabízí vnímání těla a pohyb v krajině. V článku popisujeme a reflektujeme průběh skupinových sesí tanečně pohybové terapie s klienty s psychosomatickým onemocněním. Tanečně pohybová terapie rozvíjí výrazovou, expresivní část neverbální komunikace a kultivuje uvědomění těla. Lidé s psychosomatickým onemocněním pociťují diskomfort, napětí nebo bolest v těle, zároveň však často nedokáží tělo plně vnímat a uvědomovat si, jaké potřeby jim signalizuje. Často obtížně rozeznávají, pojmenovávají a vyjadřují to, co prožívají. Znovuzískání kontaktu s tělesným Já napomáhá integrovat fyzickou, emoční a kognitivní rovinu jimi prožívané skutečnosti. Z tohoto pohledu se příroda jeví jako skvělý spojenec práce s tělem. Smyslové vnímání pomáhá klientům znovu začít vnímat tělo, a díky tělesnému naladění na krajinu a živoucí formy nacházejí bezpečnou a vitální podporu v přírodním prostředí. Skupinová setkání a jejich dynamika napomáhají rozvoji pohybového repertoáru, pohybových kvalit, a přispívají tak k vytvoření zdravějších kopingových strategií v životě klientů.

The aim of this paper is to introduce possibilities of dance movement therapy practice in nature and to describe interventions and work processes with clients suffering from psychosomatic disease. The efficacy of verbal therapy can be limited when people with psychosomatic ilnesses have difficulty recognizing, naming and expressing their feelings and emotions. Often people simply feel discomfort, tension or pain in their bodies. Many lack full awareness of their body, including the body’s innate needs and signals. Dance movement therapy enables us to cultivate the expressive part of nonverbal communication and embodied relational language in order to regain body awareness and sense of self. This, in turn, is necessary to integrate experiences on the physical, emotional and cognitive levels. From this perspective, nature is a great ally. In this article we will describe how focusing on perception and the senses while in nature can help patients find a sense of self in relation to their physical bodies. Through attunement to the landscape and its living forms, clients find safe and vital support in a group setting within a natural environment. They can experience and widen their range of movement, the quality of movement and range of emotions, which can then contribute to the creation of healthier coping strategies in their lives.

• MeSH
• environmentální psychologie * metody   MeSH
• lidé MeSH
• psychoterapie metody   MeSH
• terapie tancem MeSH
• Check Tag
• lidé MeSH

The multiyear cultivation of Miscanthus × giganteus Greef et Deu (M.×giganteus) at the soils polluted by metal(loid)s were researched. The biomass parameters and concentrations of elements: Ti, Mn, Fe, Cu, Zn, As, Sr, and Mo were determined in the plant's organs at harvest. The same metal(loid)s were monitored in the plant's leaves throughout three vegetation seasons. The principal component analysis and general linear model approaches were applied for statistical evaluation followed by Box-Cox transformation. The difference in the distribution of elements in the plant, the content of elements in the soil, various regime of uptake to the plant tissues, and the year of vegetation were analyzed as driving factors of the phytoremediation. The results showed that the leading promoter was the factor of the zone, which was the most essential for Ti, Fe, and Cu and the smallest for Mn. The factor of differences in soil pollution was essential for Zn and Mo, much less for As, Sr, and Mn, limited for Fe, and was not seen for Ti and Cu. The factor of the interrelation effects of the zone and experiment reflected the different regime of uptake for the plant tissues was seen for two elements: more prominent for Cu and smaller for Ti. While analyzing the dynamic of foliar concentrations of the metal(loid)s during 3 years, two groups were defined. Firstly, Fe, Ni, Mn, and Sr showed stable curves with limited distribution of the plant life cycle. Secondly, As, Zn, Cu, and Mo showed different fluctuations in the curves, which can be attributed to essential influence of those elements to the plant life cycle. Further research will be focused on the application of M.×giganteus to the polluted soil in a bigger scale and comparison results of laboratory and field experiments.

• MeSH
• biodegradace MeSH
• látky znečišťující půdu analýza   MeSH
• půda MeSH
• těžké kovy analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Chorvatsko MeSH

We have developed a novel simple method for effective preparing gold nanoparticles (AuNPs) intended for utilization in biomedicine. The method is based on gold sputtering into liquid poly(ethylene glycol) (PEG). The PEG was used as a basic biocompatible stabilizer of the AuNP colloid. In addition, two naturally occurring polysaccharides - Chitosan (Ch) and Methylcellulose (MC) - were separately diluted into the PEG base with the aims to enhance the yield of the sputtering without changing the sputtering parameters, and to further improve the stability and the biocompatibility of the colloid. The colloids were sterilized by steam, and their stability was measured before and after the sterilization process by dynamic light scattering and UV-Vis spectrophotometry. The results indicated a higher sputtering yield in the colloids containing the polysaccharides. The colloids were also characterized by atomic absorption spectroscopy (AAS) to reveal the composition of the prepared nanoparticles by transmission electron microscopy (TEM) to visualize the nanoparticles and to evaluate their size and clustering, and by rheometry to estimate the viscosity of the colloids. The zeta-potential of the AuNPs was also determined as an important parameter indicating the stability and the biocompatibility of the colloid. In addition, in vitro tests of antimicrobial activity and cytotoxicity were carried out to estimate the biological activity and the biocompatibility of the colloids. Antimicrobial tests were performed by a drip test on two bacterial strains - Gram-positive Staphylococcus epidermidis and Gram-negative Escherichia coli. AuNP with chitosan proved to possess the highest antibacterial activity, especially towards the Gram-positive S. epidermidis. In vitro tests on eukaryotic cells, i.e. human osteoblastic cell line SAOS-2 and primary normal human dermal fibroblasts (NHDF), were performed after a 7-day cultivation to determine the effect and the toxic dose of the colloids on human cells. The studied colloid concentrations were in the range from 0.6 μg/ml to 6 μg/ml. Toxicity of the colloids started to reappear at a concentration of 4.5 μg/ml, especially with chitosan in the colloid, where the colloid with a concentration of 6 μg/ml proved to be the most toxic, especially towards the SAOS-2 cell line. However, the PEG and PEG-MC containing colloids proved to be relatively non-toxic, even at the highest concentration, but with a slowly decreasing tendency of the cell metabolic activity.

• MeSH
• antibakteriální látky chemie   farmakologie   MeSH
• buněčné linie MeSH
• chitosan chemie   MeSH
• dynamický rozptyl světla MeSH
• Escherichia coli účinky léků   MeSH
• koloidy chemie   MeSH
• kovové nanočástice chemie   MeSH
• lidé MeSH
• methylcelulosa chemie   MeSH
• polysacharidy chemie   MeSH
• stabilita léku MeSH
• Staphylococcus epidermidis účinky léků   MeSH
• sterilizace MeSH
• velikost částic MeSH
• zlato chemie   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: The aim was to design and thoroughly characterize monodisperse Fe3O4@SiO2-Ag nanoparticles with strong antibacterial properties, which makes them a candidate for targeting bacterial infections. METHODS: The monodisperse Fe3O4 nanoparticles were prepared by oleic acid-stabilized thermal decomposition of Fe(III) oleate; the particles were coated with silica shell using a water-in-oil reverse microemulsion, involving hydrolysis and condensation of tetramethyl orthosilicate. Resulting Fe3O4@SiO2 particles were modified by (3-mercaptopropyl)trimethoxysilane to introduce 1.1 mmol SH/g. Finally, the Fe3O4@SiO2-SH nanoparticles were decorated with silver nanoclusters formed by reduction of silver nitrate with NaBH4. The particles were analyzed by FTIR, X-ray photoelectron and atomic absorption spectroscopy, dynamic light scattering and vibrating sample magnetometry. The antibacterial activity of the Fe3O4@SiO2 and Fe3O4@SiO2-Ag nanoparticles was tested against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria cultivated on Luria agar plates or in Luria broth. RESULTS: The superparamagnetic Fe3O4@SiO2-Ag nanoparticles (21 nm in diameter; saturation magnetization 26 A∙m2/kg) were successfully obtained and characterized. Inhibitory and toxic effects against bacteria were documented by incubation of the Fe3O4@SiO2-Ag nanoparticles with Staphylococcus aureus and Escherichia coli. CONCLUSIONS: The combination of magnetic properties together with bactericidal effects is suitable for the disinfection of medical instruments, water purification, food packaging, etc.

• MeSH
• antibakteriální látky chemie   farmakologie   MeSH
• Escherichia coli účinky léků   MeSH
• kyselina olejová chemie   MeSH
• magnetické nanočástice chemie   MeSH
• oxid křemičitý chemie   MeSH
• povrchové vlastnosti MeSH
• silany chemie   MeSH
• Staphylococcus aureus účinky léků   MeSH
• stříbro chemie   farmakologie   MeSH
• velikost částic MeSH
• Publikační typ
• časopisecké články MeSH