The isolation and study of fungi within specific contexts yield valuable insights into the intricate relationships between fungi and ecosystems. Unlike culture-independent approaches, cultivation methods are advantageous in this context because they provide standardized replicates, specific species isolation, and easy sampling. This study aimed to understand the ecological process using a microcosm system with pesticide concentrations similar to those found in the soil, in contrast to high doses, from the isolation of the enriched community. The atrazine concentrations used were 0.02 mg/kg (control treatment), 300 ng/kg (treatment 1), and 3000 ng/kg (treatment 2), using a 28-day microcosm system. Ultimately, the isolation resulted in 561 fungi classified into 76 morphospecies. The Ascomycota phylum was prevalent, with Purpureocillium, Aspergillus, and Trichoderma being consistently isolated, denoting robust and persistent genera. Diversity analyses showed that the control microcosms displayed more distinct fungal morphospecies, suggesting the influence of atrazine on fungal communities. Treatment 2 (higher atrazine concentration) showed a structure comparable to that of the control, whereas treatment 1 (lower atrazine concentration) differed significantly, indicating that atrazine concentration impacted community variance. Higher atrazine addition subtly altered ligninolytic fungal community dynamics, implying its potential for pesticide degradation. Finally, variations in atrazine concentrations triggered diverse community responses over time, shedding light on fungal resilience and adaptive strategies against pesticides.
- MeSH
- Atrazine * metabolism pharmacology MeSH
- Biodegradation, Environmental MeSH
- Phylogeny MeSH
- Herbicides * metabolism MeSH
- Fungi * classification isolation & purification metabolism drug effects genetics growth & development MeSH
- Soil Pollutants metabolism MeSH
- Mycobiome * drug effects MeSH
- Soil Microbiology MeSH
- Publication type
- Journal Article MeSH
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
- Cell Differentiation * physiology MeSH
- Cell Culture Techniques methods MeSH
- Cell Line MeSH
- Spheroids, Cellular * metabolism MeSH
- Mice MeSH
- Neurons metabolism MeSH
- Prion Diseases * metabolism pathology MeSH
- Prions metabolism MeSH
- Cell Culture Techniques, Three Dimensional methods MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article 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.
- Publication type
- Journal Article 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
- Ecosystem * MeSH
- Plant Diseases genetics MeSH
- Oomycetes * genetics MeSH
- Plants MeSH
- Lactuca MeSH
- Plant Breeding MeSH
- Publication type
- Journal Article MeSH
- Review 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.
- Publication type
- Journal Article 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
- Anti-Bacterial Agents chemistry pharmacology MeSH
- Biomimetic Materials chemistry pharmacology MeSH
- Escherichia coli drug effects growth & development MeSH
- Hedgehogs * MeSH
- Microbial Sensitivity Tests MeSH
- Zinc Oxide chemistry pharmacology MeSH
- Staphylococcus aureus drug effects growth & development MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article 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, Plant genetics MeSH
- Endosperm genetics MeSH
- Genetic Variation genetics MeSH
- Hordeum genetics MeSH
- Polyploidy MeSH
- Genetics, Population methods MeSH
- Seeds genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Israel 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
- Environmental Psychology * methods MeSH
- Humans MeSH
- Psychotherapy methods MeSH
- Dance Therapy MeSH
- Check Tag
- Humans 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
- Biodegradation, Environmental MeSH
- Soil Pollutants analysis MeSH
- Soil MeSH
- Metals, Heavy analysis MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Croatia 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
- Anti-Bacterial Agents chemistry pharmacology MeSH
- Cell Line MeSH
- Chitosan chemistry MeSH
- Dynamic Light Scattering MeSH
- Escherichia coli drug effects MeSH
- Colloids chemistry MeSH
- Metal Nanoparticles chemistry MeSH
- Humans MeSH
- Methylcellulose chemistry MeSH
- Polysaccharides chemistry MeSH
- Drug Stability MeSH
- Staphylococcus epidermidis drug effects MeSH
- Sterilization MeSH
- Particle Size MeSH
- Gold chemistry pharmacology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
