Herein, we investigated the anti-amoebic activity of phosphonium-chloride-based deep eutectic solvents against pathogenic Acanthamoeba castellanii of the T4 genotype. Deep eutectic solvents are ionic fluids composed of two or three substances, capable of self-association to form a eutectic mixture with a melting point lower than each substance. In this study, three distinct hydrophobic deep eutectic solvents were formulated, employing trihexyltetradecylphosphonium chloride as the hydrogen bond acceptor and aspirin, dodecanoic acid, and 4-tert-butylbenzoic acid as the hydrogen bond donors. Subsequently, all three deep eutectic solvents, denoted as DES1, DES2, DES3 formulations, underwent investigations comprising amoebicidal, adhesion, excystation, cytotoxicity, and cytopathogenicity assays. The findings revealed that DES2 was the most potent anti-amoebic agent, with a 94% elimination rate against the amoebae within 24 h at 30 °C. Adhesion assays revealed that deep eutectic solvents hindered amoebae adhesion to human brain endothelial cells, with DES2 exhibiting 88% reduction of adhesion. Notably, DES3 exhibited remarkable anti-excystation properties, preventing 94% of cysts from reverting to trophozoites. In cytopathogenicity experiments, deep eutectic solvent formulations and dodecanoic acid alone reduced amoebae-induced human brain endothelial cell death, with DES2 showing the highest effects. Lactate dehydrogenase assays revealed the minimal cytotoxicity of the tested deep eutectic solvents, with the exception of trihexyltetradecylphosphonium chloride, which exhibited 35% endothelial cell damage. These findings underscore the potential of specific deep eutectic solvents in combating pathogenic Acanthamoeba, presenting promising avenues for further research and development against free-living amoebae.
- Klíčová slova
- Acanthamoeba castellanii, Cytotoxicity, Deep eutectic solvents, Excystment, Lactate dehydrogenase,
- MeSH
- Acanthamoeba castellanii * účinky léků genetika MeSH
- amébicidy farmakologie chemie MeSH
- buněčná adheze účinky léků MeSH
- endoteliální buňky účinky léků MeSH
- genotyp * MeSH
- lidé MeSH
- organofosforové sloučeniny farmakologie chemie MeSH
- rozpouštědla * chemie farmakologie MeSH
- trofozoiti účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- amébicidy MeSH
- organofosforové sloučeniny MeSH
- rozpouštědla * MeSH
BACKGROUND: This study evaluated in vitro antigiardial activity in four Indonesian plants (Archidendron fagifolium, Diospyros sumatrana, Piper betle and Shorea sumatrana) extracted in methanol, methanol-tetrahydrofuran, and water. These plants exhibiting promising anti-parasitic activity were selected on the basis of collected behavioral data and their ability to decrease parasite load in Sumatran orangutans. Extracts of Arabidopsis thaliana, a plant routinely used as a laboratory model in research, were used as a negative control. METHODS: Plant extracts of different concentrations (400, 100, 25 and 6.25 µg/ml) and metronidazole (100 µg/ml), a standard giardicidal drug, were incubated with 25,000 trophozoites per milliliter of growth medium in 12-well tissue culture plates under anaerobic conditions for 72 h. Cultures were counted in a hemocytometer using a light microscope and then statistically evaluated. The cytotoxicity of the extracts was determined by the MTT Cell Viability Assay. RESULTS: We recorded a statistically-significant decrease in giardia trophozoites in two extracts: the methanolic extract of A. fagifolium (Kruskal-Wallis: p = 0.013) and the aqueous extract of P. betle (Kruskal-Wallis: p = 0.002). Effective concentrations of 400 µg/ml and 100 µg/ml, respectively, were revealed. Cytotoxicity was not demonstrated. CONCLUSIONS: The results of our study show that extracts of A. fagifolium and P. betle have potential as an alternative treatment of G. intestinalis infection. This is the first scientific proof of the biological activity of A. fagifolium, confirming the assumption that orangutans use this plant (as well as P. betle) for self-medication.
- Klíčová slova
- Giardia intestinalis, Antigiardial activity, Drug of choice, Natural antiparasitic, Parasite,
- MeSH
- antiprotozoální látky farmakologie MeSH
- Giardia lamblia * účinky léků MeSH
- rostlinné extrakty * farmakologie MeSH
- trofozoiti účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Indonésie MeSH
- Názvy látek
- antiprotozoální látky MeSH
- rostlinné extrakty * MeSH
Advanced imaging of microorganisms, including protists, is challenging due to their small size. Specimen expansion prior to imaging is thus beneficial to increase resolution and cellular details. Here, we present a sample preparation workflow for improved observations of the single-celled eukaryotic pathogen Giardia intestinalis (Excavata, Metamonada). The binucleated trophozoites colonize the small intestine of humans and animals and cause a diarrhoeal disease. Their remarkable morphology includes two nuclei and a pronounced microtubular cytoskeleton enabling cell motility, attachment and proliferation. By use of expansion and confocal microscopy, we resolved in a great detail subcellular structures and organelles of the parasite cell. The acquired spatial resolution enabled novel observations of centrin localization at Giardia basal bodies. Interestingly, non-luminal centrin localization between the Giardia basal bodies was observed, which is an atypical eukaryotic arrangement. Our protocol includes antibody staining and can be used for the localization of epitope-tagged proteins, as well as for differential organelle labelling by amino reactive esters. This fast and simple technique is suitable for routine use without a superresolution microscopy equipment.
- Klíčová slova
- Caltractin, Centrin, Cytoskeleton, Expansion microscopy, Giardia, Protist,
- MeSH
- Giardia lamblia * ultrastruktura cytologie MeSH
- konfokální mikroskopie * MeSH
- lidé MeSH
- organely ultrastruktura chemie MeSH
- protozoální proteiny analýza chemie MeSH
- trofozoiti ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- protozoální proteiny MeSH
Acanthamoeba is a ubiquitous genus of amoebae that can trigger a severe and progressive ocular disease known as Acanthamoeba Keratitis (AK). Furthermore, current treatment protocols are based on the combination of different compounds that are not fully effective. Therefore, an urgent need to find new compounds to treat Acanthamoeba infections is clear. In the present study, we evaluated staurosporine as a potential treatment for Acanthamoeba keratitis using mouse cornea as an ex vivo model, and a comparative proteomic analysis was conducted to elucidate a mechanism of action. The obtained results indicate that staurosporine altered the conformation of actin and tubulin in treated trophozoites of A. castellanii. In addition, proteomic analysis of treated trophozoites revealed that this molecule induced overexpression and a downregulation of proteins related to key functions for Acanthamoeba infection pathways. Additionally, the ex vivo assay used validated this model for the study of the pathogenesis and therapies of AK. Finally, staurosporine eliminated the entire amoebic population and prevented the adhesion and infection of amoebae to the epithelium of treated mouse corneas.
- Klíčová slova
- Acanthamoeba, PCD, ex vivo, mouse cornea, proteomic analysis, staurosporine,
- MeSH
- Acanthamoeba castellanii * účinky léků MeSH
- akantamébová keratitida * farmakoterapie parazitologie MeSH
- aktiny metabolismus MeSH
- modely nemocí na zvířatech * MeSH
- myši MeSH
- proteomika * metody MeSH
- rohovka * účinky léků parazitologie MeSH
- staurosporin * farmakologie MeSH
- trofozoiti účinky léků MeSH
- tubulin metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- aktiny MeSH
- staurosporin * MeSH
- tubulin MeSH
In this study, we present a high-resolution dataset and bioinformatic analysis of the proteome of Bacillus subtilis 168 trp+ (BSB1) during germination and spore outgrowth. Samples were collected at 14 different time points (ranging from 0 to 130 min) in three biological replicates after spore inoculation into germination medium. A total of 2191 proteins were identified and categorized based on their expression kinetics. We observed four distinct clusters that were analyzed for functional categories and KEGG pathways annotations. The examination of newly synthesized proteins between successive time points revealed significant changes, particularly within the first 50 min. The dataset provides an information base that can be used for modeling purposes and inspire the design of new experiments.
- Klíčová slova
- KEGG pathways analysis, protein functional analysis, time series of protein expression,
- MeSH
- Bacillus subtilis * metabolismus růst a vývoj MeSH
- bakteriální proteiny * metabolismus MeSH
- proteom * metabolismus analýza MeSH
- proteomika metody MeSH
- spory bakteriální * metabolismus růst a vývoj MeSH
- výpočetní biologie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bakteriální proteiny * MeSH
- proteom * MeSH
Chlorosplenium is a small genus comprising five species of inoperculate discomycetes in the order Helotiales (Leotiomycetes) often recognizable by their bright yellowish-green colors and gregarious growth on wood. In this study, we describe five new species-C. aotearoa, C. australiense, C. cusucoense, C. epimorsicum, and C. hawaiiense-based on a combination of recent fieldwork and examination of previously collected fungarium specimens. We use an integrative taxonomic approach to support the distinction of new species, incorporating morphology and DNA sequence data with biogeography. Macro- and micromorphological features of apothecia for all species and culture characteristics for four of the five new species are documented. A multilocus phylogeny based on nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2, partial large subunit nuc ribosomal DNA (28S nuc rDNA), and A-B regions of the largest subunit of RNA polymerase II (RPB1) gene is presented. Additionally, we report Chlorosplenium chlora from Europe for the first time and expand our knowledge of the diversity and distributions of species in this genus in America, Australia, and New Zealand.
- Klíčová slova
- 5 new taxa, Chlorospleniaceae, semicryptic species,
- MeSH
- Ascomycota * genetika klasifikace izolace a purifikace MeSH
- biodiverzita MeSH
- DNA fungální * genetika MeSH
- dřevo mikrobiologie MeSH
- fylogeneze * MeSH
- mezerníky ribozomální DNA * genetika MeSH
- molekulární sekvence - údaje MeSH
- ribozomální DNA genetika MeSH
- RNA ribozomální 28S genetika MeSH
- RNA-polymerasa II genetika MeSH
- sekvenční analýza DNA MeSH
- spory hub cytologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- DNA fungální * MeSH
- mezerníky ribozomální DNA * MeSH
- ribozomální DNA MeSH
- RNA ribozomální 28S MeSH
- RNA-polymerasa II MeSH
During sporulation, Bacillus subtilis forms an asymmetric septum, dividing the cell into two compartments, a mother cell and a forespore. The site of asymmetric septation is linked to the membrane where FtsZ and SpoIIE initiate the formation of the Z-ring and the E-ring, respectively. These rings then serve as a scaffold for the other cell division and peptidoglycan synthesizing proteins needed to build the septum. However, despite decades of research, not enough is known about how the asymmetric septation site is determined. Here, we identified and characterized the interaction between SpoIIE and RefZ. We show that these two proteins transiently colocalize during the early stages of asymmetric septum formation when RefZ localizes primarily from the mother cell side of the septum. We propose that these proteins and their interplay with the spatial organization of the chromosome play a role in controlling asymmetric septum positioning.
- Klíčová slova
- Bacillus, RefZ, SpoIIE, asymmetric cell division, chromosomes, protein-protein interaction, sporulation,
- MeSH
- Bacillus subtilis * metabolismus fyziologie MeSH
- bakteriální proteiny * metabolismus genetika MeSH
- buněčné dělení MeSH
- cytoskeletální proteiny metabolismus genetika MeSH
- spory bakteriální * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bakteriální proteiny * MeSH
- cytoskeletální proteiny MeSH
- spore-specific proteins, Bacillus MeSH Prohlížeč
PREMISE: Apomixis in ferns is relatively common and obligatory. Sterile hybrids may restore fertility via apomixis at a cost of long-term genetic stagnation. In this study, we outlined apomixis as a possible temporary phase leading to sexuality and analyzed factors relating to transitioning to and away from apomixis, such as unreduced and reduced spore formation in apomict and apo-sex hybrid ferns. METHODS: We analyzed the genome size of 15 fern species or hybrids ("taxa") via flow cytometry. The number of reduced and unreduced gametophytes was established as a proxy for viable spore formation of either type. We also calculated the spore abortion ratio (sign of reduced spores) in several taxa, including the apo-sex hybrid Dryopteris × critica and its 16 apomictically formed offspring. RESULTS: Four of 15 sampled taxa yielded offspring variable in genome size. Specifically, each variable taxon formed one viable reduced plant among 12-451 sampled gametophytes per taxon. Thus, haploid spore formation in the studied apomicts was very rare but possible. Spore abortion analyses indicated gradually decreasing abortion (haploid spore formation) over time. In Dryopteris × critica, abortion decreased from 93.8% to mean 89.5% in one generation. CONCLUSIONS: Our results support apomixis as a transitionary phase toward sexuality. Newly formed apomicts hybridize with sexual relatives and continue to form haploid spores early on. Thus, they may get the genomic content necessary for regular meiosis and restore sexuality. If the missing relative goes extinct, the lineage gets locked into apomixis as may be the case with the Dryopteris affinis complex.
- Klíčová slova
- Dryopteris, apogamy, apomixis, evolutionary biology, fern life cycle, flow cytometry, gametophyte, genome size, pteridophytes, reproduction,
- MeSH
- apomixie * genetika MeSH
- délka genomu * MeSH
- genom rostlinný * MeSH
- hybridizace genetická MeSH
- kapradiny * genetika fyziologie MeSH
- spory * fyziologie genetika MeSH
- Publikační typ
- časopisecké články MeSH
The long-term effects of the Central Atlantic Magmatic Province, a large igneous province connected to the end-Triassic mass-extinction (201.5 Ma), remain largely elusive. Here, we document the persistence of volcanic-induced mercury (Hg) pollution and its effects on the biosphere for ~1.3 million years after the extinction event. In sediments recovered in Germany (Schandelah-1 core), we record not only high abundances of malformed fern spores at the Triassic-Jurassic boundary, but also during the lower Jurassic Hettangian, indicating repeated vegetation disturbance and stress that was eccentricity-forced. Crucially, these abundances correspond to increases in sedimentary Hg-concentrations. Hg-isotope ratios (δ202Hg, Δ199Hg) suggest a volcanic source of Hg-enrichment at the Triassic-Jurassic boundary but a terrestrial source for the early Jurassic peaks. We conclude that volcanically injected Hg across the extinction was repeatedly remobilized from coastal wetlands and hinterland areas during eccentricity-forced phases of severe hydrological upheaval and erosion, focusing Hg-pollution in the Central European Basin.
- MeSH
- extinkce biologická * MeSH
- geologické sedimenty * chemie MeSH
- kapradiny * MeSH
- mutageneze MeSH
- podnebí MeSH
- rtuť * analýza MeSH
- sopečné erupce MeSH
- spory MeSH
- zkameněliny * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Německo MeSH
- Názvy látek
- rtuť * MeSH
Coccidiosis is a protozoan intestinal disease that reduces the production of the sheep industry and causes large economic losses for sheep. Although chemically synthesised drugs are routinely employed to treat coccidiosis in sheep, the anticoccidial drug resistance and drug residues in edible meat have prompted an urgent search for alternatives. Herein, the anticoccidial properties of diclazuril, a conventional anticoccidial drug, and Allium sativum, Houttuynia cordata and Portulaca oleracea were assessed. Forty 45-day-old lambs naturally infected with Eimeria spp. were selected and randomly divided into five groups. The results showed that the sheep treated for coccidiosis had considerably decreased average daily gain (ADG) during both administration and withdrawal of the drug compared to the control group. Furthermore, at days 14, 21, 28 and 35, respectively, the three herbs and diclazuril had similar anticoccidial effects, with lower oocysts per gram (OPG) than the control group. On day 78, OPG in the three herbal groups was significantly lower than in the diclazuril group. In addition, the abundance and composition of the gut microbiota were changed in sheep treated with the three herbs and diclazuril compared to the untreated sheep. Moreover, some intestinal microorganisms have a correlation with OPG and ADG when using Spearman correlation analysis. In summary, our results suggest that all three herbs produce anticoccidial effects similar to diclazuril and modulate the balance of gut microbiota in growing lambs.
- Klíčová slova
- Eimeria, Epizootiology, microbiome, prevention and treatment, sheep,
- MeSH
- Eimeria účinky léků fyziologie MeSH
- kokcidiostatika farmakologie aplikace a dávkování MeSH
- kokcidióza * veterinární farmakoterapie parazitologie MeSH
- léky rostlinné čínské farmakologie aplikace a dávkování MeSH
- nemoci ovcí * parazitologie farmakoterapie MeSH
- oocysty účinky léků MeSH
- ovce MeSH
- střevní mikroflóra * účinky léků MeSH
- triaziny farmakologie aplikace a dávkování MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kokcidiostatika MeSH
- léky rostlinné čínské MeSH
- triaziny MeSH
