Francisella tularensis influences several host molecular/signaling pathways during infection. Ubiquitination and deubiquitination are among the most important regulatory mechanisms and respectively occur through attachment or removal of the ubiquitin molecule. The process is necessary not only to mark molecules for degradation, but also, for example, to the activation of signaling pathways leading to pro-inflammatory host response. Many intracellular pathogens, including Francisella tularensis, have evolved mechanisms of modifying such host immune responses to escape degradation. Here, we describe that F. tularensis interferes with the host's ubiquitination system. We show increased total activity of deubiquitinating enzymes (DUBs) in human macrophages after infection, while confirm reduced enzymatic activities of two specific DUBs (USP10 and UCH-L5), and demonstrate increased activity of USP25. We further reveal the enrichment of these three enzymes in exosomes derived from F. tularensis-infected cells. The obtained results show the regulatory effect on ubiquitination mechanism in macrophages during F. tularensis infection.

• Klíčová slova
• DUBs, Francisella, UCH-L5, USP10, USP25, deubiquitination, exosomes, extracellular vesicles,
• MeSH
• deubikvitinasy metabolismus   MeSH
• Francisella tularensis * MeSH
• gramnegativní bakteriální infekce * metabolismus   MeSH
• lidé MeSH
• makrofágy MeSH
• signální transdukce MeSH
• thiolesterasa ubikvitinu metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• deubikvitinasy MeSH
• thiolesterasa ubikvitinu MeSH
• USP10 protein, human MeSH Prohlížeč
• USP25 protein, human MeSH Prohlížeč

Three-dimensional cell culture systems are increasingly used for biological and anticancer drug screening as they mimic the structure and microenvironment of tumors more closely than conventional two-dimensional cell models. In this study, the growth kinetics of colon adenocarcinoma-derived spheroids (HT-29 cell line) cultivated in liquid marble micro-bioreactors and nonadherent PDMS-coated well plates was investigated in detail and enabled precise control of the spheroid size by the seed cell density and cultivation time. The therapeutic effect of 5-fluorouracil and irinotecan hydrochloride in 2D monolayer cell culture and 3D tumor spheroids revealed an unexpected twist in their efficacy due to different ability to penetrate through 3D microtissue. For 5-fluorouracil, the inhibitory concentration IC50 after 48 h exposure increased from 11.3 µM for a 2D cell culture to 707.7 µM for a 3D spheroid. In the case of irinotecan, IC50 increased from 24.9 µM to 77.8 µM. Despite its higher molar weight, irinotecan appeared to penetrate the 3D spheroid structure more efficiently than 5-fluorouracil. While 5-fluorouracil mainly caused a suppression of spheroid growth from the outside, irinotecan affected the entire spheroid and caused its originally compact structure to disintegrate. The acquired results highlight the need to screen cancer chemotherapeutics on 3D tumor models, as contrasting results can be obtained compared to standard 2D cell cultures.

• Klíčová slova
• 3D Cell cultures, 5-Fluorouracil, Irinotecan hydrochloride, Liquid marbles, Polydimethylsiloxane, Tumor spheroids,
• MeSH
• adenokarcinom * MeSH
• buněčné sféroidy MeSH
• cytostatické látky * farmakologie   MeSH
• fluoruracil farmakologie   MeSH
• irinotekan farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• nádory tračníku * farmakoterapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytostatické látky * MeSH
• fluoruracil MeSH
• irinotekan MeSH

(1) Background: Huntington's disease (HD) is rare incurable hereditary neurodegenerative disorder caused by CAG repeat expansion in the gene coding for the protein huntingtin (HTT). Mutated huntingtin (mHTT) undergoes fragmentation and accumulation, affecting cellular functions and leading to neuronal cell death. Porcine models of HD are used in preclinical testing of currently emerging disease modifying therapies. Such therapies are aimed at reducing mHTT expression, postpone the disease onset, slow down the progression, and point out the need of biomarkers to monitor disease development and therapy efficacy. Recently, extracellular vesicles (EVs), particularly exosomes, gained attention as possible carriers of disease biomarkers. We aimed to characterize HTT and mHTT forms/fragments in blood plasma derived EVs in transgenic (TgHD) and knock-in (KI-HD) porcine models, as well as in HD patients' plasma. (2) Methods: Small EVs were isolated by ultracentrifugation and HTT forms were visualized by western blotting. (3) Results: The full length 360 kDa HTT co-isolated with EVs from both the pig model and HD patient plasma. In addition, a ~70 kDa mutant HTT fragment was specific for TgHD pigs. Elevated total huntingtin levels in EVs from plasma of HD groups compared to controls were observed in both pig models and HD patients, however only in TgHD were they significant (p = 0.02). (4) Conclusions: Our study represents a valuable initial step towards the characterization of EV content in the search for HD biomarkers.

• Klíčová slova
• Huntington´s disease, KI-HD, TgHD, biomarker, exosome, extracellular vesicle, fragment, huntingtin, neurodegenerative disease, pig model,
• MeSH
• biologické markery MeSH
• extracelulární vezikuly * metabolismus   MeSH
• Huntingtonova nemoc * metabolismus   MeSH
• krevní plazma metabolismus   MeSH
• lidé MeSH
• prasata MeSH
• proteiny nervové tkáně genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• proteiny nervové tkáně MeSH

Yeast glucan particles are porous polysaccharide cell walls extracted from Saccharomyces cerevisiae. Being mildly immunogenic, they are efficiently phagocytosed and have therefore been proposed as possible vehicles for drug delivery. Using curcumin as a model poorly water-soluble drug, a systematic comparison of three different physical loading methods - incipient wetness impregnation, slurry evaporation, and spray drying - was carried out and their influence on the particle morphology, encapsulation efficiency, amorphous drug content and release kinetics was evaluated. It was found that yeast glucan particles can contain up to 30% wt. of curcumin in the amorphous form when prepared by slurry evaporation. The dissolution of curcumin from glucan particles lead to a supersaturated solution in asimilar way as amorphous solid dispersions do, despite the fact that glucan particles themselves do not dissolve. Bi-phasic dissolution tests revealed up to 4-fold acceleration of curcumin dissolution rate from amorphous glucan particles compared to its crystalline form. Crucially, glucan particles were shown to retain the ability to be recognised and phagocytosed even after drug encapsulation.

• Klíčová slova
• Amorphous solid dispersion, Incipient wetness impregnation, Slurry evaporation, Spray drying, Two-phase dissolution test, β-glucan,
• MeSH
• beta-glukany chemie   MeSH
• farmaceutická chemie metody   MeSH
• kinetika MeSH
• krystalizace MeSH
• kurkumin aplikace a dávkování   chemie   MeSH
• lékové transportní systémy * MeSH
• nosiče léků chemie   MeSH
• příprava léků metody   MeSH
• rozpustnost MeSH
• uvolňování léčiv MeSH
• voda chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• beta-glukany MeSH
• kurkumin MeSH
• nosiče léků MeSH
• voda MeSH

Glucan particles (GPs) from Saccharomyces cerevisiae consist mainly of β-1,3-d-glucan. Curcumin is a phenolic compound of plant origin. A 24 h incubation with a mixture of GPs and curcumin increased the expression of the Nrf2 protein and increased the activation of the Nrf2-ARE system significantly.

• MeSH
• antioxidancia * chemie   metabolismus   farmakologie   MeSH
• buňky Hep G2 MeSH
• faktor 2 související s NF-E2 metabolismus   MeSH
• glukany chemie   MeSH
• kurkumin * chemie   farmakologie   MeSH
• lidé MeSH
• oxidační stres účinky léků   MeSH
• příprava léků MeSH
• Saccharomyces cerevisiae chemie   MeSH
• THP-1 buňky MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antioxidancia * MeSH
• faktor 2 související s NF-E2 MeSH
• glukany MeSH
• kurkumin * MeSH
• NFE2L2 protein, human MeSH Prohlížeč

Yeast glucan particles (GPs) are promising agents for the delivery of biologically active compounds as drugs. GPs possess their own biological activities and can act synergistically with their cargo. This study aimed to determine how incorporating artemisinin, ellagic acid, (-)-epigallocatechin gallate, morusin, or trans-resveratrol into GPs affects their anti-inflammatory and antioxidant potential in vitro. Two different methods - slurry evaporation and spray drying - were used to prepare composites (GPs + bioactive compound) and the anti-inflammatory and antioxidative properties of the resultant products were compared. Several of the natural compounds showed the beneficial effects of being combined with GPs. The materials prepared by spray drying showed greater activity than those made using a rotary evaporator. Natural compounds incorporated into yeast GPs showed greater anti-inflammatory potential in vitro than simple suspensions of these compounds as demonstrated by their inhibition of the activity of transcription factors NF-κB/AP-1 and the secretion of the pro-inflammatory cytokine TNF-α.

• Klíčová slova
• Drug carrier, Inflammation, Monocytes, Natural compounds, Pharmaceutical composite, β-glucan microparticles,
• MeSH
• antiflogistika farmakologie   MeSH
• antioxidancia farmakologie   MeSH
• artemisininy farmakologie   MeSH
• cytokiny MeSH
• flavonoidy chemie   farmakologie   MeSH
• glukany chemie   farmakologie   MeSH
• katechin analogy a deriváty   chemie   farmakologie   MeSH
• kyselina ellagová chemie   farmakologie   MeSH
• monocyty účinky léků   MeSH
• NF-kappa B MeSH
• resveratrol chemie   farmakologie   MeSH
• Saccharomyces cerevisiae - proteiny MeSH
• Saccharomyces cerevisiae MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiflogistika MeSH
• antioxidancia MeSH
• artemisinin MeSH Prohlížeč
• artemisininy MeSH
• cytokiny MeSH
• epigallocatechin gallate MeSH Prohlížeč
• flavonoidy MeSH
• glukany MeSH
• katechin MeSH
• kyselina ellagová MeSH
• morusin MeSH Prohlížeč
• NF-kappa B MeSH
• resveratrol MeSH
• Saccharomyces cerevisiae - proteiny MeSH

The goal of this work was to assess the usability of yeast glucan particles (GPs) as carriers for curcumin and determine the beneficial effect of a pharmacological composite of curcumin in GPs on dextran sulfate sodium induced colitis in rats. The assessment of the anti-inflammatory effect of particular substances was evaluated on the basis of the calculated disease activity index and by assessment of cytokines and enzymes from the gut tissue - tumor necrosis factor α (TNF-α), transforming growth factor β1, interleukin (IL)-1β, IL-6, IL-10, IL-17, catalase, superoxide dismutase 2, myeloperoxidase (MPO), and matrix metalloproteinase 9. Composites of GPs with incorporated curcumin showed promising results with the capability to lower symptoms of colitis and significantly decrease the production of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and the activity of MPO, as well. The anti-inflammatory effect of the composites was greater than those of pure GPs or curcumin.

• Klíčová slova
• Colitis targeting, Curcumin, Inflammation, Yeast glucan particles, β-Glucan,
• MeSH
• antiflogistika aplikace a dávkování   terapeutické užití   MeSH
• cytokiny metabolismus   MeSH
• glukany aplikace a dávkování   terapeutické užití   MeSH
• interleukiny metabolismus   MeSH
• kolitida chemicky indukované   farmakoterapie   MeSH
• krysa rodu Rattus MeSH
• kurkumin aplikace a dávkování   terapeutické užití   MeSH
• nosiče léků terapeutické užití   MeSH
• potkani Wistar MeSH
• Saccharomyces cerevisiae metabolismus   MeSH
• síran dextranu MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiflogistika MeSH
• cytokiny MeSH
• glukany MeSH
• interleukiny MeSH
• kurkumin MeSH
• nosiče léků MeSH
• síran dextranu MeSH

Geranylated flavanone diplacone is a flavanone iso- lated from Paulownia tomentosa (Thunb.) Steud. (Paulowniaceae) with anti-inflammatory and antioxidant properties, nevertheless showing high lipophilicity and low solubility in water. Diplacone was therefore used as a model molecule for incorporation into glucan particles (GPs). GPs are prepared by intensive washing of yeast (Saccharomyces cerevisiae) leading to hollow shells consisting of β-(13)/β-(16) glucan mainly. The aim of this study was to compare anti-inflammatory potential of GPs-diplacone composites with the compound itself, GPs themselves and the physical mixture of GPs and diplacone. The cell line THP1-XBlueTM-MD2-CD14 derived from human leukemic monocytes was stimulated with lipopolysaccharide (LPS) from Escherichia coli to trigger inflammatory reaction. The composites of GPs with diplacone significantly decreased the activity of pro-inflammatory transcription factors nuclear factor κB (NF-κB) and activator protein 1 (AP-1).

• Klíčová slova
• AP-1, NF-κB, diplacone, encapsulation, glucan particles, inflammation,
• MeSH
• antiflogistika farmakologie   MeSH
• flavanony farmakologie   MeSH
• glukany chemie   MeSH
• lidé MeSH
• NF-kappa B metabolismus   MeSH
• Saccharomyces cerevisiae chemie   MeSH
• THP-1 buňky MeSH
• transkripční faktor AP-1 metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiflogistika MeSH
• diplacone MeSH Prohlížeč
• flavanony MeSH
• glukany MeSH
• NF-kappa B MeSH
• transkripční faktor AP-1 MeSH

Natural compounds offer a wide spectrum of potential active substances, but often they have a poor bioavailability. To increase the bioavailability and bioactivity of the natural anti-inflammatory molecules curcumin and diplacone, we used glucan particles (GPs), hollow shells from Saccharomyces cerevisiae composed mainly of β-1,3-d-glucan. Their indigestibility and relative stability in the gut combined with their immunomodulatory effects makes them promising carriers for such compounds. This study aimed to determine how curcumin and diplacone, either alone or incorporated in GPs, affect the immunomodulatory activity of the latter by assessing the respiratory burst response and the secretion of pro-inflammatory cytokines by primary porcine innate immune cells. Incorporating curcumin and diplacone into GPs by controlled evaporation of the organic solvent substantially reduced the respiratory burst response mediated by GPs. Incorporated curcumin in GPs also reduced GPs mediated secretion of IL-1β and TNF-α by innate immune cells. The obtained results indicate a potentially beneficial effect of the incorporation of curcumin or diplacone into GPs against inflammation.

• Klíčová slova
• Blood immune cells, Curcumin, Diplacone, Glucan particles, Immunomodulatory effect, Inflammation,
• MeSH
• antiflogistika chemie   farmakologie   MeSH
• beta-glukany chemie   izolace a purifikace   farmakologie   MeSH
• flavanony chemie   farmakologie   MeSH
• imunologické faktory chemie   izolace a purifikace   farmakologie   MeSH
• interleukin-1beta metabolismus   MeSH
• kultivované buňky MeSH
• kurkumin chemie   farmakologie   MeSH
• leukocyty mononukleární účinky léků   imunologie   metabolismus   MeSH
• neutrofily účinky léků   imunologie   metabolismus   MeSH
• nosiče léků * MeSH
• příprava léků MeSH
• proteoglykany MeSH
• respirační vzplanutí účinky léků   MeSH
• rozpouštědla chemie   MeSH
• Saccharomyces cerevisiae chemie   MeSH
• Sus scrofa MeSH
• TNF-alfa metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiflogistika MeSH
• beta-glukany MeSH
• diplacone MeSH Prohlížeč
• flavanony MeSH
• imunologické faktory MeSH
• interleukin-1beta MeSH
• kurkumin MeSH
• nosiče léků * MeSH
• polysaccharide-K MeSH Prohlížeč
• proteoglykany MeSH
• rozpouštědla MeSH
• TNF-alfa MeSH

In this work, novel amorphous solid dispersions based on yeast glucan particles were produced. Yeast glucan particles are hollow and porous, and they are mainly composed of amorphous polysaccharides. We hypothesized that these particles are suitable candidates for the amorphization of drugs with low water solubility. Model drugs ibuprofen and curcumin were successfully encapsulated in glucan particles by spray drying. Different spray-drying parameters were tested to evaluate the influence of atomizing droplet size and initial solid content on encapsulation efficiency. It was shown that higher solid content and, more significantly, larger droplet sizes lead to higher encapsulation efficiencies. The encapsulation efficiency of ibuprofen (10 wt%) into glucan particles was considerably improved from 41.3 ± 0.5% to 64.3 ± 0.2% by increasing initial solid content and droplet size with the two-fluid nozzle. The spray drying process was further optimized by using the ultrasonic nozzle and it was possible to achieve complete encapsulation of ibuprofen and curcumin without any precipitation of the active compound outside of the glucan particles. Overall, it was possible to produce completely amorphous composites with outstanding wettability and dispersion properties, and with significantly faster dissolution rates when compared to the micronized crude drug.

• Klíčová slova
• Amorphous solid dispersions, Drug delivery, Poorly soluble drugs, Spray drying, Yeast glucan particles,
• MeSH
• aerosoly MeSH
• beta-glukany chemie   izolace a purifikace   MeSH
• ibuprofen chemie   MeSH
• kinetika MeSH
• kurkumin chemie   MeSH
• nosiče léků * MeSH
• příprava léků MeSH
• rozpustnost MeSH
• Saccharomyces cerevisiae chemie   MeSH
• ultrazvuk * MeSH
• uvolňování léčiv MeSH
• velikost částic MeSH
• vysoušení * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aerosoly MeSH
• beta-glukany MeSH
• ibuprofen MeSH
• kurkumin MeSH
• nosiče léků * MeSH