- MeSH
- banky biologického materiálu * organizace a řízení MeSH
- biomedicínský výzkum MeSH
- lidé MeSH
- uchovávání tkání MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
Elektrochemická detekce biomolekul se neustále posouvá a má velký potenciál využití v klinické praxi. V posledních letech je kladen důraz na přesné a rychlé stanovení biomarkerů na bázi nukleových kyselin – DNA a RNA. Značná část výzkumných projektů však nereflektuje požadavky na demonstraci vyvinutých metod na klinickém materiálu, což jejich aplikační potenciál značně snižuje. Klinický materiál vnáší do výzkumu oproti modelovým vzorkům větší variabilitu a práce s ním vyžaduje specifické podmínky. Problém získávání klinického materiálu pro výzkumné účely z velké části řeší banky biologického materiálu, které mohou nabídnout vzorky a data vhodné pro konkrétní výzkumný záměr.
Precise diagnostics of cancer or other diseases is crucial when selecting proper treatment. Personalized medicine puts high demands on the accuracy of nucleic acid biomarkers analysis, where subtle differences at the nucleotide level are often involved. Isothermal amplification techniques offer new possibilities of DNA and RNA amplification without using PCR, and their combination with electrochemistry provide a promising fast and cost-effective alternative diagnostic tool. Although electrochemical biosensors are still insufficiently applied to clinical material, thus hindering their development, recent advancements show great promise in translational research. Banks of biological material (biobanks) are specialized workplaces focused on the long-term preservation and processing of clinical material and offer a wide range of expert services, primarily for research purposes, in particular the provision of biological samples and associated pseudonymized data. Their involvement in the field of electrochemical biosensors can facilitate application of electrochemical methods into clinical laboratories and expand the portfolio of currently used diagnostic methods.
- MeSH
- banky biologického materiálu MeSH
- biosenzitivní techniky metody MeSH
- elektrochemické techniky * metody MeSH
- nádorové biomarkery * analýza klasifikace MeSH
- techniky amplifikace nukleových kyselin metody MeSH
- translační biomedicínský výzkum MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
The complexity of drug delivery mechanisms calls for the development of new transport system designs. Here, we report a robust synthetic procedure toward stable glycodendrimer (glyco-DDM) series bearing glucose, galactose, and oligo(ethylene glycol)-modified galactose peripheral units. In vitro cytotoxicity assays showed exceptional biocompatibility of the glyco-DDMs. To demonstrate applicability in drug delivery, the anticancer agent doxorubicin (DOX) was encapsulated in the glyco-DDM structure. The anticancer activity of the resulting glyco-DDM/DOX complexes was evaluated on the noncancerous (BJ) and cancerous (MCF-7 and A2780) cell lines, revealing their promising generation- and concentration-dependent effect. The glyco-DDM/DOX complexes show gradual and pH-dependent DOX release profiles. Fluorescence spectra elucidated the encapsulation process. Confocal fluorescence microscopy demonstrated preferential cancer cell internalization of the glyco-DDM/DOX complexes. The conclusions were supported by computer modeling. Overall, our results are consistent with the assumption that novel glyco-DDMs and their drug complexes are very promising in drug delivery and related applications.
- MeSH
- antitumorózní látky * farmakologie MeSH
- doxorubicin chemie farmakologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory vaječníků * MeSH
- nosiče léků chemie MeSH
- polyethylenglykoly chemie MeSH
- silany MeSH
- systémy cílené aplikace léků metody MeSH
- uvolňování léčiv MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Aquatic biotests are important tools targeting various effects in ecotoxicology, including endocrine disruption. Unintentional exposure of bioassay organisms to endocrine disruptors during cultivation or testing may interfere with assessed endpoints. We illustrate this issue on the example of laboratory phytoplankton cultivation, where possible sources of estrogenic compounds have been revealed. Fifty-four blank samples (water and fresh or cultivated growth media) were assessed by in vitro biotests for their estrogenicity, and major known estrogens originating from plastic materials, bisphenol A and alkylphenols, were analyzed in selected samples. The samples of freshly prepared growth medium elicited weak estrogenic response in bioassays and some samples of the aerated media caused responses even above the 50% of maximum of the reference compound (17β-estradiol, E2), while the samples from diverse laboratory water sources did not show significant estrogenic activity. The results identified substances contained in the growth medium as minor but reproducible contributors to estrogenicity in the cultivations. Sporadic but significant effects (up to 4.9 ng E2 equivalent/L) can be ascribed to compounds released from the used plastic materials during aeration of the cultivations. The potential sources of unintentional exposure to estrogenic compounds need to be considered in aquatic cultivations and biotests, since they could impact their outcomes, especially in arrangements assessing reproduction or whole life cycle biotests, or production of bioactive compounds by phytoplankton. The findings emphasize the necessity to assess all relevant blanks, ideally by sensitive high throughput in vitro assays that reflect also unknown pollutants and minimize all potential sources of background contamination. In vitro assays show very good applicability for this purpose since they enable to screen for any background estrogenicity of the used media and materials without the need of analyzing individual compounds, which often might not be known.
- MeSH
- akcelerometrie MeSH
- chůze (způsob) MeSH
- dítě MeSH
- ergoterapie MeSH
- kyčelní kloub MeSH
- lidé MeSH
- mladiství MeSH
- mozková obrna * rehabilitace MeSH
- neurorehabilitace MeSH
- pilotní projekty MeSH
- postižené děti rehabilitace MeSH
- posturální rovnováha MeSH
- předškolní dítě MeSH
- terapie využívající koní * MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- předškolní dítě MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Insulin-like growth factors 2 and 1 (IGF2 and IGF1) and insulin are closely related hormones that are responsible for the regulation of metabolic homeostasis, development and growth of the organism. Physiological functions of insulin and IGF1 are relatively well-studied, but information about the role of IGF2 in the body is still sparse. Recent discoveries called attention to emerging functions of IGF2 in the brain, where it could be involved in processes of learning and memory consolidation. It was also proposed that these functions could be mediated by the receptor for IGF2 (IGF2R). Nevertheless, little is known about the mechanism of signal transduction through this receptor. Here we produced His-tagged domain 11 (D11), an IGF2-binding element of IGF2R; we immobilized it on the solid support through a well-defined sandwich, consisting of neutravidin, biotin and synthetic anti-His-tag antibodies. Next, we prepared specifically radiolabeled [125I]-monoiodotyrosyl-Tyr2-IGF2 and optimized a sensitive and robust competitive radioligand binding assay for determination of the nanomolar binding affinities of hormones for D11 of IGF2. The assay will be helpful for the characterization of new IGF2 mutants to study the functions of IGF2R and the development of new compounds for the treatment of neurological disorders.
- MeSH
- insulinu podobný růstový faktor I metabolismus MeSH
- insulinu podobný růstový faktor II metabolismus MeSH
- kompetitivní vazba MeSH
- kultivované buňky MeSH
- lidé MeSH
- radioizotopy jodu MeSH
- radioligandová zkouška metody MeSH
- receptor IGF typ 2 imunologie ultrastruktura MeSH
- signální transdukce MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The C-type lectin DC-SIGN expressed on immature dendritic cells is a promising target for antiviral drug development. Previously, we have demonstrated that mono- and divalent C-glycosides based on d-manno and l-fuco configurations are promising DC-SIGN ligands. Here, we described the convergent synthesis of C-glycoside dendrimers decorated with 4, 6, 9, and 12 α-l-fucopyranosyl units and with 9 and 12 α-d-mannopyranosyl units. Their affinity against DC-SIGN was assessed by surface plasmon resonance (SPR) assays. For comparison, parent O-glycosidic dendrimers were synthesized and tested, as well. A clear increase of both affinity and multivalency effect was observed for C-glycomimetics of both types (mannose and fucose). However, when dodecavalent C-glycosidic dendrimers were compared, there was no difference in affinity regarding the sugar unit (l-fuco, IC50 17 μM; d-manno, IC50 12 μM). For the rest of glycodendrimers with l-fucose or d-mannose attached by the O- or C-glycosidic linkage, C-glycosidic dendrimers were significantly more active. These results show that in addition to the expected physiological stability, the biological activity of C-glycoside mimetics is higher in comparison to the corresponding O-glycosides and therefore these glycomimetic multivalent systems represent potentially promising candidates for targeting DC-SIGN.
- MeSH
- biomimetické materiály chemie farmakologie MeSH
- fukosa chemie MeSH
- inhibiční koncentrace 50 MeSH
- lektiny typu C antagonisté a inhibitory MeSH
- mannosa chemie MeSH
- molekuly buněčné adheze antagonisté a inhibitory MeSH
- receptory buněčného povrchu antagonisté a inhibitory MeSH
- Publikační typ
- časopisecké články MeSH
Phytoplankton can produce various bioactive metabolites, which may affect other organisms in the aquatic environment. This study provides the first information on the total retinoid-like activity associated with both intracellular and extracellular metabolites produced by selected phytoplankton species that could play a role in teratogenic effects and developmental disruption in exposed organisms. The studied species included a coccoid cyanobacteria (Microcystis aeruginosa), a filamentous cyanobacteria (Aphanizomenon gracile) and a green alga (Desmodesmus quadricauda), all of which commonly occur in freshwater bodies in Europe. Methanolic extracts from cellular material and extracellular exudates were prepared from cultures cultivated in two light-intensity variants with five replicates for each species. The retinoid-like activity was evaluated by in vitro assays along with chemical analyses of two potent retinoic acids (all-trans retinoic acid (ATRA) and 9cis-RA). The mean total retinoid-like activity of metabolites produced by the three studied species representing different phytoplankton taxonomic groups ranged from 705 to 5572ng ATRA equivalent/g dry matter corresponding to 0.064-0.234ng ATRA/106 cells. Retinoid-like activity was found in the cellular extracts of all species, while only the extracellular exudates of cyanobacteria exhibited detectable activity (41-1081ng ATRA/L). The greatest extracellular as well as total (extra- and intra- cellular together) retinoid-like activity was detected for Microcystis aeruginosa. The two potent retinoic acids studied were more frequently detected in cellular extracts than in extracellular exudates of all species. Their contribution to observed in vitro effects was relatively low for all tested samples (<10%), indicating a substantial contribution of other retinoid-like compounds to the overall activity. The results indicate possible influence of light intensity and cell density on the production of metabolites with retinoid-like activity and the cyanotoxin microcystin by the studied species. The recalculation of the results per dry weight, water volume, per 106 cells and biovolume enables a direct comparison of the retinoid-like activity distribution between extracts and exudates and the use of the data for risk assessment in water bodies.
- MeSH
- buněčné linie MeSH
- Chlorophyta chemie MeSH
- fytoplankton chemie metabolismus mikrobiologie MeSH
- Microcystis MeSH
- mikrocystiny MeSH
- retinoidy farmakologie MeSH
- rostlinné extrakty farmakologie MeSH
- sinice chemie MeSH
- tretinoin farmakologie MeSH
- viabilita buněk účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa MeSH
Cyanobacteria contain various types of bioactive compounds, which could cause adverse effects on organisms. They are released into surface waters during cyanobacterial blooms, but there is little information on their potential relevance for effects in vivo. In this study presence of bioactive compounds was characterized in cyanobacteria Microcystis aeruginosa (Chroococcales), Planktothrix agardhii (Oscillatoriales) and Aphanizomenon gracile (Nostocales) with selected in vitro assays. The in vivo relevance of detected bioactivities was analysed using transgenic zebrafish embryos tg(cyp19a1b-GFP). Teratogenic potency was assessed by analysis of developmental disorders and effects on functions of the neuromuscular system by video tracking of locomotion. Estrogenicity in vitro corresponded to 0.95-54.6 ng estradiol equivalent(g dry weight (dw))(-1). In zebrafish embryos, estrogenic effects could not be detected potentially because they were masked by high toxicity. There was no detectable (anti)androgenic/glucocorticoid activity in any sample. Retinoid-like activity was determined at 1-1.3 μg all-trans-retinoic acid equivalent(g dw)(-1). Corresponding to the retinoid-like activity A. gracile extract also caused teratogenic effects in zebrafish embryos. Furthermore, exposure to biomass extracts at 0.3 gd wL(-1) caused increase of body length in embryos. There were minor effects on locomotion caused by 0.3 gd wL(-1)M. aeruginosa and P. agardhii extracts. The traditionally measured cyanotoxins microcystins did not seem to play significant role in observed effects. This indicates importance of other cyanobacterial compounds at least towards some species or their developmental phases. More attention should be paid to activity of retinoids, estrogens and other bioactive substances in phytoplankton using in vitro and in vivo bioassays.
- MeSH
- Aphanizomenon chemie MeSH
- biotest MeSH
- dánio pruhované embryologie genetika metabolismus MeSH
- embryo nesavčí účinky léků MeSH
- endokrinní disruptory toxicita MeSH
- geneticky modifikovaná zvířata embryologie genetika metabolismus MeSH
- Microcystis chemie MeSH
- neurotoxiny toxicita MeSH
- sinice chemie MeSH
- teratogeny toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
