The moving volutin (polyphosphate) granules known as "dancing bodies" can be observed in the vacuoles of the yeast cells. The aim of work was to study the effects of cultivation conditions and influences of physico-chemical factors on the motion of vacuolar volutin granules in Saccharomyces cerevisiae cells. The motion of granules is a non-Markovian process. It does not depend on the cell cycle phase, but depends on the growth stage. The maximal number of cells with "dancing bodies" was observed under cultivation of yeast at 25-28 °C and pH 5.4-5.8. Irradiation by non-ionizing electromagnetic radiation (EMR) of extremely high frequency (61.22 GHz, 100 μW, 30 min) had no effect on granule motion. After irradiation by non-ionizing EMR of very high frequency (40.68 MHz, 30 W, 30 min) the number of cells with "dancing bodies" decreased significantly and in 2 h restored almost to the control value. The possible nature of the moving volutin granules phenomenon due to metabolic processes is discussed.

• MeSH
• elektromagnetické záření MeSH
• koncentrace vodíkových iontů MeSH
• konfokální mikroskopie MeSH
• kultivační média MeSH
• počítačové zpracování obrazu MeSH
• pohyb těles MeSH
• polyfosfáty chemie   MeSH
• Saccharomyces cerevisiae chemie   cytologie   účinky záření   MeSH
• teplota MeSH
• vakuoly chemie   účinky záření   MeSH
• Publikační typ
• časopisecké články MeSH

Synthesis of cytosine, uracil, and 7-deazaadenine 2'-deoxyribonucleosides and triphosphates (dNTPs) bearing hexamethylated phenyl-bodipy fluorophore attached at position 5 of pyrimidines or at position 7 of 7-deazapurine was developed. All the title labeled nucleosides and dNTPs displayed bright green fluorescence with very high quantum yields. The modified dNmBdpTPs were good substrates to diverse DNA polymerases and were used for in vitro enzymatic synthesis of labeled DNA by primer extension or PCR. In combination with cationic cyclodextrin-peptide-based dNTP transporter, the dNmBdpTPs were successfully used for staining of genomic DNA in living cells for applications in confocal microscopy and in flow cytometry. The best performing cytosine nucleotide dCmBdpTP was used to monitor mitosis in live cells.

• MeSH
• adenin analogy a deriváty   chemie   MeSH
• barvení a značení MeSH
• cytosin chemie   MeSH
• deoxyribonukleosidy chemie   MeSH
• DNA analýza   MeSH
• fluorescenční barviva chemie   MeSH
• lidé MeSH
• metylace MeSH
• nádorové buněčné linie MeSH
• optické zobrazování MeSH
• polyfosfáty chemie   MeSH
• průtoková cytometrie MeSH
• puriny chemie   MeSH
• sloučeniny boru chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Zika virus is a global health threat due to significantly elevated risk of fetus malformations in infected pregnant women. Currently, neither an effective therapy nor a prophylactic vaccination is available for clinical use, desperately necessitating novel therapeutics and approaches to obtain them. Here, we present a structural model of the Zika virus RNA-dependent RNA polymerase (ZIKV RdRp) in complex with template and nascent RNAs, Mg2+ ions and accessing nucleoside triphosphate. The model allowed for docking studies aimed at effective pre-screening of potential inhibitors of ZIKV RdRp. Applicability of the structural model for docking studies was illustrated with the NITD008 artificial nucleotide that is known to effectively inhibit the function of the ZIKV RdRp. The ZIKV RdRp - RNA structural model is provided for all possible variations of the nascent RNA bases pairs to enhance its general utility in docking and modelling experiments. The developed model makes the rational design of novel nucleosides and nucleotide analogues feasible and thus provides a solid platform for the development of advanced antiviral therapy.

• MeSH
• adenosin analogy a deriváty   chemie   farmakologie   MeSH
• hořčík chemie   MeSH
• infekce virem zika genetika   virologie   MeSH
• konformace proteinů účinky léků   MeSH
• lidé MeSH
• molekulární modely MeSH
• nukleosidy chemie   MeSH
• nukleotidy chemie   MeSH
• polyfosfáty chemie   MeSH
• replikace viru genetika   MeSH
• RNA-dependentní RNA-polymerasa chemie   genetika   MeSH
• RNA chemie   genetika   MeSH
• simulace molekulového dockingu MeSH
• virové nestrukturální proteiny chemie   genetika   MeSH
• virus zika chemie   genetika   patogenita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

2'-Deoxyribonucleoside triphosphates (dNTPs) containing 5-(hydroxymethyl)cytosine (5hmC) protected with photocleavable groups (2-nitrobenzyl or 6-nitropiperonyl) were prepared and studied as substrates for the enzymatic synthesis of oligonucleotides and DNA containing a photocaged epigenetic 5hmC base. DNA probes containing photocaged or free 5hmC in the recognition sequence of restriction endonucleases were prepared and used for the study of the photorelease of caged DNA by UV or visible light at different wavelengths. The nitrobenzyl-protected dNTP was a slightly better substrate for DNA polymerases in primer extension or PCR, whereas the nitropiperonyl-protected nucleotide underwent slightly faster photorelease at 400 nm. However, both photocaged building blocks can be used in polymerase synthesis and the photorelease of 5hmC in DNA.

• MeSH
• 5-methylcytosin analogy a deriváty   chemická syntéza   chemie   MeSH
• deoxyribonukleosidy chemická syntéza   chemie   MeSH
• DNA chemická syntéza   chemie   MeSH
• fotochemické procesy MeSH
• polyfosfáty chemická syntéza   chemie   MeSH
• světlo MeSH
• ultrafialové záření MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Yeasts Cryptococcus humicola accumulated cadmium, cobalt, and iron (~ 50, 17, and 4% of the content in the medium, respectively) from the medium containing glucose, phosphate, and 2 mmol/L of metal salts. The effects of metal absorption on the levels of orthophosphate (Pi) and inorganic polyphosphate (polyP) varied for the metals under study. The levels of Pi and polyP increased in the case of cadmium and cobalt, respectively. In the case of iron, no changes in the levels of Pi and polyP were observed. Multiple DAPI-stained polyP inclusions were observed in the cytoplasm of cadmium-containing cells. The intensity of DAPI staining of the cell wall especially increased in case of cobalt and iron accumulation.

• MeSH
• biomasa MeSH
• Cryptococcus metabolismus   MeSH
• dusík metabolismus   MeSH
• kadmium chemie   metabolismus   farmakokinetika   MeSH
• kobalt chemie   metabolismus   farmakokinetika   MeSH
• polyfosfáty chemie   metabolismus   farmakokinetika   MeSH
• sorpční detoxikace MeSH
• železo chemie   metabolismus   farmakokinetika   MeSH
• Publikační typ
• časopisecké články MeSH

The scope of this work was to investigate the dependence of selected textural (texture profile analysis, TPA) and viscoelastic properties of processed cheese on the composition of ternary mixtures of emulsifying salts [disodium hydrogenphosphate, DSP; tetrasodium diphosphate, TSPP; sodium salt of polyphosphate (with mean length n ≈ 20), P20; and trisodium citrate, TSC] during a 60-d storage period (6±2°C). The processed cheese samples [40% wt/wt dry matter (DM) content, 50% wt/wt fat in DM content] were manufactured using Swiss-type cheese (as the main raw material) with 4 different maturity degrees (4, 8, 12, and 16 wk of ripening). Moreover, the pH of the samples was adjusted (the target values within the range of 5.60-5.80), corresponding to the standard pH values of spreadable processed cheese. With respect to the individual application of emulsifying salts (regardless of the maturity degree of the Swiss-type cheese applied), the samples prepared with P20 were the hardest, followed by those prepared with TSPP, TSC, and DSP. Furthermore, a specific ratio of DSP:TSPP (1:1) led to a significant increase in the hardness of the samples. On the whole, the hardness of all processed cheese samples increased with the prolonging storage period, whereas their hardness significantly dropped with the rising ripening stage of the raw material utilized. In all of the cases, the trends of hardness development remained analogous, and only the absolute values differed significantly. Moreover, the findings of TPA were in accordance with those of the rheological analysis. In particular, the specific ratio of DSP:TSPP (1:1) resulted in the highest gel strength and interaction factor values, followed by P20, TSPP, TSC, and DSP (used individually), reporting the same trend which was demonstrated by TPA. The monitored values of the gel strength and interaction factor decreased with the increasing maturity degree of the Swiss-type cheese used. The intensity of the rigidity of the samples showed an analogous relationship to the intensity of the gel strength; the higher the gel strength of the sample, the more inflexible the product is expected to be.

• MeSH
• koncentrace vodíkových iontů MeSH
• manipulace s potravinami MeSH
• polyfosfáty chemie   MeSH
• soli chemie   MeSH
• sýr * MeSH
• tvrdost MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The objective of this study was to describe the dependence of textural properties (hardness, cohesiveness, and relative adhesiveness) of processed cheese spreads on the proportion of disodium phosphate (DSP), tetrasodium diphosphate (TSPP), and sodium salts of polyphosphate in ternary mixtures of emulsifying salts. Sodium salts of polyphosphate with different mean lengths (n ≈ 5, 9, 13, 20, and 28) were used. Pentasodium triphosphate (PSTP) was used instead of TSPP in the second part of the study. Products with and without pH adjustment were tested (the target pH value was 5.60-5.80). Textural properties of the processed cheese were observed after 2, 9, and 30 d of storage at 6°C. Hardness of the processed cheese with a low content of polyphosphate increased at a specific DSP:TSPP ratio (~1:1 to 3:4). This trend was the same for all the polyphosphates used; only the absolute values of texture parameters were different. The same trends were observed in the ternary mixtures with PSTP, showing lower final values of hardness compared with samples containing TSPP. Hardness and cohesiveness decreased and relative adhesiveness increased in the samples with increased pH values and vice versa; the main trend remained unchanged.

• MeSH
• difosfáty chemie   MeSH
• emulze MeSH
• fosfáty chemie   MeSH
• koncentrace vodíkových iontů MeSH
• manipulace s potravinami metody   MeSH
• polyfosfáty chemie   MeSH
• skladování potravin MeSH
• soli chemie   MeSH
• sýr analýza   MeSH
• tvrdost MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

New fluorescent nucleosides and nucleoside triphosphate (dNTPs) analogs bearing the F-Bodipy fluorophore linked through a short, flexible nonconjugate tether were synthesized. The Bodipy-labeled dNTPs were substrates for several DNA polymerases which incorporated them into DNA in primer extension, nicking enzyme amplification reaction, and polymerase chain reaction. The fluorescence of F-Bodipy is not quenched upon incorporation in DNA and can be detected both in solutions and on gels.

• MeSH
• DNA-dependentní DNA-polymerasy metabolismus   MeSH
• DNA chemická syntéza   chemie   genetika   MeSH
• fluorescenční barviva chemie   MeSH
• nukleotidy chemie   MeSH
• polyfosfáty chemie   MeSH
• sekvence nukleotidů MeSH
• sloučeniny boru chemie   MeSH
• techniky syntetické chemie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Modified nucleosides (dA(R)s and dC(R)s) bearing bipyridine or terpyridine ligands attached through an octadiyne linker were prepared by single-step aqueous-phase Sonogashira cross-coupling of 7-iodo-7-deaza-2'-deoxyadenosine and 5-iodo-2'-deoxycytidine with the corresponding bipyridine- or terpyridine-octadiynes and were triphosphorylated to the corresponding nucleoside triphosphates (dA(R)TPs and dC(R)TPs). The modified dN(R)TPs were successfully incorporated into the oligonucleotides by primer extension experiment (PEX) using different DNA polymerases and the PEX products were used for post-synthetic complexation with divalent metal cations. The complexation of these DNAs containing flexibly-tethered ligands was compared with the previously reported ones bearing rigid acetylene-linked ligands suggesting the possible formation of both inter- and intra-strand complexes with Ni(2+) or Fe(2+).

• MeSH
• alkyny chemie   MeSH
• DNA chemie   MeSH
• kationty chemie   MeSH
• ligandy MeSH
• molekulární struktura MeSH
• nukleosidy chemická syntéza   chemie   MeSH
• organokovové sloučeniny chemická syntéza   chemie   MeSH
• polyfosfáty chemická syntéza   chemie   MeSH
• přechodné kovy chemie   MeSH
• pyridiny chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this study was to monitor the antibacterial effect of seven phosphate salts on selected strains of Gram-negative and Gram-positive bacteria, which could be considered responsible for food-borne diseases (Bacillus cereus, Bacillus subtilis, Enterococcus faecalis, Micrococcus luteus, Staphylococcus aureus, Citrobacter freundii, Escherichia coli, Proteus mirabilis, Salmonella enterica ser. Enteritidis and Pseudomonas aeruginosa). For these purposes, phosphates differing in chain length were used. The tested concentrations were in the range of 0.1-2.0% (wt v(-1)) applied at the model conditions. In the majority of cases the visible inhibitory effect on the growth of observed microorganisms could be seen. Due to the chemical structure of salts and their dissociation both the pH values of cultivation broth and similarly the growth characteristics of bacterial strains were affected. The inhibition of above mentioned bacteria was apparently supported by this dissociation. Phosphates obviously made the development of most Gram-positive bacteria impossible. Especially Micrococcus luteus was extremely sensitive to the presence of these substances. On the other hand, Gram-negative bacteria seemed to be resistant to the phosphate incidence. The exemption clause from the tested salts was represented by a high alkaline trisodium phosphate. It should be pointed out that generally the most significant antibacterial effects were shown by polyphosphates HEXA68 and HEXA70, trisodium phosphate undecahydrate, tetrasodium pyrophosphate and finally trisodium phosphate. By comparing the inhibitory effects of various phosphate salts can be concluded that the antibacterial activity was not determined only by the condensation degree but there was also proved the dependence on pH values.

• MeSH
• antibakteriální látky farmakologie   MeSH
• antibiotická rezistence MeSH
• fosfáty chemie   farmakologie   MeSH
• gramnegativní bakterie účinky léků   růst a vývoj   MeSH
• grampozitivní bakterie účinky léků   růst a vývoj   MeSH
• mikrobiální testy citlivosti MeSH
• polyfosfáty chemie   farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH