The potential of nanomaterials use is huge, especially in fields such as medicine or industry. Due to widespread use of nanomaterials, their cytotoxicity and involvement in cellular pathways ought to be evaluated in detail. Nanomaterials can induce the production of a number of substances in cells, including reactive oxygen species (ROS), participating in physiological and pathological cellular processes. These highly reactive substances include: superoxide, singlet oxygen, hydroxyl radical, and hydrogen peroxide. For overall assessment, there are a number of fluorescent probes in particular that are very specific and selective for given ROS. In addition, due to the involvement of ROS in a number of cellular signaling pathways, understanding the principle of ROS production induced by nanomaterials is very important. For defense, the cells have a number of reparative and especially antioxidant mechanisms. One of the most potent antioxidants is a tripeptide glutathione. Thus, the glutathione depletion can be a characteristic manifestation of harmful effects caused by the prooxidative-acting of nanomaterials in cells. For these reasons, here we would like to provide a review on the current knowledge of ROS-mediated cellular nanotoxicity manifesting as glutathione depletion, including an overview of approaches for the detection of ROS levels in cells.

• Klíčová slova
• cell injury, fluorescence probes, glutathione, nanotoxicity, oxidative stress, reactive oxygen species,
• MeSH
• buňky účinky léků   metabolismus   MeSH
• glutathion metabolismus   MeSH
• lidé MeSH
• nanostruktury toxicita   MeSH
• oxidační stres účinky léků   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• glutathion MeSH
• reaktivní formy kyslíku MeSH

Translationally controlled tumor protein (TCTP) is a multifunctional and highly conserved protein from yeast to humans. Recently, its role in non-selective autophagy has been reported with controversial results in mammalian and human cells. Herein we examine the effect of Mmi1, the yeast ortholog of TCTP, on non-selective autophagy in budding yeast Saccharomyces cerevisiae, a well-established model system to monitor autophagy. We induced autophagy by nitrogen starvation or rapamycin addition and measured autophagy by using the Pho8Δ60 and GFP-Atg8 processing assays in WT, mmi1Δ, and in autophagy-deficient strains atg8Δ or atg1Δ. Our results demonstrate that Mmi1 does not affect basal or nitrogen starvation-induced autophagy. However, an increased rapamycin-induced autophagy is detected in mmi1Δ strain when the cells enter the post-diauxic growth phase, and this phenotype can be rescued by inserted wild-type MMI1 gene. Further, the mmi1Δ cells exhibit significantly lower amounts of reactive oxygen species (ROS) in the post-diauxic growth phase compared to WT cells. In summary, our study suggests that Mmi1 negatively affects rapamycin-induced autophagy in the post-diauxic growth phase and supports the role of Mmi1/TCTP as a negative autophagy regulator in eukaryotic cells.

• Klíčová slova
• Mmi1, TCTP, autophagy, nitrogen starvation, rapamycin, reactive oxygen species, translationally controlled tumor protein,
• MeSH
• autofagie * účinky léků   MeSH
• dusík nedostatek   MeSH
• glukosa farmakologie   MeSH
• mutace genetika   MeSH
• nádorové biomarkery chemie   MeSH
• proteiny vázající vápník metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny metabolismus   MeSH
• Saccharomyces cerevisiae cytologie   účinky léků   růst a vývoj   MeSH
• sirolimus farmakologie   MeSH
• superoxidy metabolismus   MeSH
• translačně kontrolovaný nádorový protein 1 MeSH
• zelené fluorescenční proteiny metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dusík MeSH
• glukosa MeSH
• nádorové biomarkery MeSH
• proteiny vázající vápník MeSH
• Saccharomyces cerevisiae - proteiny MeSH
• sirolimus MeSH
• superoxidy MeSH
• TMA19 protein, S cerevisiae MeSH Prohlížeč
• TPT1 protein, human MeSH Prohlížeč
• translačně kontrolovaný nádorový protein 1 MeSH
• zelené fluorescenční proteiny MeSH

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.

• MeSH
• alergologie a imunologie normy   MeSH
• fenotyp MeSH
• konsensus MeSH
• lidé MeSH
• průtoková cytometrie metody   normy   MeSH
• separace buněk metody   normy   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• DNA analýza   MeSH
• falešně pozitivní reakce MeSH
• imunofenotypizace MeSH
• imunologické techniky * MeSH
• lidé MeSH
• proliferace buněk MeSH
• průtoková cytometrie MeSH
• řízení kvality MeSH
• RNA analýza   MeSH
• separace buněk MeSH
• směrnice jako téma * MeSH
• software MeSH
• T-lymfocyty cytologie   MeSH
• výzkumný projekt MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• DNA MeSH
• RNA MeSH

Methyltriphenylphosphonium (TPMP) salts have been widely used to measure the mitochondrial membrane potential and the triphenylphosphonium (TPP+) moiety has been attached to many bioactive compounds including antioxidants to target them into mitochondria thanks to their high affinity to accumulate in the mitochondrial matrix. The adverse effects of these compounds on cellular metabolism have been insufficiently studied and are still poorly understood. Micromolar concentrations of TPMP cause a progressive inhibition of cellular respiration in adherent cells without a marked effect on mitochondrial coupling. In permeabilized cells the inhibition was limited to NADH-linked respiration. We found a mixed inhibition of the Krebs cycle enzyme 2-oxoglutarate dehydrogenase complex (OGDHC) with an estimated IC50 3.93 [3.70-4.17] mM, which is pharmacologically plausible since it corresponds to micromolar extracellular concentrations. Increasing the lipophilic character of the used TPP+ compound further potentiates the inhibition of OGDHC activity. This effect of TPMP on the Krebs cycle ought to be taken into account when interpreting observations on cells and mitochondria in the presence of TPP+ derivatives. Compounds based on or similar to TPP+ derivatives may also be used to alter OGDHC activity for experimental or therapeutic purposes.

• MeSH
• buněčné linie MeSH
• citrátový cyklus účinky léků   MeSH
• citrátsynthasa účinky léků   metabolismus   MeSH
• glutamátdehydrogenasa účinky léků   metabolismus   MeSH
• isocitrátdehydrogenasa účinky léků   metabolismus   MeSH
• ketoglutarátdehydrogenasový komplex antagonisté a inhibitory   metabolismus   MeSH
• kosterní svaly enzymologie   MeSH
• krysa rodu Rattus MeSH
• malátdehydrogenasa účinky léků   metabolismus   MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• oniové sloučeniny farmakologie   MeSH
• potkani Wistar MeSH
• pyruvátdehydrogenasový komplex účinky léků   metabolismus   MeSH
• svalové mitochondrie účinky léků   enzymologie   MeSH
• tritylové sloučeniny farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• citrátsynthasa MeSH
• glutamátdehydrogenasa MeSH
• isocitrátdehydrogenasa MeSH
• ketoglutarátdehydrogenasový komplex MeSH
• malátdehydrogenasa MeSH
• oniové sloučeniny MeSH
• pyruvátdehydrogenasový komplex MeSH
• triphenylmethylphosphonium MeSH Prohlížeč
• tritylové sloučeniny MeSH

BACKGROUND: The lipophilic positively charged moiety of triphenylphosphonium (TPP+) has been used to target a range of biologically active compounds including antioxidants, spin-traps and other probes into mitochondria. The moiety itself, while often considered biologically inert, appears to influence mitochondrial metabolism. METHODOLOGY/PRINCIPAL FINDINGS: We used the Seahorse XF flux analyzer to measure the effect of a range of alkylTPP+ on cellular respiration and further analyzed their effect on mitochondrial membrane potential and the activity of respiratory complexes. We found that the ability of alkylTPP+ to inhibit the respiratory chain and decrease the mitochondrial membrane potential increases with the length of the alkyl chain suggesting that hydrophobicity is an important determinant of toxicity. CONCLUSIONS/SIGNIFICANCE: More hydrophobic TPP+ derivatives can be expected to have a negative impact on mitochondrial membrane potential and respiratory chain activity in addition to the effect of the biologically active moiety attached to them. Using shorter linker chains or adding hydrophilic functional groups may provide a means to decrease this negative effect.

• MeSH
• buněčné linie MeSH
• elektronový transportní řetězec metabolismus   MeSH
• heterocyklické sloučeniny farmakologie   MeSH
• krysa rodu Rattus MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• organofosforové sloučeniny farmakologie   MeSH
• potkani Wistar MeSH
• spotřeba kyslíku účinky léků   MeSH
• svalové mitochondrie metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• elektronový transportní řetězec MeSH
• heterocyklické sloučeniny MeSH
• organofosforové sloučeniny MeSH
• tris(o-phenylenedioxy)cyclotriphosphazene MeSH Prohlížeč