A copper(II) tetrapyrazole-based complex of the composition of [Cu(tpyr)(H2O)(ONO2)]NO3 (1), where tpyr represents a tetradentate N-donor ligand formed by the condensation of 1H-pyrazole-5-carbaldehyde in NaOH/MeOH medium, has been prepared and characterized by elemental analysis, infrared spectroscopy, ultraviolet-visible spectroscopy, mass spectrometry, electron paramagnetic resonance and single-crystal X-ray diffraction. Spectrophotometric measurements demonstrated a remarkable peroxidase activity of the complex, which utilized hydrogen peroxide for the oxidation of phenolic compounds such as guaiacol or 3,5-dichloro-2-hydroxybenzene sulfonic acid. The optimum conditions for this reaction were found at pH 8 in ammonium bicarbonate buffer, although the activity was low but still detectable at pH 5-6 in ammonium acetate. As a peroxidase mimic, the complex exhibited enzyme-like Michaelis-Menten kinetics, showing a hyperbolic dependence of the reaction rate on hydrogen peroxide concentration. The determined Km and kcat values were 651 μmol·l-1 and 6.7 × 10-4 s-1, respectively, compared to 41 μmol·l-1 and 73 s-1 for horseradish peroxidase. EPR spectroscopy of the reaction mixture revealed no change in the copper (II) oxidation state during catalysis, suggesting that the oxidation of guaiacol may occur simultaneously with the reduction of hydrogen peroxide to water at the copper centre.

• Klíčová slova
• Copper(II), Crystal structure, MALDI-TOF, Peroxidase activity, Tetrapyrazole, XPS,
• MeSH
• biomimetické materiály * chemie   MeSH
• kinetika MeSH
• komplexní sloučeniny * chemie   chemická syntéza   MeSH
• krystalografie rentgenová MeSH
• měď * chemie   MeSH
• oxidace-redukce MeSH
• peroxid vodíku chemie   MeSH
• peroxidasa * chemie   MeSH
• pyrazoly * chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• komplexní sloučeniny * MeSH
• měď * MeSH
• peroxid vodíku MeSH
• peroxidasa * MeSH
• pyrazoly * MeSH

The glycoprotein clusterin (CLU) is involved in cell proliferation and DNA damage repair and is highly expressed in tumor cells. Here, we aimed to investigate the effects of CLU dysregulation on two human astrocytic cell lines: CCF-STTG1 astrocytoma cells and SV-40 immortalized normal human astrocytes. We observed that suppression of CLU expression by RNA interference inhibited cell proliferation, triggered the DNA damage response, and resulted in cellular senescence in both cell types tested. To further investigate the underlying mechanism behind these changes, we measured reactive oxygen species, assessed mitochondrial function, and determined selected markers of the senescence-associated secretory phenotype. Our results suggest that CLU deficiency triggers oxidative stress-mediated cellular senescence associated with pronounced alterations in mitochondrial membrane potential, mitochondrial mass, and expression levels of OXPHOS complex I, II, III and IV, indicating mitochondrial dysfunction. This report shows the important role of CLU in cell cycle maintenance in astrocytes. Based on these data, targeting CLU may serve as a potential therapeutic approach valuable for treating gliomas.

• Klíčová slova
• Astrocytes, Cellular senescence, Clusterin, Mitochondria, Oxidative stress,
• MeSH
• astrocyty * metabolismus   patologie   MeSH
• klusterin * nedostatek   metabolismus   MeSH
• lidé MeSH
• membránový potenciál mitochondrií MeSH
• mitochondrie metabolismus   MeSH
• nádorové buněčné linie MeSH
• oxidační stres MeSH
• oxidativní fosforylace MeSH
• poškození DNA MeSH
• proliferace buněk MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• stárnutí buněk * fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• CLU protein, human MeSH Prohlížeč
• klusterin * MeSH
• reaktivní formy kyslíku MeSH

Accumulating evidence suggests that manganese oxide nanoparticles (NPs) show multiple enzyme-mimicking antioxidant activities, which supports their potential in redox-targeting therapeutic strategies for diseases with impaired redox signaling. However, the systemic administration of any NP requires thorough hemocompatibility testing. In this study, we assessed the hemocompatibility of synthesized Mn3O4 NPs, identifying their ability to induce spontaneous hemolysis and eryptosis or impair osmotic fragility. Concentrations of up to 20 mg/L were found to be safe for erythrocytes. Eryptosis assays were shown to be more sensitive than hemolysis and osmotic fragility as markers of hemocompatibility for Mn3O4 NP testing. Flow cytometry- and confocal microscopy-based studies revealed that eryptosis induced by Mn3O4 NPs was accompanied by Ca2+ overload, altered redox homeostasis verified by enhanced intracellular reactive oxygen species (ROS) and reactive nitrogen species (RNS), and a decrease in the lipid order of cell membranes. Furthermore, Mn3O4 NP-induced eryptosis was calpain- and caspase-dependent.

• Klíčová slova
• calcium signaling, cytotoxicity, eryptosis, nanoparticles, oxidative stress, regulated cell death,
• MeSH
• buněčná membrána * metabolismus   účinky léků   MeSH
• eryptóza * účinky léků   MeSH
• erytrocyty účinky léků   metabolismus   MeSH
• hemolýza účinky léků   MeSH
• kalpain * metabolismus   MeSH
• kaspasy * metabolismus   MeSH
• lidé MeSH
• nanočástice * chemie   MeSH
• oxidy * farmakologie   chemie   MeSH
• reaktivní formy dusíku * metabolismus   MeSH
• reaktivní formy kyslíku * metabolismus   MeSH
• sloučeniny manganu * farmakologie   chemie   MeSH
• vápník * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kalpain * MeSH
• kaspasy * MeSH
• manganese oxide MeSH Prohlížeč
• oxidy * MeSH
• reaktivní formy dusíku * MeSH
• reaktivní formy kyslíku * MeSH
• sloučeniny manganu * MeSH
• vápník * MeSH

The objective of our in vitro study was to quantify the biochemical profile where the total polyphenol, flavonoid and phenolic acid content was determined. The antioxidant potential of microgreen extract from Trigonella foenum-graecum L., was measured molybdenum reducing power assay. Specifically, the study assessed parameters such as metabolic activity (AlamarBlueTM assay), membrane integrity (CFDA-AM assay), mitochondrial potential (JC-1 assay), as well as reactive oxygen species generation (NBT assay). In addition, the steroid hormone release in TM3 murine Leydig cells after 12 h and 24 h exposures were quantified by enzyme-linked immunosorbent assay. The gained results indicate the highest value in total flavonoid content (182.59+/-2.13 mg QE) determination, supported by a significant (108.25+/-1.27 mg TE) antioxidant activity. The effects on metabolic activity, cell membrane integrity, and mitochondrial membrane potential were found to be both time- and dose-dependent. Notably, a significant suppression in reactive oxygen species generation was confirmed at 150, 200 and 250 microg/ml after 24 h exposure. In addition, progesterone and testosterone release was stimulated up to 250 microg/ml dose of Trigonella, followed by a decline in both steroid production at 300 and 1000 microg/ml. Our results indicate, that Trigonella at lower experimental doses (up to 250 microg/ml) may positively affect majority of monitored cell parameters in TM3 Leydig cells. Overleaf, increasing experimental doses may negatively affect the intracellular parameters already after 12 h of in vitro exposure. Key words Microgreens, Trigonella foenum-graecum L., Fenugreek, Leydig cells, Male reproduction.

• MeSH
• antioxidancia farmakologie   MeSH
• buněčné linie MeSH
• fytonutrienty farmakologie   MeSH
• Leydigovy buňky * účinky léků   metabolismus   MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• myši MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rostlinné extrakty * farmakologie   MeSH
• testosteron metabolismus   MeSH
• Trigonella * chemie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antioxidancia MeSH
• fytonutrienty MeSH
• reaktivní formy kyslíku MeSH
• rostlinné extrakty * MeSH
• testosteron MeSH

To explore the effects and underlying mechanisms of Mdivi-1 on three common clinical models of acute kidney injury (AKI). Three common AKI cell models were constructed, classified into the control group (human renal tubular epithelial cells [HK-2] cells), the Iohexol group (HK-2 cells treated with Iohexol), the Genta group (HK-2 cells treated with Gentamicin), and the Cis group (HK-2 cells treated with Cisplatin). To explore the optimal protective concentration of Mdivi-1 for each AKI cell model, the experimental design consisted of the following seven groups: the control group (HK-2 cells cultured in medium), three injury groups (HK-2 cells subjected to Iohexol, Gentamicin, or Cisplatin), and the corresponding protection groups (with a certain concentration of Mdivi-1 added to each injury group). Cellular survival and apoptosis, reactive oxygen species (ROS) levels, and the expression of recombinant Sirtuin 3 (SIRT3) in each group were measured. Mitochondrial fission and fusion dynamics in cells were observed under an electron microscope. To explore relevant pathways, the changes in relevant pathway proteins were analyzed through Western blotting. The half maximal inhibitory concentration (IC50) values were 150.06 mgI/ml at 6 h in the Iohexol group, 37.88 mg/ml at 24 h in the Gentamicin group, and 13.48 microM at 24 h in the Cisplatin group. Compared with the control group, the three injury groups showed increased cell apoptosis rates and higher expressions of apoptotic proteins in HK-2 cells, with an accompanying decrease in cell migration. After the addition of corresponding concentrations of Mdivi-1, the optimal concentrations were 3 µM in the Iohexo-3 group, 1 microM in the Genta-1 group, and 5 µM in the Cis-5 group, HK-2 cells showed the highest survival rate, reduced apoptosis, decreased mitochondrial ROS and SIRT3 expression, and reduced mitochondrial fission and autophagy when compared with each injury group. Further verification with Western blot analysis after the addition of Mdivi-1 revealed a reduction in the expressions of mitochondrial fission proteins DRP1, Nrf2, SIRT3, Caspase-3, Jun N-terminal Kinase (JNK)/P-JNK, NF-kappaB, Bcl2, and autophagic protein P62, as well as reduced ROS levels. Mdivi-1 had protective effects on the three common AKI cell models by potentially reducing mitochondrial fission in cells and inhibiting the production of ROS through the mediation of the NF- B/JNK/SIRT3 signaling pathway, thereby exerting protective effects. Key words AKI, Cisplatin, Gentamicin, Iohexol, Mdivi-1.

• MeSH
• akutní poškození ledvin * metabolismus   patologie   farmakoterapie   MeSH
• apoptóza účinky léků   MeSH
• buněčné linie MeSH
• lidé MeSH
• MAP kinasový signální systém účinky léků   fyziologie   MeSH
• mitochondriální dynamika * účinky léků   fyziologie   MeSH
• NF-kappa B * metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• signální transdukce * účinky léků   MeSH
• sirtuin 3 * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• NF-kappa B * MeSH
• reaktivní formy kyslíku MeSH
• SIRT3 protein, human MeSH Prohlížeč
• sirtuin 3 * MeSH

Bone marrow stromal cells (BMSCs) serve as a valuable reservoir of multipotent stem cells important in the regulation of bone homeostasis and energy metabolism. Here, we present a protocol for isolating human BMSCs (hBMSCs) and characterizing their cellular metabolism related to hBMSC functional properties. We describe steps for bioenergetics, cell senescence, and production of reactive oxygen species (ROS), together with description of the data analysis. These assays provide information on hBMSC metabolic status valuable to regenerative medicine and therapeutic applications. For complete details on the use and execution of this protocol, please refer to Tencerova et al.1.

• Klíčová slova
• Cell Biology, Metabolism, Stem Cells,
• MeSH
• buněčné kultury metody   MeSH
• buňky kostní dřeně cytologie   metabolismus   MeSH
• energetický metabolismus fyziologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky * metabolismus   cytologie   MeSH
• reaktivní formy kyslíku * metabolismus   MeSH
• separace buněk metody   MeSH
• stárnutí buněk fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• reaktivní formy kyslíku * MeSH

Plasma-activated water (PAW) has been shown to have antimicrobial properties, making it a promising tool for surface decontamination. This study evaluated the ability of PAW generated from high voltage atmospheric cold plasma to remove Salmonella from common surfaces (stainless steel (SS), polyvinyl chloride (PVC), concrete, and wood) found in poultry houses. PAW was generated by exposing distilled water to atmospheric cold plasma in 80% humid air at 90 kV and 60 Hz for 30 min. The resulting PAW contained 1120 ppm of nitrate and 1370 ppm of hydrogen peroxide, with a pH of 1.83. PAW was then applied to coupons of SS, PVC, wood, and concrete surfaces inoculated with 7-8 log10 CFU of cocktail of Salmonella spp. (S. Typhimurium, S. Newport, S. Montevideo, and S. Enteritidis). PAW effectively reduced Salmonella levels on SS and PVC surfaces to below the detection limit within 30 s. On wood surfaces, a longer treatment time of 7.5 min was required to achieve a maximum reduction of 2.63 log10 CFU, likely due to the porosity of the wood limiting PAW contact with the bacteria. On concrete surfaces, the reduction in Salmonella levels was only 0.98 log10 CFU. This was likely due to the greater surface roughness and high alkalinity, which neutralized the PAW species.

• Klíčová slova
• Cold plasma, Decontamination, Plasma-activated water, Poultry house, Salmonella spp,
• MeSH
• bydlení zvířat * MeSH
• dekontaminace * metody   MeSH
• dezinficiencia farmakologie   MeSH
• dřevo mikrobiologie   chemie   MeSH
• drůbež * mikrobiologie   MeSH
• konstrukční materiály mikrobiologie   MeSH
• nerezavějící ocel MeSH
• peroxid vodíku farmakologie   MeSH
• plazmové plyny * farmakologie   MeSH
• počet mikrobiálních kolonií MeSH
• polyvinylchlorid chemie   MeSH
• Salmonella * růst a vývoj   MeSH
• voda * chemie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• dezinficiencia MeSH
• nerezavějící ocel MeSH
• peroxid vodíku MeSH
• plazmové plyny * MeSH
• polyvinylchlorid MeSH
• voda * MeSH

Clover yellow vein virus (ClYVV), a potyvirus that infects various dicotyledonous plants, poses a significant threat to the cultivation of legumes. Although potyviral NIa-Pro was extensively studied in viral infection cycle and host antiviral responses, the contribution of NIa-Pro protease activity to virus systemic symptoms has not yet been reported. In this study, we developed infectious clones of a ClYVV isolated from Pisum sativum. The rescued ClYVV showed robust infectivity and induced obvious systemic mosaic and necrosis symptoms in the model host Nicotiana benthamiana and natural hosts Pisum sativum and Vicia faba. Using a potato virus X (PVX) vector to express 11 ClYVV proteins in N. benthamiana ectopically, we identified that NIa-Pro is the key determinant in inducing systemic symptoms and causes higher leaf ROS levels and cell death. Further, we found that the protease-inactive mutant NIa-ProC151A causes significantly reduced systemic symptoms when expressed via the PVX vector and does not induce higher cellular ROS levels and cell death when transiently overexpressed compared to wild-type NIa-Pro. Overall, this study provides evidence supporting that the protease activity of a potyvirus protein NIa-Pro directly contributes to the virus symptoms.

• Klíčová slova
• CIYVV, Infectious clone, NIa-pro, Nicotiana benthamiana, PVX,
• MeSH
• hrách setý virologie   MeSH
• listy rostlin virologie   MeSH
• nemoci rostlin * virologie   MeSH
• Potyvirus * patogenita   genetika   fyziologie   enzymologie   MeSH
• proteasy metabolismus   genetika   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• tabák * virologie   MeSH
• Vicia faba virologie   MeSH
• virové proteiny * metabolismus   genetika   MeSH
• virulence MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• proteasy MeSH
• reaktivní formy kyslíku MeSH
• virové proteiny * MeSH

Silymarin is an extract obtained from the seeds of milk thistle (Sylibum marianum L., Asteraceae) and contains several structurally related flavonolignans and a small family of flavonoids. Mouse spleen cells represent highly sensitive primary cells suitable for studying the pharmacological potential and biofunctional properties of natural substances. Cultivation of splenocytes for 24 h under standard culture conditions (humidity, 37 °C, 5% CO2, atmospheric oxygen) resulted in decreased viability of splenocytes compared to intact cells. A cytoprotective effect of silybin (SB), silychristin (SCH) and 2,3-dehydrosilybin (DHSB) was observed at concentrations as low as 5 µmol/ml. At 50 µmol/ml, these substances restored and/or stimulated viability and mitochondrial membrane potential and had anti-apoptotic effect in the order SB > DHSB > SCH. The substances demonstrated a concentration-dependent activity in restoring the redox balance based on the changes in the concentration of reactive oxygen species (ROS), hydrogen peroxide (H2O2) and nitric oxide. This was in the order DHSB > SCH > SB, which correlated with the suppressed expression of nuclear factor erythroid 2-related factor 2 (Nrf2), catalase and glutathione peroxidase. The strong stimulation of the superoxide dismutase 1 gene converting ROS to H2O2 points to its dominant role in the maintaining redox homeostasis in splenocytes, which was disrupted by oxidative stress due to non-physiological culture conditions. Our study showed significant differences in the cytoprotective, antioxidant and anti-apoptotic activities of SB, SCH, and DHSB on splenocytes exposed to mild and AAPH-induced oxidative stress.

• Klíčová slova
• 2,3-dehydrosilybin, Apoptosis, Mouse splenocytes, Redox balance, Silybin, Silychristin, Viability,
• MeSH
• antioxidancia * farmakologie   MeSH
• apoptóza * účinky léků   MeSH
• cytoprotekce účinky léků   MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• myši inbrední BALB C * MeSH
• myši MeSH
• oxidační stres účinky léků   MeSH
• peroxid vodíku MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• silibinin * farmakologie   MeSH
• silymarin * farmakologie   MeSH
• slezina * cytologie   účinky léků   metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antioxidancia * MeSH
• peroxid vodíku MeSH
• reaktivní formy kyslíku MeSH
• silibinin * MeSH
• silychristin MeSH Prohlížeč
• silymarin * MeSH

Bilirubin (BR) is a water-insoluble product of heme catabolism in mammals. Elevated blood concentrations of BR, especially in the neonatal period, are treated with blue-green light phototherapy. The major mechanism of BR elimination during phototherapy is photoisomerization, while a minor, less studied mechanism of degradation is oxidation. In this work, we studied the oxidation of the bilirubin model tetramethyl-dipyrrinone (Z-13) by singlet oxygen in methanol using UV-vis and ESI-MS spectroscopy, resulting in propentdyopents as the main oxidation products. We also identified two additional intermediates that were formed during the reaction (hydroperoxide 21a and imine 17). The structure of the hydroperoxide was confirmed by helium-tagging IR spectroscopy. Such reaction intermediates formed during the oxidation of BR or bilirubin models have not been described so far. We believe that this work can be used as a first step in studying the complex oxidation mechanism of BR during phototherapy.

• MeSH
• bilirubin chemie   MeSH
• fotochemické procesy MeSH
• metamizol chemie   MeSH
• molekulární struktura MeSH
• oxidace-redukce * MeSH
• singletový kyslík * chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bilirubin MeSH
• metamizol MeSH
• singletový kyslík * MeSH