New bioanalytical SPE-HPLC-PDA-FL method for the determination of the neuroleptic drug tiapride and its N-desethyl metabolite was developed, validated and applied to xenobiochemical and pharmacokinetic studies in humans and animals. The sample preparation process involved solid-phase extraction of diluted plasma spiked with sulpiride (an internal standard) using SPE cartridges DSC-PH Supelco, USA. Chromatographic separation of the extracts was performed on a Discovery HS F5 250 mm × 4 mm (Supelco) column containing pentafluorophenylpropylsilyl silica gel. Mobile phase (acetonitrile-0.01 M phosphate buffer pH=3, flow rate 1 ml min(-1)) in the gradient mode was employed in the HPLC analysis. Tandem UV photodiode-array→fluorescence detection was used for the determination of the analytes. Low concentrations of tiapride and N-desethyl tiapride were determined using a more selective fluorescence detector (λ(exc.)/λ(emiss.)=232 nm/334 nm), high concentrations (500-6000 pmol ml(-1)) using a UV PDA detector at 212 nm with a linear response. Each HPLC run lasted 15 min. Lower limits of quantification (LLOQ) for tiapride (N-desethyl tiapride) were found to be 8.24 pmol ml(-1) (10.11 pmol ml(-1)). The recoveries of tiapride ranged from 89.3 to 94.3%, 81.7 to 86.8% for internal standard sulpiride and 90.9 to 91.8% for N-desethyl tiapride.

• MeSH
• extrakce na pevné fázi MeSH
• fluorescenční spektrometrie metody   MeSH
• jaterní mikrozomy metabolismus   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• limita detekce MeSH
• lineární modely MeSH
• mladý dospělý MeSH
• reprodukovatelnost výsledků MeSH
• spektrofotometrie ultrafialová metody   MeSH
• sulpirid krev   MeSH
• tiapamil-hydrochlorid analogy a deriváty   krev   farmakokinetika   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Photodynamic therapy (PDT) is an alternative method of tumour treatment. It is based on a photochemical reaction of a photosensitizer, irradiation, and O(2) which converts to cytotoxic (1)O(2) and other forms of reactive oxygen species (ROS). The comet assay (also called single-cell gel electrophoresis, SCGE) is a sensitive, simple and quantitative technique for detection of DNA damage. In our study we investigated the phototoxicity of the two porphyrin photosensitizers, TPPS4 and MgTPPS4, on HeLa cells. Three different radiation doses and six different concentrations of the photosensitizers were used. Our results show that the DNA of the cells treated with the TPPS(4) and MgTPPS(4) at the concentrations higher than 5 μM was highly fragmented indicating a strong phototoxic effect resulting in a cell apoptosis. On the base of our results we can hypothesize that even the irradiation dose of 1 J cm(-2) is sufficient enough to provoke the DNA fragmentation.

• MeSH
• apoptóza účinky léků   MeSH
• dávka záření MeSH
• fotochemoterapie metody   MeSH
• fotosenzibilizující látky aplikace a dávkování   farmakologie   MeSH
• fragmentace DNA účinky léků   MeSH
• HeLa buňky MeSH
• hořčík chemie   MeSH
• kometový test MeSH
• lidé MeSH
• metaloporfyriny aplikace a dávkování   chemie   farmakologie   MeSH
• porfyriny aplikace a dávkování   chemie   farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Photodynamic therapy (PDT) has been approved as proper and effective kind of treatment for certain types of cancer and non-malignant diseases. We tested photodynamic effects on G361 human melanoma cells sensitized by zinc-5,10,15,20-tetrakis(4-sulphonatophenyl) porphyrine (ZnTPPS(4)), chloraluminium phtalocyanine disulfonate (ClAlPcS(2)) and 5-aminolevulinic acid (ALA). In particular, we examined the PDT efficiency depending on applied light dose (0.8; 1.7; 3.3; 6.6; 13.2; 26.4Jcm(-2)). The DNA gel electrophoresis, methylthiazol tetrazolium bromide (MTT) viability test, fluorescent microscopy using calcein AM and propidium iodide (PI) staining, and rhodamine 123 mitochondrial membrane potential assay were performed to detect and evaluate the cell death process. We also measured the time course of reactive oxygen species (ROS) production and its dependence on sensitizer concentration within continuously irradiated sensitized cells. In conclusion, these results demonstrate most significant phototoxic effect of ClAlPcS(2)-PDT in spite of significantly higher ROS production induced by ZnTPPS(4)-PDT on G361 cells. On the other hand, ALA-PDT has a minimal photoeffect and induces negligible ROS formation in G361 cells at the conditions described below.

• MeSH
• barvicí látky MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• fluorescenční barviva MeSH
• fluorescenční mikroskopie MeSH
• fotochemoterapie MeSH
• fotosenzibilizující látky farmakologie   MeSH
• fragmentace DNA účinky léků   účinky záření   MeSH
• indoly farmakologie   MeSH
• kyselina aminolevulová farmakologie   MeSH
• lidé MeSH
• melanom experimentální terapie   MeSH
• membránové potenciály účinky léků   MeSH
• metaloporfyriny farmakologie   MeSH
• mitochondriální membrány účinky léků   MeSH
• nádorové buněčné linie MeSH
• organokovové sloučeniny farmakologie   MeSH
• propidium MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rhodamin 123 MeSH
• světlo MeSH
• tetrazoliové soli MeSH
• thiazoly MeSH
• vztah dávky záření a odpovědi MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Úvod a ciele: Poškodenia DNA spôsobené pôsobením reaktívnych foriem kyslíka (ROS) a nedostatočným fungovaním reparačných mechanizmov DNA sa dáva do súvislosti s nádorovými ochoreniami. Počiatočné štádium karcinogenézy sa spája so zmenami v molekule DNA, ktorými sú zlomy vlákien DNA, modifikácie dusíkatých zásad a cross-linky DNA proteíny. Hlavným produktom oxidačného poškodenia DNA je 8-oxo-7,8-dihydro-2´-deoxyguanozín (8-oxodG), ktorý je považovaný za potenciálny biomarker mutagenézy vyplývajúcej z oxidačného stresu. Cieľom tejto štúdie je sledovať oxidačné poškodenie DNA u pacientov s karcinómom prostaty a u zdravých jedincov, pochopenie mechanizmu, monitorovanie participácie oxidačného stresu v procese poškodenia biologicky dôležitých molekúl a jeho účasti v procese karcinogenézy. Materiál a metódy: Do štúdie, ktorej cieľom bola analýza hladín 8-oxodG s využitím modifikovanej kométovej metódy bolo zaradených 20 pacientov s karcinómom prostaty (priemerný vek 63,35 ± 1,8 roka) a 20 zdravých jedincov (priemerný vek 65,95 ± 1,8 roka). Štatistická analýza výsledkov sa robila pomocou Studentovho párového t-testu. Výsledky: Pacienti s karcinómom prostaty mali signifikantne vyššie hladiny 8-oxodG (p < 0,05) v porovnaní s kontrolnou skupinou. U pacientov s karcinómom prostaty sme zistili pozitívnu koreláciu medzi hladinami 8-oxodG a sérovým PSA, nie však Gleasonovým skóre. Záver: Výsledky našej štúdie poukázali na to, že zvýšený oxidačný stres môže zohrávať úlohu v patogenéze karcinómu prostaty ako dôsledok nerovnováhy oxidant/antioxidant v prospech oxidačného poškodenia DNA.

Introduction and Aims: Reactive oxygen species (ROS) mediated DNA damage in addition to ineffective DNA repair mechanisms are well established lesions common to human cancers. DNA alterations like strand breaks, base modifications and DNA-protein cross linkages are all strongly implicated in the initiation stage of carcinogenesis. Major oxidative DNA damage product 8-oxo-7,8-dihydro-2´-deoxyguanosine (8-oxodG) is regarded as a potential biomarker of mutagenesis consequent to oxidative stress. The aim of the present study was to detect the DNA oxidative damage in prostate cancer and in healthy individuals to understand the mechanism, to monitor participation of the oxidative stress in the process of damage of biologically important molecules and in the process of carcinogenesis. Material and Methods: The study included 20 prostate cancer patients (mean ± SEM: 63.35 ± 1.8 years) and 20 control subjects (mean ± SEM: 65.95 ± 1.8 years) for the analysis of 8-oxodG using a modified single cell gel electrophoresis (Comet assay). The results were analysed with the Student’s paired t-test. Results: The levels of 8-oxodG were significantly higher (p < 0.05) in prostate cancer patients than in controls. There was a positive correlation between the levels of 8-oxodG and serum PSA levels and we have found no correlation between 8-oxodG and Gleason score in prostate cancer patients. Conclusions: Our results demonstrate that an increased rate of oxidative stress might play a role in the pathogenesis of prostate cancer as evidenced by a failure in the oxidant/antioxidant balance in favour of DNA damage.

One of the main contributors to pharmaceutical pollution of surface waters are non-steroidal anti-inflammatory drugs (NSAIDs) that contaminate the food chain and affect non-target water species. As there are not many studies focusing on toxic effects of NSAIDs on freshwater fish species and specially effects after dietary exposure, we selected rainbow trout (Oncorhynchus mykiss) as the ideal model to examine the impact of two NSAIDs - diclofenac (DCF) and ibuprofen (IBP). The aim of our study was to test toxicity of environmentally relevant concentrations of these drugs together with exposure doses of 100× higher, including their mixture; and to deepen knowledge about the mechanism of toxicity of these drugs. This study revealed kidneys as the most affected organ with hyalinosis, an increase in oxidative stress markers, and changes in gene expression of heat shock protein 70 to be signs of renal toxicity. Furthermore, hepatotoxicity was confirmed by histopathological analysis (i.e. dystrophy, congestion, and inflammatory cell increase), change in biochemical markers, increase in heat shock protein 70 mRNA, and by oxidative stress analysis. The gills were locally deformed and showed signs of inflammatory processes and necrotic areas. Given the increase in oxidative stress markers and heat shock protein 70 mRNA, severe impairment of oxygen transport may be one of the toxic pathways of NSAIDs. Regarding the microbiota, an overgrowth of Gram-positive species was detected; in particular, significant dysbiosis in the Fusobacteria/Firmicutes ratio was observed. In conclusion, the changes observed after dietary exposure to NSAIDs can influence the organism homeostasis, induce ROS production, potentiate inflammations, and cause gut dysbiosis. Even the environmentally relevant concentration of NSAIDs pose a risk to the aquatic ecosystem as it changed O. mykiss health parameters and we assume that the toxicity of NSAIDs manifests itself at the level of mitochondria and proteins.

• MeSH
• antiflogistika nesteroidní metabolismus   MeSH
• biologické markery metabolismus   MeSH
• chemické látky znečišťující vodu * metabolismus   MeSH
• diklofenak metabolismus   MeSH
• dysbióza MeSH
• ekosystém MeSH
• epidemický výskyt choroby MeSH
• ibuprofen metabolismus   toxicita   MeSH
• kyslík metabolismus   MeSH
• léčivé přípravky metabolismus   MeSH
• messenger RNA metabolismus   MeSH
• Oncorhynchus mykiss * metabolismus   MeSH
• oxidační stres MeSH
• proteiny tepelného šoku HSP70 metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• střevní mikroflóra * MeSH
• voda metabolismus   MeSH
• zánět chemicky indukované   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• buňky * MeSH
• molekulární biologie MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• molekulární biologie, molekulární medicína
• NLK Publikační typ
• učebnice vysokých škol