Reactive oxygen species play a key role in cellular homeostasis and redox signaling at physiological levels, where excessive production affects the function and integrity of macromolecules, specifically proteins. Therefore, it is important to define radical-mediated proteotoxic stress in macrophages and identify target protein to prevent tissue dysfunction. A well employed, THP-1 cell line was utilized as in vitro model to study immune response and herein we employ immuno-spin trapping technique to investigate radical-mediated protein oxidation in macrophages. Hydroxyl radical formation along macrophage differentiation was confirmed by electron paramagnetic resonance along with confocal laser scanning microscopy using hydroxyphenyl fluorescein. Lipid peroxidation product, malondialdehyde, generated under experimental conditions as detected using swallow-tailed perylene derivative fluorescence observed by confocal laser scanning microscopy and high-performance liquid chromatography, respectively. The results obtained from this study warrant further corroboration and study of specific proteins involved in the macrophage activation and their role in inflammations.
Příčina neplodnosti infertilního páru v polovině případů stojí na straně muže. Oxidační stres je jedním z důvodů zhoršené plodnosti muže. Článek rozebírá etiologii a mechanizmus vzniku oxidačního stresu a shrnuje současné poznatky o možnostech diagnostiky a léčby oxidačního stresu spermií.
In half of the cases of infertility in infertile pairs is due to the pathology of the man. Oxidative stress is one of the reasons of impaired fertility in a male. This article analyses the etiology and mechanism of oxidative stress and summarizes the current state of knowledge regarding the diagnostics options and treatment of oxidative stress of sperm.
- Klíčová slova
- koenzym Q10,
- MeSH
- alfa-tokoferol terapeutické užití MeSH
- antioxidancia * terapeutické užití MeSH
- koenzymy terapeutické užití MeSH
- kyselina askorbová terapeutické užití MeSH
- lidé MeSH
- mužská infertilita * etiologie farmakoterapie MeSH
- oxidační stres * účinky léků MeSH
- potravní doplňky MeSH
- selen terapeutické užití MeSH
- spermie fyziologie účinky léků MeSH
- varikokéla komplikace MeSH
- vitamin E terapeutické užití MeSH
- volné radikály analýza MeSH
- zinek terapeutické užití MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- přehledy MeSH
x
- MeSH
- antibakteriální látky farmakologie MeSH
- dihydroxyfenylalanin * farmakologie chemická syntéza MeSH
- dopaminové látky farmakologie chemická syntéza MeSH
- elektronová paramagnetická rezonance * statistika a číselné údaje MeSH
- fixní kombinace léků MeSH
- inhibitory syntézy proteinů farmakologie MeSH
- kyslík * fyziologie MeSH
- melaniny * farmakologie chemická syntéza MeSH
- streptomycin * farmakologie MeSH
- volné radikály analýza MeSH
- Publikační typ
- práce podpořená grantem MeSH
BACKGROUND: At present time the number of implantations of joint replacements as well as their revisions increases. Higher demands are required on the quality and longevity of implants. The aim of this work was to determine the degree of oxidative degradation and the amount of free/residual radicals in selected ultra-high molecular weight polyethylene (UHMWPE) components of the joint replacements and demonstrate that the measured values are closely connected with quality and lifetime of the polymer components. METHODS: We tested both new (4 samples) and explanted (4 samples) UHMWPE polymers for total joint replacements. The samples were characterized by infrared spectroscopy (IR), electron spin resonance (ESR) and microhardness (MH) test. The IR measurements yielded the values of oxidation index and trans-vinylene index. The ESR measurements gave the free radicals concentration. RESULTS: In the group of new polyethylene components, we found oxidation index values ranging from 0.00-0.03 to 0.24. The trans-vinylene index values ranged from 0.044 to 0.080. The value of free radical concentration was zero in virgin and also in sample of Beznoska Company and non-zero in the other samples. In the group of explanted components, the measured values were associated with their history, micromechanical properties and performance in vivo. CONCLUSIONS: We demonstrated that measuring of oxidative damage may help the orthopaedic surgeon in estimating the quality of UHMWPE replacement component and thus radically to avoid early joint replacement failure due to worse polyethylene quality.
- MeSH
- biokompatibilní materiály chemie MeSH
- elektronová paramagnetická rezonance MeSH
- krystalizace MeSH
- kyčelní protézy * MeSH
- lidé MeSH
- oxidace-redukce MeSH
- polyethyleny chemie MeSH
- protézy kolene * MeSH
- reoperace MeSH
- selhání protézy MeSH
- spektrofotometrie infračervená MeSH
- testování materiálů MeSH
- tvrdost MeSH
- volné radikály analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Melanins are polymorphous and multifunctional biopolymers with a relatively high concentration of free radicals. EPR spectroscopy was used to study o-semiquinone free radicals in model eumelanins synthesized from 3,4-dihydroxyphenylalanine (DOPA) and tyrosine in the presence of tyrosinase, and melanins isolated from A-375 and G-361 human melanoma malignum cells exposed to two compounds: 5,7-dimethoxycoumarin (DMC) and valproic acid (VPA). Changes were determined in the concentrations of free radicals in the individual melanins from tumour cells treated with DMC and VPA. A strong decrease in the concentrations of free radicals characterizes melanins isolated from tumour cells treated together with DMC and VPA. Slow spin-lattice relaxation processes were noted in the melanins tested with homogeneous broadened EPR spectra. The EPR technique may be useful not only for the elucidation of free radicals in melanins from A-375 and G-361 cells treated with VPA and DMC but it could also be applied to establish the relationship between melanin type and the malignancy of melanoma malignum.
- Klíčová slova
- 5,7-dimethoxycoumarin,
- MeSH
- biopolymery analýza MeSH
- elektronová paramagnetická rezonance * metody statistika a číselné údaje využití MeSH
- kumariny farmakologie MeSH
- kyselina valproová farmakologie MeSH
- lidé MeSH
- melaniny * analýza MeSH
- melanom MeSH
- nádorové buněčné linie cytologie účinky léků MeSH
- volné radikály * analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
In the food industry, in the process of creating new agricultural plant products, and in the testing of anti-cancer drugs there is often a need to assay multiple samples of low molecular weight antioxidants, plant samples and foods rich in antioxidants, with minimal additional costs and low degrees of uncertainty. With these demands in mind, we decided to study the fully automated assay of antioxidants using not only automated sample measurements but also automated processing of samples and application of reagents. The automated pipetting system epMotion 5075 and the automated spectrophotometer BS 400 were chosen for the assay purposes. Five methods were introduced for the automation: 2-diphenyl-1-picrylhydrazyl (DPPH) test, ferric reducing antioxidant power (FRAP) method, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) based test, N,N-dimethyl-1,4-diaminobenzene (DMPD) based test and the free radicals method. Samples containing one of the four antioxidants (standard rutin, quercitrin, ferulic and gallic acid) in a range 1–1000 μg/ml were used throughout. All of the tested methods were found suitable for implementation in an automated assay. However, some of them, such as the ABTS test failed to assay all tested antioxidants. The coefficients of determination were also unequal. From the analytical point of view, FRAP methods provided the most reliable results in the automated assay; because of the capacity of the method, approximately 240 samples per hour (one sample per 15 seconds) can be assayed using the automated protocol. We were encouraged by the data received and we expect further interest in the practical performance of such automation. As a mean of testing the robustness of our method, in the next step of our study, oxidative status was assessed in model cell lines derived from prostate cancer (PC-3, PNT1A and 22RV1) that were cultured on ellipticine (0, 0.5, 1, 1.5, 2, 2.5, 5, 7.5, 10, 15 μmol/l) supplemented agar. Antioxidant activity was assessed (DPPH, ABTS, FRAP, DMPD, FR) and calculated on the phenolic antioxidant level (rutin, quercitrin, ferulic and gallic acid), and thus an estimation was formulated of the oxidative stress as a result of the impact of anti-cancer drugs. It can be demonstrated that the new method has wide applicability.
- MeSH
- antioxidancia analýza MeSH
- chemické techniky analytické metody přístrojové vybavení statistika a číselné údaje MeSH
- elipticiny analýza chemie metabolismus MeSH
- financování organizované MeSH
- FRAP MeSH
- kalibrace MeSH
- kyselina gallová analýza chemie MeSH
- kyseliny kumarové analýza chemie MeSH
- laboratorní automatizace metody přístrojové vybavení MeSH
- luminiscenční měření MeSH
- nádorové buněčné linie MeSH
- oxidační stres účinky léků MeSH
- protinádorové látky chemie metabolismus toxicita MeSH
- quercetin analogy a deriváty analýza chemie MeSH
- reprodukovatelnost výsledků MeSH
- rutin analýza chemie MeSH
- screeningové testy protinádorových léčiv metody statistika a číselné údaje MeSH
- spektrofotometrie metody přístrojové vybavení statistika a číselné údaje MeSH
- viabilita buněk účinky léků MeSH
- volné radikály analýza MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- hodnotící studie MeSH
- statistiky MeSH
- tabulky MeSH
The oxidative effects of photodynamic therapy with 5,10,15,20-tetrakis(4-methoxyphenyl) porphyrin (TMP) and Zn-5,10,15,20-tetrakis(4-methoxyphenyl) porphyrin (ZnTMP) were evaluated in Wistar rats subcutaneously inoculated with Walker 256 carcinoma. The animals were irradiated with red light (? = 685 nm; D = 50 J/cm2; 15 min) 3 h after intra-peritoneal administration of 10 mg/kg body weight of porphyrins. The presence of free radicals in tumours after photodynamic therapy with TMP and ZnTMP revealed by chemiluminescence of luminol attained the highest level at 18 h after irradiation. Lipid peroxides measured as thiobarbituric-reactive substances and protein carbonyls, which are indices of oxidative effects produced on susceptible biomolecules, were significantly increased in tumour tissues of animals 24 h after photodynamic therapy. The levels of thiol groups and total antioxidant capacity in the tumours were decreased. The activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were also increased in tumour tissues after photodynamic therapy. Increased levels of plasma lipid peroxides as well as changes in the levels of erythrocyte antioxidant enzyme activities suggest possible systemic effects of photodynamic therapy with TMP and ZnTMP.
- MeSH
- antioxidancia analýza metabolismus MeSH
- časové faktory MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky farmakologie metabolismus terapeutické užití MeSH
- karbonylace proteinů účinky záření MeSH
- karcinom Walkerův 256 farmakoterapie metabolismus MeSH
- krysa rodu rattus MeSH
- látky reagující s kyselinou thiobarbiturovou analýza metabolismus MeSH
- lipidové peroxidy analýza krev metabolismus MeSH
- luminiscence diagnostické užití MeSH
- luminol chemie MeSH
- oxidace-redukce účinky záření MeSH
- oxidoreduktasy analýza krev metabolismus MeSH
- porfyriny farmakologie metabolismus terapeutické užití MeSH
- potkani Wistar MeSH
- reaktivní formy kyslíku metabolismus MeSH
- spotřeba kyslíku účinky léků MeSH
- sulfhydrylové sloučeniny analýza krev metabolismus MeSH
- volné radikály analýza krev metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
Increasing attention has been paid to clinical manifestation of oxidative stress and reactive oxygen and nitrogen species (RONS), e.g. in atherosclerosis, diabetes mellitus, hyperlipidemia, neurodegenerative and psychiatric diseases. The most clinically significant RONS are free radicals (superoxide anion, hydroxyl and nitroxide radicals) and non-radical species (peroxynitrite, hydrogen peroxide). The aim of this study was to investigate by EPR the concentrations of hydroxyl and nitroxide radicals in blood serum of patients with hyperlipidemia and with depressions and to compare them with healthy persons. Nitroxide radical concentrations were significantly higher in the depressive patients compared with controls. The clinical significance of this finding is not quite clear, but it supports the hypothesis on the participation of oxidative stress in pathogenesis of depressive disorders.
