INTRODUCTION: Rat mesenchymal stem cells (rMSCs) labeled with 1) poly-l-lysine-coated superparamagnetic iron oxide nanoparticles or 2) silica-coated cobalt-zinc-iron nanoparticles were implanted into the left brain hemisphere of rats, to assess their effects on the levels of oxidative damage to biological macromolecules in brain tissue. METHODS: Controls were implanted with unlabeled rMSCs. Animals were sacrificed 24 hours or 4 weeks after the treatment, and the implantation site along with the surrounding tissue was isolated from the brain. At the same intervals, parallel groups of animals were scanned in vivo by magnetic resonance imaging (MRI). The comet assay with enzymes of excision DNA repair (endonuclease III and formamidopyrimidine-DNA glycosylase) was used to analyze breaks and oxidative damage to DNA in the brain tissue. Oxidative damage to proteins and lipids was determined by measuring the levels of carbonyl groups and 15-F2t-isoprostane (enzyme-linked immunosorbent assay). MRI displayed implants of labeled cells as extensive hypointense areas in the brain tissue. In histological sections, the expression of glial fibrillary acidic protein and CD68 was analyzed to detect astrogliosis and inflammatory response. RESULTS: Both contrast labels caused a similar response in the T2-weighted magnetic resonance (MR) image and the signal was clearly visible within 4 weeks after implantation of rMSCs. No increase of oxidative damage to DNA, lipids, or proteins over the control values was detected in any sample of brain tissue from the treated animals. Also, immunohistochemistry did not indicate any serious tissue impairment around the graft. CONCLUSION: Both tested types of nanoparticles appear to be prospective and safe labels for tracking the transplanted cells by MR.

• MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Isoprostanes analysis   metabolism   MeSH
• Cobalt chemistry   MeSH
• Metal Nanoparticles administration & dosage   chemistry   toxicity   MeSH
• Magnetic Resonance Imaging methods   MeSH
• Mesenchymal Stem Cells chemistry   MeSH
• Brain diagnostic imaging   drug effects   metabolism   MeSH
• Silicon Dioxide chemistry   MeSH
• Rats, Inbred Lew MeSH
• Prospective Studies MeSH
• Tissue Extracts MeSH
• Mesenchymal Stem Cell Transplantation * MeSH
• Ferric Compounds chemistry   MeSH
• Iron chemistry   MeSH
• Zinc chemistry   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Nanomaterials are currently the subject of intense research due to their wide variety of potential applications in the biomedical, optical and electronic fields. We prepared and tested cobalt zinc ferrite nanoparticles (Co0.5Zn0.5Fe2O4+γ [CZF-NPs]) encapsulated by amorphous silica in order to find a safe contrast agent and magnetic label for tracking transplanted cells within an organism using magnetic resonance imaging (MRI). Rat mesenchymal stem cells (rMSCs) were labeled for 48 h with a low, medium or high dose of CZF-NPs (0.05; 0.11 or 0.55 mM); silica NPs (Si-NPs; 0.11 mM) served as a positive control. The internalization of NPs into cells was verified by transmission electron microscopy. Biological effects were analyzed at the end of exposure and after an additional 72 h of cell growth without NPs. Compared to untreated cells, Annexin V/Propidium Iodide labeling revealed no significant cytotoxicity for any group of treated cells and only a high dose of CZF-NPs slowed down cell proliferation and induced DNA damage, manifested as a significant increase of DNA-strand breaks and oxidized DNA bases. This was accompanied by high concentrations of 15-F2t-isoprostane and carbonyl groups, demonstrating oxidative injury to lipids and proteins, respectively. No harmful effects were detected in cells exposed to the low dose of CZF-NPs. Nevertheless, the labeled cells still exhibited an adequate relaxation rate for MRI in repeated experiments and ICP-MS confirmed sufficient magnetic label concentrations inside the cells. The results suggest that the silica-coated CZF-NPs, when applied at a non-toxic dose, represent a promising contrast agent for cell labeling.

• MeSH
• Staining and Labeling MeSH
• Cell Culture Techniques MeSH
• Isoprostanes metabolism   MeSH
• Protein Carbonylation drug effects   MeSH
• Cobalt chemistry   toxicity   MeSH
• Contrast Media chemistry   toxicity   MeSH
• Rats MeSH
• Cells, Cultured MeSH
• Magnetic Resonance Imaging MeSH
• Lipid Metabolism drug effects   MeSH
• Mesenchymal Stem Cells drug effects   metabolism   ultrastructure   MeSH
• Nanoparticles chemistry   toxicity   MeSH
• Silicon Dioxide chemistry   toxicity   MeSH
• DNA Damage * MeSH
• Surface Properties MeSH
• Cell Proliferation drug effects   MeSH
• Zinc Compounds chemistry   toxicity   MeSH
• Microscopy, Electron, Transmission MeSH
• Cell Survival drug effects   MeSH
• Dose-Response Relationship, Drug MeSH
• Ferric Compounds chemistry   toxicity   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

OBJECTIVES: The aim of our study is to investigate the impact of the type of delivery - vaginal vs. cesarean section on oxidative damage determined as the lipid peroxidation (15-F2t-isoprostane (15-F2t-IsoP) in the cord blood of newborns and venous blood from mothers in two localities with different levels of air pollution: Ceske Budejovice (CB), a locality with a clean air, and Karvina, a locality with high air pollution. RESUTLS: In Karvina, the concentration of PM2.5 was higher than in CB in the summer 2013 (mean±SD: 20.41±6.28 vs. 9.45±3.62 μg/m3, p<0.001) and in the winter 2014 (mean±SD: 53.67±19.76 vs. 27.96±12.34 μg/m3, p<0.001). Similarly, the concentration of B[a]P was higher in Karvina than in CB in the summer 2013 (mean±SD: 1.16±0.91 vs. 0.16±0.26 ng/m3, p<0.001) and in the winter 2014 (5.36±3.64 vs. 1.45±1.19 ng/m3, p<0.001). Delivery procedures differed by the type of anesthesia; at the Cesarean section in CB was used general anesthesia in 73.8% vs. 20.8% in Karvina (p<0.001), epidural anesthesia in CB in 26.2% vs. 77.1% in Karvina (p<0.001), at vaginal delivery was local anesthesia used in CB in 58.9% vs. 14.1% in Karvina (p<0.001). In CB was oxidative stress higher after vaginal delivery (101.7±31.0 pg 15-F2t-isoP/ml plasma) vs. Cesarean section (83.9±26.9 pg 15-F2t-isoP/ml plasma, p<0.001), no difference between the type of delivery was observed in Karvina. CONCLUSION: No difference between the types of delivery was observed in mothers in CB as well as in Karvina. Oxidative stress in newborns in Karvina was significantly affected by the concentrations of PM2.5 and B[a]P in the polluted air.

• MeSH
• Isoprostanes metabolism   MeSH
• Air Pollutants pharmacology   MeSH
• Humans MeSH
• Infant, Newborn MeSH
• Oxidative Stress physiology   MeSH
• Lipid Peroxidation drug effects   MeSH
• Parturition physiology   MeSH
• Pregnancy MeSH
• Delivery, Obstetric * MeSH
• Check Tag
• Humans MeSH
• Infant, Newborn MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Ambient air particulate matter (PM) represents a class of heterogeneous substances that form one component of air pollution. Oxidative stress has been implicated as an important action mechanism for PM on the human organism. Oxidative damage induced by reactive oxygen species (ROS) may affect any cellular macromolecule. The aim of our study was to investigate the impact of air pollution on oxidative DNA damage [8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG)] and lipid peroxidation [15-F2t-isoprostane (15-F2t-IsoP)] in the urine and blood from mothers and newborns from two localities with different levels of air pollution: Ceske Budejovice (CB), a locality with a clean air, and Karvina, a locality with high air pollution. The samples from normal deliveries (38-41 week+) of nonsmoking mothers and their newborns were collected in the summer and winter seasons. Higher PM2.5 concentrations were found in Karvina than in CB in the summer 2013 (mean±SD: 20.41±6.28 vs. 9.45±3.62μg/m(3), P<0.001), and in the winter 2014 (mean±SD: 53.67±19.76 vs. 27.96±12.34μg/m(3), P<0.001). We observed significant differences in 15-F2t-IsoP levels between the summer and winter seasons in Karvina for newborns (mean±SD: 64.24±26.75 vs. 104.26±38.18pg/ml plasma, respectively) (P<0.001). Levels of 8-oxodG differed only in the winter season between localities, they were significantly higher (P<0.001) in newborns from Karvina in comparison with CB (mean±SD: 5.70±2.94 vs. 4.23±1.51 nmol/mmol creatinine, respectively). The results of multivariate regression analysis in newborns from Karvina showed PM2.5 concentrations to be a significant predictor for 8-oxodG excretion, PM2.5 and B[a]P (benzo[a]pyrene) concentrations to be a significant predictor for 15-F2t-IsoP levels. The results of multivariate regression analysis in mothers showed PM2.5 concentrations to be a significant predictor of 8-oxodG levels.

• MeSH
• Deoxyguanosine analogs & derivatives   urine   MeSH
• Adult MeSH
• Isoprostanes blood   MeSH
• Cotinine urine   MeSH
• Air Pollutants analysis   MeSH
• Humans MeSH
• Maternal Exposure * MeSH
• Mothers MeSH
• Young Adult MeSH
• Infant, Newborn MeSH
• Lipid Peroxidation * MeSH
• Particulate Matter analysis   MeSH
• Polycyclic Aromatic Hydrocarbons analysis   MeSH
• DNA Damage * MeSH
• Pregnancy blood   urine   MeSH
• Air Pollution analysis   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Infant, Newborn MeSH
• Pregnancy blood   urine   MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH

Isoprostanes (IsoP) are important biologically active metabolites of arachidonic and eicosapentaenoic acids in mammals and humans. IsoPs are becoming the gold-standard biomarkers for monitoring oxidative stress. Their in-vivo detected amounts can be correlated with a number of diseases. Similarly, phytoprostanes derived from ?-linolenic acid and neuroprostanes from docosahexaenoic acid are other tools for the determination of oxidative stress in plants and the human brain, respectively. To further develop the field, it becomes more and more important to develop total syntheses of these metabolites since this is the only way of obtaining reasonable amounts of pure compounds to study their biological effects and to quantify lipid oxidative stress. In this review, new synthetic strategies, developed in 2008–2013 are summarized, showing the significant advancement in the field.

• MeSH
• Models, Biological MeSH
• Cyclopentanes chemistry   MeSH
• Isoprostanes * chemical synthesis   chemistry   metabolism   MeSH
• Catalysis MeSH
• Neuroprostanes * chemical synthesis   chemistry   metabolism   MeSH
• Oxidative Stress physiology   MeSH
• Lipid Peroxidation MeSH
• Stereoisomerism MeSH
• Chemistry Techniques, Synthetic MeSH

Populations living in industrialised regions are at higher risk of a number of diseases and shortened life span. These negative effects are primarily brought about by damage to cells and macromolecules caused by environmental pollutants. In this study, we analysed the effect of exposure to benzo[a]pyrene, a particulate matter of aerodynamic diameter < 2.5 µm (PM2.5), and benzene on oxidative stress markers [including 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), 15-F(2t)-isoprostane (15-F2t-IsoP) and protein carbonyls] and cytogenetic parameters (stable and unstable chromosomal aberrations). The samples were collected from subjects living in the Ostrava region characterised by very high levels of air pollution and in Prague with comparatively lower concentrations of pollutants in three seasons (winter 2009, summer 2009 and winter 2010). Despite several-fold higher concentrations of air pollutants in the Ostrava region, the levels of stable aberrations (genomic frequency of translocations per 100 cells, percentage of aberrant cells and frequency of acentric fragments) were mostly comparable in both locations. The frequency of unstable aberrations measured as the number of micronuclei was unexpectedly significantly lower in the Ostrava region subjects in both seasons of 2009. Urinary excretion of 8-oxodG did not differ between locations in either season. Lipid peroxidation measured as levels of 15-F2t-IsoP in blood plasma was elevated in the Ostrava subjects sampled in 2009. Protein oxidation was higher in Prague samples collected in summer 2009. Multivariate analyses conducted separately in subjects from Prague and Ostrava showed a negative association between the frequency of micronuclei and concentrations of benzo[a]pyrene and PM2.5 in both regions. A positive relationship was observed between lipid peroxidation and air pollution; protein oxidation seems to be positively affected by PM2.5 in both regions.

• MeSH
• Benzo(a)pyrene toxicity   MeSH
• Biomarkers analysis   blood   urine   MeSH
• Chromosome Aberrations * MeSH
• Deoxyguanosine analogs & derivatives   urine   MeSH
• Adult MeSH
• Isoprostanes blood   MeSH
• Protein Carbonylation MeSH
• Air Pollutants adverse effects   analysis   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lymphocytes drug effects   MeSH
• Multivariate Analysis MeSH
• Oxidative Stress drug effects   MeSH
• Lipid Peroxidation drug effects   MeSH
• Particulate Matter adverse effects   MeSH
• Cities MeSH
• Air Pollution adverse effects   analysis   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH
• Cities MeSH

Vyvinutí standardizované metodiky odběru a analýzy kondenzátu vydechovaného vzduchu a moče s důrazem na studium řady parametrů, které mohou ovlivnit validitu markerů ze skupiny metabolitů arachidonové kyseliny (8-isoprostan, leukotrien LTB4, cysteinylovéleukotrieny LTC4, LTD4, LTE4). V rámci studie budou neinvazivní metodou při stanovení markerů v různých denních i ročních obdobích, za různých klimatických podmínek i kontaminace venkovního ovzduší u zdravých dobrovolníků a osob s alergickými nemocemi (astma bronchiale, alergická rýma) zjišťovány optimální technické i analytické podmínky, zaručující standardizaci získaných výsledků. Analytická část bude využívat kombinace izolace biomarkeru z komplexní biologické matrice, případně jeho zkoncentrování,s následnou vysoce selektivní a citlivou detekční metodou na principu hmotnostní spektrometrie, která dovoluje identifikovat a kvantifikovat biomarkery v koncentracích na úrovni pikogramů v mililitru biologické matrice.; Development of a standardized methodology for the collection and exact analysis of exhaled breath condensate with the emphasis on the reliable recording of a number of parameters potentially affecting the validity of the acquired concentration levels ofarachidonic acid derivatives (leukotriene ? LTB4, cysteinyl LTC4, LTD4, LTE4, 8-isoprostane). Within the framework of the project, optimal technical and analytical conditions will be assessed during the determinations of LT in different day and year periods, different climatic conditions and environmental air pollution. Markers will be studied both in healthy volunteers and subjects with allergic diseases (bronchial asthma, allergic rhinitis). Analytic methodology will consist of biomarker isolation from a complex biological matrix, followed by a highly selective and sensitive detection method based on mass spectrometry, enabling to identify and quantify biomarkers at the concentration levels in picograms in a milliliter of the biological matrix.

• MeSH
• Biomarkers MeSH
• Asthma diagnosis   MeSH
• Rhinitis, Allergic, Perennial diagnosis   MeSH
• Breath Tests MeSH
• Mass Spectrometry MeSH
• Isoprostanes analysis   blood   urine   MeSH
• Climate Change MeSH
• Clinical Laboratory Techniques MeSH
• Leukotrienes analysis   blood   urine   MeSH
• Reference Standards MeSH
• Rhinitis, Allergic, Seasonal diagnosis   MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• pneumologie a ftizeologie
• biochemie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

Alzheimerova choroba (ACH) se řadí mezi závažná neurodegenerativní onemocnění provázená oxidačním stresem. Produkty radikálových reakcí mohou difundovat z primárních míst a být detekovány v cerebrospinální tekutině (CSF) či krvi. Tyto produkty představují potenciální biochemické markery pro diagnostiku ACH. Nejvíce pozornosti je zaměřeno na analýzu CSF, protože odráží patologické změny v mozkové tkáni u ACH. V CSF byly potvrzeny zvýšené hladiny oxidačních produktů lipidů i proteinů. Hladiny vitaminů jsou snížené, nicméně existují i práce, které nenašly rozdíly oproti kontrolám. Některé studie se zaměřily na detekci produktů oxidačního stresu v krvi u ACH, ale výsledky nejsou konzistentní. Část prací ukazuje na nárůst oxidačních produktů a snížené hladiny antioxidantů v plazmě. Nicméně jiné práce tyto výsledky nepotvrdily. Z hlediska hledání diagnostického biomarkeru pro ACH má význam se zaměřit na specifické produkty peroxidace lipidů, tzv. lipofuscinoidní pigmenty (LFP), v erytrocytech. Na základě fluorescenčních analýz LFP je možné najít specifický produkt v krvi u ACH.

Alzheimer´s disease (AD) is a serious neurodegenerative disorder accompanied by oxidative stress. Products of free radical reactions diffuse from primary sites and can be detected in cerebrospinal fluid (CSF) and blood. Such products represent potential biochemical markers for diagnosis of AD. Most studies are focused on CSF since its composition reflects pathological changes in the brain in AD. Increased levels of lipid and protein oxidative products have been found in CSF. Levels of vitamins were reduced in CSF. However, there are studies which show no difference between AD and controls. There is research on free radical products in blood in AD but results are not consistent. Several studies show increased oxidative products and reduced antioxidant in plasma. Nevertheless, others did not confirm it. Considering the investigation of a diagnostic biomarker for AD, specific end-products of lipid peroxidation, so called lipofuscin-like pigments (LFP), in erythrocytes represent an important possibility. A specific product in blood in AD can be found by means of fluorescence analyses of LFP.

• Keywords
• biomarkery,
• MeSH
• Alzheimer Disease physiopathology   MeSH
• Biomarkers blood   MeSH
• Financing, Organized MeSH
• Isoprostanes diagnostic use   blood   MeSH
• Ascorbic Acid diagnostic use   blood   MeSH
• Malondialdehyde diagnostic use   blood   MeSH
• Oxidative Stress MeSH
• Lipid Peroxidation MeSH
• Glycation End Products, Advanced diagnostic use   MeSH
• Vitamin E diagnostic use   blood   MeSH

Nalezení optimální metody pro analýzu markerů oxidačního stresu v biologickém materiálu - v kondenzátu vydechovaného vzduchu, v krvi a moči (8-izoprostan, malondialdehyd, 4-hydroxy-trans-2-nonenal) u osob s předpokládanou zvýšenou přítomností aktivních forem kyslíku v plicích a dýchacích cestách, tj. u osob se silikózou plic a onemocněními z azbestu. U těchto skupin osob jsme nedávno zjistili zvýšenou koncentraci 8-izoprostanu v kondenzátu vydechovaného vzduchu, pro diferenciální diagnostiku a predikcidalšího vývoje je třeba odlišit vliv oxidačního stresu v důsledku jiných faktorů, popřípadě onemocnění. Budou vyvinuty citlivé metody detekce těchto markerů pomocí hmotnostní spektrometrie s vysokou selektivitou a citlivostí.; Finding optimum methods for analysis of markers of oxidative stress in biological material - exhaled breath condensate, in blood and urine: 8-isoprostane, 4-hydroxy-trans-2-nonenal (HNE) and malondialdehyde (MDA) in subjects with supposed increase of active oxygen species in the lungs and airways, i.e. in patients with silicosis and diseases due to asbestos. We have shown recently for the first time, that 8-isoprostane in exhaled breath condensate of these persons was significantly increased. For differential diagnosis and prediction of further development it is necessary to distinguish the influence of oxidative stress due to other factors or diseases. Sensitive methods to detect these markers will be developed by means of mass spectrometry with highselectivity and sensitivity.

• MeSH
• Asbestosis MeSH
• Biomarkers analysis   MeSH
• Diagnosis, Differential MeSH
• Mass Spectrometry methods   utilization   MeSH
• Isoprostanes MeSH
• Malondialdehyde MeSH
• Oxidative Stress MeSH
• Silicosis MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• biologie
• toxikologie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR

Oxidative damage to macromolecules may have numerous negative health consequences. We measured oxidative damage to DNA, proteins and lipids in 80 newborns and 79 mothers, analyzed the effect of mother's tobacco smoke exposure on oxidative stress, and assessed correlations between oxidative stress markers and bulky and PAH (polycyclic aromatic hydrocarbons)-specific DNA adducts. Mean levels (+/-S.D.) of 8-oxodeoxyguanosine (8-oxodG) per 10(5) dG in the placenta were 2.85+/-0.78; we did not see a difference between 8-oxodG levels in newborns born to mothers exposed and unexposed to tobacco smoke. Protein carbonyl levels, a marker of protein oxidation, were comparable in the umbilical cord and in maternal venous blood plasma (17.4+/-3.2 and 17.6+/-4.2nmol/ml plasma in newborns and mothers, respectively, p=0.66). Lipid peroxidation measured as levels of 15-F(2t)-isoprostane (15-F(2t)-IsoP) in plasma was significantly higher in newborns than in mothers (362+/-129 and 252+/-130pg/ml in newborns and mothers, respectively, p<0.001). We did not find any effect of tobacco smoke exposure on either biomarker in any group. Levels of both protein carbonyls and 15-F(2t)-IsoP in cord blood significantly correlated with those in maternal plasma (p<0.001). 8-oxodG levels positively correlated with plasma carbonyls in cord plasma, as well as with cotinine levels (marker of tobacco smoke exposure) in maternal plasma. 8-oxodG levels also correlated with bulky DNA adducts in lymphocyte DNA of newborns and mothers and with PAH-DNA adducts in the placenta. Our results showed higher lipid peroxidation in newborns than in mothers, close correlation of analyzed oxidative stress markers between newborns and mothers, and a relationship between oxidative stress and induction of DNA adducts.

• MeSH
• DNA Adducts blood   MeSH
• Biomarkers blood   MeSH
• Deoxyguanosine analogs & derivatives   metabolism   MeSH
• Adult MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• F2-Isoprostanes MeSH
• Fetal Blood chemistry   MeSH
• Protein Carbonylation MeSH
• Cotinine analysis   MeSH
• Smoking MeSH
• Blood Proteins analysis   MeSH
• Air Pollutants blood   MeSH
• Humans MeSH
• Lymphocytes drug effects   MeSH
• Maternal-Fetal Exchange MeSH
• Maternal Exposure MeSH
• Young Adult MeSH
• Infant, Newborn MeSH
• Oxidation-Reduction MeSH
• Oxidative Stress MeSH
• Lipid Peroxidation MeSH
• Placenta metabolism   MeSH
• Polycyclic Aromatic Hydrocarbons blood   MeSH
• Pregnancy MeSH
• Vitamin A analysis   MeSH
• Vitamin E analysis   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Young Adult MeSH
• Infant, Newborn MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH