Overproduction Of Hydrogen Peroxide Research Articles

Organ-specific differences in endogenous phytohormone and antioxidative responses in potato upon PSTVd infection

Although structurally simple, viroids can trigger numerous changes in host plants and cause loss of yield in agronomically important crops. This study investigated changes in the endogenous status of phytohormones and antioxidant enzyme activity in Solanum tuberosum cv. Désirée in response to Potato spindle tuber viroid (PSTVd) infection. Phytohormone analysis showed that the content of endogenous jasmonic acid (JA) and its precursor cis-OPDA significantly increased in leaves, while the content of castasterone (CS) increased in both leaves and tubers of systemically infected plants compared to mock-inoculated control plants at 8 weeks post-inoculation. The indole-3-acetic acid content moderately increased only in tubers, while no differences in salicylic acid and abscisic acid content were observed between infected and control plants. Changes in endogenous phytohormone content were associated with upregulated expression of genes involved in the biosynthesis of JA and brassinosteroids, and the metabolism of auxins. Additionally, PSTVd infection provoked overproduction of hydrogen peroxide, which coincided with increased activity of guaiacol peroxidase in leaves and ascorbate peroxidase in potato tubers. The activity of catalase decreased in leaves, while superoxide dismutase activity remained steady regardless of the treatment and organ type. Total ascorbate and glutathione did not change significantly, although a shift towards oxidized forms was observed. Results suggest the existence of organ-specific differences in phytohormone and antioxidative responses in potato upon PSTVd infection. Possible effects of the observed changes on symptom development are discussed.

A novel online fluorescence method for in-vivo measurement of hydrogen peroxide during oxidative stress produced in a temporal lobe epilepsy model.

Status epilepticus (SE) can result in an overproduction of hydrogen peroxide (H2O2), which contributes to oxidative stress and brain injury during different phases of epileptogenesis and seizures. In this study, we measured the extracellular H2O2 concentration in the rat hippocampus in a temporal lobe epilepsy model. A new fluorescent technique for measuring H2O2 in vivo simultaneously with electroencephalography recording was tested. The method consists of mixing microdialysate with an enzymatic reactor to produce a fluorescent compound. The fluorescence intensity was measured every second and was proportional to the H2O2 concentration. The results showed that H2O2 was released during SE; we detected a significant increase of up to five times over the baseline value that correlated with changes in electrical activity. We also observed that H2O2 was produced for days after SE and was associated with continuous neuronal death and seizure generation. Therefore, we monitored H2O2 48 h and 15 days after SE, observing increases of up to 96 and 124%, respectively, accompanied by changes in electrical activity with spontaneous discharges of large amplitude. These changes may reflect the oxidative stress generated during epileptogenesis that remains during the chronic period (458% increased) with the presence of large spikes, indicating that the H2O2 could also participate in the generation and maintenance of spontaneous recurrent seizures. There are no previous reports on the detection of H2O2 at this temporal resolution; thus, this study contributes a novel technique for studying and understanding epileptogenesis to develop new antioxidant strategies for the treatment of temporal lobe epilepsy.

MicroRNA-455 targets cullin 3 to activate Nrf2 signaling and protect human osteoblasts from hydrogen peroxide.

Over-production of hydrogen peroxide (H2O2) will lead to human osteoblast dysfunction and apoptosis, causing progression of osteoporosis and osteonecrosis. NF-E2-related factor 2 (Nrf2) is a well-characterized anti-oxidant signaling. Cullin 3 (Cul3) ubiquitin E3 ligase dictates Nrf2 degradation. We demonstrate that microRNA-455 (“miR-455”) is a putative Cul3-targeting microRNA. Forced-expression of miR-455 in both hFOB1. 19 osteoblast cell line and primary human osteoblasts induced Cul3 degradation and Nrf2 protein stabilization, which led to subsequent transcription of ARE (anti-oxidant response element)-dependent genes (NQO1, HO1 and GCLC). Cul3 silencing, by expressing miR-455 or targeted-shRNA, protected human osteoblasts from H2O2. Reversely, miR-455 anti-sense caused Cul3 accumulation and Nrf2 degradation, which exacerbated H2O2 damages in hFOB1. 19 cells. Moreover, forced over-expression of Cul3 in hFOB1. 19 cells silenced Nrf2 and sensitized H2O2. Together, we propose that miR-455 activated Nrf2 signaling and protected human osteoblasts from oxidative stress possibly via targeting Cul3.

Peroxyoxalate Chemiluminescent Reaction as a Tool for Elimination of Tumour Cells Under Oxidative Stress

The overproduction of hydrogen peroxide is an inherent feature of some tumour cells and inflamed tissues. We took advantage of this peculiarity to eliminate cells using chemiluminescent peroxyoxalate reaction. We designed dispersions containing polyoxalate and tetramethylhematoporhyrin (TMHP) in dimethylphthalate droplets stabilized with Pluronic L64. The porphyrin plays the dual role. On the one hand, it serves as an activator of the peroxyoxalate reaction of polyoxalate with intracellular hydrogen peroxide and experiences excitation as a result of the reaction. The light emitted in the reaction in the model system without cells was used to optimize the dispersion’s composition. On the other hand, TMHP acts as a photosensitizer (PS) causing cell damage. The formation of singlet oxygen led to cell elimination if the dispersions were used in combination with inducers of oxidative stress: hydrogen peroxide, paraquat, antitumour drug doxorubicin, or a nutritional additive menadione. The PS-induced cytotoxicity correlated with the level of intracellular ROS. The developed approach targeted to endogenous ROS is orthogonal to the classical chemotherapy and can be applied to increase its efficiency.

Comparative effect of calcium and EDTA on arsenic uptake and physiological attributes of Pisum sativum

ABSTRACTIn this study, we determined the effect of ethylenediaminetetraacetic acid (EDTA) and calcium (Ca) on arsenic (As) uptake and toxicity to Pisum sativum. Plants were treated with three levels of As (25, 125, and 250 µM) in the presence and absence of three levels of Ca (1, 5, and 10 mM) and EDTA (25, 125, and 250 µM). Exposure to As caused an overproduction of hydrogen peroxide (H2O2) in roots and leaves, which induced lipid peroxidation and decreased pigment contents. Application of both Ca and EDTA significantly reduced As accumulation by pea, Ca being more effective in reducing As accumulation. Both Ca and EDTA enhanced As-induced H2O2 production, but reduced lipid peroxidation. In the case of pigment contents, EDTA significantly reduced pigment contents, whereas Ca significantly enhanced pigment contents compared to As alone. The effect of As treatment in the presence and absence of EDTA and Ca was more pronounced in younger leaves compared to older leaves. The effect of amendments varied greatly with their applied levels, as well as type and age of plant organs. Importantly, due to possible precipitation of Ca-As compounds, the soils with higher levels of Ca ions are likely to be less prone to food chain contamination.

Mycorrhization protects Betula pubescens Ehr. from metal-induced oxidative stress increasing its tolerance to grow in an industrial polluted soil

In recent years, the use of woody plants in phytoremediation has gained popularity due to their high biomass production and their association with mycorrhizal fungi, which can improve their survival and development rates under stress conditions. In this study, mycorrhized and non-mycorrhized white birch plants (Betula pubescens Ehr.) were grown in control and a metal-polluted industrial soil. After 60days of culture, plant growth and metal accumulation, the content of photosynthetic pigments and oxidative-stress markers, as well as the enzymatic activities and gene expressions of antioxidant enzymes were measured.According to our results, mycorrhized birch plants grown in control soil showed an increased activity and gene expression of catalase and ascorbate peroxidase, along with hydrogen peroxide overproduction, which could support the importance of the reactive oxygen species as signaling molecules in the regulation of plant-fungus interactions. Additionally, in polluted soil mycorrhized plants had higher biomass but lower metal accumulation, probably because the symbiotic fungus acted as a barrier to the entrance of metals into the host plants. This behavior led to mitigation in the oxidative challenge, reduced hydrogen peroxide content and diminished activities of the antioxidant enzymes in comparison to non-mycorrhized plants.

Hydrogen Peroxide-Responsive Nanoparticle Reduces Myocardial Ischemia/Reperfusion Injury.

BackgroundDuring myocardial ischemia/reperfusion (I/R), a large amount of reactive oxygen species (ROS) is produced. In particular, overproduction of hydrogen peroxide (H2O2) is considered to be a main cause of I/R‐mediated tissue damage. We generated novel H2O2‐responsive antioxidant polymer nanoparticles (PVAX and HPOX) that are able to target the site of ROS overproduction and attenuate the oxidative stress‐associated diseases. In this study, nanoparticles were examined for their therapeutic effect on myocardial I/R injury.Methods and ResultsThe therapeutic effect of nanoparticles during cardiac I/R was evaluated in mice. A single dose of PVAX (3 mg/kg) showed a significant improvement in both cardiac output and fraction shortening compared with poly(lactic‐coglycolic acid) (PLGA) particle, a non‐H2O2‐activatable nanoparticle. PVAX also significantly reduced the myocardial infarction/area compared with PLGA (48.7±4.2 vs 14.5±2.1). In addition, PVAX effectively reduced caspase‐3 activation and TUNEL‐positive cells compared with PLGA. Furthermore, PVAX significantly decreased TNF‐α and MCP‐1 mRNA levels. To explore the antioxidant effect of PVAX by scavenging ROS, dihydroethidium staining was used as an indicator of ROS generation. PVAX effectively suppressed the generation of ROS caused by I/R, whereas a number of dihydroethidium‐positive cells were observed in a group with PLGA I/R. In addition, PVAX significantly reduced the level of NADPH oxidase (NOX) 2 and 4 expression, which favors the reduction in ROS generation after I/R.ConclusionsTaken together, these results suggest that H2O2‐responsive antioxidant PVAX has tremendous potential as a therapeutic agent for myocardial I/R injury.

Phase Transition of the Bacterium upon Invasion of a Host Cell as a Mechanism of Adaptation: a Mycoplasma gallisepticum Model.

What strategies do bacteria employ for adaptation to their hosts and are these strategies different for varied hosts? To date, many studies on the interaction of the bacterium and its host have been published. However, global changes in the bacterial cell in the process of invasion and persistence, remain poorly understood. In this study, we demonstrated phase transition of the avian pathogen Mycoplasma gallisepticum upon invasion of the various types of eukaryotic cells (human, chicken, and mouse) which was stable during several passages after isolation of intracellular clones and recultivation in a culture medium. It was shown that this phase transition is manifested in changes at the proteomic, genomic and metabolomic levels. Eukaryotic cells induced similar proteome reorganization of M. gallisepticum during infection, despite different origins of the host cell lines. Proteomic changes affected a broad range of processes including metabolism, translation and oxidative stress response. We determined that the activation of glycerol utilization, overproduction of hydrogen peroxide and the upregulation of the SpxA regulatory protein occurred during intracellular infection. We propose SpxA as an important regulator for the adaptation of M. gallisepticum to an intracellular environment.

Boron Supply and Water Deficit Consequences in Young Paricá (<i>Schizolobium parahyba</i> var.<i> amazonicum</i>) Plants

Boron (B) is a very important nutrient required by forest plants; when supplied in adequate amounts, plants can ameliorate the negative effects of abiotic stresses. The objective of this study was to (i) investigate gas exchange, (ii) measure oxidant and antioxidant compounds, and (iii) respond how B supply acts on tolerance mechanism to water deficit in young Schizolobium parahyba plants. The experiment employed a factorial that was entirely randomised, with two boron levels (25 and 250 µmol L-1, simulating conditions of sufficient B and high B, respectively) and two water conditions (control and water deficit). Water deficit induced negative modifications on net photosynthetic rate, stomatal conductance and water use efficiency, while B high promoted intensification of the effects on stomatal conductance and water use efficiency. Hydrogen peroxide and electrolyte leakage of both tissues suffered non-significant increases after B high and when applied water deficit. Ascorbate levels presented increases after water deficit and B high to leaf and root. Our results suggested that the tolerance mechanism to water deficit in young Schizolobium parahyba plants is coupled to increases in total glutathione and ascorbate aiming to control the overproduction of hydrogen peroxide and alleviates the negative consequences on electrolyte leakage and gas exchange. In relation to B supply, this study proved that sufficient level promoted better responses under control and water deficit conditions.

Boron Supply and Water Deficit Consequences in Young Paricá (<i>Schizolobium parahyba</i> var.<i> amazonicum</i>) Plants

Boron (B) is a very important nutrient required by forest plants; when supplied in adequate amounts, plants can ameliorate the negative effects of abiotic stresses. The objective of this study was to (i) investigate gas exchange, (ii) measure oxidant and antioxidant compounds, and (iii) respond how B supply acts on tolerance mechanism to water deficit in young Schizolobium parahyba plants. The experiment employed a factorial that was entirely randomised, with two boron levels (25 and 250 µmol L-1, simulating conditions of sufficient B and high B, respectively) and two water conditions (control and water deficit). Water deficit induced negative modifications on net photosynthetic rate, stomatal conductance and water use efficiency, while B high promoted intensification of the effects on stomatal conductance and water use efficiency. Hydrogen peroxide and electrolyte leakage of both tissues suffered non-significant increases after B high and when applied water deficit. Ascorbate levels presented increases after water deficit and B high to leaf and root. Our results suggested that the tolerance mechanism to water deficit in young Schizolobium parahyba plants is coupled to increases in total glutathione and ascorbate aiming to control the overproduction of hydrogen peroxide and alleviates the negative consequences on electrolyte leakage and gas exchange. In relation to B supply, this study proved that sufficient level promoted better responses under control and water deficit conditions.

Nanoparticles based on quantum dots and a luminol derivative: implications for in vivo imaging of hydrogen peroxide by chemiluminescence resonance energy transfer.

Overproduction of hydrogen peroxide is involved in the pathogenesis of inflammatory diseases such as cancer and arthritis. To image hydrogen peroxide via chemiluminescence resonance energy transfer in the near-infrared wavelength range, we prepared quantum dots functionalized with a luminol derivative.

Hydrogen peroxide-activatable antioxidant prodrug as a targeted therapeutic agent for ischemia-reperfusion injury.

Overproduction of hydrogen peroxide (H2O2) causes oxidative stress and is the main culprit in the pathogenesis of ischemia/reperfusion (I/R) injury. Suppression of oxidative stress is therefore critical in the treatment of I/R injury. Here, we report H2O2-activatable antioxidant prodrug (BRAP) that is capable of specifically targeting the site of oxidative stress and exerting anti-inflammatory and anti-apoptotic activities. BRAP with a self-immolative boronic ester protecting group was designed to scavenge H2O2 and release HBA (p-hydroxybenzyl alcohol) with antioxidant and anti-inflammatory activities. BRAP exerted potent antioxidant and anti-inflammatory activity in lipopolysaccharide (LPS)- and H2O2-stimulated cells by suppressing the generation of ROS and pro-inflammatory cytokines. In mouse models of hepatic I/R and cardiac I/R, BRAP exerted potent antioxidant, anti-inflammatory and anti-apoptotic activities due to the synergistic effects of H2O2-scavenging boronic esters and therapeutic HBA. In addition, administration of high doses of BRAP daily for 7 days showed no renal or hepatic function abnormalities. Therefore BRAP has tremendous therapeutic potential as H2O2-activatable antioxidant prodrug for the treatment of I/R injuries.

Abstract 14909: Hydrogen Peroxide-Responsive Nanoparticles Reduce Myocardial Ischemia/Reperfusion Injury

During myocardial ischemia/reperfusion (I/R), a large amount of reactive oxygen species (ROS) is produced which causes oxidative stress and leads to severe cellular and tissue damage. In particular, overproduction of hydrogen peroxide (H2O2) is considered as a main cause of I/R-mediated tissue damage by inducing inflammation and apoptosis. Therefore, rapid elimination of H2O2 and suppression of oxidative stress are thought to be reasonable strategies to treat myocardial I/R injury. Recently, we generated novel H2O2-responsive antioxidant polymer nanoparticles (PVAX and HPOX) that are able to target the site of ROS overproduction and impede the progression of oxidative stress-associated diseases. In this study, PVAX nanoparticles were selected as therapeutic agents for myocardial I/R injury based on the results of direct comparison studies in vitro and in vivo. To explore therapeutic effect of PVAX on cardiac I/R injury, cardiac function 2 weeks after reperfusion was evaluated. A single dose of PVAX (3mg/kg) showed a significant improvement in both cardiac output (8.1± 0.7 vs. 3.4±0.3) and fraction shortening (43.8± 0.9 vs. 31.2±3.8) compared with poly (lactic-co-glycolic acid) (PLGA) particle, a lacking H2O2 responsible nanoparticle. Consistent with cardiac function evaluation, PVAX significantly reduced the myocardial infarction /area at risk compared with PLGA (48.7±4.2 vs.14.5±2.1). In addition, PVAX effectively reduced caspase-3 activation (2.6±0.1 vs.1.4±0.1) and TUNEL-positive cells (8.2±1.8 vs. 2.7±0.6) compared with PLGA. Furthermore, PVAX significantly decreased TNF-α (5.5±0.4 vs. 3.0±0.3) and MCP-1 (2.3±0.1 vs. 1.4±0.1) mRNA levels. To explore the antioxidant effect of PVAX by scavenging ROS, dihydroethidium (DHE) staining was used as an indicator of ROS generation. PVAX effectively suppressed the generation of ROS caused by I/R while a number of DHE-positive cells were observed in a group of PLGA I/R. In addition, PVAX significantly reduced the level of NADPH oxidase (NOX) 2 and 4 expression, which favors the reduction in ROS generation after I/R. Taken together, these results suggest that H2O2-responsive antioxidant PVAX has tremendous potential as a therapeutic agent for myocardial I/R injury.

Rapid plant rehydration initiates permanent and adverse changes in the photosynthetic apparatus of triticale

The reasons for partial recovery after a soil drought are not fully understood and have not been studied so far. This study investigated the physiological and biochemical responses of triticale cultivars with differential recovery ability after soil water deficit. Activity of the photosynthetic apparatus under soil drought followed by rehydration was estimated. Plant antioxidant potential was determined based on the measurement of catalase and peroxidase activity. The levels of hydrogen peroxide and superoxide radical were assessed. Under rehydration, the not fully-recovered cultivar experienced further significant increase in the content of H2O2 and inhibited activity of the photosynthetic apparatus, as compared to the drought period. On 42nd day of the rehydration, the not fully-recovered cultivar showed also a reduced photosynthetic activity in the flag leaves, which resulted in a significant decrease in its grain yield. The first week of a rapid rehydration involved a decrease in total peroxidase and catalase activities. The increased content of H2O2 was compensated only when leaf water content was gradually restored in the first week of the rehydration and no further decrease in the activity of the photosynthetic apparatus was noticed. A destructive effect of the rapid rehydration was manifested in an intensification of the physiological processes associated with reactive oxygen species (ROS) overproduction. An important cause of hydrogen peroxide overproduction seems to be the electron leakage due to overloading of the electron transport chain (ETC) in the PSI and PSII.

Mitochondrial free radical overproduction due to respiratory chain impairment in the brain of a mouse model of Rett syndrome: protective effect of CNF1

Rett syndrome (RTT) is a pervasive neurodevelopmental disorder mainly caused by mutations in the X-linked MECP2 gene associated with severe intellectual disability, movement disorders, and autistic-like behaviors. Its pathogenesis remains mostly not understood and no effective therapy is available. High circulating levels of oxidative stress markers in patients and the occurrence of oxidative brain damage in MeCP2-deficient mouse models suggest the involvement of oxidative stress in RTT pathogenesis. However, the molecular mechanism and the origin of the oxidative stress have not been elucidated. Here we demonstrate that a redox imbalance arises from aberrant mitochondrial functionality in the brain of MeCP2-308 heterozygous female mice, a condition that more closely recapitulates that of RTT patients. The marked increase in the rate of hydrogen peroxide generation in the brain of RTT mice seems mainly produced by the dysfunctional complex II of the mitochondrial respiratory chain. In addition, both membrane potential generation and mitochondrial ATP synthesis are decreased in RTT mouse brains when succinate, the complex II respiratory substrate, is used as an energy source. Respiratory chain impairment is brain area specific, owing to a decrease in either cAMP-dependent phosphorylation or protein levels of specific complex subunits. Further, we investigated whether the treatment of RTT mice with the bacterial protein CNF1, previously reported to ameliorate the neurobehavioral phenotype and brain bioenergetic markers in an RTT mouse model, exerts specific effects on brain mitochondrial function and consequently on hydrogen peroxide production. In RTT brains treated with CNF1, we observed the reactivation of respiratory chain complexes, the rescue of mitochondrial functionality, and the prevention of brain hydrogen peroxide overproduction. These results provide definitive evidence of mitochondrial reactive oxygen species overproduction in RTT mouse brain and highlight CNF1 efficacy in counteracting RTT-related mitochondrial defects.

DNA hypomethylation concomitant with the overproduction of ROS induced by naphthoquinone juglone on tobacco BY-2 suspension cells

Juglone, 5-hydroxy-1,4-naphthoquinone, is a naturally occurring plant secondary metabolite with allelopathic and cytotoxic properties. The cytotoxic effect of juglone and changes at the level of DNA methylation were investigated on a plant cell model, tobacco BY-2 cell suspension culture. We found that juglone in a concentration-dependent manner inhibits cell growth and causes creation of reactive oxygen species (ROS) that were visible mainly in nucleus, nucleolus, cytoplasm, and plasma membrane. Analysis of the changes in DNA methylation level using Luminometric Methylation Assay showed that juglone at the concentrations of 60 and 80μM causes hypomethylation of DNA. Decrease in DNA methylation was confirmed by the more sensitive technique, confocal immunofluorescence microscopy, already at 30μM, and was accompanied by significant overproduction of hydrogen peroxide, hydroxyl radicals, and superoxide anion radicals. The highest percentage of the signs of programed cell death, especially apoptic-like bodies that indicate DNA fragmentation was observed at 30μM of juglone. On the basis of results we suggest that overproduction of ROS is the main mechanism that causes hypomethylation of DNA, which may contribute to the initiation of the programmed cell death.

Oxidative stress, thyroid dysfunction & Down syndrome

Down syndrome (DS) is one of the most common chromosomal disorders, occurring in one out of 700-1000 live births, and the most common cause of mental retardation. Thyroid dysfunction is the most typical endocrine abnormality in patients with DS. It is well known that thyroid dysfunction is highly prevalent in children and adults with DS and that both hypothyroidism and hyperthyroidism are more common in patients with DS than in the general population. Increasing evidence has shown that DS individuals are under unusual increased oxidative stress, which may be involved in the higher prevalence and severity of a number of pathologies associated with the syndrome, as well as the accelerated ageing observed in these individuals. The gene for Cu/Zn superoxide dismutase (SOD1) is coded on chromosome 21 and it is overexpressed (~50%) resulting in an increase of reactive oxygen species (ROS) due to overproduction of hydrogen peroxide (H2O2). ROS leads to oxidative damage of DNA, proteins and lipids, therefore, oxidative stress may play an important role in the pathogenesis of DS.

Synergistic antimicrobial activity based on the combined use of a gemini-quaternary ammonium compound and ultraviolet-A light

This study examined the utility of synergistic disinfection employing a gemini-quaternary ammonium salt (a gemini-QUAT, namely 3,3′-(2,7-dioxaoctane)bis(1-decylpyridinium bromide)), as an organic biocide in combination with irradiation by an ultraviolet-A (UV-A) light-emitting diode (LED) with a peak wavelength of 365nm. The combined system represents a novel disinfection method utilizing facilitated in situ oxidation depending on overproduction of reactive oxygen species (ROS) triggered by the initial action of the gemini-QUAT on the bacterial membrane. We demonstrate that this combination decreased the viability of pathogenic bacteria in a significant and rapid manner, and depended on doses of the gemini-QUAT and the fluence: the viability of Escherichia coli was reduced by greater than 5.0-logs by the combination procedure, but the decrease in viability was only 2.3-logs for exposure to UV at the same fluence dose in the absence of the gemini-QUAT. Adding catalase as a radical scavenger decreased bacterial inactivation by the combined disinfection procedure. Flow cytometric analysis indicated superoxide and hydrogen peroxide overproduction within cells treated with the combined disinfection procedure. The excessive superoxide, detected only in the combined system, appeared to be generated by the action of the gemini-QUAT at the bacterial membrane, leading to excessive and rapid generation of ROS in the system. Our data strongly suggested that this ROS promoted bacterial membrane peroxidation during initial treatment by the combination method, resulting in increased oxidative modification of DNA. These oxidative reactions may play an important role in the efficacy of this disinfection procedure.

Effects of pre- and post-treatment with plant polyphenols on human keratinocyte responses to solar UV

The understanding of the anti-inflammatory mechanisms of action of plant polyphenols (PPs) and clarification of the relationship between their anti-inflammatory and antioxidant properties may result in a new therapeutic approach to skin cancers. To elucidate the underlying mechanism, we analyzed the ability of PPs to attenuate inflammatory, metabolic and oxidative cellular responses to UV irradiation. Normal human epidermal keratinocytes (NHEK) were exposed to physiologically relevant dose of solar-simulated UV irradiation. Effects of pre- and post-treatment with PPs on the overproduction of peroxides and inflammatory mediators (mRNA and protein) were analyzed using real-time RT-PCR, enzyme-linked immunosorbent and fluorometric techniques. Differences between the effectiveness of pre- and post-treatment with polyphenols was found. In particular, PPs post-treatment, but not pretreatment, completely abolished overexpression of Cyp1a1 and Cyp1b1 genes and elevation of intracellular peroxides in NHEK irradiated by UV. Post-treatment with PPs also more efficiently than pretreatment prevented UV-induced overexpression of IL-1 beta, IL-6 and COX2 mRNAs. Our data strongly suggest that PPs predominantly affect delayed molecular and cellular events initiated in NHEK by solar UV rather than primary photochemical reactions. PPs may be important component in cosmetic formulations for post-sun skin care.

2012 Late-Breaking Basic Science Oral Session

### Cardiac Myosin Binding Protein C is an Ultra-early and Cardiac-specific Biomarker of Myocardial Necrosis Diederik W Kuster1, Adriana Cardenas-Ospina2, Lawson Miller2, Christian Troidl3, Holger M Nef3, Christoph Liebetrau3, Mollmann Helge3, Karen S Pieper4, Kenneth W Mahaffey4, Neal S Kleiman5, Bruno D Stuyvers2, Ali J Marian6, Sakthivel Sadayappan1; 1Loyola Univ Chicago, Maywood, IL, 2Memorial Univ, St John’s, Canada, 3Kerckhoff Heart and Thorax Cntr, Bad Neuheim, Germany, 4Duke Univ Med Cntr, Durham, NC, 5The Methodist DeBakey Heart and Vascular Cntr, Houston, TX, 6Univ of Texas Health Sciences, Houston, TX Rationale: We recently demonstrated that proteolytic cleavage fragments of cardiac myosin binding protein-C (cMyBP-C) can be detected in the serum after myocardial infarction (MI) in a rat model and in patients with MI. The findings implied that serum cMyBP-C might be useful as cardiac-specific biomarker for MI. The release kinetics of cMyBP-C in the circulation post-MI remains to be elucidated. Objective: Determine the release kinetics of cMyBP-C as an ultra-early and cardiac-specific biomarker of myocardial necrosis. Method and Results: To determine the exact timing of cMyBP-C release in the bloodstream post-acute MI, left anterior descending (LAD) coronary artery was ligated in adult swine (n=6). ECG showed significant ST elevation. Infarct size represented 12.4 ± 1.9% of total ventricular mass. Blood samples were collected before and at predetermined time points between 30 min and 14 days after LAD ligation. Plasma cMyBP-C level was quantified using a highly sensitive and rapid sandwich enzyme-linked immunosorbent assay. Compared with baseline, cMyBP-C levels were increased in post-MI serum within 45 min (0.64 ± 0.52 ng/ml) after LAD ligation and declined after 16 hrs to the baseline level (0.01 ± 0.00 ng/ml). In contrast, cardiac troponin I (cTnI) level peaked after …