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Nitrate Reductase Activity of Mitochondrial Aldehyde Dehydrogenase (ALDH-2) as a Redox Sensor for Cardiovascular Oxidative Stress

In 2002, mitochondrial aldehyde dehydrogenase (ALDH-2) was identified as an organic nitrate bioactivating enzyme. This so-called nitrate reductase activity denitrates nitroglycerin (glycerol trinitrate) to its 1,2-glycerol dinitrate metabolite and nitrite. This reac ...

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pO2 and ROS/RNS Measurements in the Microcirculation in Hypoxia

We expose methods for in vivo assessment of oxygen, nitric oxide (NO), and reactive oxygen species (ROS)/reactive nitrogen species (RNS), in the microcirculation during normoxia and hypoxia. We provide an example of the related mechanisms of ROS/RNS and oxygen level in the process of regulat ...

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Current Status of Measuring Oxidative Stress

Although the healthcare field is increasingly aware of the importance of free radicals and oxidative stress, screening and monitoring has not yet become a routine test, since, dangerously, there are no symptoms of this condition. Therefore, in very few cases is oxidative stress addressed. P ...

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Lipofuscin: Detection and Quantification by Microscopic Techniques

Since lipofuscin, the so-called “aging pigment”, turned out to play a fundamental role in the aging process, particularly in the postmitotic senescence of muscle or neuronal cells, it became a focus of aging and stress research. During normal aging, lipofuscin accumulates in a nearly linear wa ...

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Using Fluorescence-Activated Flow Cytometry to Determine Reactive Oxygen Species Formation and Membrane Lipid Peroxidation in Viable Boar Spermatozoa

Fluorescence-activated flow cytometry analyses were developed for determination of reactive oxygen species (ROS) formation and membrane lipid peroxidation in live spermatozoa loaded with, respectively, hydroethidine (HE) or the lipophilic probe 4,4-difluoro-5-(4-ph ...

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Multiphoton Redox Ratio Imaging for Metabolic Monitoring In Vivo

Metabolic monitoring at the cellular level in live tissues is important for understanding cell function, disease processes, and potential therapies. Multiphoton imaging of the relative amounts of NADH and FAD (the primary electron donor and acceptor, respectively, in the electron t ...

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Structural and Functional Changes in the Insulin Molecule Produced by Oxidative Stress

The change produced by oxidative stress on proteins (cross-links, backbone cleavage, amino acid modification) generates structural changes with a wide range of consequences such as increased propensity to the aggregation or proteolysis, altered immunogenicity and frequent ...

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Identification of ROS Using Oxidized DCFDA and Flow-Cytometry

Cells constantly generate reactive oxygen species (ROS) during aerobic metabolism. The ROS generation plays an important protective and functional role in the immune system. The cell is armed with a powerful antioxidant defense system to combat excessive production of ROS. Oxidative ...

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Cupric Ion Reducing Antioxidant Capacity Assay for Antioxidants in Human Serum and for Hydroxyl Radical Scavengers

Tests measuring the combined antioxidant effect of the nonenzymatic defenses in biological fluids may be useful in providing an index of the organism’s capability to counteract reactive species known as pro-oxidants, resist oxidative damage, and combat oxidative stress-related ...

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OXY-SCORE: A Global Index to Improve Evaluation of Oxidative Stress by Combining Pro- and Antioxidant Markers

Our group recently proposed OXY-SCORE, a summary index of oxidative stress, computed by combining plasma free and total malondialdehyde (F- and T-MDA), glutathione in disulphide/reduced forms (GSSG/GSH), α- and γ-tocopherol (TH), urine isoprostanes (iPF2α-III) levels, and plasma ind ...

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Using N,N,N,N-tetramethyl-p-phenylenediamine (TMPD) to Assay Cyclooxygenase Activity In Vitro

Prostaglandin endoperoxide synthase (PGH synthase), also known as cyclooxygenase (COX), was identified over 30 years ago and is the key enzyme in the pathway by which arachidonic acid is converted to the range of biologically active lipid mediators known as the prostanoids that participa ...

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Determination of Erythrocyte Fragility as a Marker of Pesticide-Induced Membrane Oxidative Damage

Erythrocytes are readily available cells and a good model system to study the health status of individuals with pathologic complications. It can also serve as a meaningful target to study toxicant/xenobiotic-induced damages. We have prepared different concentrations of a carbamate ...

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Measurement of Mitochondrial Membrane Potential and Proton Leak

The major component of mitochondrial electrochemical potential gradient of protons is the mitochondrial membrane potential (ΔΨ), and hence it is a suitable parameter for assessment of mitochondrial function. Dissipation of the mitochondrial membrane potential causes unco ...

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Electron Paramagnetic Resonance Oximetry and Redoximetry

Reactive oxygen/nitrogen species (ROS/RNS) have been increasingly recognized as important mediators and play a number of critical roles in cell injury, metabolism, disease pathology, diagnosis, and clinical treatment. Electron paramagnetic resonance (EPR) spectroscopy e ...

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Measurement of 8-Isoprostane in Exhaled Breath Condensate

Oxidative stress is functionally involved in the pathophysiology of lung diseases including asthma and chronic obstructive pulmonary disease. 8-Isoprostane, which is derived from free radical-catalyzed peroxidation of arachidonic acid, is one of the most reliable biomarke ...

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Analysis of Antioxidant Activities in Vegetable Oils and Fat Soluble Vitamins and Biofactors by the PAO-SO Method

Accumulated evidences indicate that reactive oxygen species (ROS) are involved in the pathophysiology of aging process. Antioxidants are believed to play an important role in the defense system to counteract ROS in the body. While excess hydrophilic antioxidants can be excreted easi ...

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Measuring Antioxidant Capacity Using the ORAC and TOSC Assays

Recent epidemiological studies have shown that there may be a link between oxidative stress and the development of several types of chronic diseases. Studies have also shown that diets rich in fruits and vegetables may decrease the incidence of cancer and other chronic diseases. The antioxi ...

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Assessing the Neuroprotective Effect of Antioxidant Food Factors by Application of Lipid-Derived Dopamine Modification Adducts

Advances in understanding the neurodegenerative pathologies are creating new opportunities for the development of neuroprotective therapies, such as antioxidant food factors, lifestyle modification and drugs. However, the biomarker by which the effect of the agent on neurod ...

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LIBS-Based Detection of Antioxidant Elements: A New Strategy

The present study deals with the scientific evaluation of antioxidant potential of aqueous extract of Trichosanthes dioica fruits on diabetes-induced oxidative stress of diabetic rats. The most effective dose of mg/kg bw of fruit aqueous extract was given orally to diabetic rats for 30 da ...

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A Method for Evaluation of Antioxidant Activity Based on Inhibition of Free Radical-Induced Erythrocyte Hemolysis

There are many in vitro methods for evaluating antioxidant activity. In this chapter, we describe an operationally simple cell-based assay, oxidative hemolysis inhibition assay (OxHLIA). OxHLIA is based on inhibition of free radical-induced membrane damage in erythrocytes by ant ...

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