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Meta-Analysis: Drawing Conclusions When Study Results Vary

Low-dose aspirin has been suggested to positively impact a number of clinical outcomes associated with oxidative stress; however, results of clinical trials surrounding its effect on a woman’s ability to achieve and sustain pregnancy have been inconclusive. A meta-analysis is an advan ...

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Mapping of Oxidative Stress Response Elements of the Caveolin-1 Promoter

According to the “free radical theory” of aging, normal aging occurs as the result of tissue damages inflicted by reactive oxygen species (ROS). ROS are known to induce cellular senescence, and senescent cells are believed to contribute to organismal aging. The molecular mechanisms that med ...

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HPLC Purification of Adenoviral Vectors

Adenoviruses are attractive vectors for gene therapy where short-term transgene expression is required. In order to meet the clinical requirements of adenovirus for use beyond the laboratory, advanced methods are required for the purification and quantitation of recombinant ad ...

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Generation of Antioxidant Adenovirus Gene Transfer Vectors Encoding CuZnSOD, MnSOD, and Catalase

Replication-deficient adenovirus gene transfer vectors are very useful for the experimental delivery of genes into cells and are widely used both in vitro and in vivo to determine the effects of transgene expression. Having a broad cell tropism, these vectors allow efficient transduct ...

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Separation of Phenylpropanoids and Evaluation of Their Antioxidant Activity

Phenylpropanoids are a group of natural products with a wide range of biological and pharmacological importance. They have been isolated from a large number of plants by utilizing a diverse range of chromatographic techniques. We describe here the utilization of normal, reverse phase, and ...

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Activation of Erythrocyte Plasma Membrane Redox System Provides a Useful Method to Evaluate Antioxidant Potential of Plant Polyphenols

Plant polyphenols are known to possess antioxidant acitivities. In recent years, there have been numerous reports confirming the efficacy of these compounds to improve plasma antioxidant capacity in humans. Current methods to evaluate the antioxidant potential of polyphenols a ...

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Direct Determination of Tissue Aminothiol, Disulfide, and Thioether Levels Using HPLC-ECD with a Novel Stable Boron-Doped Diamond Working Electrode

This chapter describes two different methods using reversed-phase HPLC with electrochemical detection on a boron-doped diamond (BDD) working electrode for the direct, routine, sensitive and simultaneous measurement of a number of aminothiols, disulfides, and thioethers, in ei ...

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Characterization of the Antioxidant Properties of Pentaerithrityl Tetranitrate (PETN)-Induction of the Intrinsic Antioxidative System Heme Oxygenase-1

Organic nitrates are among the oldest and yet most commonly employed drugs in the chronic therapy of coronary artery disease and congestive heart failure. While they have long been used in clinical practise, our understanding of their mechanism of action and of their side effects remains inco ...

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Design and Synthesis of Antioxidant -Lipoic Acid Hybrids

The design and synthesis of hybrid molecules encompassing two pharmacophores in one molecular scaffold is a well-established approach to the synthesis of more potent drugs with dual activity. In this chapter, we will present the most important synthetic methodologies we have applied f ...

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Measurement of Polyphosphoinositides in Cultured Mammalian Cells

The seven phosphorylated derivatives of phosphatidylinositol (PtdIns), often collectively referred to as polyphosphoinositides (PPIn), are a minor component of eukaryotic cell membranes. Nevertheless, their synthesis is needed for an ever-increasing spectrum of cellu ...

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Lipidomic Analysis of Phospholipids and Related Structures by Liquid Chromatography-Mass Spectrometry

Lipids are highly dynamic molecules with many different roles ranging from structural to signaling. Alterations in particular lipid levels change cellular behavior. In humans, inappropriate changes may manifest as diseases such as Alzheimer’s, atherosclerosis, diabetes, ob ...

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Eicosanoids: Generation and Detection in Mammalian Cells

Eicosanoids are 20-carbon lipids generated by the oxidation of arachidonic acid that are involved in physiological signaling in virtually all organ systems. Three primary enzymatic pathways are responsible for their synthesis in mammalian cells: lipoxygenase, cyclooxygena ...

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Proteomic Analysis of the Sterol-Mediated Signaling Pathway in Caenorhabditis elegans

Since Caenorhabditis elegans is incapable of de novo cholesterol biosynthesis, it must utilize other nonpermissive sterols that are present in the environment by converting them into cholesterol for cellular function. The inhibition of sterol conversion to cholesterol in C. eleg ...

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Effects of Sterols on the Development and Aging of Caenorhabditis elegans

Although Caenorhabditis elegans lacks several components of the de novo sterol biosynthetic pathway, it requires sterols as essential nutrients. Supplemental cholesterol undergoes extensive enzymatic modification in C. elegans to form certain sterols of unknown function. ...

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Ceramide-Induced Transbilayer (Flip-Flop) Lipid Movement in Membranes

Lipids in biological membranes are asymmetrically distributed across the bilayer. The choline-containing lipids, phosphatidylcholine (PtdCho) and sphingomyelin (SM), are more abundant in the external leaflet. In contrast, the amino-containing glycerophospholipids, ...

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Inositol Lipid-Dependent Functions in Saccharomyces cerevisiae: Analysis of Phosphatidylinositol Phosphates

Inositol phospholipids regulate many cellular processes, including cell survival, membrane trafficking, and actin polymerization. Quantification of inositol lipids is one of the essential techniques needed for studies that aim to decipher inositol lipid-dependent cell ...

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Lipid Quantification and Structure Determination of Nuclear Envelope Precursor Membranes in the Sea Urchin

Nuclear envelope assembly is a fundamental cellular process normally taking place once in every cell cycle in eukaryotes. The timing of fusion of nuclear membrane precursors to form the complete double membrane surrounding the chromosomes is tightly controlled, but much remains uncl ...

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Solution and Solid-State NMR of Lipids

Lipid structure and dynamics are of first importance for cellular function. Lipids such as phosphatidyl inositol (PtdIns) are essential in signaling pathways, as they are recognition sites at the membrane surface. Their head-group or chain structure appears to be crucial for such a signa ...

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Revealing Signaling in Single Cells by Single- and Two-Photon Fluorescence Lifetime Imaging Microscopy

Lipids are actively involved in many cellular processes. Their roles pivot toward determining membrane structure, compartment targeting, and membrane fusion but also regulation of cell signaling via their interactions with proteins and the production of second messengers. As th ...

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Preparation of Membrane Rafts

The concept that biological membranes contain microdomains of specialized lipid and protein composition has attracted great attention in recent years. Initially, the focus in the field was very much on the characterization of cholesterol-and sphingolipid-rich plasma membrane ...

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