生物化学技术

Oxidative damage has been strongly implicated in the pathogenesis of a number of neurodegenerative diseases including Alzheimer’s disease (AD), Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS), and Huntington’s disease (HD) (1–9). Several years ago, we reported the di ...

The nitric oxide free radical (•NO) is formed by the enzymatic oxidation of arginine in a reaction catalyzed by various isoforms of the enzyme nitric oxide synthase (NOS). •NO serves myriad physiologic functions, including actions as a vasodilator. Under conditions of inflammation, macr ...

Protein oxidation is a recognized component of aging and a consequence of severe or prolonged exposure to reactive oxygen species (ROS). Direct attack of protein by ROS causes formation of protein-bound carbonyl groups (1). These carbonyl functions represent a variety of site-specific m ...

Reactive oxygen species (ROS) are a family of molecules including molecular oxygen and its derivatives produced in all aerobic cells. Extensive production of ROS has been implicated in the oxidation of biological macromolecules, such as DNA, proteins, carbohydrates, and lipids. This c ...

The biological importance of hydrogen peroxide and other “reactive oxygen species” (ROS) has become greatly appreciated in recent years. It has become apparent that certain ROS, in particular hydrogen peroxide (H2O2) and nitric oxide (•NO), are ubiquitously used as intra- or intercellu ...

A role of free radicals has been implicated in the pathogenesis of a number of neurological disorders, including amyotrophic lateral sclerosis, Huntington’s disease (HD), Parkinson’s disease (PD), and Alzheimer’s disease (AD) (1–9). Furthermore, reactive aldehydes produced as prod ...

The single-cell gel electrophoresis or comet assay was initially reported in 1984 as a novel technique to directly visualize DNA damage in individual cells (1). The method was modified by adding strong alkaline conditions to denature genomic DNA and detect alkali labile lesions, greatly im ...

In 1968, one of us began a quest to develop a general method for the synthesis of RNA sequences. The major problems associated with the chemical assembly of ribonucleotide chains were already well established. RNA sequences are very sensitive to chemical and enzymatic degradation. Conseque ...

Ribonucleoside H-phosphonates were introduced by Todd in 1957 to prepare diribonucleotide phosphate diesters (1). The H-phosphonates were activated for condensation with diphenyl chlorophosphate, and this activator was later extended to the synthesis of deoxyribonucle ...

The solid-phase strategy for oligonucleotide synthesis has been responsible for much of the widespread utilization of synthetic oligonucleotides. Indeed, without this approach, the chemical synthesis of oligonucleotides would have remained a difficult and tedious task sui ...

Synthetic DNA of defined sequences are commonly termed “oligonucleotides,” which are primarily composed of four different types of nucleosides linked through well-defined deoxyribose phosphate. Over the past ten years, because of refinement in synthesis chemistries (1a–1e) a ...

Solution-phase methods have been used for the first synthesis of an internucleotidic bond (1) and, some 20 years ago, had a glorious period during the first gene synthesis. At that time, their present-day competitor, the polymer support technique, was also developed and saw some first applicat ...

Nonionic oligonucleotide analogs have attracted much interest in recent years as potential candidates for oligonucleotide-based therapeutics. Compared to their natural parent molecules, they are expected to have the advantage of: 1. Being stable against degrading nucleases,

The development of deoxyribonucleoside phosphoramidite derivatives for the synthesis of oligodeoxyribonucleotides was first described by Beaucage and Caruthers (1) in 1981. The conceptual basis of this methodology emerged as a modification of the “phosphorodichloridi ...

As demonstrated by this volume, analogs of oligonucleotides retaining the molecular recognition properties of natural oligonucleotides, but having altered physical, chemical, and biological properties are synthetic targets of some interest. Much of the interest comes from t ...

As has been emphasized in corresponding chapters of this book, isotopic or elemental replacement of any of two nonbridging oxygens attached to internucleotide phosphorus atom(s) by substituent X (Fig. 1) creates, by virtue of asymmetry, new center(s) of chirality and results in the formati ...

Monomeric units of nucleic acids DN A or RN A are characterized by a β configuration at the anomeric center of the sugar moiety (Fig. 1). Natural constitutive nucleosides of this configuration are readily commercially available, and from such starting building blocks, numerous phosphate-b ...

Fluoro derivatives of phosphorus are of great importance in the chemistry and biochemistry of both elements. The incorporation of fluorine into biomolecules has frequently resulted in a remarkable change of biological properties. Simple phosphofluoridates and their structu ...

A number of short modified synthetic oligodeoxynucleotides have shown promise as agents for gene therapy (1–8). Among these, two classes of oligonucleotides with modified backbones (the phosphorothioates and methylphosphonates ) have emerged as primary candidates. We have syn ...

The common interest in the synthesis of oligo(nucleoside O-alkyl phosphates) was posed first by the generally accepted view that alkylation of DNA by several alkylating agents may also occur at internucleotide phosphate nonbridging oxygen atom(s) inducing DNA misfunction (1). Pert ...