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The Use of Consensus PCR and Direct Sequence Analysis for the Identification of HPV

The human papillomaviruses (HPVs) form a large group of small double-stranded DNA viruses of about 7900 bp. The viruses infect mucosal and cutaneous epithelia and are associated with a number of benign and malignant lesions (1–4) HPVs originally have been described to be present in skin warts. St ...

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PCR Detection of HIV

The human immunodeficiency viruses type 1 and 2 (HIV-1, HIV-2) are members of the lentivirus subfamily of the retroviruses. As typical for all retroviruses, viral RNA replication requires the synthesis of a DNA intermediate, which is stably integrated into the host cell DNA (termed provirus). ...

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Improved PCR Methods for Identification of Phytopathogenic Viruses

Knowledge of the nucleotide sequence of a viral genome enables the design of specific oligonucleotides for use as primers for selective amplification of a target nucleic acid from a pool of complex template by a polymerase chain reaction (PCR) driven by Taq, a thermostable DNA polymerase (1,2). ...

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The Identification of Dengue Virus Using PCR

Dengue virus is a member of the family Flaviviridae with a single-stranded positive sense RNA genome. The virus particle consists of a lipid bilayer with envelope (E) and membrane (M) proteins surrounding a spherical nucleocapsid composed of a genome RNA and capsid (C) proteins. The virion stru ...

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Differential Expression of TIMP-3 During Neoplastic Progression in the Mouse JB6 Model System

The differential display technique has proved to be of particular value in our laboratory in identifying genes whose transcription is deregulated as part of the carcinogenesis process. The power of this technique, when applied to biologically relevant model systems for cancer develo ...

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Identification of Genes Associated with Resistance to Photodynamic Therapy-Mediated Oxidative Stress

Photodynamic therapy (PDT) is an experimental treatment for malignant diseases (1,2). This procedure involves the systemic administration of a tumor-localizing photosensitizer and subsequent exposure of the malignant lesion to tissue penetrating red light (3). Properties of ...

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The Application of Differential Display to the Brain: Adaptations for the Study of Heterogeneous Tissue

One of the main advantages of using differential display is the ability to examine simultaneously gene expression in multiple mRNA populations. In other techniques, such as differential screening or subtractive hybridization, only two mRNA populations can be easily examined at the s ...

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RAP-PCR Using RNA from Tissue Microdissection

Cell culture provides powerful models for studying cell biology, but many pathologic states cannot be appropriately studied without tissue-based approaches. We have developed a tissue-based microdissection method for molecular analysis and have chosen premalignant breast ...

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Analysis of Gene Expression in Hypothalamus in Obese and Normal Mice Using Differential Display

Obesity may result from increased calorie consumption, decreased energy expenditure, or both (1). These processes are controlled, at least in part, by the hypothalamus. Hypothalamic dysfunction is important in the development of obesity and it has long been known that lesions in the ventr ...

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Immobilization of Enzymes by Selective Adsorption on Biotinylaminopropyl Celite or Glass

Use of immobilized enzymes in bioprocesses offers many advantages, including greater productivity because the same enzyme molecules can be used over a long period of time, there is more precise control of the extent of reaction, there is the capability of automation and continuous operati ...

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Preparation of Immobilized Subunits of a Multisubunit Enzyme

It is well established that many intracellular enzymes are oligomeric with dimers and tetramers being the most common of the possible subunit structures (1,2). Evaluation of the functional significance of the quaternary structure is an important part of the fundamental studies on the ca ...

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Calcium Alginate Film Formed on a Stainless Steel Mesh

Viable cells can be immobilized in calcium alginate to improve the volumetric productivity of the fermentation process by increasing the cell numbers per unit of bioreactor volume and using the cells in their most productive growth phase. Also, in a continuous process, immobilization pr ...

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Enzyme Immobilization Using Chitosan-Xanthan Complexes

The immobilization of enzymes is a technique extensively studied since the late 1960s (1), and the knowledge accumulated on enzyme immobilization studies has grown steadily since then (2–6). Hydrogels are a particular category of support material that can be used for convenient immobil ...

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Covalent Immobilization of Enzymes to Graphitic Particles

The use of carbon as a support material for enzymes is receiving increasing attention, particularly in the field of enzyme electrodes, in which use is made both of the conducting nature of carbon and the degree of surface chemical reactivity associated with the material. This final considerat ...

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Cell Immobilization with Phosphorylated Polyvinyl Alcohol (PVA) Gel

Phosphorylated polyvinyl alcohol (PVA) gel is nontoxic and can be economically feasible for industrial scale applications. The use of PVA for cell immobilization has recently attracted much attention (1–5). In this chapter, we provide details of a novel cell immobilization technique b ...

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Immobilization of Liposomes and Proteoliposomes in Gel Beads

The lipid bilayers of liposomes are structurally similar to those of biological membranes and are often used as membrane models. Solubilized membrane proteins (transporters, receptors, enzymes, and so on) can be reconstituted into the bilayers (1–3). Interactions between the recons ...

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Immobilized Hepatocytes in Xenobiotic Biotransformation Studies

In Chapter 21, we described a method for hepatocyte immobilization in agarose threads that enables perfusion of the cells and facilitates varied biomedical studies on the biochemical properties of hepatocytes. Since the liver is the most important and unique organ in metabolic process ...

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Hepatocyte Immobilization in Agarose and Functional Integrity Testing

Isolated liver cells are extensively used in various studies and find wide applications in the fields of biochemistry, physiology, molecular biology, pharmacotoxicology, testing chemical pollutants, mutagenicity, carcinogenicity, and in other biomedical studies. The imp ...

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Immobilization of Cells Using Electrostatic Droplet Generation

A major concern in cell and bioactive agent immobilization has been the production of very small microbeads to minimize the mass transfer resistance problem associated with large diameter beads (i.e., 1000 �m) (1). Conventional technology involves production of alginate beads with dia ...

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Primary Cell Cultures for the Study of Myelination

Tissue culture allows the separate establishment of neurons and glia under a variety of conditions of substrate and media. By subsequent recombination of relatively pure cell populations, a number of questions of neuronal development and neuronal-glia interactions, specifical ...

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