Methods in molecular biology
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Methylation of cytosines is a very important epigenetic modification of genomic DNA in many different eukaryotes, and it is frequently involved in transcriptional regulation of genes. In plants, DNA methylation is regulated by a complex interplay between several methylating and demethylating enzymes. ⋯ Subsequent PCR and sequence analysis of individual amplicons displays the degree, position, and sequence context of methylation of every cytosine residue in individual genomic sequences. We describe the application of bisulfite sequencing for the analysis of DNA methylation at defined individual sequences of plant genomic DNA.
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The UV-B pain model utilizes ultraviolet light to induce a small area of inflammation allowing assessment of mechanical and thermal thresholds. Pharmacologic testing has mainly focused on reduction of primary hyperalgesia, although the effect of analgesics on secondary hyperalgesia has also been investigated. The model requires an instrument to precisely generate controlled UV-B tissue hyperalgesia. ⋯ Tissue is then assessed for inflammation using color Doppler imaging or flare measurements. Heat pain thresholds and pain tolerance are often evaluated using a commercially available thermal sensory testing device. Analgesics can be administered to determine the influence on these clinical endpoints.
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Standard therapies for many common cancers remain toxic and are often ineffective. Cellular immunotherapy has the potential to be a highly targeted alternative, with low toxicity to normal tissues but a high capacity to eradicate tumor. ⋯ Many of these approaches are proving successful in hematologic malignancy and in melanoma. In this chapter we discuss the advantages and limitations of each and how over the next decade investigators will attempt to broaden their reach, increase their efficacy, and simplify their application.
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The study of procedural sedation and analgesia has experienced significant development recently. As specific procedural sedation and analgesia agents have been developed and introduced into clinical practice, safety and efficacy studies have been conducted. ⋯ As procedural sedation and analgesia research has expanded, measurement techniques have been refined to allow for precise comparisons between smaller groups of subjects to improve the capacity to compare these procedures. We have used capnography, bispectral EEG analysis, and subject perceptions of pain and recall as surrogate predictors of adverse events in order to compare agents and procedural techniques in procedural sedation and analgesia.
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Animal models of tissue injury have been used to investigate the mechanisms of pain. Here, we describe a variety of animal models that have been used to mimic acute surgical pain in human subjects, which include the plantar, tail, and gastrocnemius incision models. We also provide discussion on animal models of laparotomy, thoracotomy, visceral pain, and bone injury. Preclinical studies using these models have provided insights into the mechanisms and causes of acute surgical pain as well as the treatment options to control postsurgical pain.