Cancer Prevention: Dietary Factors and Pharmacology (Methods in Pharmacology and Toxicology)
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Focused on the discovery of precise molecular targets for the development of the cancer preventive agents, Cancer Prevention: Dietary Factors and Pharmacology provides researchers and non-researchers with practical methodologies for developing and validating small molecule and phytochemical-derived drug discovery and mechanisms by which these compounds can modulate distinct target proteins involved in oncogenic signaling. While this volume is primarily focused toward cancer prevention research, the range of techniques demonstrated in the book also provides an introduction of cancer prevention research methods to researchers outside the field. Chapters deal with a critical discussion of both laboratory and clinical topics, with each chapter containing both a discursive section along with a detailed methods section. As part of the Methods in Pharmacology and Toxicology series, this meticulous volume includes the kind of key implementation advice that seeks to ensure successful results in the lab.
Practical and authoritative, Cancer Prevention: Dietary Factors and Pharmacology aims to guide research toward identifying molecular targets and conducting human studies with phytochemicals which would, ideally, provide an enhanced approach to the goal of personalized cancer prevention.
achieved by fluorescence labeling of the nuclei of cells in suspension and then analyzing the fluorescence properties of each cell in the population. Nucleic acid dyes like propidium iodide (PI) are applied for DNA assays . Quiescent and G1 phase cells with only one DNA copy will have 1× fluorescence intensity. Cells in G2–M phase of the cell cycle have two DNA copies and accordingly have 2× intensity. Because cells in S phase are synthesizing DNA, they populate in a range between 1× and 2×
Transcription factors such as activator protein-1 (AP-1), nuclear factor-kappaB (NF-κB), p53, nuclear factor of activated T cells (NFAT), and cAMP response element-binding (CREB) protein have been shown to play a critical role in carcinogenesis and all are regulated by the mitogen-activated protein (MAP) kinase cascades. The activation of these or other transcription factors results in transcription of genes that encode proteins that regulate a multitude of cellular responses including apoptosis,
a picture of the 3D structure of the protein. The Hormel Institute’s High Performance Computing facility consists of two supercomputers completely dedicated to molecular modeling and docking experiments. The computational power of the two systems together clocks in at approximately 35 TFLOPS (teraflops). The original system installed in 2008 is a single rack IBM BlueGene/L with 1,024 dual core Power PC CPUs. System management and storage are implemented with five Linux servers and a DS4200
added during the nucleic acid protein binding reaction (referred to 170 Ha-Na Lee et al. as a supershift assay). Here, the method for checking DNA binding activity of NF-κB is described. 3.8.1 Preparation of Radiolabelled DNA Probes ( Note 8) 1. Mix 3 μL of oligonucleotide containing the NF-κB binding domains (Promega), 2 μL of 5× T4 polynucleotide buffer (Promega), 1 μL of T4 kinase (10 unit/μL; Promega) and 11 μL of DDW. 2. Incubate for 10 min on ice. 3. Add 3 μL of the mixture. P-γATP
50 μL of mammalian cell lysis solution to 100 μL of cell suspension/inhibitors per well, including the standards and blanks, and shake the plate for 5 min in a platform shaker at 300 rpm. 5. Reconstitute one lyophilized substrate solution bottle by adding 5 mL of substrate buffer solution. Agitate gently until the solution is homogeneous. 6. Using a multichannel pipette, add 50 μL of reconstituted substrate solution to the wells, including the standards and blanks. 7. Wrap the plate in aluminum