MedWorm.com provides a medical RSS filtering service. Over 6000 RSS medical sources are combined and output via different filters. This feed contains the latest items from the 'Springer protocols feed by Cancer Research' source. Fri, 19 Mar 2010 13:59:37 +0100 http://www.medworm.com/index.php?rid=3351522&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_8 Gold nanorods (GNRs) are strongly absorbing at near-infrared (NIR) frequencies and can be employed as multifunctional agents for biological imaging and theragnostics. GNRs can support nonlinear optical microscopies based on two-photon-excited luminescence and can enhance the contrast of biomedical imaging modalities such as optical coherence tomography and photoacoustic tomography. GNRs are also efficient at mediating the conversion of NIR light energy into heat and can generate localized photothermal effects. However, future clinical applications will require the rigorous removal of CTAB, a micellar surfactant used in GNR synthesis, and reliable methods of surface functionalization for cell-selective targeting and for minimizing nonspecific uptake into cells. This can be accomplished by u... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351522 Thu, 11 Mar 2010 13:51:55 +0100 3351522 http://www.medworm.com/index.php?rid=3351521&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_7 Cancer is a leading cause of death in the United States and contributes to yearly rising health care costs. Current methods of treating cancer involve surgical removal of easily accessible tumors, radiation therapy, and chemotherapy. These methods do not always result in full treatment of the cancer and can in many cases damage healthy cells both surrounding the tissue area and systemically. Nanoshells are optically tunable core/shell nanoparticles that can be fabricated to strongly absorb in the near-infrared (NIR) region where light transmits deeply into tissue. When injected systemically, these particles have been shown to accumulate in the tumor due to the enhanced permeability and retention (EPR) effect and induce photothermal ablation of the tumor when irradiated with an NIR laser. T... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351521 Thu, 11 Mar 2010 13:51:55 +0100 3351521 http://www.medworm.com/index.php?rid=3351520&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_6 Gold nanocages are hollow nanostructures with porous walls that can be simply prepared via the galvanic replacement reaction between silver nanocubes and chloroauric acid. Their optical resonance peaks can be precisely tuned into the near-infrared region, in which the adsorption caused by blood or soft tissue is essentially negligible. Significantly, the strong absorption of gold nanocages makes them attractive as a novel class of contrast enhancement and photothermal agents for cancer detection and treatment. The well-established chemistry for gold also allows them to target specific cells by functionalizing their surface with various moieties such as antibodies, peptides, and DNAs. In this chapter, we focus on their use as a photothermal agent for the ablation of cancer cells and as a co... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351520 Thu, 11 Mar 2010 13:51:55 +0100 3351520 http://www.medworm.com/index.php?rid=3351519&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_5 A wide variety of bioimaging techniques (e.g., ultrasound, computed X-ray tomography, magnetic resonance imaging (MRI), and positron emission tomography) are commonly employed for clinical diagnostics and scientific research. While all of these methods use a characteristic “energy–matter” interaction to provide specific details about biological processes, each modality differs from another in terms of spatial and temporal resolution, anatomical and molecular details, imaging depth, as well as the desirable material properties of contrast agents needed for augmented imaging. On many occasions, it is advantageous to apply multiple complimentary imaging modalities for faster and more accurate prognosis. Since most imaging modalities employ exogenous contrast agents to improv... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351519 Thu, 11 Mar 2010 13:51:54 +0100 3351519 http://www.medworm.com/index.php?rid=3351518&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_4 Nanotechnology is actively being used to develop promising diagnostics and therapeutics tools for the treatment of cancer and many other diseases. The unique properties of nanomaterials offer an exciting frontier of possibilities for biomedical researchers and scientists. Because existing knowledge of macroscopic materials does not always allow for adequate prediction of the characteristics and behaviors of nanoscale materials in controlled environments, much less in biological systems, careful nanoparticle characterization should accompany biomedical applications of these materials. Informed correlations between adequately characterized nanomaterial properties and reliable biological endpoints are essential for guiding present and future researchers toward clinical nanotechnology-based so... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351518 Thu, 11 Mar 2010 13:51:54 +0100 3351518 http://www.medworm.com/index.php?rid=3351517&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_3 Effective cancer therapy remains one of the most challenging tasks to the scientific community, with little advancement on overall cancer survival landscape during the last two decades. A major limitation inherent to most conventional anticancer chemotherapeutic agents is their lack of tumor selectivity. One way to achieve selective drug targeting to solid tumors is to exploit abnormalities of tumor vasculature, namely hypervascularization, aberrant vascular architecture, extensive production of vascular permeability factors stimulating extravasation within tumor tissues, and lack of lymphatic drainage. Due to their large size, nano-sized macromolecular anticancer drugs administered intravenously (i.v.) escape renal clearance. Being unable to penetrate through tight endothelial junctions o... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351517 Thu, 11 Mar 2010 13:51:54 +0100 3351517 http://www.medworm.com/index.php?rid=3351516&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_2 In order to stop malignant tumor growth, >90% of a critical biochemical pathway needs to be blocked. Due to extraordinary advances in molecular biology, there is an increased understanding of rationale and relevant molecular targets in cancer. However, due to the heterogeneity of the molecular abnormalities in multiple tumor types, strategies designed to interfere with multiple molecular abnormalities will be necessary to impact survival. Nanoparticles have the potential to provide therapies not possible with other drug modalities. Researchers and clinicians must take advantage of these opportunities in order for nanotechnology to make an impact in the diagnosis and treatment of malignancy. A discussion of relevant targets either on the cell surface or the cytoplasm and strategies to ac... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351516 Thu, 11 Mar 2010 13:51:54 +0100 3351516 http://www.medworm.com/index.php?rid=3351515&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_26 Liposomal doxorubicin and nab-paclitaxel are nanoparticle formulations of traditional cancer chemotherapy drugs which have ample clinical experience both pre- and post-nanoparticle modification. The alterations in pharmacokinetics, pharmacodynamics, efficacy, and toxicity compared with their parent compounds are instructive for future development of nanoparticle-based therapies. In this article we review the current status of these agents, emphasizing the alterations in clinical behavior resulting from the nanoparticle formulation of the parent compound. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351515 Thu, 11 Mar 2010 13:51:54 +0100 3351515 http://www.medworm.com/index.php?rid=3351514&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_25 Since their initial description in 1857, gold nanoparticles have been used extensively in the fields of diagnostics and therapeutics. Now, gold nanoparticles are engineered to target the delivery of potent anti-cancer therapeutics to solid tumors to improve either their safety or efficacy or both. Described in this chapter is the development of one such nanotherapeutic, termed CYT-6091, that targets the delivery of tumor necrosis factor alpha (TNF) to solid tumors. Outlined in the presentation is a discussion of nanoparticles and specifically colloidal gold, an historical review on the biology of TNF and its limited use in the clinic when administered systemically, and finally, how gold nanoparticles bound with TNF may improve the safety and efficacy profiles of TNF. (Source: Springer prot... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351514 Thu, 11 Mar 2010 13:51:54 +0100 3351514 http://www.medworm.com/index.php?rid=3351513&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_24 Shortwave (MHz range) radiofrequency (RF) energy is nonionizing, penetrates deeply into biological tissues with no adverse side effects, and heats metallic nanoparticles efficiently. Targeted delivery of these nanoparticles to cancer cells should result in hyperthermic cytotoxicity upon exposure to a focused, noninvasive RF field. We have demonstrated that gold nanoparticles conjugated with cetuximab (C225) are quickly internalized by Panc-1 (pancreatic adenocarcinoma) and Difi (colorectal adenocarcinoma) cancer cells overexpressing epidermal growth factor receptor (EGFR). Panc-1 or Difi cells treated with naked gold nanoparticles or nonspecific IgG-conjugated gold nanoparticles demonstrated minimal intracellular uptake of gold nanoparticles by transmission electron microscopy (TEM). In co... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351513 Thu, 11 Mar 2010 13:51:54 +0100 3351513 http://www.medworm.com/index.php?rid=3351512&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_23 This chapter describes the application of gold nanorods in biomedical imaging and photothermal therapy. The photothermal properties of gold nanorods are summarized and the synthesis as well as antibody conjugation of gold nanorods is outlined. Biomedical applications of gold nanorods include cancer imaging using their enhanced scattering property and photothermal therapy using their enhanced nonradiative photothermal property. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351512 Thu, 11 Mar 2010 13:51:54 +0100 3351512 http://www.medworm.com/index.php?rid=3351511&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_22 Apoptosis, or programmed cell death, is a morphologically and biochemically distinct form of cell death, which together with proliferation plays an important role in tissue development and homeostasis. Insufficient apoptosis is important in the pathology of various disorders such as cancer and autoimmune diseases, whereas a high apoptotic activity is associated with myocardial infarction, neurodegenerative diseases, and advanced atherosclerotic lesions. Consequently, apoptosis is recognized as an important therapeutic target, which should be either suppressed, e.g., during an ischemic cardiac infarction, or promoted, e.g., in the treatment of cancerous lesions. Imaging tools to address location, amount, and time course of apoptotic activity non-invasively in vivo are therefore of great cli... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351511 Thu, 11 Mar 2010 13:51:54 +0100 3351511 http://www.medworm.com/index.php?rid=3351510&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_21 Nanotechnology is the key to a new, noninvasive photoacoustic imaging technique that could detect early stages of disease tissues. The combination of photoacoustic imaging with nanotechnology holds promise for determining the structural and functional properties of tissues with enhanced sensitivity and specificity and for monitoring the treatment of diseases. In this chapter, we described in detail photoacoustic reconstruction methods and imaging systems. We also review the recent advances in nanoparticles and their in vivo applications in the field of photoacoustic imaging. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351510 Thu, 11 Mar 2010 13:51:53 +0100 3351510 http://www.medworm.com/index.php?rid=3351509&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_20 Immunoliposomes generated by coupling of antibodies to the liposomal surface allow for an active tissue targeting, e.g., through binding to tumor cell-specific receptors. Instead of whole antibodies, single-chain Fv fragments (scFv), which represent the smallest part of an antibody containing the entire antigen-binding site, find increasing usage as targeting moiety. Here we provide protocols for the preparation of type II scFv immunoliposomes by the conventional coupling method as well as the post-insertion method. Furthermore protocols to analyze binding of these immunoliposomes to antigen-expressing cells as well as internalization through receptor-mediated endocytosis are included. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351509 Thu, 11 Mar 2010 13:51:53 +0100 3351509 http://www.medworm.com/index.php?rid=3351508&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_1 Cancer nanotechnology has the potential to dramatically improve current approaches to cancer detection, diagnosis, imaging, and therapy while reducing toxicity associated with traditional cancer therapy (1, 2). In this overview, we will define cancer nanotechnology, consider issues related to application of nanotechnology for cancer imaging and therapy, and broadly consider implications for continued development in nanotechnology for the future of clinical cancer care. These considerations will place in perspective the methodological approaches in cancer nanotechnology and subject reviews outlined in this volume. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351508 Thu, 11 Mar 2010 13:51:53 +0100 3351508 http://www.medworm.com/index.php?rid=3351507&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_19 Synthesis and evaluation of a novel cancer cell’s receptor-targeted internally quaternized and surface neutral poly(amidoamine) (PAMAM) generation four dendrimer as well as PAMAM–paclitaxel conjugate are described. The advantages of developed nanocarriers include but are not limited to (1) internal cationic charges for the complexation with small interfering RNA or antisense oligonucleotides and their protection from the degradation in systemic circulation; (2) neutral-modified surface for low cytotoxicity of empty unloaded dendrimers; (3) efficient internalization by cancer cells; and (4) preferential accumulation in the tumor and the prevention of adverse side effects of chemotherapy. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351507 Thu, 11 Mar 2010 13:51:53 +0100 3351507 http://www.medworm.com/index.php?rid=3351506&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_18 Inhalation of aerosols represents the most frequently used drug delivery method for the treatment of lung diseases. To further improve drug efficacy in the lungs, it may be advantageous to control aerosol deposition and target aerosols to diseased or disease-causing lung tissue and cellular structures in order to maximize drug potency and minimize side effects in unaffected tissue. We have recently investigated a novel method which brings aerosol delivery to an advanced level of specificity by making use of magnetic gradient fields to direct magnetizable aerosol droplets containing superparamagnetic iron oxide nanoparticles (SPION) specifically to desired regions of the lungs in mice. In this chapter, we will present a detailed description of this procedure for application in mice. (Source... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351506 Thu, 11 Mar 2010 13:51:53 +0100 3351506 http://www.medworm.com/index.php?rid=3351505&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_17 Nanoparticulate medicines offer the advantage of allowing delivery of large quantities of unmodified drug within the same particle. Nanoparticle uptake by cancer cells can, however, be compromised due to the large size and hydrophilicity of the particle. To circumvent cell penetration problems and simultaneously improve tumor specificity, nanoparticulate medicines have been linked to targeting ligands that bind to malignant cell surfaces and enter cells by receptor-mediated endocytosis. In this chapter, we summarize multiple methods for delivering nanoparticles into cancer cells by folate receptor-mediated endocytosis, devoting special emphasis to folate-targeted liposomes. Folate receptor-mediated endocytosis has emerged as an attractive strategy for nanoparticle delivery due to both over... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351505 Thu, 11 Mar 2010 13:51:53 +0100 3351505 http://www.medworm.com/index.php?rid=3351504&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_16 Aptamers are DNA or RNA oligonucleotide sequences that selectively bind to their target with high affinity and specificity. They are obtained using an iterative selection protocol called SELEX. Several small molecules and proteins have been used as targets. Recently, a variant of this methodology, known as cell-SELEX, has been developed for a new generation of aptamers, which are capable of recognizing whole living cells. We have used this methodology for the selection of aptamers, which show high affinity and specificity for several cancer cells. In this chapter, we describe (1) the process followed for the generation of aptamers capable of recognizing acute leukemia cells (CCRF–CEM cells) and (2) the method of enhancing the selectivity and sensitivity of these aptamers by conjugati... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351504 Thu, 11 Mar 2010 13:51:53 +0100 3351504 http://www.medworm.com/index.php?rid=3351503&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_15 In this chapter, we outline the protocols for PEGylation of some drug carriers, such as dendrimer, polymeric nanoparticles, liposomes, for systemic delivery. PEGylation simply refers to the modification of particle surface by covalently grafting, entrapping, or adsorbing PEG chains of vivid length. However, limitation of simple adsorption being easy displacement of the coating layer in vivo, covalent mode for PEGylation of nanoparticles is mostly preferred, and outlined herein. Derivatization and activation of polyethylene glycol is an important step during PEGylation and its chemistry chiefly relies on availability as well as type of functional groups on carrier periphery. A summarized set of protocols for PEGylation of widely explored nanocarriers for systemic delivery is presented. (Sou... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351503 Thu, 11 Mar 2010 13:51:53 +0100 3351503 http://www.medworm.com/index.php?rid=3351502&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_14 During nanoparticle system in drug delivery, liposomes were perhaps the best characterized and one of the first to be developed. Stealth liposomes (SLs), containing polyethylene glycol-conjugated lipid, which can form a hydro-layer around liposomes bilayer, have a long circulation time and hence result in enhanced drug efficiency. Doxorubicin (DOX), an effective anticancer drug, can be loaded into liposomes by transmembrane pH gradient method to get high encapsulation efficiency with high drug/lipid ratio. Liposomal doxorubicin is a successful clinical formulation, and also a perfect model drug system for cancer-therapy research. Here we described the preparation of SLs via extrusion, DOX loading by transmembrane pH gradient method, and characterization analysis, including phospholipid con... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351502 Thu, 11 Mar 2010 13:51:53 +0100 3351502 http://www.medworm.com/index.php?rid=3351501&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_13 Pancreatic cancer is generally detected at later stages with a poor prognosis and a high-mortality rate. Development of theranostic imaging agents that noninvasively target pancreatic cancer by gene expression and deliver therapies directly to malignant cells could greatly improve therapeutic outcomes. Small-peptide ligands that bind cell-surface proteins and are conjugated to imaging moieties have demonstrated efficacy in cancer imaging. Identification of cancer-specific targets is a major bottleneck in the development of such agents. Herein, a method is presented that uses DNA microarray expression profiling of large sets of normal and cancer tissues to identify targets expressed in cancer but not expressed in relevant normal tissues. Identified targets are subsequently validated for pro... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351501 Thu, 11 Mar 2010 13:51:53 +0100 3351501 http://www.medworm.com/index.php?rid=3351500&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_12 This chapter describes the methodology by which mAb-F19-conjugated gold nanoparticles were prepared and used to label human pancreatic adenocarcinoma. Specifically, gold nanoparticles were coated with dithiol bearing hetero-bifunctional PEG (polyethylene glycol), and cancer-specific mAb F19 was attached by means of NHS-EDC coupling chemistry taking advantage of a carboxylic acid group on the heterobifunctional PEG. These conjugates were completely stable and were characterized by a variety of methods, including UV–Vis absorbance spectrometry, darkfield microscopy, DLS (dynamic light scattering), TEM (transmission electron microscopy), SEC (size-exclusion chromatography), and confocal microscopy. Nanoparticle bioconjugates were used to label sections of healthy and cancerous human pan... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351500 Thu, 11 Mar 2010 13:51:52 +0100 3351500 http://www.medworm.com/index.php?rid=3351499&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_11 The use of biodegradable polymeric nanoparticles (NPs) for controlled drug delivery has shown significant therapeutic potential. Concurrently, targeted delivery technologies are becoming increasingly important as a scientific area of investigation. In cancer, targeted polymeric NPs can be used to deliver chemotherapies to tumor cells with greater efficacy and reduced cytotoxicity on peripheral healthy tissues. In this chapter, we describe the methods of (1) preparation and characterization of drug-encapsulated polymeric NPs formulated with biocompatible and biodegradable poly(D,L-lactic-co-glycolic acid)-poly(ethylene glycol) (PLGA-b-PEG) copolymers; (2) surface functionalization of the polymeric NPs with the A10 2'-fluoropyrimidine ribonucleic acid (RNA) aptamers that recognize the prosta... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351499 Thu, 11 Mar 2010 13:51:52 +0100 3351499 http://www.medworm.com/index.php?rid=3351498&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-609-2_10 In recent years, fluorescent silica nanoparticles (FSNPs) received immense interest in cancer imaging. FSNPs are a new class of engineered optical probes consisting of silica NPs loaded with fluorescent dye molecules. These probes exhibit some attractive features, such as photostability and brightness, which allow sensitive imaging of cancer cells. In general, FSNPs are chemically synthesized in solution using appropriate silane-based precursors. Fluorescent dye molecules are entrapped during the synthesis process. The synthetic process involves hydrolysis and condensation reactions of silane precursors. Stöber’s sol–gel and water-in-oil (W/O) microemulsion methods are two popular chemical methods that have been used for synthesizing FSNPs. Silica matrix is capable of carr... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3351498 Thu, 11 Mar 2010 13:51:52 +0100 3351498 http://www.medworm.com/index.php?rid=3090247&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_8 Multidrug resistance (MDR) mediated by overexpression of P-glycoprotein (Pgp) is one of the best characterized transporter-mediated barriers to successful chemotherapy in cancer patients and is also a rapidly emerging target in the progression of neurodegenerative disorders such as Alzheimer’s and Parkinson’s diseases. Therefore, strategies capable of delivering chemotherapeutic agents into drug-resistant tumors and targeted radiopharmaceuticals acting as ultrasensitive molecular imaging probes for detecting functional Pgp expression in vivo could be expected to play a vital role in systemic biology as personalized medicine gains momentum in the twenty-first century. While targeted therapy could be expected to deliver optimal doses of chemotherapeutic drugs into the desired tar... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3090247 Fri, 23 Oct 2009 00:00:00 +0100 3090247 http://www.medworm.com/index.php?rid=3068416&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_14 The multidrug resistance (MDR) phenotype exhibited by cancer cells is believed to be the major barriers to successful chemotherapy in cancer patients. The major form of MDR phenotype is contributed by a group of ATP-binding cassette (ABC) drug transporters which include P-glycoprotein, multidrug resistance-associated protein 1, and breast cancer resistance protein. There has been intense search for compounds which can act to reverse MDR phenotype in cultured cells, in animal models, and ultimately in patients. The ongoing search for MDR modulators, compounds that act directly on the ABC transporter proteins to block their activity, has led to three generations of drugs. Some of the third-generation MDR modulators have demonstrated encouraging results compared to earlier generation MDR modu... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068416 Fri, 23 Oct 2009 00:00:00 +0100 3068416 http://www.medworm.com/index.php?rid=3068415&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_12 Breast cancer resistance protein (BCRP/ABCG2) was discovered in multidrug resistant breast cancer cells having an ATP-dependent transport-based resistance phenotype. This ABC transporter functions (at least in part) as a xenobiotic protective mechanism for the organism: in the gut and biliary tract, it prevents absorption and enhances elimination of potentially toxic substances. As a placental barrier, it protects the fetus; similarly, it serves as a component of blood-brain and blood-testis barrier; BCRP is expressed in stem cells and may protect them from potentially harmful agents. Therefore, BCRP could influence cancer outcomes by (a) endogenous BCRP affecting the absorption, distribution, metabolism, and elimination of anticancer drugs; (b) BCRP expression in cancer cells may directly... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068415 Fri, 23 Oct 2009 00:00:00 +0100 3068415 http://www.medworm.com/index.php?rid=3068414&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_11 Millions of new cancer patients are diagnosed each year and over half of these patients die from this devastating disease. Thus, cancer causes a major public health problem worldwide. Chemotherapy remains the principal mode to treat many metastatic cancers. However, occurrence of cellular multidrug resistance (MDR) prevents efficient killing of cancer cells, leading to chemotherapeutic treatment failure. Over-expression of ATP-binding cassette transporters, such as P-glycoprotein, breast cancer resistance protein and/or multidrug resistance-associated protein 1 (MRP1), confers an acquired MDR due to their capabilities of transporting a broad range of chemically diverse anticancer drugs across the cell membrane barrier. In this review, the molecular mechanism of ATP-dependent solute transpo... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068414 Fri, 23 Oct 2009 00:00:00 +0100 3068414 http://www.medworm.com/index.php?rid=3068413&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_10 ATP-binding cassette (ABC) drug efflux transporters limit intracellular concentration of their substrates by pumping them out of cell through an active, energy dependent mechanism. Several of these proteins have been originally associated with the phenomenon of multidrug resistance; however, later on, they have also been shown to control body disposition of their substrates. P-glycoprotein (Pgp) is the first detected and the best characterized of ABC drug efflux transporters. Apart from tumor cells, its constitutive expression has been reported in a variety of other tissues, such as the intestine, brain, liver, placenta, kidney, and others. Being located on the apical site of the plasma membrane, Pgp can remove a variety of structurally unrelated compounds, including clinically relevant dr... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068413 Fri, 23 Oct 2009 00:00:00 +0100 3068413 http://www.medworm.com/index.php?rid=3068412&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_13 A newly identified form of multidrug resistance (MDR) in tumor cells is presented, pertaining to the commonly encountered resistance of cancer cells to anticancer drug combinations at discrete drug:drug ratios. In vitro studies have revealed that whether anticancer drug combinations interact synergistically or antagonistically can depend on the ratio of the combined agents. Failure to control drug ratios in vivo due to uncoordinated pharmacokinetics could therefore lead to drug resistance if tumor cells are exposed to antagonistic drug ratios. Consequently, the most efficacious drug combination may not occur at the typically employed maximum tolerated doses of the combined drugs if this leads to antagonistic ratios in vivo after administration and resistance to therapeutic effects of the d... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068412 Fri, 23 Oct 2009 00:00:00 +0100 3068412 http://www.medworm.com/index.php?rid=3068411&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_15 Chemotherapy remains the mainstay in the treatment and management of many cancers. However, this treatment modality is fraught with difficulties associated with toxicity and also the emergence of chemotherapy resistance is a considerable problem. Cancer scientists and oncologists have worked together for some time to find ways of understanding anticancer drug resistance and also to develop pharmacological strategies to overcome that resistance. The greatest focus has been on the reversal of the multidrug resistance (MDR) phenotype by inhibition of the ATP-binding cassette (ABC) drug transporters. Inhibitors of ABC transporters – termed MDR modulators – have in the past been numerous and have occupied industry and academia in drug discovery programs. The field has been fraught w... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068411 Fri, 23 Oct 2009 00:00:00 +0100 3068411 http://www.medworm.com/index.php?rid=3068410&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_8 Multidrug resistance (MDR) mediated by overexpression of P-glycoprotein (Pgp) is one of the best characterized transporter-mediated barriers to successful chemotherapy in cancer patients and is also a rapidly emerging target in the progression of neurodegenerative disorders such as Alzheimer’s and Parkinson’s diseases. Therefore, strategies capable of delivering chemotherapeutic agents into drug-resistant tumors and targeted radiopharmaceuticals acting as ultrasensitive molecular imaging probes for detecting functional Pgp expression in vivo could be expected to play a vital role in systemic biology as personalized medicine gains momentum in the twenty-first century. While targeted therapy could be expected to deliver optimal doses of chemotherapeutic drugs into the desired tar... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068410 Fri, 23 Oct 2009 00:00:00 +0100 3068410 http://www.medworm.com/index.php?rid=3068409&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_7 There are several ways to detect proteins on cells. One quite frequently used method is flow cytometry. This method needs fluorescently labeled antibodies that can attach selectively to the protein to be investigated for flow cytometric detection. Flow cytometry scans individual cells, virtually without their surrounding liquid, and can scan many cells in a very short time. Because of this advantage of flow cytometry, it was adapted to investigate transport proteins on normal and cancerous human cells and cell lines. These transport proteins play important roles in human metabolism. Absorption in the intestine, excretion at the kidney, protection of the CNS compartment and the fetus from xenobiotics, and other vital functions depend on these transporters. However, several transporters are ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068409 Fri, 23 Oct 2009 00:00:00 +0100 3068409 http://www.medworm.com/index.php?rid=3068408&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_6 Drug resistance is a severe limitation of chemotherapy of various malignancies. In particular efflux transporters of the ATP-binding cassette family such as ABCB1 (P-glycoprotein), the ABCC (multidrug resistance-associated protein) family, and ABCG2 (breast cancer resistance protein) have been identified as major determinants of chemoresistance in tumor cells. Bioavailability depends not only on the activity of drug metabolizing enzymes but also to a major extent on the activity of drug transport across biomembranes. They are expressed in the apical membranes of many barrier tissues such as the intestine, liver, blood–brain barrier, kidney, placenta, testis, and in lymphocytes, thus contributing to plasma, liquor, but also intracellular drug disposition. Since expression and function... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068408 Fri, 23 Oct 2009 00:00:00 +0100 3068408 http://www.medworm.com/index.php?rid=3068407&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_5 Multidrug resistance (MDR) remains one of the key determinants in chemotherapeutic success of cancer patients. Often, acquired resistance is mediated by the overexpression of ATP-binding cassette (ABC) drug transporters. To study the mechanisms involved in the MDR phenotype, investigators have generated a variety of in vitro cell culture models using both multi-step and single-step drug selections. Sublines produced from multi-step selections have led to the discovery of several crucial drug transporters including ABCB1, ABCC1, and ABCG2. Additionally, a number of mechanisms causing gene overexpression have been elucidated. To more closely mimic in vivo conditions, investigators have also established MDR sublines with single-step drug selections. Here, we examine some of the multi-step and... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068407 Fri, 23 Oct 2009 00:00:00 +0100 3068407 http://www.medworm.com/index.php?rid=3068406&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_4 The development of multidrug resistance (MDR) to chemotherapy remains a major challenge in the treatment of cancer. Resistance exists against every effective anticancer drug and can develop by numerous mechanisms including decreased drug uptake, increased drug efflux, activation of detoxifying systems, activation of DNA repair mechanisms, evasion of drug-induced apoptosis, etc. In the first part of this chapter, we briefly summarize the current knowledge on individual cellular mechanisms responsible for MDR, with a special emphasis on ATP-binding cassette transporters, perhaps the main theme of this textbook. Although extensive work has been done to characterize MDR mechanisms in vitro, the translation of this knowledge to the clinic has not been crowned with success. Therefore, identifyin... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068406 Fri, 23 Oct 2009 00:00:00 +0100 3068406 http://www.medworm.com/index.php?rid=3068405&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_3 The generation of genetically engineered mouse models (GEMMs) that mimic breast cancer in humans provides new tools to investigate mechanisms of drug resistance in vivo. The advantages are manifold: inbred mice do not have the genomic heterogeneity seen in patients; mammary tumors are superficial and therefore easily accessible for measurement and sampling pre- and posttreatment; tumors can be transplanted orthotopically into syngeneic, immunocompetent animals; and tumor cells can be modified in vitro (e.g., gene overexpression, shRNA knockdown, insertional mutagenesis) prior to transplantation. Here, we provide an overview with experimental details of various approaches to study mechanisms of drug resistance in GEMMs for breast cancer. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068405 Fri, 23 Oct 2009 00:00:00 +0100 3068405 http://www.medworm.com/index.php?rid=3068404&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_2 Resistance of tumor cells to several structurally unrelated classes of natural products, including anthracyclines, taxanes, and epipodophyllotoxines, is often referred as multidrug resistance (MDR). This is associated with ATP-binding cassette transporters, which function as drug efflux pumps such as P-glycoprotein (Pgp) and multidrug resistance-associated protein 1 (MRP1). Because of the hypothesis in the early eighties that blockade of these efflux pumps by modulators would improve the effect of chemotherapy, extensive effort has been put to visualize these pumps using nuclear imaging with several specific tracers, using both SPECT and PET techniques. The methods and possibilities to visualize these pumps in both the tumor and the blood–brain barrier will be discussed. Because of t... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068404 Fri, 23 Oct 2009 00:00:00 +0100 3068404 http://www.medworm.com/index.php?rid=3068403&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_21 Patient relapse and metastasis of malignant cells is very common after standard cancer treatment with surgery, radiation, and/or chemotherapy. Chemotherapy, a cornerstone in the development of present day cancer therapy, is one of the most effective and potent strategies to treat malignant tumors. However, the resistance of cancer cells to the drugs remains a significant impediment to successful chemotherapy. An additional obstacle is the inability of chemotherapeutic drugs to selectively target tumor cells. Almost all the anticancer agents have severe side effects on normal tissues and organs. The toxicity of currently available anticancer drugs and the inefficiency of chemotherapeutic treatments, especially for advanced stages of the disease, have limited the optimization of clinical dru... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068403 Fri, 23 Oct 2009 00:00:00 +0100 3068403 http://www.medworm.com/index.php?rid=3068402&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_20 The ATP-binding cassette (ABC)-transporter P-glycoprotein (Pgp, also known as ABCB1) is the best characterized factor involved in multidrug resistance (MDR) of cancer cells. Pgp, which is encoded by the MDR1 gene, acts as a membrane-embedded drug extrusion pump for multiple structurally unrelated cytotoxic drugs. Inhibition of the pump activity of Pgp by low-molecular weight pharmacologically active compounds as a method to reverse MDR in cancer patients has been studied extensively, but so far clinical trials have generally been disappointing. Thus, experimental strategies for overcoming MDR are under investigation. These approaches include the application of the RNA interference (RNAi) technology. RNAi is a physiological mechanism triggered by small double-stranded RNA molecules resultin... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068402 Fri, 23 Oct 2009 00:00:00 +0100 3068402 http://www.medworm.com/index.php?rid=3068401&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_1 It is becoming increasingly clear that the proliferation of human tumours is driven by a small proportion of cells, termed tumour stem cells, which have the properties of self-renewal. On analogy with stem cells for normal tissues, there are likely to be multiple mechanisms, involving both intrinsic cellular properties and microenvironmental factors, which enable tumour stem cells to resist potentially genotoxic agents. Intrinsic properties include maintenance of cells in a predominantly non-cycling state, expression of transport proteins such as P-glycoprotein, protection from induced apoptosis or other forms of cell death, and limitation of diffusion of potential cytotoxins from the bloodstream. In addition, tumour stem cells are likely to contain multiple genetic changes that will poten... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068401 Fri, 23 Oct 2009 00:00:00 +0100 3068401 http://www.medworm.com/index.php?rid=3068400&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_19 Multidrug-resistance (MDR) is the major reason for failure of cancer therapy. ATP-binding cassette (ABC) transporters contribute to drug resistance via ATP-dependent drug efflux. P-glycoprotein (Pgp), which is encoded by MDR1 gene, confers resistance to certain anticancer agents. The development of agents able to modulate MDR mediated by Pgp and other ABC transporters remained a major goal for the past 20 years. The calcium blocker verapamil was the first drug shown to be a modulator of Pgp, and since many different chemical compounds have been shown to exert the same effect in vitro by blocking Pgp activity. These included particularly immunosuppressors. Cyclosporin A (CSA) was the first immunosuppressor that have been shown to modulate Pgp activity in laboratory models and entered very e... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068400 Fri, 23 Oct 2009 00:00:00 +0100 3068400 http://www.medworm.com/index.php?rid=3068399&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_18 The prominent role for the drug efflux pump ABCB1 (P-glycoprotein) in mediating resistance to chemotherapy was first suggested in 1976 and sparked an incredible drive to restore the efficacy of anticancer drugs. Achieving this goal seemed inevitable in 1982 when a series of calcium channel blockers were demonstrated to restore the efficacy of chemotherapy agents. A large number of other compounds have since been demonstrated to restore chemotherapeutic sensitivity in cancer cells or tissues. Where do we stand almost three decades since the first reports of ABCB1 inhibition? Unfortunately, in the aftermath of extensive fundamental and clinical research efforts the situation remains gloomy. Only a small handful of compounds have reached late stage clinical trials and none are in routine clin... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068399 Fri, 23 Oct 2009 00:00:00 +0100 3068399 http://www.medworm.com/index.php?rid=3068398&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_17 With the advent of polytherapy for cancer treatment it has become prudent to minimize, as much as possible, the potential for drug–drug interactions (DDI). Toward this end, the metabolic and transporter pathways involved in the disposition of a drug candidate (phenotyping) and potential for inhibition and induction of drug-metabolizing enzymes and transporters are evaluated in vitro. Such in vitro human data can be made available prior to human dosing and enable in vitro to in vivo-based predictions of clinical outcomes. Despite some success, however, in vitro systems are not dynamic and sometimes fail to predict drug–drug interactions for a variety of reasons. In comparison, relatively less effort has been made to evaluate predictions based on data derived from in vivo animal ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068398 Fri, 23 Oct 2009 00:00:00 +0100 3068398 http://www.medworm.com/index.php?rid=3068397&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_16 Modulation of P-glycoprotein (Pgp)-mediated transport has significant pharmacokinetic implications for Pgp substrates. Pharmacokinetic alterations may be at the systemic (blood concentrations), regional (organ or tissue concentrations), or local (intracellular concentrations) level. Regardless of the particular location of Pgp modulation, changes in substrate pharmacokinetics will have the potential to alter the magnitude and duration of pharmacologic effect (pharmacodynamics). It is important to understand each of the aspects of Pgp modulation for a given Pgp substrate in order to predict the degree to which Pgp modulation may affect that substrate, to minimize untoward effects associated with that modulation, or to exploit that modulation for specific therapeutic advantage. (Source: Spri... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068397 Fri, 23 Oct 2009 00:00:00 +0100 3068397 http://www.medworm.com/index.php?rid=3068396&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60761-416-6_9 Methylation of CpG dinucleotides is one of the major epigenetic processes involved in the regulation of gene expression. Catalyzed by DNA methyltransferases, hypermethylation of CpG islands in promoter regions is typically associated with gene silencing. DNA methylation plays an important role in normal differentiation, development, and maintenance of genomic stability, with aberrant CpG methylation being linked with a number of disease states. Three CpG islands within a 1.15- kb region characterize the chromatin landscape surrounding the transcriptional start site of the multidrug resistance 1 (MDR1) gene. We and others have demonstrated that hypermethylation of this region is correlated with MDR1 gene silencing and the inability of chemotherapeutic agents to activate MDR1 transcription. ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=3068396 Fri, 23 Oct 2009 00:00:00 +0100 3068396 http://www.medworm.com/index.php?rid=2960380&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_9 Recently, the analysis and functional elucidation of CpG island methylation has become a focus area of genomic research. Deviations from the normal parental imprinting pattern have been shown to cause developmental defects associated with serious symptoms. Aberrant DNA methylation of tumor suppressor and other functional genes, especially when found in 5′ untranslated regions and early exons, has been associated with tumorigenesis. In the context of applying DNA methylation analysis for the molecular characterization of cancer and other diseases, standardized protocols enabling parallel genome-wide methylation profiling of numerous samples are required. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960380 Tue, 15 Sep 2009 00:00:00 +0100 2960380 http://www.medworm.com/index.php?rid=2960379&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_8 Establishing a gene expression profile of defined subtypes of cells within an organ is still challenging because it frequently requires microdissection and subsequent amplification of the limited amount of messenger RNA (mRNA) isolated from the microdissected tissue in order to be able to proceed with comprehensive gene expression analyses via microarray or serial analysis of gene expression (SAGE) technology. Here we describe a manual microdissection strategy for the isolation of high-quality RNA. Furthermore, a strategy for combining linear amplification of RNA with longSAGE is described that allows the use of antisense RNA (aRNA) generated via the well-established linear amplification of RNA procedure together with the conventional SAGE or longSAGE technology. (Source: Springer protocol... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960379 Tue, 15 Sep 2009 00:00:00 +0100 2960379 http://www.medworm.com/index.php?rid=2960378&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_7 Since its invention in 1992, differential display (DD) has become the most commonly used technique for identifying differentially expressed genes because of its many advantages over competing technologies such as DNA microarray, serial analysis of gene expression (SAGE), and subtractive hybridization. A large number of these publications have been in the field of cancer, specifically on p53 target genes. Despite the great impact of the method on biomedical research, there had been a lack of automation of DD technology to increase its throughput and accuracy for systematic gene expression analysis. Many previous DD work has taken a “shotgun” approach of identifying one gene at a time, with a limited number of polymerase chain reactions (PCRs) set up manually, giving DD a low-tec... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960378 Tue, 15 Sep 2009 00:00:00 +0100 2960378 http://www.medworm.com/index.php?rid=2960377&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_6 Originally established in the beginning of the 1990s as a direct route to gene finding, expressed sequence tags (ESTs) still lend themselves as a means to analyze gene expression in almost all human tissues. The type of questions that can be addressed using public EST libraries ranges from tissue-specific gene profiling to the comparison between tissues in diseased and healthy states. Thanks to a multitude of web-based online bioinformatics resources, mining in EST libraries is not restricted to experts in the field of data analysis, but can readily be performed by the medical or life scientist. In this chapter, a couple of cases studies are presented that guide the scientist to the most useful online resources so that they can conduct their own research. (Source: Springer protocols feed b... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960377 Tue, 15 Sep 2009 00:00:00 +0100 2960377 http://www.medworm.com/index.php?rid=2960376&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_5 Cancer gene profiling has greatly profited from the progress in high-throughput technologies, including microarray-, sequencing-, and bioinformatics-based methods. The flood of data generated during the last decade has provoked a panel of “-omics” fields that significantly changed our understanding of malignant diseases. However, while the terms “-omics” and “-ome” in principle refer to the completeness of a genetic approach, we are in fact far from a complete understanding of cancer progression. We may understand gene expression patterns better and successfully use gene signatures for outcome prediction and prognosis, but truly promising molecular targets still have to find their way into novel therapeutic concepts. In this chapter, we will show how mor... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960376 Tue, 15 Sep 2009 00:00:00 +0100 2960376 http://www.medworm.com/index.php?rid=2960375&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_4 Modern array technologies allow for the simultaneous screening of virtually all human genes on the DNA and RNA level. Studies using such techniques have lead to the identification of hundreds of genes with a potential role in cancer or other diseases. The validation of all of these candidate genes requires in situ analysis of high numbers of clinical tissues samples. The tissue microarray (TMA) technology greatly facilitates such analysis. In this method, minute tissue samples (0.6 mm in diameter) from up to 1,000 different tissues can be analyzed on one microscope glass slide. All in situ methods suitable for histological studies can be applied to TMAs without major changes of protocols, including immunohistochemistry, fluorescence in situ hybridization, or RNA in situ hybridization. Beca... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960375 Tue, 15 Sep 2009 00:00:00 +0100 2960375 http://www.medworm.com/index.php?rid=2960374&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_3 Gene expression analysis requires a sound basis of cell material to obtain realistic results. Tissue, however, consists of diverse types of cells, which often differentially express target genes, so that cell populations need to be selected. If tissue diversity is moderate and negligible, manual microdissection can be the cost-efficient method of choice. In contrast, the advantage of laser microdissection is a very exact selection down to the level of a single cell, but often with a considerable time needed to get enough material for the following analyses. The latter issue and the method of tissue preparation needed for laser microdissection are the main problems to solve if RNA, highly sensitive to degradation, shall be analyzed. This method focuses on optimized laser microdissection pro... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960374 Tue, 15 Sep 2009 00:00:00 +0100 2960374 http://www.medworm.com/index.php?rid=2960373&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_2 The new opportunities of modern assays of molecular biology can only be exploited fully if the results can be accurately correlated to the tissue phenotype under investigation. This is a general problem of non-in situ techniques, whereas results from in situ techniques are often difficult to quantitate. The use of bulk tissue, which is not precisely characterized in terms of histology, has long been the basis for molecular analysis. It has, however, become apparent, that this simple approach is not sufficient for a detailed analysis of molecular alterations, which might be restricted to a specific tissue phenotype (e.g., tumor or healthy tissue, stromal or epithelial cells). Microdissection is a method to provide minute amounts of histologically characterized tissues for molecular analysis... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960373 Tue, 15 Sep 2009 00:00:00 +0100 2960373 http://www.medworm.com/index.php?rid=2960372&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_22 Target identification of novel therapeutic drugs is pivotal for the establishment of (1) new anticancer regimens, (2) to control side effects of the drugs, and (3) to identify appropriate combinations with established drugs. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960372 Tue, 15 Sep 2009 00:00:00 +0100 2960372 http://www.medworm.com/index.php?rid=2960371&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_21 DNA microarray profiles are plagued by the issue of large number of variables but small number of samples and are often notorious for their low signal-to-noise ratio for clinical applications. Therefore, a great need for meta-analysis techniques is emerging to yield more valid and informative results than each experiment separately. By exploring the power of several studies in one single analysis, meta-analysis of many cancer gene-profiling data increases the statistical power to detect differentially expressed genes and allows assessment of heterogeneity. OrderedList is such a method that was specially proposed for cancer gene expression data meta-analysis. It is superior to other methods in that it does not rely on strong effects of differential gene expression in a single study but on c... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960371 Tue, 15 Sep 2009 00:00:00 +0100 2960371 http://www.medworm.com/index.php?rid=2960370&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_20 We present our hook method, a new calibration approach that is based on a graphical summary of the actual hybridization characteristics of a particular microarray. Although single-chip related, hook performs as well as the multi-chip-related gcRMA, presently one of the best state-of-the-art methods for estimating expression values. The hook method, in addition, provides a set of chip summary characteristics that evaluate the performance of a given hybridization. The algorithm of the method is briefly described and its performance is exemplified. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960370 Tue, 15 Sep 2009 00:00:00 +0100 2960370 http://www.medworm.com/index.php?rid=2960369&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_1 The objectives of tissue repositories also vary based on the types of tissues provided (e.g., fresh tissue aliquots, fixed paraffin-embedded tissue, paraffin tissue sections, etc.) and how the tissues are to be used in research. For example, the potential use of tissues affects the need for extensive annotation of the specimen including both clinical information (e.g., clinical outcomes) and demographics. Specifically, if the tissues are to be used for extraction of proteins or basic studies of disease processes, less clinical information, if any, may be needed than if the tissues are to be used for the correlation of an aspect of the disease process with clinical outcome or response to a specific therapy. In this review, we describe, based on our experience, the major issues that should b... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960369 Tue, 15 Sep 2009 00:00:00 +0100 2960369 http://www.medworm.com/index.php?rid=2960368&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_19 The introduction of new high-throughput methodologies such as DNA microarrays constitutes a major breakthrough in cancer research. The unprecedented amount of data produced by such technologies has opened new avenues for interrogating living systems although, at the same time, it has demanded of the development of new data analytical methods as well as new strategies for testing hypotheses. A history of early successful applications in cancer boosted the use of microarrays and fostered further applications in other fields. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960368 Tue, 15 Sep 2009 00:00:00 +0100 2960368 http://www.medworm.com/index.php?rid=2960367&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_18 Within the past decade, the field of gene expression analysis has constantly evolved, with numerous technologies being available for RNA quantification, including differential display, serial analysis of gene expression (SAGE), quantitative real-time (qRT) polymerase chain reaction (PCR), and microarrays. Although every technique has its specific application, the high levels of accuracy, reproducibility, sensitivity, and specificity have established qRT-PCR as a standard method for detection and quantification of gene expression. In this chapter, all steps of the qRT-PCR procedure, including purification of total RNA from animal tissues, reverse transcription to complementary DNA (cDNA), and quantification of relative gene expression are discussed. We chose qRT-PCR analysis of CD39 in panc... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960367 Tue, 15 Sep 2009 00:00:00 +0100 2960367 http://www.medworm.com/index.php?rid=2960366&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_17 We describe the correlation between the EGFR genotype, including the KRAS mutation, and the consequences of the resulting genotype for anti-EGFR therapy in colorectal cancer. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960366 Tue, 15 Sep 2009 00:00:00 +0100 2960366 http://www.medworm.com/index.php?rid=2960365&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_16 Preoperative treatment strategies are now recommended for a variety of human cancers. Unfortunately, the response of individual tumors to a preoperative treatment is not uniform, and ranges from complete regression to resistance. This poses a considerable clinical dilemma, because patients with a priori resistant tumors could either be spared exposure to radiation or DNA-damaging drugs, i.e., they could be referred to primary surgery or dose-intensified protocols could be pursued. Because the response of an individual tumor as well as therapy-induced side effects represent the major limiting factors of current treatment strategies, identifying molecular markers of response or for treatment toxicity have become exceedingly important. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960365 Tue, 15 Sep 2009 00:00:00 +0100 2960365 http://www.medworm.com/index.php?rid=2960364&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_15 Gene profiling and expression analysis using microarrays have made a significant impact on our biological understanding of prostate cancer. The procedures for generating high-quality expression data from prostate cancer cell lines and tumors are not trivial. However, during the past 9 years, methods by which to process samples for gene profiling have been developed. In this chapter, techniques to process prostate cancer specimens either en bloc (macrodissection) or using laser capture microdissection are presented in detail along with extensive technical notes. Although we focus on prostate cancer and discuss the specific methods utilized in our lab, the processes discussed are generalizable to other tumors and amenable to the substitution of alternative instruments and/or commercially ava... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960364 Tue, 15 Sep 2009 00:00:00 +0100 2960364 http://www.medworm.com/index.php?rid=2960363&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_14 High levels of RNases present in the normal pancreas and the abundance of desmoplastic stroma of most pancreatic cancers have traditionally caused difficulty in the extraction of high-quality RNA and gene expression profiling from pancreatic tissues. However, a variety of innovative strategies have made it possible to successfully perform a molecular analysis of pancreatic cancer, and the expression profiles that have been generated have provided tremendous insight into the nature of this aggressive disease. Here, we describe some of these techniques. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960363 Tue, 15 Sep 2009 00:00:00 +0100 2960363 http://www.medworm.com/index.php?rid=2960362&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_13 During the last decade, gene expression microarrays and array-based comparative genomic hybridization (array–CGH) have unraveled the complexity of human tumor genomes more precisely and comprehensively than ever before. More recently, the simultaneous assessment of global changes in messenger RNA (mRNA) expression and in DNA copy number through “integrative oncogenomic” analyses has allowed researchers the access to results uncovered through the analysis of one-dimensional data sets, thus accelerating cancer gene discovery. In this chapter, we discuss the major contributions of DNA microarrays to the study of hematological malignancies, focusing on the integrative oncogenomic approaches that correlate genomic and transcriptomic data. We also present the basic aspects of t... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960362 Tue, 15 Sep 2009 00:00:00 +0100 2960362 http://www.medworm.com/index.php?rid=2960361&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_12 During the development of tumors, autoantibodies against aberrant or overexpressed autoantigens can be induced. Several hundreds of tumor-associated autoantibodies (TAAB) with more or less specificity for tumors have been found until now by molecular cloning and proteomics technologies. Many TAAB are detectable in preclinical stages of the disease and may be indicators of tumor development. The screening for autoantibody responses in tumor patients may lead to new diagnostic tumor markers and may be a simple and effective way to identify concomitantly cytotoxic T-lymphocyte (CTL) reactivity. However, most of the TAAB lack sufficient sensitivity and specificity for use as biomarkers in the clinical practice. For further use TAAB should be selected for their specificity regarding malignancie... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960361 Tue, 15 Sep 2009 00:00:00 +0100 2960361 http://www.medworm.com/index.php?rid=2960360&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_11 In the post-genomic era, proteomic strategies are at the forefront of cancer research. By studying the complement of all expressed genes, proteomics aims to provide knowledge of biomarkers indicative of the physiological state of cancer cells at a specific time, enabling screening, early diagnosis, monitoring the course of cancer development/progression, and gauging the efficacy and safety of novel therapeutic agents. Onco-proteomics thus has the ability to revolutionise oncology practice by delivering highly selective and individualised clinical care. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960360 Tue, 15 Sep 2009 00:00:00 +0100 2960360 http://www.medworm.com/index.php?rid=2960359&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-59745-545-9_10 Identification of hereditary factors that predispose to cancer allows targeted cancer screening and better quantification of environmental risk factors. The ability to identify which single nucleotide polymorphisms (SNPs) are associated with cancer or segregate with disease in families allows high-risk loci to be identified. In this chapter, two platforms for analysing SNPs are discussed, the Affymetrix and Illumina systems. Application of both platforms requires the same principles of good laboratory practice but there are important differences in materials and methods, which will be discussed. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2960359 Tue, 15 Sep 2009 00:00:00 +0100 2960359 http://www.medworm.com/index.php?rid=2713730&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_9 Hormones are chemical messengers produced in one part of the body and released into the blood to trigger or regulate particular functions of the body in another part. Hormone actions vary widely, but can include stimulation or inhibition of growth, induction or suppression of apoptosis, activation or inhibition of the immune system, regulating metabolism, and preparation for a new activity or phase of life. There is a growing interest in the role that hormones may play in the development and progression of various cancers. Recent research suggests that hormone levels may explain differences in risk for some of the most commonly diagnosed cancers, including breast, ovarian, and others; however, additional studies utilizing novel hormone measurement technologies are needed to investigate the... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713730 Tue, 03 Feb 2009 00:00:00 +0100 2713730 http://www.medworm.com/index.php?rid=2713729&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_8 The identification and eventual application of tumor markers in cancer screening, early detection, diagnosis, and prognosis is a continuing focus of significant translational cancer research. While many new candidate markers have been discovered and at least partly characterized, very few have found widespread clinical application limited presently to the use of CA-125 in ovarian cancer, CEA, primarily in colon cancer, and PSA in prostate cancer screening and patient monitoring. The rapidly emerging field of cancer genomics and proteomics, and their clinical translation as “molecular diagnosis” and “molecular medicine” are already beginning to transform the field, and the accelerating growth of information and technology in this research area will undoubtedly transf... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713729 Tue, 03 Feb 2009 00:00:00 +0100 2713729 http://www.medworm.com/index.php?rid=2713728&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_7 Analysis of the genome provides important information about the somatic genetic changes existing in the tissue; however, it is the proteins that do the work of the cell. Diseases such as cancer are caused by derangements in cellular protein molecular networks and cell signaling pathways. These pathways contain a large and growing collection drug targets, governing cellular survival, proliferation, invasion, and cell death. The clinical utility of reverse-phase protein microarrays (RPPA), a new technology invented in our laboratory, lies in its ability to generate a functional map of known cell signaling networks or pathways for an individual patient obtained directly from a biopsy specimen. Coupled with laser capture microdissection (LCM), the RPPA platform, the entire cellular proteome is... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713728 Tue, 03 Feb 2009 00:00:00 +0100 2713728 http://www.medworm.com/index.php?rid=2713727&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_6 Disease-related changes in serum proteins are reasonable targets for early detection particularly due to the noninvasive approach in obtaining samples. Glycoproteins specifically have been implicated in a variety of disease types ranging from immune diseases to cancers. High-throughput screening methods that can assess glycosylation states of all serum proteins in normal and diseased sample groups can facilitate early detection as well as shed light on disease progression mechanisms. Outlined here is a combination of liquid separation, protein microarray, and mass spectrometry approach to highlight candidate proteins involved in diseases through glycosylation mechanisms. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713727 Tue, 03 Feb 2009 00:00:00 +0100 2713727 http://www.medworm.com/index.php?rid=2713726&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_5 Changes in N-linked glycosylation are known to occur during the development of cancer. For example, increased branching of oligosaccharides has been associated with metastasis and has been correlated to tumor progression in human cancers of the breast, colon, and melanomas. Increases in core fucosylation have also been associated with the development of hepatocellular carcinoma (HCC). To a large extent, the proteins to which these N-linked glycans are attached have been unknown. However, with the advent of sensitive glycan analysis and proteomic technologies, the ability to comprehensively identify all the fucosylated proteins in a given population is now a possibility. This method, generally referred to as targeted glycoproteomics, is shown as applied to the detection of proteins present ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713726 Tue, 03 Feb 2009 00:00:00 +0100 2713726 http://www.medworm.com/index.php?rid=2713725&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_4 We describe those methods along with the methods for preparing and treating serum samples, running the experiments, and designing and interpreting the experiments. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713725 Tue, 03 Feb 2009 00:00:00 +0100 2713725 http://www.medworm.com/index.php?rid=2713724&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_3 Development of humoral and cellular immunity against self-cellular proteins in cancer patients is a phenomenal observation. The ability of immune system to sense the presence of the disease and to fight of the disease by generating autoantibodies against tumor antigens makes it a natural biosensor. Several screening technologies have been employed for the identification of tumor-specific antibodies in cancer patients. We have developed a multidimensional approach for the identification of diagnostic antigens that utilizes a combination of high-throughput antigen cloning and protein microarray-based serological detection of complex panels of antigens by exploiting the serum autoantibody repertoire directed toward tumor-associated antigens in cancer patients. Furthermore, validation of these... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713724 Tue, 03 Feb 2009 00:00:00 +0100 2713724 http://www.medworm.com/index.php?rid=2713723&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_2 We describe in this chapter a simple implementation of a serological monitoring platform for autoantibody measurement in cancer patients, from production of recombinant antigens to ELISA testing and interpretation. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713723 Tue, 03 Feb 2009 00:00:00 +0100 2713723 http://www.medworm.com/index.php?rid=2713722&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_22 The comprehensive work of both clinical and basic science colleagues has demonstrated a clear proof of concept for “in vitro discovered- in vivo validated” biomarkers in translational metabolic profiling research using magnetic resonance techniques. Major tissue metabolites (initially discovered by high-resolution in vitro techniques on cancer specimens) can be translated into in vivo protocols based on noninvasive magnetic resonance spectroscopy (MRS). Using 1H- and 31P-MRS on living animals or patients, a decrease in citrate and polyamines in prostate cancer, an increase of cholines in breast cancer, as well as a decreased NAA and an increased lactate in gliomas during cancer progression can be assessed noninvasively. MRS can be used to follow up conventional cytotoxic as wel... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713722 Tue, 03 Feb 2009 00:00:00 +0100 2713722 http://www.medworm.com/index.php?rid=2713721&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_21 The purpose of this chapter is to provide an introduction to magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) of human brain tumors, including the primary applications and basic terminology involved. Readers who wish to know more about this broad subject should seek out the referenced books (1. Tofts (2003) Quantitative MRI of the brain. Measuring changes caused by disease. Wiley; Bradley and Stark (1999) 2. Magnetic resonance imaging, 3rd Edition. Mosby Inc; Brown and Semelka (2003) 3. MRI basic principles and applications, 3rd Edition. Wiley-Liss) or reviews (4. Top Magn Reson Imaging 17:127–36, 2006; 5. JMRI 24:709–724, 2006; 6. Am J Neuroradiol 27:1404–1411, 2006). (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713721 Tue, 03 Feb 2009 00:00:00 +0100 2713721 http://www.medworm.com/index.php?rid=2713720&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_20 Metabolomics, one of the “omic” sciences in systems biology, is the global assessment and validation of endogenous small-molecule biochemicals (metabolites) within a biologic system. Initially, putative quantitative metabolic biomarkers for cancer detection and/or assessment of efficacy of anticancer treatment are usually discovered in a preclinical setting (using animal and human cell cultures), followed by translational validation of these biomarkers in biofluid or tumor tissue. Based on the tumor origin, various biofluids, such as blood, urine, and expressed prostatic secretions, can be used for validating metabolic biomarkers noninvasively in cancer patients. Metabolite detection and quantification is usually carried out by nuclear magnetic resonance (NMR) spectroscopy, whi... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713720 Tue, 03 Feb 2009 00:00:00 +0100 2713720 http://www.medworm.com/index.php?rid=2713719&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_1 Many studies demonstrated that cancer sera contain antibodies which react with autologous cellular antigens generally known as tumor-associated antigens (TAAs). In our laboratories, the approach used in the identification of TAAs has involved initially examining the sera of cancer patients using extracts of tissue culture cells as source of antigens in Western blotting and by indirect immunofluorescence on whole cells. With these two techniques, we identify sera which have high-titer fluorescent staining or strong signals to cell extracts on Western blotting and subsequently use these sera as probes in immunoscreening cDNA expression libraries, and also in proteomic approaches to isolate and identify targeted antigens which might potentially be involved in malignant transformation. In this... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713719 Tue, 03 Feb 2009 00:00:00 +0100 2713719 http://www.medworm.com/index.php?rid=2713718&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_19 The development of a successful classifier from multiple predictors (analytes) is a multistage process complicated typically by the paucity of the data samples when compared to the number of available predictors. Choosing an adequate validation strategy is key for drawing sound conclusions about the usefulness of the classifier. Other important decisions have to be made regarding the type of prediction model to be used and training algorithm, as well as the way in which the markers are selected. This chapter describes the principles of the classifier development and underlines the most common pitfalls. A simulated dataset is used to illustrate the main concepts involved in supervised classification. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713718 Tue, 03 Feb 2009 00:00:00 +0100 2713718 http://www.medworm.com/index.php?rid=2713717&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_18 Identification of germline mutations that may modulate individual risk of developing cancer is a rapidly developing field. Over the last few decades, germline mutations in p53, BRCA1, BRCA2, APC, MLH1, MSH2, and MSH6 have been identified in families with a large number of relatives who have been diagnosed with particular types of cancer. These mutations are rare but substantially increase the risk of cancer in carriers, and account for a small fraction of cancer cases diagnosed in the general population. The search for common mutations that correlate with a very modest increased risk of developing cancer is ongoing. With the completion of the Human Genome Project, a large array of methods to identify these genes and their variants are under development. The following chapter describes meth... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713717 Tue, 03 Feb 2009 00:00:00 +0100 2713717 http://www.medworm.com/index.php?rid=2713716&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_17 encompasses several major areas including the identification and analysis of variations of DNA and RNA that affect the efficacy and toxicity of drug therapy. It represents an integration of analytical approaches including DNA and RNA detection and quantitation which may be applied to either candidate genes or a global genome analysis. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713716 Tue, 03 Feb 2009 00:00:00 +0100 2713716 http://www.medworm.com/index.php?rid=2713715&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_16 Analysis of genetic altera tion in circulating DNA can have clinical utility in predicting disease outcome. Loss of heterozygosity (LOH) of DNA microsatellites has been shown to occur commonly among all chromosomes in various cancers, such as melanoma, breast cancer, and lung cancer. In this protocol, we focused on the utility of LOH of microsatellite biomarkers for detection of analyzing circulating DNA. The protocol describes how PCR is performed on each patient’s paired DNA samples (normal lymphocyte DNA and serum DNA) using specific microsatellite biomarkers followed by post-PCR product analysis using capillary array electrophoresis (CAE). The utility of CAE is due to its digitalization and accuracy of the post-PCR product results. (Source: Springer protocols feed by Cancer Resea... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713715 Tue, 03 Feb 2009 00:00:00 +0100 2713715 http://www.medworm.com/index.php?rid=2713714&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_15 Dideoxy DNA sequencing is routinely used in research and, increasingly, in clinical care for the detection of DNA sequence variants, single nucleotide changes, or small insertions or deletions, when the spectrum of DNA variation is unknown. DNA sequence variation can be present in tumor tissue that is not present in the normal tissue from the same individual. This somatic DNA sequence variation is often the cause of abnormal cell growth and/or regulation and, ultimately, tumorigenesis. Identification of these oncogenic DNA sequence variants has successfully led to the development of cancer therapies, since the abnormal protein products created from genomic DNA containing mutations can serve as targets for pharmacologic inhibition. Somatic DNA sequence analysis will continue to be a valuabl... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713714 Tue, 03 Feb 2009 00:00:00 +0100 2713714 http://www.medworm.com/index.php?rid=2713713&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_14 Numerous innovative and high-throughput techniques have been established to identify human disease genes. However, DNA sequencing of candidate genes still remains as a major limitation in the identification of causative mutations. Much of this limitation is due to the time and labor needed for the polymerase chain reaction (PCR) optimization and reaction setup. Toward this end, we have established a simplified protocol that utilizes a single PCR amplification condition. PCR purification is accomplished via enzymatic digestion and all products can be sequenced using universal primers. This combination of a single amplification condition, single-step purification, and sequencing setup using universal primers all contribute to a simple and high-throughput mutation screen. (Source: Springer pr... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713713 Tue, 03 Feb 2009 00:00:00 +0100 2713713 http://www.medworm.com/index.php?rid=2713712&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_13 Clear identification among early-stage cancer patients of those at highest risk of having metastatic disease would be of great benefit in treatment planning and management. Considerable additional benefit would accrue to high-risk patients if their responses to specific therapeutic alternatives could be predicted. Molecular biomarkers in the form of gene expression profiles are proving to be more effective tools for both prognostic and predictive patient stratification than more traditional methods such as patient demographics and histopathology indicators. Such biomarkers must be clinically validated before they can be effectively used to manage patients in clinical studies or clinical practice. This can be most efficiently accomplished by analyzing archived clinical samples with well-cha... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713712 Tue, 03 Feb 2009 00:00:00 +0100 2713712 http://www.medworm.com/index.php?rid=2713711&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_12 As companion diagnostics grow in prevalence and importance, the need for accurate assessment of in situ protein concentrations has increased. Traditional immunohistochemistry (IHC), while valuable for assessment of context of expression, is less valuable for quantification. The lack of rigorous quantitative potential of traditional IHC led to our development of an immunofluorescence-based method now commercialized as the AQUA® technology. Immunostaining of tissue samples, image acquisition, and use of AQUA software allow investigators to quickly, efficiently, and accurately measure levels of expression within user-defined subcellular or architectural compartments. IHC analyzed by AQUA shows high reproducibility and demonstrates protein measurement accuracy similar to ELISA assays. The ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713711 Tue, 03 Feb 2009 00:00:00 +0100 2713711 http://www.medworm.com/index.php?rid=2713710&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_11 Tissue microarrays are a platform of condensed histopathology that has revolutionized the translation of basic science to clinical utility. Tissue microarrays have resulted in a paradigm shift from histopathology to immunopathology and moved analysis of small selected samples sets of tens of specimens to a high-throughput environment of hundreds of specimens. Tissue microarrays have influenced validation strategies, but have a role in discovery as well, allowing a pathways approach to analysis of tumors. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713710 Tue, 03 Feb 2009 00:00:00 +0100 2713710 http://www.medworm.com/index.php?rid=2713709&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-811-9_10 Enzyme-linked immunosorbent assay (ELISA) microarrays promise to be a powerful tool for the detection and validation of disease biomarkers. ELISA microarrays are capable of simultaneous detection of many proteins using a small sample volume. Although there are many potential pitfalls to the use of ELISA microarrays, these can be avoided by careful planning of experiments. In this chapter we describe a high-throughput protocol for processing ELISA microarrays that will result in reliable and reproducible data. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2713709 Tue, 03 Feb 2009 00:00:00 +0100 2713709 http://www.medworm.com/index.php?rid=2359492&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_9 Despite the current availability of an impressive in vitro assay battery developed to quantitatively analyze the broad panel of small compounds and macromolecules that possess the inflammatory potential, little methodology exists nowadays that affords a researcher or clinician to quantify the ultimate output on the level of cell signaling response caused by inflammatory pathway stimulation. As a matter of fact, majority of analytical tools measure bona fide inflammatory substances (e.g., cytokines or chemokines) by their direct binding to secondary reagents such as specific antibodies or other selectively affine substrates with the final readout generated via quantification of the resulting complexes. Although specific and highly reproducible, this approach provides no discrimination betwe... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359492 Thu, 01 Jan 2009 05:00:00 +0100 2359492 http://www.medworm.com/index.php?rid=2359491&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_8 Nitric oxide (NO) is a free radical that is involved in the inflammatory process and carcinogenesis. There are four nitric oxide synthase enzymes involved in NO production: induced nitric oxide synthase (iNOS), endothelial NO synthase (eNOS), neural NO synthase (nNOS), and mitochondrial NOS. iNOS is an inducible and key enzyme in the inflamed tissue. Recent literatures indicate that NO as well as iNOS and eNOS can modulate cancer-related events including nitro-oxidative stress, apoptosis, cell cycle, angio-genesis, invasion, and metastasis. This chapter focuses on linking NO/iNOS/eNOS to inflammation and carcinogenesis from experimental evidence to potential targets on cancer prevention and treatment. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359491 Thu, 01 Jan 2009 05:00:00 +0100 2359491 http://www.medworm.com/index.php?rid=2359490&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_7 Activation of the transcription factor, nuclear factor of κB (NF-κB), has recently emerged as the critical link between inflammation and cancer. NF-κB is activated by many signals including DNA damage and microbial pathogen recognition. Although microbial infections account for an estimated 15% of all cancer related deaths, NF-κB is constitutively activated in various types of cancer as a direct result of chromosomal translocations, deletions and mutations affecting both NF-κB and its regulators. NF-κB exerts the transcriptional activation and repression of inflammatory and immune response genes as well as genes mediating cell survival and proliferation, and thus stands to impact various cellular processes. The decision is made through the integration of... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359490 Thu, 01 Jan 2009 05:00:00 +0100 2359490 http://www.medworm.com/index.php?rid=2359489&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_6 Global gene expression profiling studies conducted over the last couple of years have shown that molecular profiling of breast cancers can be used to identify clinically and genetically significant subtypes of breast carcinomas and subgroups of patients with different prognosis or disease outcome, and to predict therapeutic response to both endocrine and chemotherapeutic drugs. We studied one particularly aggressive form of locally advanced breast cancer, namely inflammatory breast cancer (IBC) using cDNA micro-arrays. We demonstrated that IBC is characterized by a different gene expression profile, not related to any of the previously identified breast cancer subtypes. This gene expression profile revealed potential therapeutic targets, such as the transcription factor NF-κB. (Sourc... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359489 Thu, 01 Jan 2009 05:00:00 +0100 2359489 http://www.medworm.com/index.php?rid=2359488&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_5 It is now apparent that epigenetic abnormalities, in particular altered DNA methylation, play a crucial role in the development and progression of human cancers. DNA hypermethylation at promoter CpG islands is now recognized as a third mechanism by which inactivation of tumor suppressor genes occurs. Aberrant CpG island hypermethylation is also frequently observed in chronic inflammation and precancerous lesions, which suggests that it is an early event in tumorigenesis that could serve as a useful tumor marker. A variety of screening techniques have been developed for genome-wide screening of methylation status. Of those, transcriptome analysis coupled with pharmacological unmasking has emerged as a powerful tool for revealing DNA methylation patterns in cancer cells and identifying new t... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359488 Thu, 01 Jan 2009 05:00:00 +0100 2359488 http://www.medworm.com/index.php?rid=2359487&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_4 NF-κB plays a pivotal role in immunity and inflammation and is considered to be a promising candidate for drug development. However, global suppression of NF-κB may have undesirable side-effects. Our data and the results of others suggest that each of the five NF-κB subunits may have a specific function in controlling the expression of inflammatory mediators in immune cells. Identifying the role for each NF-κB subunit in primary human immune cells will allow a more targeted approach to inhibiting NF-κB subunit-specific cellular functions. However, results obtained with primary human cells can often be inconsistent due to donor heterogeneity. Therefore one possible approach could be to generate human immune cell lines with stably inhibited expression of specifi... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359487 Thu, 01 Jan 2009 05:00:00 +0100 2359487 http://www.medworm.com/index.php?rid=2359486&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_3 Current dogma suggests that the positive correlation between obesity and cancer is driven by white adipose tissue that accompanies obesity, possibly through excess secretion of adipokines. However, recent studies in fatless A-Zip/F-1 mice, which have undetectable adipokine levels but display accelerated tumor formation, suggest that adipokines are not required for the enhanced tumor development. The A-Zip/F-1 mice are also diabetic and display elevated circulating levels of other molecules frequently associated with obesity and carcinogenesis: insulin, insulin-like growth factor-1, and inflammatory cytokines. Therefore, we postulate that the pathways associated with insulin resistance and inflammation, rather than adipocyte-derived factors, may represent key prevention or therapeutic targe... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359486 Thu, 01 Jan 2009 05:00:00 +0100 2359486 http://www.medworm.com/index.php?rid=2359485&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_22 Peptide YY (PYY) orchestrates the functions of the gut and the pancreas by regulating growth, digestion and absorption. In addition to its physiological role, PYY exhibits immune and antitrophic properties in the pancreas by decreasing cytokine and amylase release. Although the exact mechanism(s) of action are still incompletely understood, PYY interacts at the acinar level with numerous intracellular transcription factors. In addition to ameliorating pancreatic inflammation, novel synthetic analogs of PYY have been developed that are potent inhibitors of pancreatic cancer proliferation, in vitro and in vivo. Additionally, PYY and its analogs have been shown to inhibit the growth of breast, esophagus, and gastric cancer in vitro. We, herein, plan to review some of the methods employed in t... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359485 Thu, 01 Jan 2009 05:00:00 +0100 2359485 http://www.medworm.com/index.php?rid=2359484&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_21 Histone deacetylase inhibitors are potent inducers of growth arrest and apoptotic cell death and are currently in clinical studies for solid tumors. In addition, recent studies from our own group provide evidence for an anti-inflammatory potency of HDACi in vitro and in vivo by using various models of experimental colitis. Since inflammatory bowel disease in humans is associated with an increased risk of developing colorectal cancer, a therapeutic approach combining anti-inflammatory as well as antiprolif-erative properties is highly intriguing. Consequently, methods to further characterize the mechanisms involved include the direct analysis of the acetylation/deacetylation as well as the regulatory effect of histone deacetylase inhibitors on defined cell populations. (Source: Springer pro... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359484 Thu, 01 Jan 2009 05:00:00 +0100 2359484 http://www.medworm.com/index.php?rid=2359483&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_20 Selective delivery of drugs or imaging dyes to tumor is the central challenge for improving existing therapy and diagnosis of cancer. Phage display of random peptides has been used to identify homing peptides that are specific to tumor cells or tumor blood vessels. These homing peptides can be conjugated with imaging agents or therapeutic drugs and may be a promising tool for affinity-based targeted delivery of imaging agents and drugs. This chapter describes the procedures for phage display selection of peptides that selectively bind to bladder tumor cells and home to tumor tissues in vivo. It also describes the application of the tumor-specific peptide to the detection of bladder cancer cells in the patient urine. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359483 Thu, 01 Jan 2009 05:00:00 +0100 2359483 http://www.medworm.com/index.php?rid=2359482&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_2 Genetic instability is a hallmark of human cancers. It is the driving force for tumor development as it facilitates the accumulation of mutations in genes that regulate cell death and proliferation and therefore promotes malignant transformation. Chronic inflammation is a common underlying condition for human tumor development, accounting for approximately 20% of human cancers. TNFα is an important inflammation cytokine and is crucial to the development of inflammation-associated cancers. We have shown that TNFα can cause DNA damages through reactive oxygen species (ROS). TNFα treatment in cultured cells resulted in increased gene mutations, gene amplification, micronuclei formation and chromosomal instability. Antioxidants significantly reduced TNFα-induced genetic... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359482 Thu, 01 Jan 2009 05:00:00 +0100 2359482 http://www.medworm.com/index.php?rid=2359481&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_19 The objective of imaging is to provide a follow-up of these killer cells in real time, in order to gain a better understanding of the mechanisms and action modes of lymphocytes on the tumor. Magnetic resonance imaging (MRI) has the advantage of the innocu-ousness of the applied magnetic field. Moreover, it has an exceptional spatial resolution allowing the visualization of anatomical areas without in-depth limitations. These features make MRI particularly adapted for cellular imaging. The use of “ (ultrasmall) superparamagnetic iron oxide ” particles [(U) SPIO] offers the adequate sensitivity required for cellular imaging. To promote a sufficient capture of these particles in nonphagocytic cells and make the cell of interest “ detectable ” by MRI after its injection... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359481 Thu, 01 Jan 2009 05:00:00 +0100 2359481 http://www.medworm.com/index.php?rid=2359480&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_18 Within most human and murine cancers there is an abundant macrophage population, attracted to the tumor microenvironment by cytokines and chemokines such as CSF-1 (M-CSF) and CCL2 (MCP-1) (Cell 124:263–266, 2006). Despite their intrinsic antitumor activity there is usually, but not always, a positive association between the extent of the macrophage infiltrate in tumors and a bad prognosis (Cell 124:263–266, 2006; Nat Rev Cancer 4:71–78, 2004). According to Condeelis and Pollard (Nat Rev Cancer 4:71–78, 2004), tumor-associated macrophages are obligate partners for malignant cell migration, invasion, and metastasis. These conclusions are based not only on association studies, but also on experiments demonstrating that ablation of macrophage function, or their infiltra... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359480 Thu, 01 Jan 2009 05:00:00 +0100 2359480 http://www.medworm.com/index.php?rid=2359479&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_17 Lung carcinoma remains one of the most common malignant tumors in the world despite recent advancements in the development of new chemotherapeutic agents for its treatment. Therefore, novel approaches for drug target discovery play an important role in the effort to help extend its dismal 5-year survival rate (<15%). Many mechanisms contribute to oncogenic transformation in carcinoma cells in the lung and recent evidence indicates that the overproduction of prostaglandin E2 (PGE2), and the prostag-landin E2 receptor subtype, EP4, promote the growth and progression of human nonsmall cell lung carcinoma (NSCLC), the most common lung carcinoma. Peroxisome proliferator-activated receptor beta/ delta (PPARβ/δ), one of the nuclear hormone ligand-dependent transcription factors, has... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359479 Thu, 01 Jan 2009 05:00:00 +0100 2359479 http://www.medworm.com/index.php?rid=2359478&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_16 Non-steroidal anti-inflammatory drugs (NSAIDs) and selective cyclooxygenase-2 (COX-2) inhibitors (COXIBs) induce cancer cell apoptosis via several signaling pathways. There is evidence that they induce colon cancer cell apoptosis by suppressing peroxisome proliferator-activated receptor δ (PPARδ) through inhibition of COX-2-derived prostacyclin (PGI2). PGI2 activates PPARδ resulting in binding of PPARδ to specific PPAR response elements (PPRE) of target genes. We have identified 14-3-3ε as one of the genes that are upregulated by PPARδ. Elevated 14-3-3ε proteins in cytosol enhance sequestration of Bad and reduce mitochondrial damage by Bad and thereby control apoptosis. NSAIDs and COXIBs block PGI2 production, thereby reducing PPARδ DNA b... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359478 Thu, 01 Jan 2009 05:00:00 +0100 2359478 http://www.medworm.com/index.php?rid=2359477&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_15 Focal Adhesion Kinase (FAK) is implicated in a wide array of cellular processes and also involved in the production of matrix metalloproteinases (MMPs) which degrade extracellular matrix (ECM). We have shown that FAK plays a critical role in MMP-9 production and subsequent invasion of the cholangiocarcinoma activated by an inflammatory cytokine, TNF-α. By nature, cholangiocarcinoma is frequently associated with hepatolithiasis that causes recurrent inflammation. As degradation of the ECM is a prerequisite step for the invasion and metastasis of cancer cells, we used an assay of gelatin-degrading MMPs by Zymography to clarify the characteristic feature of the matrix degrading systems of the cancer cells. Immunoprecipitation and western blot analysis together with site specific phospho... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359477 Thu, 01 Jan 2009 05:00:00 +0100 2359477 http://www.medworm.com/index.php?rid=2359476&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_14 The Janus kinases (JAK) and signal transducer and activator of transcription (STAT) signaling are strongly activated in many tumors. STAT proteins are activated by phosphorylation at the tyrosine residue, then dimerize, translocate to the nucleus and bind DNA, initiating the transcription of target genes. Activation of JAK-STAT pathway is implicated in the regulation of cell growth, differentiation, survival and cross-talk between cancer and immune cells. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359476 Thu, 01 Jan 2009 05:00:00 +0100 2359476 http://www.medworm.com/index.php?rid=2359475&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_13 Tumor cells recruit inflammatory cells to the tumor site and transform them into tumor-supportive cells which in turn release numerous cytokines, including Transforming Growth Factor-β that enhances tumor proliferation, invasion, angiogenesis and induces immune paralysis. Activation of JNK/c-Jun signaling pathway by various stimuli often leads to a formation of the AP-1 transcriptional complex, which is a critical regulator of a complex program of gene expression that defines the invasive phenotype. Recent studies on JNK/c-Jun phosphorylation have been carried out using phospho-specific antibodies, which have greatly facilitated analysis of signal transduction. The electrophoretic mobility shift assay (EMSA, gel shift) helps in determining the transcription factor activation and is ba... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359475 Thu, 01 Jan 2009 05:00:00 +0100 2359475 http://www.medworm.com/index.php?rid=2359474&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_12 Rheumatoid arthritis (RA) is a chronic inflammatory disease characterised by pain, swelling and progressive destruction of synovial joints. The synovial membranes of the affected joints markedly increase in size due to infiltration of several cell types, of which macrophages, lymphocytes and fibroblasts are most abundant. These cell types are activated and release a plethora of inflammatory mediators, such as cytokines, chem-okines and matrix metalloproteinases (MMPs). Synovial membranes can be removed from the joints of RA patients (most commonly when the respective joint is undergoing replacement therapy) and enzymati-cally digested, analyzed or cultured ex vivo. Analysing the cytokine profile of distinct populations of ex vivo RA-patients derived synoviocytes can provide an insight into... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359474 Thu, 01 Jan 2009 05:00:00 +0100 2359474 http://www.medworm.com/index.php?rid=2359473&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_11 We describe in this chapter three separate methods to determine the effects of this NEMO-binding domain (NBD) peptide on pro-inflammatory NF-κB signaling in response to tumor necrosis factor (TNF). (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359473 Thu, 01 Jan 2009 05:00:00 +0100 2359473 http://www.medworm.com/index.php?rid=2359472&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_10 NF-κB transcription factors marshal innate and adaptive immunity and inflammation. NF-κB also counters programmed cell death (PCD) induced by the proinflammatory cytokine tumor necrosis factor (TNF)α, and this activity of NF-κB is crucial for organismal physiology, chronic inflammation, and tumorigenesis. Indeed, whereas NF-κB contributes to many aspects of oncogenesis, it is now clear that its suppressive action on PCD is central to this process. Notably, recent studies indicate that NF-κB represents a crucial link in the well-established association between inflammation and carcinogenesis. In this link, NF-κB promotes synthesis of inflammatory mediators (e.g. TNFα) that stimulate growth of cancer cells, and upregulates genes that protect th... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359472 Thu, 01 Jan 2009 05:00:00 +0100 2359472 http://www.medworm.com/index.php?rid=2359471&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_1 Chronic inflammation is induced by various infectious/infected agents and by many physical, chemical and immunological factors. Many malignancies arise from areas of infection and inflammation. Reactive oxygen species and reactive nitrogen species are considered to play the key role in inflammation-associated carcinogenesis by causing oxidative and nitrative DNA damage. 8-Nitroguanine is a mutagenic nitrative DNA lesion formed during inflammation. Development of a detection method for 8-nitroguanine would provide an insight into the mechanism of inflammation-associated carcinogenesis and the assessment of carcinogenic risk in patients with inflammatory diseases. We established the method to produce highly sensitive and specific anti-8-nitroguanine rabbit polyclonal antibody, and detect 8-n... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2359471 Thu, 01 Jan 2009 05:00:00 +0100 2359471 http://www.medworm.com/index.php?rid=2331412&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_9 Despite the current availability of an impressive in vitro assay battery developed to quantitatively analyze the broad panel of small compounds and macromolecules that possess the inflammatory potential, little methodology exists nowadays that affords a researcher or clinician to quantify the ultimate output on the level of cell signaling response caused by inflammatory pathway stimulation. As a matter of fact, majority of analytical tools measure bona fide inflammatory substances (e.g., cytokines or chemokines) by their direct binding to secondary reagents such as specific antibodies or other selectively affine substrates with the final readout generated via quantification of the resulting complexes. Although specific and highly reproducible, this approach provides no discrimination betwe... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331412 Thu, 01 Jan 2009 05:00:00 +0100 2331412 http://www.medworm.com/index.php?rid=2331411&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_8 Nitric oxide (NO) is a free radical that is involved in the inflammatory process and carcinogenesis. There are four nitric oxide synthase enzymes involved in NO production: induced nitric oxide synthase (iNOS), endothelial NO synthase (eNOS), neural NO synthase (nNOS), and mitochondrial NOS. iNOS is an inducible and key enzyme in the inflamed tissue. Recent literatures indicate that NO as well as iNOS and eNOS can modulate cancer-related events including nitro-oxidative stress, apoptosis, cell cycle, angio-genesis, invasion, and metastasis. This chapter focuses on linking NO/iNOS/eNOS to inflammation and carcinogenesis from experimental evidence to potential targets on cancer prevention and treatment. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331411 Thu, 01 Jan 2009 05:00:00 +0100 2331411 http://www.medworm.com/index.php?rid=2331410&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_7 Activation of the transcription factor, nuclear factor of κB (NF-κB), has recently emerged as the critical link between inflammation and cancer. NF-κB is activated by many signals including DNA damage and microbial pathogen recognition. Although microbial infections account for an estimated 15% of all cancer related deaths, NF-κB is constitutively activated in various types of cancer as a direct result of chromosomal translocations, deletions and mutations affecting both NF-κB and its regulators. NF-κB exerts the transcriptional activation and repression of inflammatory and immune response genes as well as genes mediating cell survival and proliferation, and thus stands to impact various cellular processes. The decision is made through the integration of... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331410 Thu, 01 Jan 2009 05:00:00 +0100 2331410 http://www.medworm.com/index.php?rid=2331409&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_6 Global gene expression profiling studies conducted over the last couple of years have shown that molecular profiling of breast cancers can be used to identify clinically and genetically significant subtypes of breast carcinomas and subgroups of patients with different prognosis or disease outcome, and to predict therapeutic response to both endocrine and chemotherapeutic drugs. We studied one particularly aggressive form of locally advanced breast cancer, namely inflammatory breast cancer (IBC) using cDNA micro-arrays. We demonstrated that IBC is characterized by a different gene expression profile, not related to any of the previously identified breast cancer subtypes. This gene expression profile revealed potential therapeutic targets, such as the transcription factor NF-κB. (Sourc... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331409 Thu, 01 Jan 2009 05:00:00 +0100 2331409 http://www.medworm.com/index.php?rid=2331408&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_5 It is now apparent that epigenetic abnormalities, in particular altered DNA methylation, play a crucial role in the development and progression of human cancers. DNA hypermethylation at promoter CpG islands is now recognized as a third mechanism by which inactivation of tumor suppressor genes occurs. Aberrant CpG island hypermethylation is also frequently observed in chronic inflammation and precancerous lesions, which suggests that it is an early event in tumorigenesis that could serve as a useful tumor marker. A variety of screening techniques have been developed for genome-wide screening of methylation status. Of those, transcriptome analysis coupled with pharmacological unmasking has emerged as a powerful tool for revealing DNA methylation patterns in cancer cells and identifying new t... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331408 Thu, 01 Jan 2009 05:00:00 +0100 2331408 http://www.medworm.com/index.php?rid=2331407&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_4 NF-κB plays a pivotal role in immunity and inflammation and is considered to be a promising candidate for drug development. However, global suppression of NF-κB may have undesirable side-effects. Our data and the results of others suggest that each of the five NF-κB subunits may have a specific function in controlling the expression of inflammatory mediators in immune cells. Identifying the role for each NF-κB subunit in primary human immune cells will allow a more targeted approach to inhibiting NF-κB subunit-specific cellular functions. However, results obtained with primary human cells can often be inconsistent due to donor heterogeneity. Therefore one possible approach could be to generate human immune cell lines with stably inhibited expression of specifi... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331407 Thu, 01 Jan 2009 05:00:00 +0100 2331407 http://www.medworm.com/index.php?rid=2331406&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_3 Current dogma suggests that the positive correlation between obesity and cancer is driven by white adipose tissue that accompanies obesity, possibly through excess secretion of adipokines. However, recent studies in fatless A-Zip/F-1 mice, which have undetectable adipokine levels but display accelerated tumor formation, suggest that adipokines are not required for the enhanced tumor development. The A-Zip/F-1 mice are also diabetic and display elevated circulating levels of other molecules frequently associated with obesity and carcinogenesis: insulin, insulin-like growth factor-1, and inflammatory cytokines. Therefore, we postulate that the pathways associated with insulin resistance and inflammation, rather than adipocyte-derived factors, may represent key prevention or therapeutic targe... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331406 Thu, 01 Jan 2009 05:00:00 +0100 2331406 http://www.medworm.com/index.php?rid=2331405&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_22 Peptide YY (PYY) orchestrates the functions of the gut and the pancreas by regulating growth, digestion and absorption. In addition to its physiological role, PYY exhibits immune and antitrophic properties in the pancreas by decreasing cytokine and amylase release. Although the exact mechanism(s) of action are still incompletely understood, PYY interacts at the acinar level with numerous intracellular transcription factors. In addition to ameliorating pancreatic inflammation, novel synthetic analogs of PYY have been developed that are potent inhibitors of pancreatic cancer proliferation, in vitro and in vivo. Additionally, PYY and its analogs have been shown to inhibit the growth of breast, esophagus, and gastric cancer in vitro. We, herein, plan to review some of the methods employed in t... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331405 Thu, 01 Jan 2009 05:00:00 +0100 2331405 http://www.medworm.com/index.php?rid=2331404&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_21 Histone deacetylase inhibitors are potent inducers of growth arrest and apoptotic cell death and are currently in clinical studies for solid tumors. In addition, recent studies from our own group provide evidence for an anti-inflammatory potency of HDACi in vitro and in vivo by using various models of experimental colitis. Since inflammatory bowel disease in humans is associated with an increased risk of developing colorectal cancer, a therapeutic approach combining anti-inflammatory as well as antiprolif-erative properties is highly intriguing. Consequently, methods to further characterize the mechanisms involved include the direct analysis of the acetylation/deacetylation as well as the regulatory effect of histone deacetylase inhibitors on defined cell populations. (Source: Springer pro... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331404 Thu, 01 Jan 2009 05:00:00 +0100 2331404 http://www.medworm.com/index.php?rid=2331403&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_20 Selective delivery of drugs or imaging dyes to tumor is the central challenge for improving existing therapy and diagnosis of cancer. Phage display of random peptides has been used to identify homing peptides that are specific to tumor cells or tumor blood vessels. These homing peptides can be conjugated with imaging agents or therapeutic drugs and may be a promising tool for affinity-based targeted delivery of imaging agents and drugs. This chapter describes the procedures for phage display selection of peptides that selectively bind to bladder tumor cells and home to tumor tissues in vivo. It also describes the application of the tumor-specific peptide to the detection of bladder cancer cells in the patient urine. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331403 Thu, 01 Jan 2009 05:00:00 +0100 2331403 http://www.medworm.com/index.php?rid=2331402&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_2 Genetic instability is a hallmark of human cancers. It is the driving force for tumor development as it facilitates the accumulation of mutations in genes that regulate cell death and proliferation and therefore promotes malignant transformation. Chronic inflammation is a common underlying condition for human tumor development, accounting for approximately 20% of human cancers. TNFα is an important inflammation cytokine and is crucial to the development of inflammation-associated cancers. We have shown that TNFα can cause DNA damages through reactive oxygen species (ROS). TNFα treatment in cultured cells resulted in increased gene mutations, gene amplification, micronuclei formation and chromosomal instability. Antioxidants significantly reduced TNFα-induced genetic... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331402 Thu, 01 Jan 2009 05:00:00 +0100 2331402 http://www.medworm.com/index.php?rid=2331401&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_19 The objective of imaging is to provide a follow-up of these killer cells in real time, in order to gain a better understanding of the mechanisms and action modes of lymphocytes on the tumor. Magnetic resonance imaging (MRI) has the advantage of the innocu-ousness of the applied magnetic field. Moreover, it has an exceptional spatial resolution allowing the visualization of anatomical areas without in-depth limitations. These features make MRI particularly adapted for cellular imaging. The use of “ (ultrasmall) superparamagnetic iron oxide ” particles [(U) SPIO] offers the adequate sensitivity required for cellular imaging. To promote a sufficient capture of these particles in nonphagocytic cells and make the cell of interest “ detectable ” by MRI after its injection... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331401 Thu, 01 Jan 2009 05:00:00 +0100 2331401 http://www.medworm.com/index.php?rid=2331400&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_18 Within most human and murine cancers there is an abundant macrophage population, attracted to the tumor microenvironment by cytokines and chemokines such as CSF-1 (M-CSF) and CCL2 (MCP-1) (Cell 124:263–266, 2006). Despite their intrinsic antitumor activity there is usually, but not always, a positive association between the extent of the macrophage infiltrate in tumors and a bad prognosis (Cell 124:263–266, 2006; Nat Rev Cancer 4:71–78, 2004). According to Condeelis and Pollard (Nat Rev Cancer 4:71–78, 2004), tumor-associated macrophages are obligate partners for malignant cell migration, invasion, and metastasis. These conclusions are based not only on association studies, but also on experiments demonstrating that ablation of macrophage function, or their infiltra... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331400 Thu, 01 Jan 2009 05:00:00 +0100 2331400 http://www.medworm.com/index.php?rid=2331399&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_17 Lung carcinoma remains one of the most common malignant tumors in the world despite recent advancements in the development of new chemotherapeutic agents for its treatment. Therefore, novel approaches for drug target discovery play an important role in the effort to help extend its dismal 5-year survival rate (<15%). Many mechanisms contribute to oncogenic transformation in carcinoma cells in the lung and recent evidence indicates that the overproduction of prostaglandin E2 (PGE2), and the prostag-landin E2 receptor subtype, EP4, promote the growth and progression of human nonsmall cell lung carcinoma (NSCLC), the most common lung carcinoma. Peroxisome proliferator-activated receptor beta/ delta (PPARβ/δ), one of the nuclear hormone ligand-dependent transcription factors, has... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331399 Thu, 01 Jan 2009 05:00:00 +0100 2331399 http://www.medworm.com/index.php?rid=2331398&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_16 Non-steroidal anti-inflammatory drugs (NSAIDs) and selective cyclooxygenase-2 (COX-2) inhibitors (COXIBs) induce cancer cell apoptosis via several signaling pathways. There is evidence that they induce colon cancer cell apoptosis by suppressing peroxisome proliferator-activated receptor δ (PPARδ) through inhibition of COX-2-derived prostacyclin (PGI2). PGI2 activates PPARδ resulting in binding of PPARδ to specific PPAR response elements (PPRE) of target genes. We have identified 14-3-3ε as one of the genes that are upregulated by PPARδ. Elevated 14-3-3ε proteins in cytosol enhance sequestration of Bad and reduce mitochondrial damage by Bad and thereby control apoptosis. NSAIDs and COXIBs block PGI2 production, thereby reducing PPARδ DNA b... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331398 Thu, 01 Jan 2009 05:00:00 +0100 2331398 http://www.medworm.com/index.php?rid=2331397&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_15 Focal Adhesion Kinase (FAK) is implicated in a wide array of cellular processes and also involved in the production of matrix metalloproteinases (MMPs) which degrade extracellular matrix (ECM). We have shown that FAK plays a critical role in MMP-9 production and subsequent invasion of the cholangiocarcinoma activated by an inflammatory cytokine, TNF-α. By nature, cholangiocarcinoma is frequently associated with hepatolithiasis that causes recurrent inflammation. As degradation of the ECM is a prerequisite step for the invasion and metastasis of cancer cells, we used an assay of gelatin-degrading MMPs by Zymography to clarify the characteristic feature of the matrix degrading systems of the cancer cells. Immunoprecipitation and western blot analysis together with site specific phospho... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331397 Thu, 01 Jan 2009 05:00:00 +0100 2331397 http://www.medworm.com/index.php?rid=2331396&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_14 The Janus kinases (JAK) and signal transducer and activator of transcription (STAT) signaling are strongly activated in many tumors. STAT proteins are activated by phosphorylation at the tyrosine residue, then dimerize, translocate to the nucleus and bind DNA, initiating the transcription of target genes. Activation of JAK-STAT pathway is implicated in the regulation of cell growth, differentiation, survival and cross-talk between cancer and immune cells. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331396 Thu, 01 Jan 2009 05:00:00 +0100 2331396 http://www.medworm.com/index.php?rid=2331395&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_13 Tumor cells recruit inflammatory cells to the tumor site and transform them into tumor-supportive cells which in turn release numerous cytokines, including Transforming Growth Factor-β that enhances tumor proliferation, invasion, angiogenesis and induces immune paralysis. Activation of JNK/c-Jun signaling pathway by various stimuli often leads to a formation of the AP-1 transcriptional complex, which is a critical regulator of a complex program of gene expression that defines the invasive phenotype. Recent studies on JNK/c-Jun phosphorylation have been carried out using phospho-specific antibodies, which have greatly facilitated analysis of signal transduction. The electrophoretic mobility shift assay (EMSA, gel shift) helps in determining the transcription factor activation and is ba... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331395 Thu, 01 Jan 2009 05:00:00 +0100 2331395 http://www.medworm.com/index.php?rid=2331394&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_12 Rheumatoid arthritis (RA) is a chronic inflammatory disease characterised by pain, swelling and progressive destruction of synovial joints. The synovial membranes of the affected joints markedly increase in size due to infiltration of several cell types, of which macrophages, lymphocytes and fibroblasts are most abundant. These cell types are activated and release a plethora of inflammatory mediators, such as cytokines, chem-okines and matrix metalloproteinases (MMPs). Synovial membranes can be removed from the joints of RA patients (most commonly when the respective joint is undergoing replacement therapy) and enzymati-cally digested, analyzed or cultured ex vivo. Analysing the cytokine profile of distinct populations of ex vivo RA-patients derived synoviocytes can provide an insight into... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331394 Thu, 01 Jan 2009 05:00:00 +0100 2331394 http://www.medworm.com/index.php?rid=2331393&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_11 We describe in this chapter three separate methods to determine the effects of this NEMO-binding domain (NBD) peptide on pro-inflammatory NF-κB signaling in response to tumor necrosis factor (TNF). (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331393 Thu, 01 Jan 2009 05:00:00 +0100 2331393 http://www.medworm.com/index.php?rid=2331392&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_10 NF-κB transcription factors marshal innate and adaptive immunity and inflammation. NF-κB also counters programmed cell death (PCD) induced by the proinflammatory cytokine tumor necrosis factor (TNF)α, and this activity of NF-κB is crucial for organismal physiology, chronic inflammation, and tumorigenesis. Indeed, whereas NF-κB contributes to many aspects of oncogenesis, it is now clear that its suppressive action on PCD is central to this process. Notably, recent studies indicate that NF-κB represents a crucial link in the well-established association between inflammation and carcinogenesis. In this link, NF-κB promotes synthesis of inflammatory mediators (e.g. TNFα) that stimulate growth of cancer cells, and upregulates genes that protect th... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331392 Thu, 01 Jan 2009 05:00:00 +0100 2331392 http://www.medworm.com/index.php?rid=2331391&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-530-9_1 Chronic inflammation is induced by various infectious/infected agents and by many physical, chemical and immunological factors. Many malignancies arise from areas of infection and inflammation. Reactive oxygen species and reactive nitrogen species are considered to play the key role in inflammation-associated carcinogenesis by causing oxidative and nitrative DNA damage. 8-Nitroguanine is a mutagenic nitrative DNA lesion formed during inflammation. Development of a detection method for 8-nitroguanine would provide an insight into the mechanism of inflammation-associated carcinogenesis and the assessment of carcinogenic risk in patients with inflammatory diseases. We established the method to produce highly sensitive and specific anti-8-nitroguanine rabbit polyclonal antibody, and detect 8-n... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2331391 Thu, 01 Jan 2009 05:00:00 +0100 2331391 http://www.medworm.com/index.php?rid=2074144&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_9 Tobacco smoke and alcohol are major risk factors for a variety of cancer sites, including those of the gastrointestinal tract. Tobacco smoke contains a great number of mutagenic and carcinogenic compounds, including polycyclic carbohydrates, nitrosamines, and nicotine, while ethanol per se has only weak carcinogenic potential, but its first metabolite, acetaldehyde, is a mutagen and carcinogen, since it forms stable adducts with DNA. The possibility of proto-oncogene mutation in gastrointestinal mucosa cells may be associated with tobacco smoking-induced cancers through the formation of unfavorable DNA adducts. Individuals with defective DNA repair mechanisms and unfavorable genetic make-up for carcinogen metabolism may be at increased risk for gastrointestinal cancers. Individuals with a ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074144 Fri, 12 Sep 2008 04:00:00 +0100 2074144 http://www.medworm.com/index.php?rid=2074143&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_8 Energy intake, physical activity, and obesity are modifiable lifestyle factors. This chapter reviews and summarizes the epidemiologic evidence on the relation of energy intake, physical activity, and obesity to cancer. High energy intake may increase the risk of cancers of colon —rectum, prostate (especially advanced prostate cancer), and breast. However, because physical activity, body size, and metabolic efficiency are highly related to total energy intake and expenditure, it is difficult to assess the independent effect of energy intake on cancer risk. There are sufficient evidences to support a role of physical activity in preventing cancers of the colon and breast, whereas the association is stronger in men than in women for colon cancer and in postmenopausal than in premenopaus... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074143 Fri, 12 Sep 2008 04:00:00 +0100 2074143 http://www.medworm.com/index.php?rid=2074142&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_7 Cancer of the breast is a significant health problem for women from the time of diagnosis through the treatment and survivorship trajectory. The disease and treatments are an assault to a woman's body, resulting in sequelae that can be debilitating. Although women diagnosed with breast cancer are living longer, concerns about functional limitations, recurrence, and survival remain paramount. Physical activity and exercise are preventative and rehabilitative measures that can be employed at various points along the breast cancer trajectory. Current research supports the beneficial role that physical activity and exercise play in reducing the risk for developing breast cancer and preventing or attenuating disease and treatment-related impairments. (Source: Springer protocols feed by Cancer R... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074142 Fri, 12 Sep 2008 04:00:00 +0100 2074142 http://www.medworm.com/index.php?rid=2074141&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_6 Exposure to metallic compounds is ubiquitous, with its widespread use in industry and its presence, mostly in trace amounts, in the environment. This paper reviews the epidemiologic evidence of the relation between lung cancer and exposure to metallic compounds by building on and updating the corresponding International Agency for Research on Cancer (IARC) assessments. Given that most of the well-identified human populations with given metal exposure are in occupational settings, this review is mostly based on results in occupational epidemiology. The epidemiological evidence is shortly reviewed for accepted carcinogens: chromium, nickel, beryllium, cadmium, arsenic, and silicon, highlighting what is still unclear. We then review in more detail metals for which the evidence is less clear: ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074141 Fri, 12 Sep 2008 04:00:00 +0100 2074141 http://www.medworm.com/index.php?rid=2074140&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_5 Childhood leukemia is the most common cancer among children, representing 31% of all cancer cases occurring in children younger than the age of 15 years in the USA. There are only few known risk factors of childhood leukemia (sex, age, race, exposure to ionizing radiation, and certain congenital diseases, such as Down syndrome and neurofibromatosis), which account for only 10% of the childhood leukemia cases. Several lines of evidence suggest that childhood leukemia may be more due to environmental rather than genetic factors, although genes may play modifying roles. Human and animal studies showed that the development of childhood leukemia is a two-step process that requires a prenatal initiating event(s) plus a postnatal promoting event(s). (Source: Springer protocols feed by Cancer Rese... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074140 Fri, 12 Sep 2008 04:00:00 +0100 2074140 http://www.medworm.com/index.php?rid=2074139&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_4 Genetic epidemiology studies in hereditary non-polyposis colorectal cancer (HNPCC) have the potential to radically improve assessment of disease risk such that more individualised information can be provided to patients susceptible to developing disease. Studies of HNPCC initially focused on disease associations and the definition of the disease and its association with different cancers within the context of an inherited predisposition. With the identification of the genetic basis of HNPCC, new insights into the disease have been forthcoming and many advances in our understanding have been made. There have been many reports examining potential modifier genes in HNPCC, yet the results remain controversial as many findings have not been replicated and therefore no clear consensus as to the ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074139 Fri, 12 Sep 2008 04:00:00 +0100 2074139 http://www.medworm.com/index.php?rid=2074138&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_3 This chapter posits that cancer is a complex and multifactorial process as demonstrated by the expression and production of key endocrine and steroid hormones that intermesh with lifestyle factors (physical activity, body size, and diet) in combination to heighten cancer risk. Excess weight has been associated with increased mortality from all cancers combined and for cancers of several specific sites. The prevalence of obesity has reached epidemic levels in many parts of the world; more than 1 billion adults are overweight with a body mass index (BMI) exceeding 25. Overweight and obesity are clinically defined indicators of a disease process characterized by the accumulation of body fat due to an excess of energy intake (nutritional intake) relative to energy expenditure (physical activit... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074138 Fri, 12 Sep 2008 04:00:00 +0100 2074138 http://www.medworm.com/index.php?rid=2074137&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_23 Despite a major decline in incidence and mortality over several decades, stomach cancer is still the fourth most common cancer and the second most common cause of cancer death in the world. There is a 10-fold variation in incidence between populations at the highest and lowest risk. The incidence is particularly high in East Asia, Eastern Europe, and parts of Central and South America, and it is about twice as high among men than among women. Prognosis is generally rather poor, with 5-year relative survival below 30% in most countries. The best established risk factors for stomach cancer are Helicobacter pylori infection, the by far strongest established risk factor for distal stomach cancer, and male sex, a family history of stomach cancer, and smoking. While some factors related to diet ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074137 Fri, 12 Sep 2008 04:00:00 +0100 2074137 http://www.medworm.com/index.php?rid=2074136&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_22 Prostate cancer is the number one killer in the United States. A comparison of US and Japanese population is discussed in this chapter to identify risk factors for prostate cancer. Screening of prostate cancer is common among Americans, but not among Japanese. A comparison of survival in different populations is also presented; and prognosis after hormonal treatment in different populations is included. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074136 Fri, 12 Sep 2008 04:00:00 +0100 2074136 http://www.medworm.com/index.php?rid=2074135&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_21 In conclusion, there are multiple factors that impact prostate cancer outcome and that may be responsible for ethnic disparity. These factors are discussed in this chapter. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074135 Fri, 12 Sep 2008 04:00:00 +0100 2074135 http://www.medworm.com/index.php?rid=2074134&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_20 Ovarian cancer represents the sixth most commonly diagnosed cancer among women in the world, and causes more deaths per year than any other cancer of the female reproductive system. Despite the high incidence and mortality rates, the etiology of this disease is poorly understood. Established risk factors for ovarian cancer include age and having a family history of the disease, while protective factors include increasing parity, oral contraceptive use, and oophorectomy. Lactation, incomplete pregnancies, and surgeries such as hysterectomy and tubal ligation may confer a weak protective effect against ovarian cancer. Infertility may contribute to ovarian cancer risk among nulliparous women. Other possible risk factors for ovarian cancer include postmenopausal hormone-replacement therapy and... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074134 Fri, 12 Sep 2008 04:00:00 +0100 2074134 http://www.medworm.com/index.php?rid=2074133&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_2 It is estimated that almost 1.5 million people in the USA are diagnosed with cancer every year. However, due to the substantial effect of modifiable lifestyle factors on the most prevalent cancers, it has been estimated that 50% of cancer is preventable. (Source: Springer protocols feed by Cancer Research) Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074133 Fri, 12 Sep 2008 04:00:00 +0100 2074133 http://www.medworm.com/index.php?rid=2074132&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_19 The 20th century witnessed the scourge of lung cancer as the disease rapidly rose the ranks to become the commonest cause of cancer mortality in the world. Epidemiological evidence conclusively associated cigarette smoking with the causation of lung cancer in the 1950s. Since then and the few decades after, lung cancer of the squamous cell or small cell histological subtypes was mainly diagnosed among male smokers in developed countries. As we move into the 21st century, the incidence of lung cancer is unlikely to abate but the burden will shift from the developed to the less-developed countries. Other epi-demiological changes of lung cancer include the narrowing of the gap between men and women affected by the disease, predominance of the adenocarcinoma histological subtypes as well as mo... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074132 Fri, 12 Sep 2008 04:00:00 +0100 2074132 http://www.medworm.com/index.php?rid=2074131&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_18 Primary fallopian tube carcinoma (PFTC) is a rare malignancy, but its incidence has been rising during the last decades and varies between 2.9/1,000,000 and 5.7/1,000,000. The epidemiology of PFTC has been sparsely studied. In Finland, the incidence rate has been rising during the last decades. The rise has been highest in the cities, in higher social classes, and in certain specific occupations. Parity gives protection against this disease, as does a previous sterilization procedure. Earlier thoughts of a previous salpingitis as a possible promoter of PFTC seem not to hold. Previous infections such as Chlamydia trachomatis infections or human papillomavirus infections cannot be regarded as risk factors. In this chapter, we clarify the possible epidemiological factors behind this disease. ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074131 Fri, 12 Sep 2008 04:00:00 +0100 2074131 http://www.medworm.com/index.php?rid=2074130&cid=s_37120_6_f&fid=37120&url=http%3A%2F%2Fwww.springerprotocols.com%2FAbstract%2Fdoi%2F10.1007%2F978-1-60327-492-0_17 Colonic carcinogenesis is characterized by progressive accumulations of genetic and epigenetic derangements. These molecular events are accompanied by histological changes that progress from mild cryptal architectural abnormalities in small adenomas to eventual invasive cancers. The transition steps from normal colonic epithelium to small adenomas are little understood. In experimental models of colonic carcinogenesis aberrant crypt foci (ACF), collections of abnormal appearing colonic crypts, are the earliest detectable abnormality and precede adenomas. Whether in fact ACF are precursors of colon cancer, however, remains controversial. Recent advances in magnification chromoendoscopy now allow these lesions to be identified in vivo and their natural history ascertained. (Source: Springer ... Springer protocols feed by Cancer Research info http://www.medworm.com/rss/comments.php?id=2074130 Fri, 12 Sep 2008 04:00:00 +0100 2074130