OLFM4, KNG1 and Sec24C identified by proteomics and immunohistochemistry as potential markers of early colorectal cancer stages
Florence Quesada‐CalvoCharlotte MassotVirginie BertrandRémi LonguespéeNoëlla BlétardJoan SomjaGabriel MazzucchelliNicolas SmargiassoDominique BaiwirMarie‐Claire De Pauw‐GilletPhilippe DelvenneMichel MalaiseCarla Coimbra MarquesMarc PolusEdwin De PauwMarie‐Alice MeuwisÉdouard Louis
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Despite recent advances in colorectal cancer (CRC) diagnosis and population screening programs, the identification of patients with preneoplastic lesions or with early CRC stages remains challenging and is important for reducing CRC incidence and increasing patient’s survival. We analysed 76 colorectal tissue samples originated from early CRC stages, normal or inflamed mucosa by label-free proteomics. The characterisation of three selected biomarker candidates was performed by immunohistochemistry on an independent set of precancerous and cancerous lesions harbouring increasing CRC stages. Out of 5258 proteins identified, we obtained 561 proteins with a significant differential distribution among groups of patients and controls. KNG1, OLFM4 and Sec24C distributions were validated in tissues and showed different expression levels especially in the two early CRC stages compared to normal and preneoplastic tissues. We highlighted three proteins that require further investigations to better characterise their role in early CRC carcinogenesis and their potential as early CRC markers.This chapter contains sections titled: Introduction Proteomics of Plasma and Serum of Animals Proteomics of the Liver of Animals Proteomics of the Immune System of Animals Proteomics of the CNS in Animals Proteomics of Muscle in Animals Proteomics of Milk and the Mammary Gland of Animals Proteomics of the Intestine in Animals Proteomics of Skin in Animals Proteomics of the Respiratory Organs in Animals Proteomics of the Visual Apparatus in Animals Proteomics of Bone and Cartilage in Animals Proteomics of Endocrinology in Animals Proteomics of Kidney and Urine in Animals Conclusions References
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Immunohistochemistry currently is the most powerful tool to identify specific molecules in situ. Moreover, antibodies can detect modifications of cellular polypeptides that occur naturally or as a result of toxic or other injuries. Here we report on a variety of parameters (fixation, tissue pretreatments, chromagen intensification, etc.) that we found to enhance immunohistochemical staining. Further, the use of well characterized antibodies and concomitant immunoblotting help avoid false positive results in immunohistochemistry. Finally, the sensitivity of the immunohistochemical procedure, regional differences in immunostaining within the central nervous system, as well as the recognition of equivalent molecules across different animal species are discussed.
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Immunohistochemistry (IHC) is the technique of assessing the expressed protein in vivo through antigen-antibody reaction. In this technique, a more expressed protein is darkly stained than a less expressed one. Immunohistochemistry (IHC) is a technique that involves histological, immunological, and biochemical techniques. IHC with both direct (involving primary antibody) and indirect (involving both primary and secondary antibodies) methods was discussed. A detailed protocol for IHC has been discussed with practical examples of IHC as conducted in our laboratory of growth hormone in pituitary gland, and CD4 and CD8 expressions in uterus. The basic steps for IHC include fixing and embedding the tissue; cutting and mounting the section; deparaffinizing and rehydrating the section; antigen retrieval; immunohistochemical staining; counterstaining (if desired); dehydrating and stabilizing with mounting medium; and viewing the staining under the microscope.
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Immunohistochemistry has become an important tool in research and in surgical pathology. Rapid growth of a new methods in immunohistochemistry supplement traditional histochemical and histological light microscopy investigations. Immunohistochemistry has given pathologists a chance to location different antigens on the cell surface as well as in the cell compartments. The expression of antigens are mostly influenced by factors connected with tissue processing; fixation and embedding. The aim of present article is to show the role of these factors and their influence on some immunohistochemical staining results. Not all the problems are discussed here, the main goal which authors would like to obtain is to show the way how to solve problems which can occur during immunohistochemical staining procedure. They want also to delineate the importance of standardization in immunohistochemistry to make the results more reliable between different laboratories.
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Immunohistochemistry [IHC] is an important application of monoclonal as well as polyclonal antibodies to determine the tissue distribution of an antigen [protein or lipid] by specific antigen/antibody reaction tagged with a visible label. Immunohistochemistry has an expanding role in diagnostic and research laboratories. This article highlights the various applications of IHC in health and diseases and gives more information in the Future directions of immunohistochemistry.
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Objective To discuss the application o immunohistochemistry autostainer in immunohistochemistry.Methods More than 30 kinds of first antibodies of the Pathology Department,such as AAT,CerBb-2,CK,HBcAg,ER,PR,Ki-67,and unified second antibodies were selected.Immunohistochemistry autostainer and manual operation were applied to the staining of the antibodies,and then the results by the above methods were compared.Results The antibodies stained by immunohistochemistry autostainer,gifted with clear background,no edge effect,uniform staining and accurate positive results,were all better than those by manual operation from all aspects.Conclusion Immunohistochemistry autostainer is highly automatic,time-saving,manpower-saving,repeatable and highly standardized.
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In the diagnosis of clinical pathology,immunohistochemistry(IHC) is a very important technology and tools.The immunohistochemical technique used in the pathological diagnosis started the 1970s,pathologist diagnosis of tumor classification,prognosis had a huge impact,but also extends the understanding of the people for various diseases and tumor formationpathological diagnosis and research..However,with the extensive application of immunohistochemistry,the immunohistochemical technique have some limitations.In-depth study of the principles and techniques of immunohistochemistry,familiar with all kinds of really positive antibody reaction site,and inter-laboratory standardization of immunohistochemistry,play the largest role in support of immunohistochemistry in the pathological diagnosis.This article on the above issues as follows.
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Immunohistochemistry (IHC) is an important application of monoclonal as well as polyclonal antibodies to determine the tissue distribution of an antigen of interest in health and disease. IHC is widely used for diagnosis of cancers; specific tumor antigens are expressed de novo or up-regulated in certain cancers. This article deals with the various applications of IHC in diagnosis of diseases, with IHC playing an important role in diagnostic and research laboratories.
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