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DNA microarrays is a method used for the determination of gene expression level, detection of single nucleotide polymorphism (SNP) and quantification of the exact number of viral gene copies per one cell. The technique has a broad application in studies on tumorous diseases of humans and animals. Microarrays are also applied in studies of drug influence on the cell metabolism and the molecular silencing of genes. The main advantage of DNA and protein microarrays is the possibility to analyse a number of complex cellular processes in progress of a tumoral disease during a single run of analysis. In the case of a tumorous disease the microarray technique makes it possible to quickly identify the main genes responsible for the oncogenesis and to determine the type of the disease. In spite of a high costs of microarray production as well as the need to synthesize probes and substrates for specific analyses, microarrays become increasingly popular in numerous research laboratories.
The article presents the current knowledge on the microarray technique and its applications in medical sciences and parasitology. The first part of the article is focused on the technical aspects (microarray preparation, different microarray platforms, probes preparation, hybridization and signal detection). The article also describes possible ways of proceeding during laboratory work on organism of which the genome sequence is not known or has been only partially sequenced. The second part of the review describes how microarray technique have been, or possibly will be, used for better understanding parasite life cycles and development, host-parasite relationship, comparative genomics of virulent organisms, develpoment vaccines against the most virulent parasites and host responses to infection.
The aim of this review is to explain fundamentals of DNA microarray technique and indicate possibilities of its use in veterinary sciences. DNA microarray technology is a powerful tool for functional and structural genomics. It allows the analysis of an entire genome expression (transcriptomic profile) on one glass or plastic slide in a short period of time. It gives the opportunity to establish genes expression patterns characteristic for different physiological and pathological conditions and allows its use as a diagnostic tool. The use of expression microarrays gives the possibility to examine the influence of new treatment strategies in particular diseases and enables the assessment of advantages and disadvantages of this type of therapy. It also allows finding new targets (key proteins, enzymes) for newly developed drugs and makes possible the adaptation of a drug therapy to individual patients. By using DNA microarray technique it is possible to make a new hypothesis and to validate current ones. In this review the origin of the DNA microarray technique, its molecular basics, different kinds of microarray slides and analysis platform as well as the methodology of labeling and hybridization were described. This review may be very useful for people who are planning to use or are have already begun using DNA microarray technique.
The review presents a summary of knowledge on canine genome research using the DNA microarray technique - a new tool in molecular biology. Canine genome was sequenced in 2005, and since then numerous studies on gene expression in various tissues and organs as well as in different physiological or pathological conditions have been published. Canine DNA microarray has been used in research on neoplastic diseases (e.g. brain tumor), kidney diseases (Alport syndrome, hypertension in obese dogs), cardiac diseases (atrial fibrillation, dilation of the heart) and cartilage diseases. This technique has also enabled the analysis of gene expression in acute phase response and in renal tubular epithelial cell line (MDCK) stimulated by hepatocyte growth factor.
The developement of DNA microarray technique has opened the possibility of large-scale studies on genomic determinants of growth and maliganancy of neoplasms. It enables a wide analysis of cancer gene expression in a short period of time. Such defined and specific profiles of gene expression are a molecular signature (“molecular portrait”), which in the future could be used as a reliable marker of cancer malignancy and prognosis. Molecular identification of breast cancer sub-types and their relation with prognosis was one of the most important microarray studies. Unfortunately, in contrast to medical oncogenomics, the development of veterinary oncogenomics is not so dynamic. The first oncogenomic studies on canine mammary gland tumor cell lines were performed at Utrecht University. They successfully identified deregulated pathways pertaining to cell line phenotype similar to the published human breast cancer data sets. The results presented in this paper show that high proliferative and anti-apoptotic potential of canine mammary gland cancer cells may be associated with an enhanced expression of genes involved in the Ca²⁺ signaling pathway (clamodulin 1, 2 and 3, and ryanodine receptor) and growth hormone cellular pathway. Moreover, a comparative study of gene expression in canine mammary tumor tissue and primary cell lines derived from those tumors was conducted. The obtained results show that the transcriptional profile of the primary cell culture resembles that in tumor tissue, and prove that cell cultures can be used as a reliable model for oncogenomic and pharmacogenomic studies. These promising results make it possible to anticipate a progressive development of veterinary oncogenomics.
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