Browsing by Subject "Genomics"

Now showing 1 - 3 of 3
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    Laser capture microdissection: Big data from small samples
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2015) Datta, Soma; Malhotra, Lavina; Dickerson, Ryan; Chaffee, Scott; Sen, Chandan K.; Roy, Sashwati
    Any tissue is made up of a heterogeneous mix of spatially distributed cell types. In response to any (patho) physiological cue, responses of each cell type in any given tissue may be unique and cannot be homogenized across cell-types and spatial co-ordinates. For example, in response to myocardial infarction, on one hand myocytes and fibroblasts of the heart tissue respond differently. On the other hand, myocytes in the infarct core respond differently compared to those in the peri-infarct zone. Therefore, isolation of pure targeted cells is an important and essential step for the molecular analysis of cells involved in the progression of disease. Laser capture microdissection (LCM) is powerful to obtain a pure targeted cell subgroup, or even a single cell, quickly and precisely under the microscope, successfully tackling the problem of tissue heterogeneity in molecular analysis. This review presents an overview of LCM technology, the principles, advantages and limitations and its down-stream applications in the fields of proteomics, genomics and transcriptomics. With powerful technologies and appropriate applications, this technique provides unprecedented insights into cell biology from cells grown in their natural tissue habitat as opposed to those cultured in artificial petri dish conditions.
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    Structure and Enzymatic Properties of an Unusual Cysteine Tryptophylquinone-Dependent Glycine Oxidase from Pseudoalteromonas luteoviolacea
    (ACS Publications, 2018) Andreo-Vidal, Andres; Mamounis, Kyle J.; Sehanobish, Esha; Avalos, Dante; Campillo-Brocal, Jonatan C.; Sanchez-Amat, Antonio; Yukl, Erik T.; Davidson, Victor L.; Genética y Microbiología
    Glycine oxidase from Pseudoalteromonas luteoviolacea (PlGoxA) is a cysteine tryptophylquinone (CTQ)-dependent enzyme. Sequence and phylogenetic analysis place it in a newly designated subgroup (Group IID) of a recently identified family of LodA-like proteins, which are predicted to possess CTQ. The crystal structure of PlGoxA reveals that it is a homo-tetramer. It possesses an N-terminal domain with no close structural homologues in the Protein Data Bank. The active site is quite small due to intersubunit interactions, which may account for the observed cooperativy towards glycine. Steady-state kinetic analysis yielded values of kcat=6.0±0.2 s−1, K0.5=187±18 μM and h=1.77±0.27. In contrast to other quinoprotein amine dehydrogenases and oxidases that exhibit anomalously large primary kinetic isotope effects on the rate of reduction of the quinone cofactor by the amine substrate, no significant primary kinetic isotope effect was observed for this reaction of PlGoxA. The absorbance spectrum of the glycine-reduced PlGoxA exhibits features in the 400-650 nm range that have not previously been seen in other quinoproteins. Thus, in addition to the unusual structural features of PlGoxA, the kinetic and chemical reaction mechanisms of the reductive half-reaction of PlGoxA appear to be distinct from those of other amine dehydrogenases and amine oxidases that use tryptophylquinone and tyrosylquinone cofactors.
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    The hepatic stellate cell in the post-genomic era
    (Murcia : F. Hernández, 2002) Okuyama, H.; Shimahara, Y.; Kawada, N.
    The draft human genome sequence was published on February 15, 2001, which will provide a huge amount of information on human genetics, human disease, and human cell biology. Now, medical scientists and cell biologists are turning their attention to illustrating gene expression pattern using gene microarray and to identifying the functions and the expression patterns of proteins encoded by the genes. Hepatic stellate cell is one of the sinusoidconstituent cells that play multiple roles in the liver p a t h o p h y s i o l o g y. Transformation of stellate cells from the vitamin A-storing phenotype to the “myofibroblastic” one closely correlates to hepatic fibrosis during chronic liver trauma. Analyses of the molecular mechanisms of stellate cell activation have made a great progress, in particular, in the field of intracellular signal transduction of transforming growth f a c t o r-ß and platelet-derived growth factor, integrin signaling related to cell-adhesion, and cell motilityassociated Rho and focal-adhesion kinase. Accumulation of the information on the stellate cell activation would shed light on the establishment of a novel therapeutic strategy against fibrosis of human liver disease.