196,115 research outputs found
Cesare Borgia di Francia Gonfaloniere di Santa Romana Chiesa (1498-1503). Conquiste effimere e progettualità statale, Atti del convegno di studi, Urbino 4-5-6 dicembre 2003
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Tra ragione e azzardo: la secolarizzazione del cardinale Valentino
No full textIl contributo illustra le motivazioni strutturali che spinsero Cesare Borgia ad abbandonare la condizione di cardinale per avventurarsi in quella di principe secolare alla ricerca di uno stato territoriale. La scelta, della quale viene comunque messa in risalto una insopprimibile componente psicologica, viene ricondotta alle particolari condizioni della curia romana e del Sacro Collegio a fine Quattrocento, in modo particolare per quanto riguardava il complesso rapporto tra la famiglia Borgia e i Re cattolici di Spagna
Murine and Human Mammary Cancer Cell Lines: Functional Tests
No full textThe biological characterization of mammary cancer cells is a prerequisite that helps the scientist understand some aspect of tumor biology. Once isolated from the tumor, cells are subjected to multiple tests that dissect their ability to growth, migrate, degrade the surrounding strome, produce 3-dimensional structures and differentiate. Targeted inhibitors, when added to these tests, are used to unravel how specific growth factors, receptors, and intracellular translational pathways promote the ability of mammary tumor cells to achieve their biological behavior. Herein we describe a set of techniques used to put in focus the biological capacities of mammary cancer cells. When the characterization of a biological trait (e.g., proliferation) is assessable by multiple assays, we will limit the description to only one technique, possibly the easier to manage and that requires minimal laboratory equipment
MicroRNAs as Biomarkers for Animal Health and Welfare in Livestock
No full textMicroRNAs (miRNAs) are small and highly conserved non-coding RNA molecules that orchestrate a wide range of biological processes through the post-transcriptional regulation of gene expression. An intriguing aspect in identifying these molecules as biomarkers is derived from their role in cell-to-cell communication, their active secretion from cells into the extracellular environment, their high stability in body fluids, and their ease of collection. All these features confer on miRNAs the potential to become a non-invasive tool to score animal welfare. There is growing interest in the importance of miRNAs as biomarkers for assessing the welfare of livestock during metabolic, environmental, and management stress, particularly in ruminants, pigs, and poultry. This review provides an overview of the current knowledge regarding the potential use of tissue and/or circulating miRNAs as biomarkers for the assessment of the health and welfare status in these livestock species
Determination of sound properties on acoustic panels made from recycled cigarette filters
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A Sorting Strategy For Bovine Mammary Progenitors
No full textThe mammary gland is a bilayered system organized into a series of branching ducts that terminate in secretory alveoli. Their epithelium is composed of an inner layer of cytokeratin(CK)18+ luminal cells, which secrete milk, and an outer layer of contractile CK14+ myoepithelial cells, which contribute to milk ejection during lactation. The existence of mammary renewable stem cells (MaSCs) has been deduced from the profound regenerative ability of the mammary epithelium. It undergoes different morphogenetic changes during puberty and during the subsequent pregnancies. The knowledge of MaSCs properties and functions should represent an interesting opportunity in the field of animal production as the appropriate regulation of these cells should benefit milk yeld. The aim of our study is to characterize the MaSCs population of bovine species and set up a sorting strategy to purify different primitive populations. The Colony-Forming Cell (CFC) assay performed on bovine mammary samples obtained from virgin, two years old cows revealed the presence in the mammary gland of both myoepithelial and luminal-restricted progenitor cells. In order to select and isolate the different types of progenitors we have performed a flow cytometric analysis of cells dissociated from bovine mammary tissue. Cells were then stained with antibodies directed to CD49f, whose expression at high levels is associated with progenitor and stem cell activity, and p-cadherin, that is normally expressed in the myoepithelial/basal layer of the mammary epithelium. After depletion of hematopoietic cells (using an anti-CD45 antibody) we identified four different subpopulations. They were characterized by different levels of CD49f/P-cad expression: CD49f/P-cad- (15,6%), CD49f++/P-cad- (25,4%), CD49f+/P-cad+ (27,1%), CD49f+/P-cad++ (10,2%). After sorting, the CFC assay performed with the cells of the different subpopulations showed that CD49f++/P-cad- population was able to generate colonies with an higher frequency compared to the others, revealing an enrichment in stem cells. In particular, 35,7% of colonies showed a myoepithelial phenotype and 64,3% a luminal one. In order to analyze whether these cells also had an in vivo tissue regenerative activity, the four subpopulation were implanted in collagen gels under the kidney capsule of NOD/SCID immunodeficient mice in numbers proportional to their frequency in the CD45- population. Immunohistochemical analysis of gels recovered 4 weeks later revealed that only CD49f++/P-cad- subpopulation was able to regenerate in vivo mammary alveolar structures characterized by a lumen surrounded by two layers of cells: an inner K18+ layer and an outer K14+ layer. The CFC assay performed with single cell suspension obtained from recovered gels showed a higher colony-forming ability of CD49f++/P-cad- population compared to the others. Therefore the structures produced in these gels contained clonogenic progenitors, that were able to form the same types of colonies observed from freshly isolated cells (36,2% of myoepithelial colonies and 63,8% of luminal ones). By using a novel sorting strategy we show that the mammary myoepithelial cell layer can be divided in different subpopulations. Each one has a different progenitor content and only in one fraction (CD49f++/P-Cad-) we were able to detect a subset of adult stem cells able to regenerate a mammary epithelium. This sorting strategy might prove useful for understanding the stem cells population dynamics during the different phases that the mammary gland undergoes. Besides, given the similar organization of the mammary tissue in the bovine and the human species, the cow might represent a much better animal model than the mouse
Functional effect of mir-27b on myostatin expression: a relationship in Piedmontese cattle with double-muscled phenotype
No full textAbstract
Background: In Piedmontese cattle the double-muscled phenotype is an inherited condition associated to a point
mutation in the myostatin (MSTN) gene. The Piedmontese MSTN missense mutation G938A is translated to C313Y myostatin protein. This mutation alters MSTN function as a negative regulator of muscle growth, thereby inducing muscle hypertrophy. MiRNAs could play a role in skeletal muscle hypertrophy modulation by down-regulating gene
expression.
Results: After identifying a 30-UTR consensus sequence of several negative and positive modulator genes involved
in the skeletal muscle hypertrophy pathway, such as IGF1, IGF1R, PPP3CA, NFATc1, MEF2C, GSK3b, TEAD1 and
MSTN, we screened miRNAs matching to it. This analysis led to the identification of miR-27b, miR-132, miR-186 and
miR-199b-5p as possible candidates. We collected samples of longissimus thoracis from twenty Piedmontese and
twenty Friesian male bovines. In Piedmontese group miR-27b was up-regulated 7.4-fold (p < 0.05). Further, we
report that the level of MSTN mRNA was about 5-fold lower in Piedmontese cattle vs Friesian cattle (p < 0.0001)
and that less mature MSTN protein was detected in the Piedmontese one (p < 0.0001). Cotransfection of miR-27b
and psi-check2 vector with the luciferase reporter gene linked to the bovine wild-type 30-UTR of MSTN strongly
inhibited the luciferase activity (79%, p < 0.0001).
Conclusions: These data demonstrate that bovine MSTN is a specific target of miR-27b and that miRNAs contribute
to explain additive phenotypic hypertrophy in Piedmontese cattle selected for the MSTN gene mutation, possibly
outlining a more precise genetic signature able to elucidate differences in muscle conformation
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