Rechtliche Aspekte von Sterben und Tod

Tables S1-S8. Demonstrating genes from the groups A-H and their functional annotations. (PDF 63 kb

El ingreso de estudiantes en situación de discapacidad a la UNLP : Apoyos, políticas y desafíos

La presente ponencia es producto de los debates, interrogantes y reflexiones, que surgen de la labor en la Comisión Universitaria sobre Discapacidad de la UNLP (en adelante, CUD). Desde este espacio de trabajo colectivo e interdisciplinario, se trabaja en la planificación, ejecución y evaluación de políticas destinadas a garantizar, entre otras acciones, la accesibilidad académica de estudiantes en situación de discapacidad. En esta oportunidad, se desarrollan las estrategias implementadas con los estudiantes ingresantes a nuestra alta casa de estudios.Eje 2: Nuevas experiencias y trayectorias estudiantiles. Desafíos para la inclusión educativa en la universidad. Reflexiones y debates en torno de la inclusión educativa en la universidadSecretaría de Asuntos Académico

Indirect immunofluorescence.

<p>Kera5, MaFi132, 308 and NIH 3T3 cells were grown on cover slips, fixed with acetone, stained with keratin 14 or vimentin specific antibodies and detected with AlexaFluor488 or AlexaFluor594-conjugated secondary antibodies, respectively. Murine keratinocytes (308 cells) and fibroblasts (NIH 3T3) were used as controls. Green fluorescence indicates keratin 14 expression only in Kera5 and 308 cells, whereas red fluorescence shows vimentin expression only in MaFi132 and NIH 3T3 (original magnification: 200x).</p

MYCNAMP NB4 70, a novel miRNA-like sequence resulting from a duplication event

<p><b>Copyright information:</b></p><p>Taken from "New miRNAs cloned from neuroblastoma"</p><p>http://www.biomedcentral.com/1471-2164/9/52</p><p>BMC Genomics 2008;9():52-52.</p><p>Published online 29 Jan 2008</p><p>PMCID:PMC2254388.</p><p></p> Short sequences flanking MYCNAMP_NB4_70 were extracted from the human genome, as well as homologuous sequences from chimpanzee, macaque, mouse, rat and dog genomes and aligned. The bar indicates the MYCNAMP_NB4_70 sequence. The duplication is marked by a box

Predicted secondary structures of the putative novel human miRNA precursors

<p><b>Copyright information:</b></p><p>Taken from "New miRNAs cloned from neuroblastoma"</p><p>http://www.biomedcentral.com/1471-2164/9/52</p><p>BMC Genomics 2008;9():52-52.</p><p>Published online 29 Jan 2008</p><p>PMCID:PMC2254388.</p><p></p> Human genomic sequences upstream and downstream of the novel miRNAs were folded with the computer program RNAfold. Grey areas represent the cloned miRNA. a) miRNA with canonical hairpin. b) borderline structures

Establishment of an Immortalized Skin Keratinocyte Cell Line Derived from the Animal Model <i>Mastomys coucha</i>

<div><p>In the present report we describe the establishment of a spontaneous immortalized skin keratinocyte cell line derived from the skin of the multimammate rodent <i>Mastomys coucha</i>. These animals are used in preclinical studies for a variety of human diseases such as infections with nematodes, bacteria and papillomaviruses, especially regarding cutaneous manifestations such as non-melanoma skin cancer. Here we characterize the cells in terms of their origin and cytogenetic features. Searching for genomic signatures, a spontaneous mutation in the splicing donor sequence of <i>Trp53</i> (G to A transition at the first position of intron 7) could be detected. This point mutation leads to alternative splicing and to a premature stop codon, resulting in a truncated and, in turn, undetectable form of p53, probably contributing to the process of immortalization. <i>Mastomys coucha</i>-derived skin keratinocytes can be used as an <i>in vitro</i> system to investigate molecular and immunological aspects of infectious agent interactions with their host cells.</p></div

Calcium-induced differentiation of Kera5.

<p>Kera5 cells at passage 139 were grown on cover slips. After 24 h, the dKSFM (<0.1 mM Ca²⁺) was additionally supplemented with 0.35 mM, 0.7 mM or 1.05 mM Ca²⁺ to induce differentiation. After additional 24 h incubation, the cells were fixed with acetone, stained with an involucrin antibody and detected with an AlexaFluor488 secondary antibody, respectively. Green fluorescence indicates elevated involucrin expression in a dose-dependent manner (original magnification: 200x).</p

Cell morphology.

<p><b>(A)</b><i>Mastomys coucha</i> keratinocytes at passage 6. <b>(B)</b> Kera5 at passage 175, showing a typical cobblestone phenotype and an increased cell size (arrowheads mark ongoing mitoses; magnification: 200x).</p

Sequencing of the <i>Mastomys</i> p53 gene (<i>Trp53</i>) and partial alignment with mouse and rat sequences.

<p><b>(A)</b> Sequencing of <i>Trp53</i> in Kera5 (passages 8 and 103) shows a G>A transition at the first position of intron 7 (underlined). Freshly isolated primary keratinocytes (Kera5, p0) as well as five individual <i>Mastomys</i> samples do not harbor this mutation and are similar to murine and rat sequences at this position. Numbers refer to the sequences of murine <i>Trp53</i> and rat <i>Tp53</i>. For intron 7, only 5´-start and 3´-ends are shown. Exon 7: green, exon 8: grey, insertion: orange, frames: splicing signals; “a” indicates the original splicing donor, “b” alternative splicing donor signal in intron 7, “c”: splicing acceptor. <b>(B)</b> Sequencing chromatograms of <i>Trp53</i> reveal the G>A transition in a subpopulation of Kera5 cells at p8 which is not present at p0. A switch of the major peak from G to A from p8 to p103 occurs, suggesting the outgrowth of a single cell colony. At p146 only the A peak is left, revealing homozygosity. The arrows indicate the position of the mutation.</p

p53 cDNA sequence and transcriptional analyses.

<p><b>(A)</b> The relevant nucleotides of the mutated p53 cDNA are shown. At passage 13, an insertion of 19 nt is detectable in the cDNA of p53 (exon 7: green, exon 8: grey, insertion: orange). <b>(B)</b> Comparison of translated wildtype and mutant sequences derived from the cDNA. The insertion leads to a premature stop codon (*) that results in a truncated form of p53. <b>(C)</b> Semi-quantitative RT-PCR of <i>Trp53</i> at different passages. <i>GAPDH</i> was used as reference gene. <b>(D)</b> Semi-quantitative RT-PCR to detect the insertion within the p53 cDNA, resulting in a slower-migrating PCR product at higher passages (p27-p105). <i>GAPDH</i> was used as reference gene. <b>(E)</b> Western blotting. Left panel: lack of p53 expression in Kera5. Kera5 (p155) were exposed to UVB (40 or 250 mJ/cm²) or treated with adriamycin (1.0 μg/ml) and harvested as indicated. 50 μg protein/lane were loaded. To control the specificity of the p53 antibody, MaFi132 were transiently transfected with pPK-CMV-E3 containing the cDNA for wildtype (p53wt) or the truncated (p53trunc)) form of p53. Since only 25 μg of protein were loaded for transfected MaFi132, a longer exposure time was chosen for actin. Right panel: NIH 3T3 cells were used as reference for stabilization of p53 after cells damage with UV or Adriamycin, respectively. NIH 3T3 cells were exposed to UVB (20 or 100 mJ/cm²) or treated with Adriamycin (0.75 μg/ml) and harvested as indicated. 50 μg protein/lane were loaded. After low dose UVB, p53 stabilization occurs slowly, whereas after high dose UVB the stabilization is fast, but transient. Adriamycin permanently stabilizes p53 already after 7h. Actin was used as loading control.</p