Análisis teórico-práctico de los requisitos y presupuestos de la Ley 100 de 1993 y Ley 797 de 2003

Artículo de investigaciónEste trabajo hace un análisis detallado del Sistema Pensional Colombiano, partiendo de la multiplicidad de regímenes de transición que se han presentado derivadas de las reformas a través del tiempo y la problemática que ello a generado, para luego detallar los requisitos necesarios para obtener las diferentes modalidades de pensión, tanto en el Régimen de Ahorro Individual con Solidaridad RAIS, como en el Régimen de Prima Media con Prestación Definida RPM. Finalmente, contiene un breve análisis del Acto Legislativo 01 de 2005.48 p.INTRODUCCIÓN 1. PLANTEAMIENTO DEL PROBLEMA 2. OBJETO DE LA INVESTIGACION 3. REGIMEN DE TRANSICION 4. LEY 100 DE 1993 5. LEY 797 DE 2003 6. ACTO LEGISLATIVO 01 DE 2005 CONCLUSIONESPregradoAbogad

HLM006474 IC₅₀ values across various cell types.

<p>The IC₅₀ of various cell lines was determined as described in the methods. STDEV refers to the standard deviation of indicated values calculated using Excel STDEV function. The average IC₅₀s of the NSCLC and SCLC groups are calculated in bold along with STDEVs. The differences are not statistically significant.</p

Hypothetical model to explain the role of Trim28 in the stages of lung adenocarcinoma.

<p>This model explains this work and our previous work demonstrating a tumor suppressor role for Trim28 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101040#pone.0101040-Chen1" target="_blank">[9]</a>.</p

Trim28 expression is induced by TGF-β.

<p><i>A</i> and <i>B</i>, A549 and H358 stably expressing non-silencing shRNA and Trim28 shRNA were treated with TGF-β (2 ng/ml) for 3 to 10 days. Whole cell lysates and RNA extracts were subjected to western blotting using the indicated antibodies (A) and real-time PCR (B). <i>Asterisks</i> represent significant <i>p</i> values, *: <i>p</i><0.05.</p

Expression of common E2F-regulated genes increases in early time point treatments with HLM006474.

<p><i>A</i>. The NSCLC cell lines H1299 and H292 were treated with 60 µM HLM006474 for 0, 3, 6, 9, 12, and 24 hours, then harvested and examined through Western blots. Modest increases in E2F1 and more dramatic increases in E2F3a and b protein levels were noted at early time point in both cell lines. <i>B–D</i>. mRNA was harvested from cells that were treated as described in 1A, converted to cDNA using RT-PCR, and analyzed for expression levels of E3F3 (<i>B</i>), E2F1 (<i>C</i>), and E2F4 (<i>D</i>) using real-time PCR and tubulin as a control. Short treatments with 6474 altered mRNA expression levels of E2F3 and E2F1, but not E2F4. <i>E–G.</i> mRNA expression of well-known genes commonly regulated by E2Fs were analyzed in a manner similar to the previous description in 1B–D. Expression levels of MCM10 (<i>E</i>), MCM2 (<i>F</i>), and CCNE2 (<i>G</i>) mRNA were analyzed with tubulin as a control and were all noted to be more highly expressed following short treatments with 6474. Notes: n.s. represents not significant, * represents p<0.05, ** represents p<0.01.</p

Trim28 regulates histone modifications of E-cadherin and N-cadherin promoters.

<p><i>A</i>, ChIP assays were conducted in control and Trim28 knockdown A549 cells. Quantitative PCR was performed to examine the occupancy of IgG, acetylated histone 3 K9, dimethylated histone 3 K9, and trimethylated histone 3 K9 on E-cadherin and N-cadherin promoters. B, ChIP assays were conducted in control and Trim28 stably expressed A549 cells. Quantitative PCR was performed to examine the occupancy of IgG, acetylated histone 3 K9, dimethylated histone 3 K9, and trimethylated histone 3 K9 on E-cadherin and N-cadherin promoters.</p

Trim28 knockdown attenuates cell migration.

<p><i>A</i>, control and Trim28 knockdown H358 cells were subjected to wound-healing assay. Cells were photographed right after scratch, and 24 hrs and 48 hrs after scratch. <i>B</i>, control and Trim28 knockdown A549 cells were subjected to wound healing assay in the absence or presence of serum. Cells were photographed right after scratch and 24 hrs later. <i>C</i>, control and Trim28 knockdown cells were plated in triplicate on the top wells of Matrigel-coated or non-coated chambers. After 18 hours, cells that had invaded through the Matrigel layer were fixed and stained. The percentage of invasive cells is shown in the bar graph. <i>Asterisks</i> represent significant <i>p</i> values, *: <i>p</i><0.05. <i>D</i>, control and Trim28 knockdown A549 cells were cultured in a three-dimensional model and treated with TGF-β (5 ng/ml) for 72 hrs. RNA was extracted from the cells and subjected to real-time PCR. The expression of target genes was determined.</p

Trim28 is required for TGF-β-induced EMT.

<p><i>A</i>, control and Trim28 knockdown cells were treated with TGF-β or left untreated. Cells were photographed at 40× magnification. <i>B</i>, RNA was extracted from control and Trim28 knockdown A549 cells treated with TGF-β as above or left untreated. Real-time PCR was performed and the expression of target genes was determined. <i>C</i>, control and Trim28 knockdown A549 and H358 cells were treated with TGF-β (2 ng/ml) until morphology changes were observed or left untreated. Whole cell lysates were subjected to western blotting using antibodies as indicated.</p

Model of HLM006474 synergy with paclitaxel.

<p>In untreated cells, E2F/dimerization partner (DP)/Rb complexes predominate, and thus, E2F3 levels are relative low (as well as many other E2F-regulated genes). Treatment with HLM006474 disrupts the E2F/DP/Rb repressive complexes from binding DNA, resulting in increased transcription of E2F3 (as well as many other E2F-regulated genes). In this model, the elevated levels of E2F3 sensitize cells to taxane treatment, as previously demonstrated, through an unknown mechanism.</p

Depletion of E2F3 alters sensitivity to paclitaxel.

<p><i>A</i>. H1299 cells were transiently transfected with 200 pmol control, E2F3a, or E2F3b siRNA and harvested after 24 hours. Western blots of these lysates demonstrate the extent of E2F knockdown. <i>B</i>. MTS assays were used to determine the sensitivity of each cell line to 50 nM paclitaxel. Cells with diminished E2F3 levels were more viable in the presence of paclitaxel than control cells. Notes: * represents p<0.05, ** represents p<0.01.</p