Designer Exons Inform a Biophysical Model for Exon Definition

Pre-mRNA molecules in humans contain mostly short internal exons flanked by long introns. To explain the removal of such introns, recognition of the exons instead of recognition of the introns has been proposed. This thesis studies this exon definition mechanism using a bottom-up approach. To reduce the complexity of the system under study, this exon definition mechanism was addressed using designer exons made up of prototype sequence modules of our own design (including an exonic splicing enhancer or silencer). Studies were performed in vitro with a set of DEs obtained from random combinations of the exonic splicing enhancer and the exonic splicing silencer modules. The results showed considerable variability both in terms of the composition and size of the DEs and in terms of their inclusion level. To understand how different DEs generated different inclusion levels, the problem was divided into understanding separately parameters varied between DEs. Subsequent studies focused on each of three parameters: size, ESE composition and ESS composition. The final objective was to be able to combine their effects to predict the inclusion levels obtained for the "random" DEs mentioned previously. To complement this experimental approach an equation was generated in two stages. First a general "framework" equation was obtained modeling a necessary exon definition complex that enabled commitment to inclusion. This equation used rates for the formation and dissociation of this complex without elaborating on the details for how those rates came about. In the second stage, however, formation and dissociation were modeled using novel but intuitive ideas and these models were combined into a final equation. This equation using the single-parameter perturbation data obtained previously performed well in predicting the inclusion levels of the "random" DEs. Additionally, both the final equation and the mechanisms proposed align well with results published by other groups. In order to make these results more accessible and to open more opportunities to extend them, an initial attempt is presented to identify the proteins involved in the functionality observed for each of the sequences used

Identification of individualization techniques for criminal records in sanction lists

Using efficient searching techniques on sanctions lists and press articles allows a better filtering on individuals and entities to establish a commercial relationship with, including those who are going to have access to confidential information belonging to the company, in order to minimize the risk of leakage or information mismanagement. That process of filtering on individuals or entities could be automated by using individualization algorithms, searching techniques based on string comparisons, artificial intelligence, and facial recognition. Diverse methods were examined to be applied on each mentioned technique in order to identify which ones are ideal to its application on individualization due to their characteristics, in order to obtain agile and reliable results; taking into account that different methods are complementary and not exclusive, and that their combination allows to minimize human interaction in the classification of information, avoiding analysis of irrelevant data for that particular search. Keywords: Criminal records second False positives Filters Sanctions list Verification method

Saturation mutagenesis reveals manifold determinants of exon definition.

To illuminate the extent and roles of exonic sequences in the splicing of human RNA transcripts, we conducted saturation mutagenesis of a 51-nt internal exon in a three-exon minigene. All possible single and tandem dinucleotide substitutions were surveyed. Using high-throughput genetics, 5560 minigene molecules were assayed for splicing in human HEK293 cells. Up to 70% of mutations produced substantial (greater than twofold) phenotypes of either increased or decreased splicing. Of all predicted secondary structural elements, only a single 15-nt stem-loop showed a strong correlation with splicing, acting negatively. The in vitro formation of exon-protein complexes between the mutant molecules and proteins associated with spliceosome formation (U2AF35, U2AF65, U1A, and U1-70K) correlated with splicing efficiencies, suggesting exon definition as the step affected by most mutations. The measured relative binding affinities of dozens of human RNA binding protein domains as reported in the CISBP-RNA database were found to correlate either positively or negatively with splicing efficiency, more than could fit on the 51-nt test exon simultaneously. The large number of these functional protein binding correlations point to a dynamic and heterogeneous population of pre-mRNA molecules, each responding to a particular collection of binding proteins

Saturation mutagenesis reveals manifold determinants of exon definition.

To illuminate the extent and roles of exonic sequences in the splicing of human RNA transcripts, we conducted saturation mutagenesis of a 51-nt internal exon in a three-exon minigene. All possible single and tandem dinucleotide substitutions were surveyed. Using high-throughput genetics, 5560 minigene molecules were assayed for splicing in human HEK293 cells. Up to 70% of mutations produced substantial (greater than twofold) phenotypes of either increased or decreased splicing. Of all predicted secondary structural elements, only a single 15-nt stem-loop showed a strong correlation with splicing, acting negatively. The in vitro formation of exon-protein complexes between the mutant molecules and proteins associated with spliceosome formation (U2AF35, U2AF65, U1A, and U1-70K) correlated with splicing efficiencies, suggesting exon definition as the step affected by most mutations. The measured relative binding affinities of dozens of human RNA binding protein domains as reported in the CISBP-RNA database were found to correlate either positively or negatively with splicing efficiency, more than could fit on the 51-nt test exon simultaneously. The large number of these functional protein binding correlations point to a dynamic and heterogeneous population of pre-mRNA molecules, each responding to a particular collection of binding proteins

Plan de negocio de servicios de cuidados paliativos domiciliarios para pacientes oncol?gicos de Lima Este

El objetivo del estudio fue implementar un plan de negocio de cuidados paliativos domiciliario para pacientes oncol?gicos de Lima Este. En consecuencia, se valor? a los diferentes proveedores del servicio de cuidados paliativos domiciliario a trav?s de un an?lisis de los posibles competidores, se estableci? un an?lisis estrat?gico y una estrategia de posicionamiento de la marca y canales de venta de un servicio de cuidados paliativos domiciliario, se elabor? un plan de operaciones, se estableci? los recursos humanos, administrativos, log?sticos y financieros necesarios para el desarrollo eficiente del servicio de cuidados paliativos domiciliario y se evalu? la viabilidad del plan de negocios de servicios de cuidados paliativos domiciliario, mediante la elaboraci?n de un plan financiero

Infiltrating CD16 +

Our aim was to characterize glomerular monocytes (Mo) infiltration and to correlate them with peripheral circulating Mo subsets and severity of lupus nephritis (LN). Methods. We evaluated 48 LN biopsy samples from a referral hospital. Recognition of Mo cells was done using microscopic view and immunohistochemistry stain with CD14 and CD16. Based on the number of cells, we classified LN samples as low degree of diffuse infiltration (<5 cells) and high degree of diffuse infiltration (≥5 cells). Immunophenotyping of peripheral Mo subsets was done using flow cytometry. Results. Mean age was 34.0±11.7 years and the mean SLEDAI was 17.5±6.9. The most common SLE manifestations were proteinuria (91%) and hypocomplementemia (75%). Severe LN was found in 70% of patients (Class III, 27%; Class IV, 43%). Severe LN patients and patients with higher grade of CD16+ infiltration had lower levels of nonclassical (CD14+CD16++) Mo in peripheral blood. Conclusions. Our results might suggest that those patients with more severe forms of LN had a higher grade of CD14+CD16+ infiltration and lower peripheral levels of nonclassical (CD14+CD16++) Mo and might reflect a recruitment process in renal tissues. However, given the small sample, our results must be interpreted carefully

Genetic diversity and antimicrobial resistance of Salmonella serotypes recovered throughout the beef production chain and from patients with salmonellosis

Salmonella is one of the major foodborne pathogens worldwide. The antimicrobial resistance (AMR) of this foodborne pathogen has raised a great concern in recent years. Studies on the frequency and characterization of Salmonella serotypes can help to improve our knowledge on the epidemiology of this pathogen. The purpose of this study was to compare the serotypes, AMR and genetic profiles of Salmonella isolates recovered from raw beef throughout the beef production chain and from human feces associated with clinical cases of salmonellosis. The serotype, AMR and pulsed-field gel electrophoresis profile of 243 Salmonella enterica isolates recovered from beef carcasses (n = 78), ground beef (n = 135), and human feces from clinical cases of salmonellosis (n = 30) were compared. Forty-three different Salmonella serotypes were identified and regardless of the source, the top five corresponded to Typhimurium, Give, Group B (partially serotyped), Infantis and Anatum. Twelve serotypes from beef carcasses were also found in ground beef, showing their presence throughout the beef production chain. Salmonella Typhimurium, Infantis, Anatum and Montevideo were the only serotypes identified in all sample types. Resistance to tetracyclines was the most frequent (41.2%) followed by resistance to aminoglycosides (37%), folate pathway inhibitors (21%), quinolones (20.2%), phenicols (17.1%), penicillins (15.6%) and cephems (7%). Multidrug resistance was observed in 28.8% of the isolates, and those from human feces showed resistance to a larger number of antimicrobials. Great concern arises from the resistance and reduced susceptibility observed to quinolones and cephalosporins because these drugs are the first line of treatment for invasive Salmonella infections. Twenty-seven distinct pulse-types were detected among 238 isolates. Clustering analysis for the most frequent serotypes identified groups of isolates with similar AMR profiles. Multidrug resistance spreading throughout the food production chain should be continually monitored and its importance emphasized