A Computational Module Assembled from Different Protease Family Motifs Identifies PI PLC from Bacillus cereus as a Putative Prolyl Peptidase with a Serine Protease Scaffold

Proteolytic enzymes have evolved several mechanisms to cleave peptide bonds. These distinct types have been systematically categorized in the MEROPS database. While a BLAST search on these proteases identifies homologous proteins, sequence alignment methods often fail to identify relationships arising from convergent evolution, exon shuffling, and modular reuse of catalytic units. We have previously established a computational method to detect functions in proteins based on the spatial and electrostatic properties of the catalytic residues (CLASP). CLASP identified a promiscuous serine protease scaffold in alkaline phosphatases (AP) and a scaffold recognizing a beta-lactam (imipenem) in a cold-active Vibrio AP. Subsequently, we defined a methodology to quantify promiscuous activities in a wide range of proteins. Here, we assemble a module which encapsulates the multifarious motifs used by protease families listed in the MEROPS database. Since APs and proteases are an integral component of outer membrane vesicles (OMV), we sought to query other OMV proteins, like phospholipase C (PLC), using this search module. Our analysis indicated that phosphoinositide-specific PLC from Bacillus cereus is a serine protease. This was validated by protease assays, mass spectrometry and by inhibition of the native phospholipase activity of PI-PLC by the well-known serine protease inhibitor AEBSF (IC50 = 0.018 mM). Edman degradation analysis linked the specificity of the protease activity to a proline in the amino terminal, suggesting that the PI-PLC is a prolyl peptidase. Thus, we propose a computational method of extending protein families based on the spatial and electrostatic congruence of active site residues.

Evidence for classification of c.1852_1853AA > GC in MLH1 as a neutral variant for Lynch syndrome

Background: Lynch syndrome (LS) is an autosomal dominant inherited cancer syndrome characterized by early onset cancers of the colorectum, endometrium and other tumours. A significant proportion of DNA variants in LS patients are unclassified. Reports on the pathogenicity of the c.1852_1853AA>GC (p.Lys618Ala) variant of the MLH1 gene are conflicting. In this study, we provide new evidence indicating that this variant has no significant implications for LS. Methods: The following approach was used to assess the clinical significance of the p.Lys618Ala variant: frequency in a control population, case-control comparison, co-occurrence of the p.Lys618Ala variant with a pathogenic mutation, co-segregation with the disease and microsatellite instability in tumours from carriers of the variant. We genotyped p.Lys618Ala in 1034 individuals (373 sporadic colorectal cancer [CRC] patients, 250 index subjects from families suspected of having LS [revised Bethesda guidelines] and 411 controls). Three well-characterized LS families that fulfilled the Amsterdam II Criteria and consisted of members with the p.Lys618Ala variant were included to assess co-occurrence and co-segregation. A subset of colorectal tumour DNA samples from 17 patients carrying the p.Lys618Ala variant was screened for microsatellite instability using five mononucleotide markers. Results: Twenty-seven individuals were heterozygous for the p.Lys618Ala variant; nine had sporadic CRC (2.41%), seven were suspected of having hereditary CRC (2.8%) and 11 were controls (2.68%). There were no significant associations in the case-control and case-case studies. The p.Lys618Ala variant was co-existent with pathogenic mutations in two unrelated LS families. In one family, the allele distribution of the pathogenic and unclassified variant was in trans, in the other family the pathogenic variant was detected in the MSH6 gene and only the deleterious variant co-segregated with the disease in both families. Only two positive cases of microsatellite instability (2/17, 11.8%) were detected in tumours from p.Lys618Ala carriers, indicating that this variant does not play a role in functional inactivation of MLH1 in CRC patients. Conclusions: The p.Lys618Ala variant should be considered a neutral variant for LS. These findings have implications for the clinical management of CRC probands and their relatives.

Evolución de los pacientes de leucemia linfocítica crónica : estudio de la concentración de hemoglobina, el recuento de plaquetas y el número de linfocitos

379 p. : il.

Desarrollo de una hoja de procesos para mecanizado de una cabeza de pistón de automóvil

[ES]El proyecto a realizar consiste en el desarrollo y elaboración de una hoja de procesos para mecanizado de un componente de automoción. Para ello se seleccionarán las herramientas adecuadas y calcularán los parámetros y tiempos de mecanizado

Mercados financieros: análisis fundamental y técnico

Explicación de conceptos claves relacionados con los mercados financieros desde una perspectiva simple y dirigida a personas sin grandes conocimientos de la materia.

Mercados financieros: análisis técnico

Sin tener conocimiento alguno de cual sería nuestro futuro en un mercado tan castigado por la crisis, comenzamos la carrera universitaria y lo hicimos apostando porque la situación mejoraría con el paso del tiempo y aspirando a poder mejorarla nosotros mismos. A día de hoy, siguen produciéndose consecuencias en todos los ámbitos de la vida socio-económica mundial a causa de la gran crisis de 2007. Una crisis que estalló debido en gran parte a la especulación de valores y productos financieros en las bolsas más importantes del mundo. En este trabajo se manejarán conceptos básicos e importantes sobre los mercados bursátiles, con el fin de explicar el funcionamiento de éstos identificando algunas claves y mecanismos que ayudarán a la hora de invertir en los productos financieros que se ofertan en dichos mercados

Justicia penal restaurativa y justicia penal negociada, retórica y práctica. Estudio empírico del programa de mediación intrajudicial penal en Bizkaia

633 p.

Distrofia miotonikoa duten pazienteen eta euren senideen bizi-kalitatearen azterketa

Helburuak. Distrofia miotonikoan ematen diren sintomarik esanguratsuenak bildu, gaixoen eta senideen bizi-kalitatea aztertu eta osasunaren aldetik eman daitezkeen esku hartzeak aztertzea. Metodologia. Pubmed, Cochrane, ScienceDirect, Canadian Journal of Neuroscience Nursing (CJNN) eta hainbat erakunderen web orrialdeetan burututako bilaketa bibliografiko baten bidez, eta erreferentziazko pertsonekin kontaktuan jarriz, 14 artikulu, tesi bat, bost liburu eta hainbat web orrialderen bilaketa egin da. Konklusioak. Beharrezkoa da gaixotasun honen inpaktuaren azterketa sakonago bat burutzea. Alde fisiopatologikoa nahiko garatua dagoen arren, gaixotasuna pazienteen ikuspegitik aztertzeak atentziorako datu garrantzitsuak eman ditzake. Gaixotasun honen jarraipenerako gidak garatzen ari diren arren, oraindik asko dago gai honen inguruan egiteko.

Distribución de automóviles y restricciones verticales a la libre competencia

[ES] Analisis del mercado de distribución de automoviles en la UE y las diferencias de precios existentes, tratando de determinar las posibles causas.

Lymphangioleiomyomatosis Biomarkers Linked to Lung Metastatic Potential and Cell Stemness

Lymphangioleiomyomatosis (LAM) is a rare lung-metastasizing neoplasm caused by the proliferation of smooth muscle-like cells that commonly carry loss-of-function mutations in either the tuberous sclerosis complex 1 or 2 (TSC1 or TSC2) genes. While allosteric inhibition of the mechanistic target of rapamycin (mTOR) has shown substantial clinical benefit, complementary therapies are required to improve response and/or to treat specific patients. However, there is a lack of LAM biomarkers that could potentially be used to monitor the disease and to develop other targeted therapies. We hypothesized that the mediators of cancer metastasis to lung, particularly in breast cancer, also play a relevant role in LAM. Analyses across independent breast cancer datasets revealed associations between low TSC1/2 expression, altered mTOR complex 1 (mTORC1) pathway signaling, and metastasis to lung. Subsequently, immunohistochemical analyses of 23 LAM lesions revealed positivity in all cases for the lung metastasis mediators fascin 1 (FSCN1) and inhibitor of DNA binding 1 (ID1). Moreover, assessment of breast cancer stem or luminal progenitor cell biomarkers showed positivity in most LAM tissue for the aldehyde dehydrogenase 1 (ALDH1), integrin-beta 3 (ITGB3/CD61), and/or the sex-determining region Y-box 9 (SOX9) proteins. The immunohistochemical analyses also provided evidence of heterogeneity between and within LAM cases. The analysis of Tsc2-deficient cells revealed relative over-expression of FSCN1 and ID1; however, Tsc2-deficient cells did not show higher sensitivity to ID1-based cancer inhibitors. Collectively, the results of this study reveal novel LAM biomarkers linked to breast cancer metastasis to lung and to cell stemness, which in turn might guide the assessment of additional or complementary therapeutic opportunities for LAM.