Inhibition of the anaphase-promoting complex by the Xnf7 ubiquitin ligase.

Degradation of specific protein substrates by the anaphase-promoting complex/cyclosome (APC) is critical for mitotic exit. We have identified the protein Xenopus nuclear factor 7 (Xnf7) as a novel APC inhibitor able to regulate the timing of exit from mitosis. Immunodepletion of Xnf7 from Xenopus laevis egg extracts accelerated the degradation of APC substrates cyclin B1, cyclin B2, and securin upon release from cytostatic factor arrest, whereas excess Xnf7 inhibited APC activity. Interestingly, Xnf7 exhibited intrinsic ubiquitin ligase activity, and this activity was required for APC inhibition. Unlike other reported APC inhibitors, Xnf7 did not associate with Cdc20, but rather bound directly to core subunits of the APC. Furthermore, Xnf7 was required for spindle assembly checkpoint function in egg extracts. These data suggest that Xnf7 is an APC inhibitor able to link spindle status to the APC through direct association with APC core components.

Control of Emi2 activity and stability through Mos-mediated recruitment of PP2A.

Before fertilization, vertebrate eggs are arrested in meiosis II by cytostatic factor (CSF), which holds the anaphase-promoting complex (APC) in an inactive state. It was recently reported that Mos, an integral component of CSF, acts in part by promoting the Rsk-mediated phosphorylation of the APC inhibitor Emi2/Erp1. We report here that Rsk phosphorylation of Emi2 promotes its interaction with the protein phosphatase PP2A. Emi2 residues adjacent to the Rsk phosphorylation site were important for PP2A binding. An Emi2 mutant that retained Rsk phosphorylation but lacked PP2A binding could not be modulated by Mos. PP2A bound to Emi2 acted on two distinct clusters of sites phosphorylated by Cdc2, one responsible for modulating its stability during CSF arrest and one that controls binding to the APC. These findings provide a molecular mechanism for Mos action in promoting CSF arrest and also define an unusual mechanism, whereby protein phosphorylation recruits a phosphatase for dephosphorylation of distinct sites phosphorylated by another kinase.

Pharmacogenomics in early-phase clinical development.

Pharmacogenomics (PGx) offers the promise of utilizing genetic fingerprints to predict individual responses to drugs in terms of safety, efficacy and pharmacokinetics. Early-phase clinical trial PGx applications can identify human genome variations that are meaningful to study design, selection of participants, allocation of resources and clinical research ethics. Results can inform later-phase study design and pipeline developmental decisions. Nevertheless, our review of the clinicaltrials.gov database demonstrates that PGx is rarely used by drug developers. Of the total 323 trials that included PGx as an outcome, 80% have been conducted by academic institutions after initial regulatory approval. Barriers for the application of PGx are discussed. We propose a framework for the role of PGx in early-phase drug development and recommend PGx be universally considered in study design, result interpretation and hypothesis generation for later-phase studies, but PGx results from underpowered studies should not be used by themselves to terminate drug-development programs.

Mitochondrial mutations in adenoid cystic carcinoma of the salivary glands.

BACKGROUND: The MitoChip v2.0 resequencing array is an array-based technique allowing for accurate and complete sequencing of the mitochondrial genome. No studies have investigated mitochondrial mutation in salivary gland adenoid cystic carcinomas. METHODOLOGY: The entire mitochondrial genome of 22 salivary gland adenoid cystic carcinomas (ACC) of salivary glands and matched leukocyte DNA was sequenced to determine the frequency and distribution of mitochondrial mutations in ACC tumors. PRINCIPAL FINDINGS: Seventeen of 22 ACCs (77%) carried mitochondrial mutations, ranging in number from 1 to 37 mutations. A disproportionate number of mutations occurred in the D-loop. Twelve of 17 tumors (70.6%) carried mutations resulting in amino acid changes of translated proteins. Nine of 17 tumors (52.9%) with a mutation carried an amino acid changing mutation in the nicotinamide adenine dinucleotide dehydrogenase (NADH) complex. CONCLUSIONS/SIGNIFICANCE: Mitochondrial mutation is frequent in salivary ACCs. The high incidence of amino acid changing mutations implicates alterations in aerobic respiration in ACC carcinogenesis. D-loop mutations are of unclear significance, but may be associated with alterations in transcription or replication.

Genome-wide association study of perioperative myocardial infarction after coronary artery bypass surgery.

OBJECTIVES: Identification of patient subpopulations susceptible to develop myocardial infarction (MI) or, conversely, those displaying either intrinsic cardioprotective phenotypes or highly responsive to protective interventions remain high-priority knowledge gaps. We sought to identify novel common genetic variants associated with perioperative MI in patients undergoing coronary artery bypass grafting using genome-wide association methodology. SETTING: 107 secondary and tertiary cardiac surgery centres across the USA. PARTICIPANTS: We conducted a stage I genome-wide association study (GWAS) in 1433 ethnically diverse patients of both genders (112 cases/1321 controls) from the Genetics of Myocardial Adverse Outcomes and Graft Failure (GeneMAGIC) study, and a stage II analysis in an expanded population of 2055 patients (225 cases/1830 controls) combined from the GeneMAGIC and Duke Perioperative Genetics and Safety Outcomes (PEGASUS) studies. Patients undergoing primary non-emergent coronary bypass grafting were included. PRIMARY AND SECONDARY OUTCOME MEASURES: The primary outcome variable was perioperative MI, defined as creatine kinase MB isoenzyme (CK-MB) values ≥10× upper limit of normal during the first postoperative day, and not attributable to preoperative MI. Secondary outcomes included postoperative CK-MB as a quantitative trait, or a dichotomised phenotype based on extreme quartiles of the CK-MB distribution. RESULTS: Following quality control and adjustment for clinical covariates, we identified 521 single nucleotide polymorphisms in the stage I GWAS analysis. Among these, 8 common variants in 3 genes or intergenic regions met p<10(-5) in stage II. A secondary analysis using CK-MB as a quantitative trait (minimum p=1.26×10(-3) for rs609418), or a dichotomised phenotype based on extreme CK-MB values (minimum p=7.72×10(-6) for rs4834703) supported these findings. Pathway analysis revealed that genes harbouring top-scoring variants cluster in pathways of biological relevance to extracellular matrix remodelling, endoplasmic reticulum-to-Golgi transport and inflammation. CONCLUSIONS: Using a two-stage GWAS and pathway analysis, we identified and prioritised several potential susceptibility loci for perioperative MI.

Universal Quake Statistics: From Compressed Nanocrystals to Earthquakes.

Slowly-compressed single crystals, bulk metallic glasses (BMGs), rocks, granular materials, and the earth all deform via intermittent slips or "quakes". We find that although these systems span 12 decades in length scale, they all show the same scaling behavior for their slip size distributions and other statistical properties. Remarkably, the size distributions follow the same power law multiplied with the same exponential cutoff. The cutoff grows with applied force for materials spanning length scales from nanometers to kilometers. The tuneability of the cutoff with stress reflects "tuned critical" behavior, rather than self-organized criticality (SOC), which would imply stress-independence. A simple mean field model for avalanches of slipping weak spots explains the agreement across scales. It predicts the observed slip-size distributions and the observed stress-dependent cutoff function. The results enable extrapolations from one scale to another, and from one force to another, across different materials and structures, from nanocrystals to earthquakes.

A miR-34a-Numb Feedforward Loop Triggered by Inflammation Regulates Asymmetric Stem Cell Division in Intestine and Colon Cancer.

Emerging evidence suggests that microRNAs can initiate asymmetric division, but whether microRNA and protein cell fate determinants coordinate with each other remains unclear. Here, we show that miR-34a directly suppresses Numb in early-stage colon cancer stem cells (CCSCs), forming an incoherent feedforward loop (IFFL) targeting Notch to separate stem and non-stem cell fates robustly. Perturbation of the IFFL leads to a new intermediate cell population with plastic and ambiguous identity. Lgr5+ mouse intestinal/colon stem cells (ISCs) predominantly undergo symmetric division but turn on asymmetric division to curb the number of ISCs when proinflammatory response causes excessive proliferation. Deletion of miR-34a inhibits asymmetric division and exacerbates Lgr5+ ISC proliferation under such stress. Collectively, our data indicate that microRNA and protein cell fate determinants coordinate to enhance robustness of cell fate decision, and they provide a safeguard mechanism against stem cell proliferation induced by inflammation or oncogenic mutation.

El mercado de carbono y el mecanismo de desarrollo limpio : caso Granja Tres Arroyos S.A. : un análisis desde la Nueva Economía Institucional

El calentamiento global consiste en el aumento de la temperatura de la tierra debido a la acumulación de los gases de efecto invernadero (GEI) en la atmósfera. Estos gases son producidos por actividades de generación de energía, el transporte, el uso del suelo, la industria y el manejo de los residuos. El aumento de GEI en la atmósfera provoca cambios climáticos e impactos en un sinfín de actividades humanas, en la productividad de la agricultura y ganadería, en la infraestructura y turismo, y también daños en la salud. La comunidad científica considera que el aumento de la temperatura para el fin del siglo debería ubicarse en los 2° C, para de esta forma poder limitar los impactos del cambio climático. Ello implicaría restringir las concentraciones de los GEI en valores cercanos a los 450 ppm (partes por millón). El problema económico del cambio climático subyace en que las emisiones de GEI constituyen una externalidad global. Una externalidad ocurre cuando la producción o consumo de un bien afecta a terceros que no participan directamente en su producción, venta o compra. Cuando hay presencia de externalidades, los precios de mercado no reflejan todos los costos ni beneficios sociales asociados a la producción de un bien. En el caso puntual del cambio climático, los emisores de GEI no asumen el costo de emitir gases a la atmósfera. Existen diferentes instrumentos de política ambiental que influyen en la percepción del recurso ambiental por parte del agente económico y que por ende, se reflejan en las decisiones económicas que ellos toman. Todos ellos tienen por objetivo asignar un precio/costo al recurso ambiental. El objetivo de los mercados de emisiones es asignar un precio al carbono. En ellos, se intercambian derechos a emitir cierta cantidad de GEI. El mecanismo de desarrollo limpio (MDL) es un instrumento de mercado definido en el Acuerdo de Marrakech bajo el marco institucional del Protocolo de Kyoto (PK). El MDL establece que un país Anexo I (país desarrollado), con compromisos de reducción de emisiones, invierta en proyectos de reducción o captación de emisiones en un país No Anexo I (país en desarrollo sin compromisos de reducción), mediante la compra de reducciones certificadas de emisiones (RCEs) generados a partir de la implementación de los proyectos. Argentina ratificó el PK a través de la ley nacional 25.438 en el 2001. Como el país se encuentra comprendido en las Partes No Anexo I, sólo puede participar como país anfitrión de un proyecto MDL y ser oferente de RCEs. Hasta la fecha, Argentina desarrolló 65 proyectos que se encuentran en distinta etapa de aprobación nacional o registro internacional. La mayoría corresponden a proyectos vinculados con rellenos sanitarios y energías. Bajo este marco de desarrollo en el país, resulta valiosa la exploración de un caso implementado en la Argentina. El objetivo de la investigación consiste en analizar el mercado de carbono a través del mecanismo de desarrollo limpio y su implementación en una empresa agroindustrial argentina, Granja Tres Arroyos S.A., bajo el enfoque teórico de la Nueva Economía y los Negocios Agroalimentarios.

El mercado de carbono y el mecanismo de desarrollo limpio : caso Granja Tres Arroyos S.A. : un análisis desde la Nueva Economía Institucional

El calentamiento global consiste en el aumento de la temperatura de la tierra debido a la acumulación de los gases de efecto invernadero (GEI) en la atmósfera. Estos gases son producidos por actividades de generación de energía, el transporte, el uso del suelo, la industria y el manejo de los residuos. El aumento de GEI en la atmósfera provoca cambios climáticos e impactos en un sinfín de actividades humanas, en la productividad de la agricultura y ganadería, en la infraestructura y turismo, y también daños en la salud. La comunidad científica considera que el aumento de la temperatura para el fin del siglo debería ubicarse en los 2° C, para de esta forma poder limitar los impactos del cambio climático. Ello implicaría restringir las concentraciones de los GEI en valores cercanos a los 450 ppm (partes por millón). El problema económico del cambio climático subyace en que las emisiones de GEI constituyen una externalidad global. Una externalidad ocurre cuando la producción o consumo de un bien afecta a terceros que no participan directamente en su producción, venta o compra. Cuando hay presencia de externalidades, los precios de mercado no reflejan todos los costos ni beneficios sociales asociados a la producción de un bien. En el caso puntual del cambio climático, los emisores de GEI no asumen el costo de emitir gases a la atmósfera. Existen diferentes instrumentos de política ambiental que influyen en la percepción del recurso ambiental por parte del agente económico y que por ende, se reflejan en las decisiones económicas que ellos toman. Todos ellos tienen por objetivo asignar un precio/costo al recurso ambiental. El objetivo de los mercados de emisiones es asignar un precio al carbono. En ellos, se intercambian derechos a emitir cierta cantidad de GEI. El mecanismo de desarrollo limpio (MDL) es un instrumento de mercado definido en el Acuerdo de Marrakech bajo el marco institucional del Protocolo de Kyoto (PK). El MDL establece que un país Anexo I (país desarrollado), con compromisos de reducción de emisiones, invierta en proyectos de reducción o captación de emisiones en un país No Anexo I (país en desarrollo sin compromisos de reducción), mediante la compra de reducciones certificadas de emisiones (RCEs) generados a partir de la implementación de los proyectos. Argentina ratificó el PK a través de la ley nacional 25.438 en el 2001. Como el país se encuentra comprendido en las Partes No Anexo I, sólo puede participar como país anfitrión de un proyecto MDL y ser oferente de RCEs. Hasta la fecha, Argentina desarrolló 65 proyectos que se encuentran en distinta etapa de aprobación nacional o registro internacional. La mayoría corresponden a proyectos vinculados con rellenos sanitarios y energías. Bajo este marco de desarrollo en el país, resulta valiosa la exploración de un caso implementado en la Argentina. El objetivo de la investigación consiste en analizar el mercado de carbono a través del mecanismo de desarrollo limpio y su implementación en una empresa agroindustrial argentina, Granja Tres Arroyos S.A., bajo el enfoque teórico de la Nueva Economía y los Negocios Agroalimentarios.

Novel inclusion complex of ibuprofen tromethamine with cyclodextrins: Physico-chemical characterization

Guest-host interactions of ibuprofen tromethamine salt (Ibu.T) with native and modified cyclodextrins (CyDs) have been investigated using several techniques, namely phase solubility diagrams (PSDs), proton nuclear magnetic resonance (H-1 NMR), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffractometry (XRPD). scanning-electron microscopy (SEM) and molecular mechanics (MM). From the analysis of PSD data (A(L)-type) it is concluded that the anionic tromethamine salt of ibuprofen (pK(a) = 4.55) forms 1: 1 soluble complexes with all CyDs investigated in buffered water at pH 7.0, while the neutral form of Ibu forms an insoluble complex with beta-CyD (B-S-type) in buffered water at pH 2.0. Ibu.T has a lower tendency to complex with beta-CyD (K-11 = 58 M-1 at pH 7.0) compared with the neutral Ibu (K-11 = 4200 M (1)) in water. Complex formation of Ibu.T with beta-CyD (Delta G degrees = -20.4 kJ/mol) is enthalpy driven (Delta H degrees = -22.9 kJ/mol) and is accompanied by a small unfavorable entropy (Delta S degrees = -8.4 J/mol K) change. H-1 NMR studies and MM computations revealed that, on complexation, the hydrophobic central benzene ring of lbu.T and part of the isobutyl group reside within the beta-CyD cavity leaving the peripheral groups (carboxylate, tromethamine and methyl groups) located near the hydroxyl group networks at either rim of beta-CyD. PSD, H-1 NMR, DSC, FT-IR, XRPD, SEM and MM studies confirmed the formation of Ibu.T/beta-CyD inclusion complex in solution and the solid state. (C) 2009 Elsevier B.V. All rights reserved.