The Novel Tubulin-Targeting Agent Pyrrolo-1,5-Benzoxazepine-15 Induces Apoptosis in Poor Prognostic Subgroups of Chronic Lymphocytic Leukemia

Pyrrolo-1,5-benzoxazepine-15 (PBOX-15) is a novel microtubule depolymerization agent that induces cell cycle arrest and subsequent apoptosis in a number of cancer cell lines. Chronic lymphocytic leukemia (CLL) is characterized by clonal expansion of predominately nonproliferating mature B cells. Here, we present data suggesting PBOX-15 is a potential therapeutic agent for CLL. We show activity of PBOX-15 in samples taken from a cohort of CLL patients (n = 55) representing both high-risk and low-risk disease. PBOX-15 exhibited cytotoxicity in CLL cells (n = 19) in a dose-dependent manner, with mean IC(50) of 0.55 mu mol/L. PBOX-15 significantly induced apoptosis in CLL cells (n = 46) including cells with poor prognostic markers: unmutated IgV(II) genes, CD38 and zeta-associated protein 70 (ZAP-70) expression, and fludarabine-resistant cells with chromosomal deletions in 17p. In addition, PBOX-15 was more potent than fludarabine in inducing apoptosis in fludarabine-sensitive cells. Pharmacologic inhibition and small interfering RNA knockdown of caspase-8 significantly inhibited PBOX-15-induced apoptosis. Pharmacologic inhibition of c-jun NH(2)-terminal kinase inhibited PBOX-15-induced apoptosis in mutated IgV(II) and ZAP-70(-) CLL cells but not in unmutated IgV(II) and ZAP-70(+) cells. PBOX-15 exhibited selective cytotoxicity in CLL cells compared with normal hematopoietic cells. Our data suggest that PBOX-15 represents a novel class of agents that are toxic toward both high-risk and low-risk CLL cells. The need for novel treatments is acute in CLL, especially for the subgroup of patients with poor clinical outcome and drug-resistant disease. This study identifies a novel agent with significant clinical potential.

Distinct poor prognostic subgroups of acute myeloid leukaemia, FLT3-ITD and P-glycoprotein-positive, have contrasting levels of FOXO1

Regulation of ABCB1 (P-glycoprotein/Pgp) in AML was investigated. In a historical cohort with Pgp and transcriptional regulator expression profiling data available (n=141), FOXO1 correlated with Pgp protein expression. This was confirmed in an independent cohort (n=204). Down-regulation (siRNA) or hyperactivation (nicotinamide) of FOXO1 led to corresponding changes in Pgp. Low FOXO1 expression correlated with FLT3-ITDs (p

DNA methylation subgroups and the CpG island methylator phenotype in gastric cancer: a comprehensive profiling approach

BACKGROUND: Methylation-induced silencing of promoter CpG islands in tumor suppressor genes plays an important role in human carcinogenesis. In colorectal cancer, the CpG island methylator phenotype (CIMP) is defined as widespread and elevated levels of DNA methylation and CIMP+ tumors have distinctive clinicopathological and molecular features. In contrast, the existence of a comparable CIMP subtype in gastric cancer (GC) has not been clearly established. To further investigate this issue, in the present study we performed comprehensive DNA methylation profiling of a well-characterised series of primary GC.

METHODS: The methylation status of 1,421 autosomal CpG sites located within 768 cancer-related genes was investigated using the Illumina GoldenGate Methylation Panel I assay on DNA extracted from 60 gastric tumors and matched tumor-adjacent gastric tissue pairs. Methylation data was analysed using a recursively partitioned mixture model and investigated for associations with clinicopathological and molecular features including age, Helicobacter pylori status, tumor site, patient survival, microsatellite instability and BRAF and KRAS mutations.

RESULTS: A total of 147 genes were differentially methylated between tumor and matched tumor-adjacent gastric tissue, with HOXA5 and hedgehog signalling being the top-ranked gene and signalling pathway, respectively. Unsupervised clustering of methylation data revealed the existence of 6 subgroups under two main clusters, referred to as L (low methylation; 28% of cases) and H (high methylation; 72%). Female patients were over-represented in the H tumor group compared to L group (36% vs 6%; P = 0.024), however no other significant differences in clinicopathological or molecular features were apparent. CpG sites that were hypermethylated in group H were more frequently located in CpG islands and marked for polycomb occupancy.

CONCLUSIONS: High-throughput methylation analysis implicates genes involved in embryonic development and hedgehog signaling in gastric tumorigenesis. GC is comprised of two major methylation subtypes, with the highly methylated group showing some features consistent with a CpG island methylator phenotype.

Discovering subgroups of patients from DNA copy number data using NMF on compacted matrices

In the study of complex genetic diseases, the identification of subgroups of patients sharing similar genetic characteristics represents a challenging task, for example, to improve treatment decision. One type of genetic lesion, frequently investigated in such disorders, is the change of the DNA copy number (CN) at specific genomic traits. Non-negative Matrix Factorization (NMF) is a standard technique to reduce the dimensionality of a data set and to cluster data samples, while keeping its most relevant information in meaningful components. Thus, it can be used to discover subgroups of patients from CN profiles. It is however computationally impractical for very high dimensional data, such as CN microarray data. Deciding the most suitable number of subgroups is also a challenging problem. The aim of this work is to derive a procedure to compact high dimensional data, in order to improve NMF applicability without compromising the quality of the clustering. This is particularly important for analyzing high-resolution microarray data. Many commonly used quality measures, as well as our own measures, are employed to decide the number of subgroups and to assess the quality of the results. Our measures are based on the idea of identifying robust subgroups, inspired by biologically/clinically relevance instead of simply aiming at well-separated clusters. We evaluate our procedure using four real independent data sets. In these data sets, our method was able to find accurate subgroups with individual molecular and clinical features and outperformed the standard NMF in terms of accuracy in the factorization fitness function. Hence, it can be useful for the discovery of subgroups of patients with similar CN profiles in the study of heterogeneous diseases.

La rehabilitación del subnormal profundo.

Demostrar cuál es el primer método a aplicar en la rehabilitación del subnormal profundo y a partir de este seguir con otros más sofisticados, a fin de que estos niños puedan alcanzar una autonomía relativa, con arreglo a sus deficiencias y al mismo tiempo puedan vivir felices dentro de su larga infancia, al comprobar su realización personal con las diversas actividades. Diez niños subnormales profundos, acogidos en un pequeño centro de tipo experimental, en un ambiente familiar, con muy pocos medios, y en el que los niños sólo han recibido reeducación con métodos aislados. Las edades de los niños oscilan de 4.3 a 26 años. Mide las aptitudes y hace una evaluación inicial proporcionando datos de la edad mental grupal e individual así como de otras aptitudes como conducta física, lenguaje, integración espacial, etc, estableciendo medias aritméticas. Deduce el CI relacionando la edad mental con la edad cronológica. Aplica una reeducación individual, partiendo del método sensorio-motriz-imagen corporal y completándolo con otros métodos. Después de 11 meses de haber empezado la reeducación, comprueba los resultados (resultados iniciales y finales) y reafirma la hipótesis de que el metodo sensorio-motriz hace posible la rehabilitación. Para medir las aptitudes y la evaluación inicial de los sujetos utiliza la prueba de desarrollo Kirk y Karnes y la prueba de funcionalidad básica de ida. Utiliza índices estadísticos de tendencia central como la media y la mediana. Comparación de medias para comparar resultados iniciales y finales. Se presentan las siguientes variaciones: aumento de la edad cronológica en 11 meses; la edad mental pasa de 10,2 meses a 18,6 meses; la conducta física pasa de 13,2 meses a 22,5 meses; el cuidado de si mismo pasa de 16,8 meses a 21,9 meses; las relaciones sociales pasan de 13,9 meses a 22,2 meses. También se observan grandes cambios en las pruebas: la psicomotriz pasa de 6,3 meses a 14,7 meses; la perceptiva pasa de 7,1 meses a 15,6 meses; la de integración espacial (mayor incremento) de 5,8 meses a 14,1 meses. A/ Para conseguir una rehabilitación integral es necesario alcanzar un desarrollo físico, psíquico y social. B/ el progreso alcanzado tras una rehabilitación esta en relación directa con el CI de los sujetos, alcanzando más desarrollo el que tiene mayor CI. C/ El alcanzar los objetivos está relacionado con la edad cronológica, alcanzando mejores resultados en los primeros años de vida, y menores a partir de esa edad, en los sujetos mayores. D/ Condiciona la rehabilitación, la adaptación del sujeto en el centro y la superación de la separación familiar. E/ El desarrollo alcanzado en un grupo está determinado por factores individuales y sociales.

Metodo Ramain : naturaleza y aplicabilidad al mundo del subnormal.

Proyectar el método Ramain al campo de los deficientes. 30 sujetos, divididos en dos grupos, normales de 7 y 8 años de edad y subnormales de 6 a 8 años. Explica la historia y situación del método Ramain, expone las bases del mismo, analiza los principios y fundamentos del método, lo que es el ejercicio, el papel del educador y los prolongamientos del método, y después realiza un trabajo experimental aplicando algunos de los ejercicios del método Ramain para confrontar los resultados entre un grupo de sujetos normales y otro de sujetos subnormales. Método Ramain. El grupo de normales realizó los siguientes ejercicios del método Ramain : dibujo copiado, laberinto, dictados mudos, ejercicios de destreza, gráficos musculares y trabajos con alambre. Mientras que al grupo de subnormales se le aplicaron los siguientes : dibujo copiado, trabajo con alambres, ejercicios de destreza y ejercicios gráficos musculares. 1) En el ejercicio de dibujo copiado, el grupo de normales realiza una estructuración aceptable para su edad, aunque hay algunos ejercicios que no llegan a captar ni a encuadrar el dibujo. Mientras que el grupo de subnormales no tiene en cuenta la cuadrícula de la hoja y por tanto la realización del ejercicio no se hace por puntos. La estructuración espacial necesitaría de bastante ejercicio para desarrollarla en lo que se refiere a las medidas que van de punto a punto. 2) En el ejercicio de trabajos con alambres, el grupo de normales lo realizó de forma satisfactoria y los niños lograron doblar el alambre con bastante perfección. Lo cual indica dominio de un útil como es el alambre, apreciación visual y buena transposición de la figura realizada al papel. Mientras que en el grupo de subnormales, se observa más tendencia a cerrar las figuras y a distorsionar los trazos debido al menor dominio de manos para doblar el alambre, la apreciación visual es más inferior que en el grupo normal. 3) En los ejercicios de destreza, el grupo de normales no sigue el espacio de las cuadrículas, los trazos están un poco distorsionados pero se pueden considerar como normales para su edad. En algunos casos no llegan a captar la estructuración del óvalo llegando a juntar los trazos. Mientras que en el grupo de subnormales, los sujetos no llegan a captar la amplitud del trazo, acabando a veces en un simple garabato, no siguen la línea del cuadro y los trazos se montan unos en otros, lo que significa que no tienen un dominio motriz en las manos ni una clara ideas de las distancias entre trazo y trazo. 4) En los ejercicios gráficos, el grupo de normales al utilizar la mano derecha obtiene una mayor fijación del trazo, mayor dominio, rapidez y calidad del mismo y existe una mayor comprensión de hacer el trazo guiándose por la cuadrícula, mientras que con la mano izquierda el trazo se ve más difuso, hay menor dominio en el movimiento psicomotriz al mover el lápiz, y como consecuencia, el trazo se ve menos fijo y con menor calidad que la mano derecha. En el grupo de los subnormales también se aprecia la diferencia entre manos, pero menos que en el grupo normal, pues la mayoría de los de esta muestra tienen igual desarrollo en ambas manos pero no llegan a captar como los normales el relieve exacto del ejercicio. 1) El método Ramain es un método de preformación educativa de base, ya que prepara al individuo para una posterior realización de trabajos o aptitudes. El centro del método es la idea de estructuración recíproca del sujeto y el mundo que le rodea. 2) El método Ramain posibilita una preparación al aprendizaje e intenta conseguir una progresiva conquista de sí mismo. En la realización del ejercicio importa sobre todo el carácter personal de la experiencia. 3) El papel del educador consiste en ayudar al alumno a ponerse en la situación determinada, a facilitar el desarrollo de su experiencia y a estructurarla poco a poco. 4) Simone Ramain, el creador del citado método, ha utilizado una pedagogía que individualiza la educación utilizando la presencia del grupo. 5) El método Ramain puede utilizarse en la reeducación de subnormales, sin olvidar que el cociente intelectual condiciona en gran parte el dominio de los ejercicios.

Intake of total and subgroups of fat minimally affect the associations between selected single nucleotide polymorphisms in the PPARγ pathway and changes in anthropometry among European adults from cohorts of the DiOGenes study

BACKGROUND: Although the peroxisome proliferator-activated receptor γ (PPARγ) pathway is central in adipogenesis, it remains unknown whether it influences change in body weight (BW) and whether dietary fat has a modifying effect on the association. OBJECTIVES: We examined whether 27 single nucleotide polymorphisms (SNPs) within 4 genes in the PPARγ pathway are associated with the OR of being a BW gainer or with annual changes in anthropometry and whether intake of total fat, monounsaturated fat, polyunsaturated fat, or saturated fat has a modifying effect on these associations. METHODS: A case-noncase study included 11,048 men and women from cohorts in the European Diet, Obesity and Genes study; 5552 were cases, defined as individuals with the greatest BW gain during follow-up, and 6548 were randomly selected, including 5496 noncases. We selected 4 genes [CCAAT/enhancer binding protein β (CEBPB), phosphoenolpyruvate carboxykinase 2, PPARγ gene (PPARG), and sterol regulatory element binding transcription factor 1] according to evidence about biologic plausibility for interactions with dietary fat in weight regulation. Diet was assessed at baseline, and anthropometry was followed for 7 y. RESULTS: The ORs for being a BW gainer for the 27 genetic variants ranged from 0.87 (95% CI: 0.79, 1.03) to 1.12 (95% CI: 0.96, 1.22) per additional minor allele. Uncorrected, CEBPB rs4253449 had a significant interaction with the intake of total fat and subgroups of fat. The OR for being a BW gainer for each additional rs4253449 minor allele per 100 kcal higher total fat intake was 1.07 (95% CI: 1.02, 1.12; P = 0.008), and similar associations were found for subgroups of fat. CONCLUSIONS: Among European men and women, the influence of dietary fat on associations between SNPs in the PPARγ pathway and anthropometry is likely to be absent or marginal. The observed interaction between rs4253449 and dietary fat needs confirmation.

The three-dimensional structure of bothropasin, the main hemorrhagic factor from Bothrops jararaca venom: Insights for a new classification of snake venom metalloprotease subgroups

Bothropasin is a 48 kDa hemorrhagic PIII snake venom metalloprotease (SVMP) isolated from Bothrops jararaca, containing disintegrin/cysteine-rich adhesive domains. Here we present the crystal structure of bothropasin complexed with the inhibitor POL647. The catalytic domain consists of a scaffold of two subdomains organized similarly to those described for other SVMPs, including the zinc and calcium-binding sites. The free cysteine residue Cys(189) is located within a hydrophobic core and it is not available for disulfide bonding or other interactions. There is no identifiable secondary structure for the disintegrin domain, but instead it is composed mostly of loops stabilized by seven disulfide bonds and by two calcium ions. The ECD region is in a loop and is structurally related to the RGD region of RGD disintegrins, which are derived from I`ll SVMPs. The ECD motif is stabilized by the Cys(117)_Cys(310) disulfide bond (between the disintegrin and cysteine-rich domains) and by one calcium ion. The side chain of Glu(276) of the ECD motif is exposed to solvent and free to make interactions. In bothropasin, the HVR (hyper-variable region) described for other Pill SVMPs in the cysteine-rich domain, presents a well-conserved sequence with respect to several other Pill members from different species. We propose that this subset be referred to as PIII-HCR (highly conserved region) SVMPs. The differences in the disintegrin-like, cysteine-rich or disintegrin-like cysteine-rich domains may be involved in selecting target binding, which in turn could generate substrate diversity or specificity for the catalytic domain. (C) 2008 Elsevier Ltd. All rights reserved.

The classification and the conjugacy classes of the finite subgroups of the sphere braid groups

Let n >= 3. We classify the finite groups which are realised as subgroups of the sphere braid group B(n)(S(2)). Such groups must be of cohomological period 2 or 4. Depending on the value of n, we show that the following are the maximal finite subgroups of B(n)(S(2)): Z(2(n-1)); the dicyclic groups of order 4n and 4(n - 2); the binary tetrahedral group T*; the binary octahedral group O*; and the binary icosahedral group I(*). We give geometric as well as some explicit algebraic constructions of these groups in B(n)(S(2)) and determine the number of conjugacy classes of such finite subgroups. We also reprove Murasugi`s classification of the torsion elements of B(n)(S(2)) and explain how the finite subgroups of B(n)(S(2)) are related to this classification, as well as to the lower central and derived series of B(n)(S(2)).

On Groups Whose Maximal Cyclic Subgroups Are Maximal

We consider the problem of classifying those groups whose maximal cyclic subgroups are maximal. We give a complete classification of those groups with this property and which are either soluble or residually finite.

Classification of the virtually cyclic subgroups of the pure braid groups of the projective plane

We classify the ( finite and infinite) virtually cyclic subgroups of the pure braid groups P(n)(RP(2)) of the projective plane. The maximal finite subgroups of P(n)(RP(2)) are isomorphic to the quaternion group of order 8 if n = 3, and to Z(4) if n >= 4. Further, for all n >= 3, the following groups are, up to isomorphism, the infinite virtually cyclic subgroups of P(n)(RP(2)): Z, Z(2) x Z and the amalgamated product Z(4)*(Z2)Z(4).