Progression to Adrenocortical Tumorigenesis in Mice and Humans through Insulin-Like Growth Factor 2 and beta-Catenin

Dysregulation of the WNT and insulin-like growth factor 2 (IGF2) signaling pathways has been implicated in sporadic and syndromic forms of adrenocortical carcinoma (ACC). Abnormal beta-catenin staining and CTNNB1 mutations are reported to be common in both adrenocortical adenoma and ACC, whereas elevated IGF2 expression is associated primarily with ACC. To better understand the contribution of these pathways in the tumorigenesis of ACC, we examined clinicopathological and molecular data and used mouse models. Evaluation of adrenal tumors from 118 adult patients demonstrated an increase in CTNNB1 mutations and abnormal beta-catenin accumulation in both adrenocortical adenoma and ACC. In ACC, these features were adversely associated with survival. Mice with stabilized beta-catenin exhibited a temporal progression of increased adrenocortical hyperplasia, with subsequent microscopic and macroscopic adenoma formation. Elevated Igf2 expression alone did not cause hyperplasia. With the combination of stabilized beta-catenin and elevated Igf2 expression, adrenal glands were larger, displayed earlier onset of hyperplasia, and developed more frequent macroscopic adenomas (as well as one carcinoma). Our results are consistent with a model in which dysregulation of one pathway may result in adrenal hyperplasia, but accumulation of a second or multiple alterations is necessary for tumorigenesis. (Ant J Pathol 2012, 181:1017-1033; http://dx.doi.org/10.1016/j.ajpath.2012.05.026)

Increased IGF-II protein affects p57kip2 expression in vivo and in vitro: Implications for Beckwith–Wiedemann syndrome

In both human and mouse, the Igf2 gene, localized on chromosomes 11 and 7, respectively, is expressed from the paternally inherited chromosome in the majority of tissues. Insulin-like growth factor-II (IGF-II) plays an important role in embryonic growth, and aberrant IGF2 expression has been documented in several human pathologies, such as Beckwith–Wiedemann syndrome (BWS), and a wide variety of tumors. Human and mouse genetic data strongly implicate another gene, CDKN1C (p57kip2), located in the same imprinted gene cluster on human chromosome II, in BWS. p57KIP2 is a cyclin-dependent kinase inhibitor and is required for normal mouse embryonic development. Mutations in CDKN1C (p57kip2) have been identified in a small proportion of patients with BWS, and removal of the gene from mice by targeted mutagenesis produces a phenotype with elements in common with this overgrowth syndrome. Patients with BWS with biallelic expression of IGF2 or with a CDKN1C (p57kip2) mutation, as well as overlapping phenotypes observed in two types of mutant mice, the p57kip2 knockout and IGF-II-overexpressing mice, strongly suggest that the genes may act in a common pathway of growth control in situations where Igf2 expression is abnormal. Herein, we show that p57kip2 expression is reduced on IGF-II treatment of primary embryo fibroblasts in a dose-dependent manner. In addition, p57kip2 expression is down-regulated in mice with high serum levels of IGF-II. These data suggest that the effects of increased IGF-II in BWS may, in part, be mediated through a decrease in p57kip2 gene expression.

The 12-Item General Health Questionnaire (GHQ-12): Reliability,external validity and factor structure in the Spanish population

The purpose of this study was to analyze the internal consistency and the external and structure validity of the 12-Item General Health Questionnaire (GHQ-12) in the Spanish general population. A stratified sample of 1001 subjects, ages between 25 and 65 years, taken from the general Spanish population was employed. The GHQ-12 and the Inventory of Situations and Responses of Anxiety-ISRA were administered. A Cronbach’s alpha of .76 (Standardized Alpha: .78) and a 3-factor structure (with oblique rotation and maximum likelihood procedure) were obtained. External validity of Factor I (Successful Coping) with the ISRA is very robust (.82; Factor II, .70; Factor III, .75). The GHQ-12 shows adequate reliability and validity in the Spanish population. Therefore, the GHQ-12 can be used with efficacy to assess people’s overall psychological well-being and to detect non-psychotic psychiatric problems. Additionally, our results confirm that the GHQ-12 can best be thought of as a multidimensional scale that assesses several distinct aspects of distress, rather than just a unitary screening measure.

Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin

Defensins are a group of cationic antimicrobial peptides which play an important role in the innate immune system by exerting their antimicrobial activity against pathogens. In this study, we cloned a novel beta-defensin cDNA from medaka (Oryzias latipes) by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA consists of 480 bp, and the open reading frame (CRF) of 189 bp encodes a polypeptide of 63 amino acids (aa) with a predicted molecular weight of 7.44 kDa. Its genomic organization was analyzed, and Southern blot detection confirmed that only one copy of beta-defensin exists in the medaka HNI strain. RT-PCR, Western blot and immunohistochemistry detections showed that the beta-defensin transcript and protein could be detected in eyes, liver, kidney, blood, spleen and gill, and obviously prevalent expression was found in eyes. Antimicrobial activity of the medaka beta-defensin was evaluated, and the antibacterial activity-specific to Gram-negative bacteria was revealed. Furthermore, the lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, was demonstrated to be able to induce about 13-fol up-regulation of the beta-defensin within first 12 h. In addition, promoter and promoter mutagenesis analysis were performed in the medaka beta-defensin. A proximal 100 base pair(bp) sequence (+26 to -73)and the next 1700 bp sequence (-73 to -1755) were demonstrated to be responsible for the basal promoter activity and for the transcription regulation. Three nuclear factor kappa B (NF-kappa B) cis-elements and a Sp1 cis-element were revealed by mutagenesis analysis to exist in the 5' flanking sequence, and they were confirmed to be responsible for the up-regulation of medaka beta-defensin stimulated by LPS. And, the Sp1 cis-element was further revealed to be related to the basal promoter activity, and transcriptional factor II D (TFIID) was found to be in charge of the gene transcription initiation. All the obtained data suggested that the novel medaka beta-defensin should have antimicrobial activity-specific to Gram-negative bacteria, and the antibacterial immune function should be modulated by NF-kappa B and Sp1. (C) 2008 Elsevier Ltd. All rights reserved.

Functional domains and the antiviral effect of the double-stranded RNA-dependent protein kinase PKR from Paralichthys olivaceus

The double-stranded RNA (dsRNA)-dependent protein kinase PKR is thought to mediate a conserved antiviral pathway by inhibiting viral protein synthesis via the phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2 alpha). However, little is known about the data related to the lower vertebrates, including fish. Recently, the identification of PKR-like, or PKZ, has addressed the question of whether there is an orthologous PKR in fish. Here, we identify the first fish PKR gene from the Japanese flounder Paralichthys olivaceus (PoPKR). PoPKR encodes a protein that shows a conserved structure that is characteristic of mammalian PKRs, having both the N-terminal region for dsRNA binding and the C-terminal region for the inhibition of protein translation. The catalytic activity of PoPKR is further evidence that it is required for protein translation inhibition in vitro. PoPKR is constitutively transcribed at low levels and is highly induced after virus infection. Strikingly, PoPKR overexpression increases eIF2 alpha phosphorylation and inhibits the replication of Scophthalmus maximus rhabdovirus (SMRV) in flounder embryonic cells, whereas phosphorylation and antiviral effects are impaired in transfected cells expressing the catalytically inactive PKR-K421R variant, indicating that PoPKR inhibits virus replication by phosphorylating substrate eIF2 alpha. The interaction between PoPKR and eIF2 alpha is demonstrated by coimmunoprecipitation assays, and the transfection of PoPKR-specific short interfering RNA further reveals that the enhanced eIF2 alpha phosphorylation is catalyzed by PoPKR during SMRV infection. The current data provide significant evidence for the existence of a PKR-mediated antiviral pathway in fish and reveal considerable conservation in the functional domains and the antiviral effect of PKR proteins between fish and mammals.

Proposed role for COUP-TFII in regulating fetal Leydig cell steroidogenesis, perturbation of which leads to masculinization disorders in rodents

Reproductive disorders that are common/increasing in prevalence in human males may arise because of deficient androgen production/action during a fetal 'masculinization programming window'. We identify a potentially important role for Chicken Ovalbumin Upstream Promoter-Transcription Factor II (COUP-TFII) in Leydig cell (LC) steroidogenesis that may partly explain this. In rats, fetal LC size and intratesticular testosterone (ITT) increased ~3-fold between e15.5-e21.5 which associated with a progressive decrease in the percentage of LC expressing COUP-TFII. Exposure of fetuses to dibutyl phthalate (DBP), which induces masculinization disorders, dose-dependently prevented the age-related decrease in LC COUP-TFII expression and the normal increases in LC size and ITT. We show that nuclear COUP-TFII expression in fetal rat LC relates inversely to LC expression of steroidogenic factor-1 (SF-1)-dependent genes (StAR, Cyp11a1, Cyp17a1) with overlapping binding sites for SF-1 and COUP-TFII in their promoter regions, but does not affect an SF-1 dependent LC gene (3β-HSD) without overlapping sites. We also show that once COUP-TFII expression in LC has switched off, it is re-induced by DBP exposure, coincident with suppression of ITT. Furthermore, other treatments that reduce fetal ITT in rats (dexamethasone, diethylstilbestrol (DES)) also maintain/induce LC nuclear expression of COUP-TFII. In contrast to rats, in mice DBP neither causes persistence of fetal LC COUP-TFII nor reduces ITT, whereas DES-exposure of mice maintains COUP-TFII expression in fetal LC and decreases ITT, as in rats. These findings suggest that lifting of repression by COUP-TFII may be an important mechanism that promotes increased testosterone production by fetal LC to drive masculinization. As we also show an age-related decline in expression of COUP-TFII in human fetal LC, this mechanism may also be functional in humans, and its susceptibility to disruption by environmental chemicals, stress and pregnancy hormones could explain the origin of some human male reproductive disorders.

Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro

Development of continuous cell lines from shrimp is essential to investigate viral pathogens. Unfortunately, there is no valid cell line developed from crustaceans in general and shrimps in particular to address this issue. Lack of information on the requirements of cells in vitro limits the success of developing a cell line, where the microenvironment of a cell culture, provided by the growthmedium, is of prime importance. Screening and optimization of growth medium components based on statistical experimental designs have been widely used for improving the efficacy of cell culture media. Accordingly, we applied Plackett–Burman design and response surface methodology to study multifactorial interactions between the growth factors in shrimp cell culture medium and to identify the most important ones for growth of lymphoid cell culture from Penaeus monodon. The statistical screening and optimization indicated that insulin like growth factor-I (IGF-I) and insulin like growth factor-II (IGF-II) at concentrations of 100 and 150 ng ml-1, respectively, could significantly influence the metabolic activity and DNA synthesis of the lymphoid cells. An increase of 53 % metabolic activity and 24.8 % DNA synthesis could be obtained, which suggested that IGF-I and IGFII had critical roles in metabolic activity and DNA synthesis of shrimp lymphoid cells

Propaganda y enseñanza : los aspectos formales de la propaganda y su posible aplicación a la tecnología didáctica.

Se trata de comprobar si los factores de comunicación verbo-icónica en los carteles de propaganda política son similares a los que en su momento se detectaron como características de la publicidad comercial en la investigación de Rodríguez Diéguez 'Publicidad y Enseñanza'. La muestra se compone de 323 carteles de propaganda política, recogidos en diapositivas, pertenecientes a las campañas electorales, efectuadas en el País Valenciano por los distintos partidos con motivo de las elecciones generales de 1977, 79 carteles; elecciones generales de 1979, 79 carteles; referéndum constitucional de 1978, 76 carteles; y elecciones municipales de 1979, 92 carteles. Para comprobar el objetivo, se valora la muestra de carteles políticos con la misma escala que se utilizó para la publicidad comercial que recogía los siguientes factores: factor icónico, factor verbal, factor de función dominante, factor de convergencia formal de contenidos y factor de determinación informativa. Escala de publicidad y enseñanza de Rodríguez Diéguez. Propaganda política. Análisis factorial de componentes principales, rotación Varimax. Análisis de correlación. La matriz factorial es sometida a dos rotaciones más que incrementen la invarianza. Programas de ordenador. Representaciones gráficas. Tras las rotaciones efectuadas se observa que el factor I en ambos casos tienen un alto nivel de concomitancia, aunque el factor resultante de este trabajo es más amplio, dándose una convergencia de aspectos formales de carácter icónico y verbales, por lo que se le denomina factor de determinación cualitativa de la información. El factor II en ambos casos. Tienen una correlación casi perfecta, a pesar de lo cual se le denomina lecturabilidad. El factor III presenta variaciones sustantivas, ya que no entran en la composición, las variables que le dan su sentido de función dominante al original, sino sólo las que aluden a aspectos formales, por lo cual se le denomina factor técnico o de diseño. El factor IV, convergencia formal de contenidos, tiene un sentido equivalente al de la hipótesis. El factor V, queda descartado. La hipótesis global queda aceptada en su mayor parte.

Motivación y personalidad : Estudio experimental a partir del MIM y MMPI.

Estudiar los conceptos más importantes de la dinámica del comportamiento humano para conocer las relaciones entre motivación y personalidad. Para la aplicación del Método de Instrucción Motivacional (MIM) son 237 religiosas en período de formación y para aplicar el Inventario Multifásico de la Personalidad de Minnesota (MMPI) son 379 sujetos. El estudio de la correlación entre los dos cuestionarios utiliza a 216 sujetos. Analiza el concepto y los niveles de integración de la personalidad, explica las dimensiones y variables fundamentales de la misma y examina las teorías existentes. Después estudia el concepto y las teorías relativas a la motivación, y expone la utilidad y el significado de los instrumentos de la investigación. Posteriormente ofrece los resultados obtenidos con la aplicación de los instrumentos y examina la relación entre los mismos . El Método de Inducción Motivacional (MIM) y el Inventario Multifásico de la Personalidad de Minnesota (MMPI). El MIM es un método que utiliza unas categorías para clasificar el contenido motivacional, éstas son: Yo, Realización, Autorealización, Contacto, Exploración, Trascendencia, Posesión y Diversión. mientras que el MMPI tiene como objetivo proporcionar una evaluación objetiva de algunas de las más importantes dimensiones de la personalidad relacionadas con la adaptación personal y social del sujeto. 1) En el contenido motivacional, MIM: a) se diferencian dos zonas, una zona del yo, donde se integran las frecuencias más altas en las categorías del Yo, de la Realización y Autorealización.Y la otra es la zona del Contacto, que recoge las frecuencias significativas en las categorías del Contacto. Por lo cual, podemos hablar de una dirección egótica motivacional en la zona del yo. Resalta la dimensión estática y contemplativa y el aspecto de dinamismo interior; b) Un aspecto que hay que considerar es la dirección alternante referente a las motivaciones de Contacto, ya que la relación interpersonal puede tomar una dirección centrífuga y otra centrípeta. Además, existe otra dirección denominada intencional que pertenece a las motivaciones de trascendencia. Se podría afirmar que las motivaciones centrífugas, con tendencia a las relaciones con las personas, predominando un altruismo y una apertura al mundo y a las cosas, reflejan personalidades equilibradas, abiertas, maduras y sólidas. Sin embargo, las motivaciones centrífugas, que manifiestan cómo los individuos se quedan más en sí mismo, en su limitación, sin abrirse y salir hacia fuera, reflejan personalidades más inmaduras, menos ricas y menos comunicativas. El horizonte abierto a la comunicación en general con personas, ambientes, cosas y a la acción subraya ser más eficaz y constructivo. 2) En el MMPI, el perfil de los sujetos está definido por los tres factores encontrados en el análisis factorial: a) El factor I, que se define como personalidad problemática incontrolada, presenta un comportamiento problemático en la lima de desviación de la personalidad; b) El factor II está definido como introversión social y clínicamente representa la disposición psicológica de la personalidad; c) El factor III es el definido como depresión social que en límites clínicos se puede dar una introversión psicotécnica y esquizofrénica. 3) La interrelación de los dos instrumentos MMI y MMPI se da en dos direcciones: a) Se puede hablar de una estabilización del yo contra las desviaciones de personalidad, sugeridas por los extremos clínicos de las escalas del MMPI. Se pueden agrupar en el núcleo antipsicótico y antineurótico. Las escalas de motivaciones de estabilización del yo son Realización, Autorealización y Contacto centrífugo. b) Por otro lado, se puede hablar de una debilidad psicótica y neurótica, propia de las categorías con dimensión más egocéntrica y pasiva. Se manifiesta en las categorías Contacto centrípedo, Exploración y el yo según sus orientaciones estáticas y pasivas. 1) Se da convergencia de formas motivacionales y de estructura de personalidad. 2) El hecho de que los impulsos motivacionales vienen a dar una orientación determina el tipo de personalidad. Es decir, la personalidad queda expresada con sus características determinadas en los contenidos y en las formas motivacionales.

La evaluación de la Enseñanza Universitaria en relación con los objetivos del profesorado : la Enseñanza Universitaria vista por los alumnos : un estudio para la evaluación de los cursos de Enseñanza Superior.

Investigación que forma parte de un programa más general que pretende una evaluación formativa del proceso docente, ofreciendo una información al profesor que favorezca cambios en la calidad docente. Intentar obtener datos más generales sobre la Enseñanza Universitaria y construir un instrumento de evaluación adecuado. Intentar encontrar criterios de validez para el cuestionario, con especial interés por los criterios empáticos. Muestra no especificada de alumnos de la Universidad Autónoma de Madrid (16069 encuestas). Muestra de profesores no especificada de la misma universidad (123 encuestas). Diseño correlacional-factorial en el que se evalúa la percepción de la calidad docente por parte de los alumnos a la vez que se estudia la fiabilidad, validez y composición factorial de una versión mejorada de un cuestionario desarrollado anteriormente. Cada alumno evalúa varios profesores (con el orden de presentación balanceado) en una serie de dimensiones. Como criterio de validez se utiliza un ítem en el que el sujeto da una valoración global (grado en el que considera que se han alcanzado los objetivos docentes). El cuestionario incluye 3 ítems de sugestión para controlar el efecto de esta variable en la evaluación de la calidad.. Cuestionario (27 ítems referidos al profesor, 10 a los objetivos didácticos y 6 a la asignatura). Fiabilidad: técnica de las dos mitades. Validez: correlación con el ítem criterio. Análisis factorial (rotación ortogonal). La fiabilidad de los ítems es adecuada. No se observan efectos del orden de cumplimentación de las encuestas. La validez es superior a la observada en el primer cuestionario. No se observa una relación muy fuerte entre los objetivos y los métodos didácticos, aunque sí entre determinados ítems y los objetivos a conseguir. El análisis factorial confirma las dimensiones obtenidas en el trabajo anterior (factor I: interacción con el grupo y preocupación por los resultados, factor II: claridad de exposición, organización y preocupación por la enseñanza, factor III: entusiasmo y dominio de la asignatura). Respecto al factor de sugestión los resultados no son claros, ya que puede indicar bien un efecto de sugestión o bien una habilidad para motivar. Por último en el análisis por Facultades se obtienen perfiles heterogéneos, aunque hay ciertas diferencias entre Ciencias y Derecho (factores I, II y III). Sin embargo, las diferencias del tamaño de las muestras relativizan los resultados.

Insulin modulates norepinephrine-mediated melatonin synthesis in cultured rat pineal gland

The mammalian pineal gland synthesizes melatonin in a circadian manner, peaking during the dark phase. This synthesis is primarily regulated by sympathetic innervations via noradrenergic fibers, but is also modulated by many peptidergic and hormonal systems. A growing number of studies reveal a complex role for melatonin in influencing various physiological processes, including modulation of insulin secretion and action. In contrast, a role for insulin as a modulator of mclatonin synthesis has not been investigated previously. The aim of the current study was to determine whether insulin modulates norepinephrine (NE)-mediated melatonin synthesis. The results demonstrate that insulin (10(-8)M) potentiated norepinephrine-mediated melatonin synthesis and tryptophan hydroxylase (TPOH) activity in ex vivo incubated pineal glands. When ex vivo incubated pineal glands were synchronized (12h NE-stimulation, followed by 12h incubation in the absence of NE), insulin potentiated NE-mediated melatonin synthesis and arylalkylamine-N-acetyltransferase (AANAT) activity. Insulin did not affect the activity of hydroxyindole-O-methyltranferase (HIOMT), nor the gene expression of tpoh, aanat, or hiomt, under any of the conditions investigated. We conclude that insulin potentiates NE-mediated melatonin synthesis in cultured rat pineal gland, potentially through post-transcriptional events. (C) 2007 Elsevier Inc. All rights reserved.

Internet - lacuna jurídica ou inferno legal

A rápida evolução da Internet, permitindo a troca de mercadorias e serviços, pode ser comparada ao período da revolução industrial. A ausência total de fronteiras na Internet suscita tanto a admiração quanto o receio. Na verdade, o crescimento da Internet baseia-se em três fatores distintos e complementares: (i) o comercial; (ii) o tecnológico; e, (iii) o jurídico. A Internet não tem um comando jurídico uniformizado e harmonizado. Daí resulta que ela se traduz num mosaico organizacional que, hoje, está permitindo o seu funcionamento. A Internet é independente, entretanto não pode subsistir sem a influência de um sistema legal. De fato, as leis nacionais de um determinado país tem vocação a se aplicar às transações comerciais celebradas via Internet, dada a inexistência de uma regulamentação única e abrangente. Contudo, a aplicação não poderá ser imediata, adaptações serão necessárias para atender aos novos modelos jurídicos utilizados na Internet. Esta pesquisa é a primeira parte de um estudo mais amplo: a influência da Internet no direito brasileiro. Por tratar-se de um primeiro estudo bibliográfico, foram examinadas as principais áreas do direito que estão sendo influenciadas pelo desenvolvimento rápido e crescente da Internet. É, portanto, a primeira etapa deste estudo a pesquisa bibliográfica, propriamente dita. Nessa etapa, foram identificadas e comparadas as posições doutrinárias tanto de países sob a influência do sistema jurídico de common law como do romano-germânico. A partir de então, na segunda etapa, foram classificados os tópicos de maior interesse para a doutrina, através de palavras-chave. Partindo assim do presente estudo, o estudo mais amplo pretenderá delinear os alicerces de uma proposta jurídica comparada, entre os sistemas de common law e romano germânico, para então examinar a influência da Internet no direito brasileiro.

Fusão de dados multisensor para a identificação e o mapeamento de ambientes flúvio-estuarinos da Amazônia

A fusão de imagens multisensor tem sido um procedimento amplamente utilizado em função da natureza complementar dos vários conjuntos de dados. Este artigo compara o desempenho de quatro métodos diferentes para fusão de imagens Landsat-7 ETM+ e RADARSAT-1 W1. A comparação foi baseada nas características espectrais das imagens, utilizando-se de análise estatística e visual dos produtos gerados. Quatro métodos foram usados para a fusão das imagens Landsat-7 ETM+ e RADARSAT-1 W1: i) fusão do SAR (radar de abertura sintética) com a tríade selecionada pelo OIF (Optimum Index Factor); ii) realce por decorrelação da tríade selecionada pelo OIF, seguida da fusão com SAR; iii) ACP (Análise por Componentes Principais) para as seis bandas ETM+ do espectro refletido (1, 2, 3, 4, 5 e 7) e posterior fusão das três primeiras componentes principais (1CP; 2CP; 3CP) com o SAR; iv) SPC-SAR (Principal Componente Seletivo - SAR). O produto SPC-SAR mostrou melhor desempenho na identificação das feições costeiras e permitiu o realce mais efetivo dos diferentes ambientes.

Imprinted gene expression in in vivo- And in vi/ro-produced bovine embryos and chorio-allantoic membranes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fatores de risco relacionados à perda de produtividade laboral e aumento dos custos ambulatoriais de pacientes atendidos por unidades básicas de saúde

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)