Dissertations / Theses on the topic 'ACSL4'

Estudamos uma família com cinco homens (dois falecidos) afetados por deficiência mental (DM) não-sindrômica em duas gerações, num padrão de herança ligada ao cromossomo X. A análise do padrão de inativação do cromossomo X, com base na metilação do gene AR, evidenciou que a mulher portadora obrigatória tinha desvio completo de inativação nos leucócitos, uma característica freqüente em portadoras de mutações do cromossomo X relacionadas com DM. Para o mapeamento da DM, genotipamos 28 locos de microssatélites ao longo do cromossomo X e delimitamos um segmento de cerca de 32 Mb, entre os marcadores DXS986 e DXS8067, compartilhado pelos afetados e pela portadora obrigatória, mas não pelo homem normal ou pelas possíveis portadoras que não tinham desvio do padrão de inativação do cromossomo X. Na busca do gene mutado, analisamos, por seqüenciamento direto, genes mapeados no intervalo compartilhado e já relacionados a DM ou que tivessem expressão em cérebro e leucócitos. Nos afetados e na portadora obrigatória, encontramos a mutação c.845C→T no gene ACSL4, que resulta na substituição do aminoácido histidina, conservado na família de sintetases de acil-CoA humanas e em diversos outros organismos, por tirosina (p.H323Y da isoforma cérebro-específica). Tratando-se de mutação que altera um aminoácido evolutivamente conservado em gene já relacionado com DM, que segregava com a DM na família, não tendo sido encontrada em amostra controle de 160 indivíduos do sexo masculino, concluímos que era a causa da DM na família. Mutações de ponto no gene ACSL4 foram relacionadas com a DM não sindrômica em três famílias descritas na literatura. O gene ACSL4 codifica a acil-coA sintetase 4 da família das sintetases de cadeia longa, que catalisa a formação de ésteres acil-coA a partir de ácidos graxos de cadeia longa. Sua expressão já foi documentada em vários tecidos, incluindo o cérebro e dados recentes mostraram que a proteína é essencial para a formação normal de espinhos dendríticos. A nova mutação do gene ACSL4 que descrevemos como causa de DM vem reforçar a relação alterações desse gene e a DM de herança ligada ao X. O padrão de inativação do X totalmente desviado foi mais uma vez observado em mulher portadora da mutação, indicando a importância da expressão desse gene em leucócitos. A presença de dificuldades de aprendizado na portadora da mutação concorda com o observado nas três famílias da literatura em que o estudo das portadoras foi relatado, indicando o efeito de mutações do gene ACSL4 sobre a função intelectual mesmo em heterozigose. A ausência de correlação entre o padrão de inativação do cromossomo X em células do sangue periférico e o comprometimento intelectual foi confirmada. Na família estudada, a identificação da mutação permitiu o aconselhamento genético.
We studied a family with five men (two of them deceased) affected by nonsyndromic mental retardation in two generations, in a pattern of X-linked inheritance (MRX). The study aimed at identifying the causative mutation. The obligate female carrier showed completely skewed inactivation of the X chromosome, based on the methylation status of the AR gene in peripheral blood in leukocytes, a common feature in carriers of X-linked mutations that cause mental retardation. We genotyped 28 microsatellite loci mapped throughout the X chromosome and delimited a 32 Mb segment, between markers DXS986 and DXS8067, that was shared by the affected males and obligate carrier, but was not present in a normal man or in two women who did not show skewed X-inactivation. We searched for the causative mutation by sequencing genes mapped to this candidate interval that had been associated with MR and/or were expressed in brain and leukocytes. In the affected men and obligate carrier, we found a c.845C→T mutation in the ACSL4 gene, resulting in the amino acid tyrosine substituting for a histidine (p.H323Y in brain isoform), which is conserved in the acyl-CoA synthetase family in humans and others organisms. This mutation was not found in a control sample of 160 men. Previously, point mutations in the ACSL4 gene had been identified as the cause of MRX in three families. ACSL4 encodes the acyl-CoA synthetase long-chain family member 4, which catalyzes the formation of acyl-CoA esters from long-chain fatty acids. It is expressed in several tissues, and in brain it is essential for the normal formation of dendritic spines. The novel mutation here described confirmed the causal association of ACSL4 mutations with non-syndromic mental retardation. The completely skewed Xinactivation, also observed in the previously described carriers, supported a functional role for this gene in peripheral blood leukocytes. The intellectual impairment present in the carrier in the family here reported is in accordance with previous findings pointing to the effect on intellectual abilities of ACSL4 mutations in heterozygosis. The absence of correlation between the pattern of X-inactivation in leukocytes and mental status was confirmed.

Recent developments in neurobiology make it clear that glia play fundamental and active roles, in the adult and in development. Many hereditary cognitive disorders have been linked to developmental defects, and in at least two cases, Rett Syndrome and Fragile X Mental Retardation, glia are important in pathogenesis. However, most studies of developmental disorders, in particular intellectual disability, focus on neuronal defects. An example is intellectual disability caused by mutations in ACSL4, a metabolic enzyme that conjugates long-chain fatty acids to Coenzyme A (CoA). Depleting ACSL4 in neurons is associated with defects in dendritic spines, a finding replicated in patient tissue, but the etiology of this disorder remains unclear. In a genetic screen to discover genes necessary for visual function, I identified the Drosophila homolog of ACSL4, Acsl, as a gene important for the magnitude of neuronal transmission, and found that it is required in glia. I determined that Acsl is required in a specific subtype of glia in the Drosophila optic lobe, and that depletion of Acsl from this population causes morphological defects. I demonstrated that Acsl is required in development, and that the phenotype can be rescued by human ACSL4. Finally, I discovered that ACSL4 is expressed in astrocytes in the mouse hippocampus. This study is highly significant for understanding glial biology and neurodevelopment. It provides information on the role of glia in development, substantiates a novel role for Acsl in glia, and advances our understanding of the potential role that glia play in the pathogenesis of intellectual disability.

Recent developments in neurobiology make it clear that glia play fundamental and active roles, in the adult and in development. Many hereditary cognitive disorders have been linked to developmental defects, and in at least two cases, Rett Syndrome and Fragile X Mental Retardation, glia are important in pathogenesis. However, most studies of developmental disorders, in particular intellectual disability, focus on neuronal defects. An example is intellectual disability caused by mutations in ACSL4, a metabolic enzyme that conjugates long-chain fatty acids to Coenzyme A (CoA). Depleting ACSL4 in neurons is associated with defects in dendritic spines, a finding replicated in patient tissue, but the etiology of this disorder remains unclear. In a genetic screen to discover genes necessary for visual function, I identified the Drosophila homolog of ACSL4, Acsl, as a gene important for the magnitude of neuronal transmission, and found that it is required in glia. I determined that Acsl is required in a specific subtype of glia in the Drosophila optic lobe, and that depletion of Acsl from this population causes morphological defects. I demonstrated that Acsl is required in development, and that the phenotype can be rescued by human ACSL4. Finally, I discovered that ACSL4 is expressed in astrocytes in the mouse hippocampus. This study is highly significant for understanding glial biology and neurodevelopment. It provides information on the role of glia in development, substantiates a novel role for Acsl in glia, and advances our understanding of the potential role that glia play in the pathogenesis of intellectual disability.

This MS thesis proposes the use of DYMOLA, an object-oriented language for modeling hierarchically structured systems, to generate ACSL simulation programs for continuous system analysis. An ACSL model described in terms of time dependent non-linear differential equations or transfer functions can be generated from a hierarchical model description of the system using DYMOLA. The model description in DYMOLA can be an equation description or a non-linear hierarchical bond graph abstraction to describe the system under investigation. The interface provides an automated method to generate ACSL simulation programs, hence eliminating the need for manual coding. The provision to specify an experiment description for run-time analysis and additional model statements is implemented. The implementation of the compiler's code generator includes parsing, error checking and system dependent file handling routines. Implementation techniques, model and control file specifications, and validation with examples in several application areas are described.

Sectional study, to identify the consumption of meat and meat products of elderly residents in geriatric institution of Belo Horizonte. Random sample (n = 52). Nutritional assessment was made by anthropometry, dietary and MAN. The process of acquisition, receipt, storage, handling of meat and meat products was observed. Was also carried out a cohort study to determine the anthropometric development. The results showed the need to reassess the process of buying, receiving and preparation of foodstuffs since there was a delivery of meat inappropriate amount and form of receipt, which contributed to a correction factor and per capita inadequate. The sample studied, 82.7% were women. The prevalence of overweight (BMI) was 46.1% and 23.1% underweight. According to MAN the prevalence of risk for malnutrition was 67.3%. According to the dietary analysis, the consumption of fat, polyunsaturated fatty acid and potassium had been insufficient in 100% of the elderly. The intake of B12 and zinc was inadequate in 44.2% and 82.7%, respectively, suggesting a low consumption of meat products. There was a prevalence of overweight and older people at high risk of diseases associated with obesity. However, from the MAN, there was a high rate of elderly people at risk for malnutrition
Estudo seccional, objetivo de identificar o consumo de carnes e derivados de idosos residentes em Instituição Geriátrica de Belo Horizonte. Amostra aleatória simples (n=52). A avaliação nutricional foi composta pela antropometria, MAN e dietética. O processo de aquisição, recepção, armazenamento, manipulação da carne e derivados foi observado. Realizou-se também estudo de coorte para verificar a evolução antropométrica. Os resultados mostraram a necessidade de reavaliar o processo de compra, recebimento e preparo dos gêneros alimentícios já que se observou uma entrega inapropriada das carnes em quantidade e forma de recebimento, o que contribuía para um fator de correção e per capita inadequados. Da amostra estudada, 82,7% eram mulheres. A prevalência de sobrepeso (IMC) foi de 46,1% e de baixo peso 23,1%. Segundo a MAN a prevalência de risco para desnutrição foi 67,3%. Segundo a análise dietética, o consumo de lipídeo, ácido graxo poliinsaturado e potássio apresentaram-se insuficiente em 100% dos idosos. A ingestão de B12 e zinco foi insuficiente em 44,2% e 82,7%, respectivamente, o que sugere um baixo consumo de produtos cárneos. Observou-se uma prevalência de idosos com sobrepeso e com risco elevado de doenças associadas à obesidade. Em contrapartida, a partir da MAN, observou-se um alto índice de idosos com risco para desnutrição

Skeletal muscle is one of the largest tissues in humans and provides many pivotal functions to support life. Abnormality in skeletal muscle functions can lead to disease. For example, insulin resistance in skeletal muscle leads to type II diabetes. The underlying mechanisms that control energy balance in skeletal muscle remain largely elusive, especially at the genetic level. Here in the second chapter, I showed that MyoD mediated the transcriptional regulation of ACSL5, a mitochondrial protein, in C2C12 myoblasts via two E-box elements. A SNP rs2419621 (T) created a de novo E-box that together with the two pre-existing proximal E-boxes strongly enhances ACSL5 expression in both CV1 and C2C12 cells. In the third chapter, I identified a novel VGLL4-interacting protein IRF2BP2 and verified the interaction with co-immunoprecipitation and mammalian two-hybrid assays. Functionally, overexpression of IRF2BP2 and transcription factor TEAD1 activates mouse VEGF-A promoter in CV1 cells and enhances the biosynthesis of VEGF-A in C2C12 myoblasts. In vivo studies showed that ischemia induced the expression of IRF2BP2 by more than three fold, suggesting that IRF2BP2 could play a pivotal role during tissue ischemia. IRF2BP2 is a nuclear protein in both mouse cardiac myocytes and C2C12 myoblasts as demonstrated by immunohistochemistry and immunocytochemistry, respectively. Therefore, I sought to delineate the mechanism for the nuclear shuttling of IRF2BP2 in the fourth chapter. With various DNA alternations, I mapped the NLS to an evolutionarily conserved sequence 354ARKRKPSP361 in IRF2BP2. Deletion of the positively charged amino acids resulted in the abolishment of the NLS signal. Next, I showed that phosphorylation of serine 360 (S360) mediates the nuclear import of the protein. Whereas an alanine substitution (S360A) at the site resulted in perinuclear accumulation of the protein, an aspartic acid substitution (S360D) forced the nuclear accumulation. Nevertheless, the forced accumulation of the S360D mutant did not enhance the activation of VEGF-A promoter in CV1 cells as did the wild-type protein. My studies revealed two novel mechanisms by which skeletal muscle could harvest energy, thus providing new insight into the energy metabolism in skeletal muscle

Le problème général de la preuve de propriétés de programmes impératifs est indécidable. Pour deslangages de programmation et de propriétés plus restrictifs, des sous-problèmes décidables sontconnus. En pratique, grâce à des heuristiques, les outils de preuve de programmes automatisent despreuves qui sortent du cadre théorique de ces sous-problèmes décidables connus. Nous illustronscette réussite pratique en construisant un catalogue de preuves, pour des programmes et despropriétés de nature similaire et de complexité croissante. Ces programmes sont principalementdes générateurs de cartes combinatoires.Ainsi, ce travail contribue aux domaines de recherche de la combinatoire énumérative et dugénie logiciel. Nous distribuons une bibliothèque C de générateurs exhaustifs bornés de tableauxstructurés, formellement spécifiés en ACSL et vérifiés avec le greffon WP de la plateforme d’analyseFrama-C. Nous proposons également une méthodologie de test qui facilite la preuve interactive enCoq, une étude formelle des cartes originale, et de nouveaux résultats en combinatoire énumérative
The general problem of proving properties of imperative programs is undecidable. Some subproblems– restricting the languages of programs and properties – are known to be decidable. Inpractice, thanks to heuristics, program proving tools sometimes automate proofs for programs andproperties living outside of the theoretical framework of known decidability results. We illustrate thisfact by building a catalog of proofs, for similar programs and properties of increasing complexity. Mostof these programs are combinatorial map generators.Thus, this work contributes to the research fields of enumerative combinatorics and softwareengineering. We distribute a C library of bounded exhaustive generators of structured arrays, formallyspecified in ACSL and verified with the WP plugin of the Frama-C analysis platform. We also proposea testing-based methodology to assist interactive proof in Coq, an original formal study of maps, andnew results in enumerative combinatorics

El presente trabajo de investigación se trató sobre la situación actual que atraviesa la empresa “ACSL S.R.L. Corredores de Seguros”, empresa fundada en 1978 con más de 34 años de trayectoria autorizada por la Superintendencia de Banca y Seguros. La Empresa, cuenta con una buena aceptación de mercado, tiene una demanda ya ganada y también presenta una ubicación estratégica obteniendo buenos resultados en todos estos años. Con el transcurrir de los años ha surgido nuevos competidores que han hecho ver muchas falencias ocasionando una serie de problemas que conllevaron a que no se desarrolle una buena integridad organizacional, sus colaboradores buscan sólo su beneficio personal (cultura individualista); Hugo Benzaquen actual administrador en Chiclayo, ha realizado una administración empírica tratando de resolver de la mejor manera cualquier dificultad de la empresa, pero percibe una crisis interna, que afectan directamente al Clima Laboral y deficiencia de sus colaboradores. Ante esta situación el fin de esta investigación fue encontrar bases teóricas, que fundamentaran la propuesta de implementar un plan para la mejora del Clima Organizacional de la empresa basada en la Teoría Antropológica de la Motivación de Pérez López. Para la investigación se elaboró un cuestionario que se dividió en 3 variables, 13 indicadores y 36 ítems con el cual se recogió la información que permitió dar respuestas a las preguntas contenidas en el problema de la investigación y rechazar o no nuestra hipótesis; para obtener la información necesitada fue necesario encuestar a los 13 trabajadores de la empresa.

El presente trabajo de investigación se trató sobre la situación actual que atraviesa la empresa “ACSL S.R.L. Corredores de Seguros”, empresa fundada en 1978 con más de 34 años de trayectoria autorizada por la Superintendencia de Banca y Seguros. La Empresa, cuenta con una buena aceptación de mercado, tiene una demanda ya ganada y también presenta una ubicación estratégica obteniendo buenos resultados en todos estos años. Con el transcurrir de los años ha surgido nuevos competidores que han hecho ver muchas falencias ocasionando una serie de problemas que conllevaron a que no se desarrolle una buena integridad organizacional, sus colaboradores buscan sólo su beneficio personal (cultura individualista); Hugo Benzaquen actual administrador en Chiclayo, ha realizado una administración empírica tratando de resolver de la mejor manera cualquier dificultad de la empresa, pero percibe una crisis interna, que afectan directamente al Clima Laboral y deficiencia de sus colaboradores. Ante esta situación el fin de esta investigación fue encontrar bases teóricas, que fundamentaran la propuesta de implementar un plan para la mejora del Clima Organizacional de la empresa basada en la Teoría Antropológica de la Motivación de Pérez López. Para la investigación se elaboró un cuestionario que se dividió en 3 variables, 13 indicadores y 36 ítems con el cual se recogió la información que permitió dar respuestas a las preguntas contenidas en el problema de la investigación y rechazar o no nuestra hipótesis; para obtener la información necesitada fue necesario encuestar a los 13 trabajadores de la empresa.
Tesis

Vérifier formellement des programmes concurrents est une tâche difficile. S’il existe différentes techniques pour la réaliser, très peu sont effectivement mises en oeuvre pour des programmes écrits dans des langages de programmation réalistes. En revanche, les techniques de vérification formelles de programmes séquentiels sont utilisées avec succès depuis plusieurs années déjà, et permettent d’atteindre de hauts degrés de confiance dans nos systèmes. Cette thèse propose une alternative aux méthodes d’analyses dédiées à la vérification de programmes concurrents consistant à transformer le programme concurrent en un programme séquentiel pour le rendre analysable par des outils dédiés aux programmes séquentiels. Nous nous plaçons dans le contexte de FRAMA-C, une plate-forme d’analyse de code C spécifié avec le langage ACSL. Les différentes analyses de FRAMA-C sont des greffons à la plate-forme, ceux-ci sont à ce jour majoritairement dédiés aux programmes séquentiels. La méthode de vérification que nous proposons est appliquée manuellement à la vérification d’un code concurrent issu d’un hyperviseur. Nous automatisons la méthode à travers un nouveau greffon à FRAMA-C qui permet de produire automatiquement, depuis un programme concurrent spécifié, un programme séquentiel spécifié équivalent. Nous présentons les bases de sa formalisation, ayant pour but d’en prouver la validité. Cette validité n’est valable que pour la classe des programmes séquentiellement consistant. Nous proposons donc finalement un prototype de solveur de contraintes pour les modèles mémoire faibles, capable de déterminer si un programme appartient bien à cette classe en fonction du modèle mémoire cible
Formal verification of concurrent programs is a hard task. There exists different methods to perform such a task, but very few are applied to the verification of programs written using real life programming languages. On the other side, formal verification of sequential programs is successfully applied for many years, and allows to get high confidence in our systems. As an alternative to dedicated concurrent program analyses, we propose a method to transform concurrent programs into sequential ones to make them analyzable by tools dedicated to sequential programs. This work takes place within the analysis framework FRAMA-C, dedicated to the analysis of C code specified with ACSL. The different analyses provided by FRAMA-C are plugins to the framework, which are currently mostly dedicated to sequential programs. We apply this method to the verification of a concurrent code taken from an hypervisor. We describe the automation of the method implemented by a new plugin to FRAMAC that allow to produce, from a specified concurrent program, an equivalent specified sequential program. We present the basis of a formalization of the method with the objective to prove its validity. This validity is admissible only for the class of sequentially consistent programs. So, we finally propose a prototype of constraint solver for weak memory models, which is able to determine whether a program is in this class or not, depending on the targeted hardware

It is estimated by the World Health Organization (WHO) that 422 million people had diabetes worldwide in 2014, including 30.3 million people in the US. The cost of treating the disease is has tripled from 2003-2013 due to the increased number of patients. One of the genes strongly associated with type 2 diabetes (T2D) is the transcription factor 7 like 2 (TCF7L2). A single nucleotide polymorphism (SNP) of the TCF7L2 results in increased expression of long chain acyl-CoA synthetase 5 (ACSL5) while deletion of this part of the TCF7L2 gene reduces ACSL5 mRNA level. The regulation of ACSL5 gene expression by the high risk TCF7L2 allele highlights the importance of investigating the role of ACSL5 in T2D. ACSL5 is one of a family of enzymes that activates FA to its CoA ester and is required for FA metabolism within cells. Mice lacking this protein have reduced fat mass and are more insulin sensitive. Chronic exposure of clonal pancreatic ß-cells to excess nutrients has been shown to result in increased intrinsic lipid droplets, reduced insulin content, a left-shift in glucose dose-dependent insulin secretion curve characterized by basal insulin hypersecretion (IH) and blunted glucose stimulated insulin secretion (GSIS). We tested the hypothesis that the use of a putative ACSL5 inhibitor (Adipo C) can reduce accumulated lipid droplets, rescue insulin content and reverse the left-shift in glucose dose-dependent insulin secretion curve. INS-1 (823/13) cells were cultured in either 4 mM or 11 mM glucose media representing physiological and excess nutrients environment. Adipo C (10-25 µM) was added to cells to both acutely (2 hrs) and chronically (72 hrs) inhibit ACSL5 activity. Thin layer chromatography with C11 Bodipy fatty acid (BFA) was used to detect acute fatty acid incorporation into neutral lipids. Nile red was used to visualize intrinsic lipid droplets inside cells. Intracellular Ca2+ activity was detected using fura 2. Insulin assay was measured by HTRF. Acute fatty acid incorporation and lipid accumulation were reduced in cells exposed to Adipo C. An Adipo C concentration dependent right shift of glucose dose-dependent insulin release and increased insulin content were observed. 11 mM glucose cells cultured in 25 µM Adipo C showed decreased intracellular Ca2+ activity at 3 mM glucose and increased Ca2+ activity at 12 mM glucose, which are characteristic of cells cultured in 4 mM glucose having reduced lipid stores. These results all indicate possible protective effects on -cells exposed to excess nutrients. Islets of T2D patients who have a physiologically elevated blood glucose level are exposed to a similar excess nutrient environment. Therefore, the results illustrated here warrant further research on Adipo C compound to explore its therapeutic potential on T2D.