The purpose of this dissertation was to determine the usefulness of the linguistic processes of Distortion, Deletion, Generalization, and Semantic Ill-Formedness as constructs which differentiate the verbal communication of families who express dissatisfaction with their current intrafamilial relationships from families expressing satisfaction with their current relationships. Specifically, it was hypothesized that dissatisfied families would use these linguistic structures to a greater extent in their interaction than would satisfied families. Thirty-one family triads (father, mother, and child) were obtained by asking families randomly selected from the local high school student directory to participate. The families were given a Revealed Differences questionnaire which they subsequently discussed together and a questionnaire regarding their satisfaction with their intrafamilial relationships. The discussions were recorded and transcribed. Each of 150 Surface Structures (a complete thought, usually a grammatical sentence) per family was scored for 11 subcategories of Distortion, Deletion, Generalization, and Semantic Ill-Formedness. Interrater reliabilities ranged from .86 to .98. A mean was computed for the questionnaire pertaining to satisfaction with family relationships. Six families who scored at least one half standard deviation below the mean comprised the "dissatisfied" family group, and six families who scored at least one half standard deviation above the mean comprised the "satisfied" family group. It was found that the dissatisfied families used significantly more Deletion (p The linguistic process of Deletion is theorized to result in impoverishing the speaker's model of the world and the behavioral choices available to the speaker. Similarily, the listener(s) who must respond to the impoverished model is limited in his response and behavioral options. Since all members of the dissatisfied families used this form of language, they perpetuate the impoverishing model of the world and the limitations on their behavior. It was concluded that, while not establishing an etiologic link between the use of Deletion and family dissatisfaction, Deletion is part of the current verbal interaction of families who express dissatisfaction. Further research involving families in which a member is symptomatic is warranted based on the findings of this study. Language may provide at least one form of explanation regarding the process by which families maintain homeostasis in the face of symptom development. The use of linguistic concepts shows promise as an intermediate link in family interaction theory as well as a form of intervention available to therapists.

A Distrofia Muscular de Duchenne (DMD) é causada por mutações no gene distrofina que codifica a proteína distrofina, responsável pela manutenção da membrana da fibra muscular. Além do comprometimento muscular, a doença tem sido associada a déficits cognitivos e problemas de comportamento. A presente pesquisa teve como objetivos: avaliar sintomas do transtorno do espectro do autismo (TEA) em uma amostra formada por pacientes com DMD de acordo com os critérios diagnósticos do DSM V; identificar a proporção de pacientes com indício de deficiência intelectual (DI); examinar a possibilidade de prejuízos das funções executivas (flexibilidade e planejamento) nesses pacientes e verificar possiveis associações das mutações downstream ao exon 45 (inicial da isoforma cerebral da distrofina Dp140) a sintomas de autismo, deficiencia intelectual ou déficits nas funções executivas. O estudo seguiu metodologia de pesquisa exploratória. Participaram do estudo 67 pacientes com DMD com idades de 5 a 17 anos (Média= 10,74 e DP= 3,2) e 19 controles com idades de 04 a 14 anos (Média=8,73 e DP= 2,94). A bateria de avaliação incluiu a Escala de Avaliação do Autismo na Infância (CARS) para avaliação dos sintomas de autismo, o Teste das Matrizes Progressivas Coloridas de Raven para exame de inteligencia não verbal e subtestes da bateria Cambridge Neuropsychological Test Automated Battery (CANTAB) para avaliação das funções executivas de flexibilidade (Intra-Extra Dimensional Set Shift - IED) e planejamento (Stockings of Cambridge - SOC). A Escala de Brooke foi utilizada para avaliação do comprometimento motor dos membros superiores dos pacientes com DMD. Dentre os pacientes submetidos à escala CARS e ao teste de Raven, 20% atingiram o ponto de corte para autismo (7% com risco para TEA somente e 13% com risco para TEA e DI) e 19% apresentou classificação de inteligência indicativa de deficiência intelectual (sem risco para TEA). Em análise dos grupos de pacientes com DMD com e sem risco para autismo, observou-se diferença significativa em 14 das 15 questões da escala CARS (p<0,05, teste t). Mediante análise qualitativa, verificou-se que o grupo com risco para TEA apresentou médias mais altas em relação aos grupos com risco para DI e risco para DI e TEA nas questões referentes à resposta emocional, comunicação verbal, resposta intelectual e às impressões gerais do avaliador da escala CARS. O grupo sem risco para TEA apresentou escores mais elevados quanto às respostas emocional e intelectual. No grupo sem risco para TEA, observou-se correlação negativa entre idade e a questão de comunicação verbal da escala CARS, evidenciando melhora desta capacidade conforme o aumento da idade (p<0,05, correlação de Pearson). Em análise subseqüente com três grupos (TEA, DI, TEA + DI), o grupo com risco para TEA apresentou escores mais elevados (indicativos de maior intensidade de sintomas de autismo) na questão referente a medo e nervosismo, enquanto o grupo com risco para TEA e DI obteve maiores pontuações nas questões de resposta auditiva e comunicação verbal da escala CARS (p<0,05, teste de Kruskal-Wallis). O grupo com risco para DI obteve escores mais baixos (indicativos de menor intensidade de sintomas de autismo) nas questões referentes à imitação, resposta emocional, uso corporal, uso de objetos, paladar, olfato e tato, comunicação não verbal e impressões gerais do examinador (p<0,05). Conforme as análises de mutações genéticas, o grupo com mutações downstream ao exon 45 apresentou maior propensão para deficiência intelectual conforme a questão referente à resposta intelectual da escala CARS. Quanto à análise de funções executivas, o grupo com DMD apresentou prejuízos nas capacidades de flexibilidade cognitiva (teste IED) e planejamento (teste SOC) em relação ao grupo controle (p<0,05, teste de Mann-Whitney). Os déficits de planejamento não foram atribuídos a dificuldades motoras, uma vez que não houve IX diferença significativa entre grupos quanto aos tempos de resposta no teste SOC. O estudo mostrou evidencias da associação da Distrofia Muscular de Duchenne ao Transtorno do Espectro Autista, à deficiência intelectual e a prejuízos em funções executivas. Os resultados suportam achados prévios da associação entre mutações distais do gene distrofina e prejuízo intelectual. A escala CARS mostrou-se sensível na diferenciação entre casos de autismo e deficiência intelectual em pacientes com DMD. Estudos futuros são necessários para a elucidação de características de autismo em pacientes com DMD com e sem o diagnóstico desse transtorno do desenvolvimento / Duchenne Muscular Dystrophy (DMD) is caused by mutations in the dystrophin gene that encodes the dystrophin protein, which is responsible for maintaining the integrity of the muscle fiber membrane. In addition to muscle impairment, the disease has been associated to cognitive deficits and behavioral problems. The objectives of the present study were: to evaluate patients with DMD for symptoms of autism spectrum disorder (ASD) according to the DSM-V criteria; to identify the proportion of patients with evidence of intellectual disability (ID) in this sample; to examine the possibility of impairment of executive functions (flexibility and planning) in these patients and to verify possible associations between mutations downstream from exon 45 (first exon for the brain isoform D140) and symptoms of autism, intellectual disability or deficits in executive functions. The study followed exploratory research methodology. Sixty seven patients with DMD with ages ranging from 5 to 17 years (mean = 10.74 and SD = 3.2) and 19 controls aged 04 to 14 years (Mean = 8.73 and SD = 2.94) participated in the study. The assessment battery included the the Childhood Autism Rating Scale (CARS) for assessing symptoms of autism, the Raven\'s Colored Progressive Matrices Test for nonverbal intelligence evaluation, and subtests of the Cambridge Neuropsychological Test Automated Battery (CANTAB) for assessment of flexibility (Intra-Extra Dimensional Set Shift - IED) and planning (Stockings of Cambridge - SOC). The Brooke Scale was used to assess motor impairment in the upper limbs of DMD group. Among the patients who were evaluated with the CARS scale and the Raven\'s test, 20% reached the cut-off point for autism (7% at risk for only ASD and 13% at risk for ASD and ID). 19% presented evidence of intellectual disability (no risk for ASD). Comparison analysis between patients with and without risk for autism revealed significant difference in 14 of the 15 CARS scale itens (p<0.05, t-test). Qualitative analysis showed that the group with risk for ASD presented higher means in relation to the groups at risk for ID and risk for ID and ASD with higher CARS scores in the scalesitens related to emotional response, verbal communication, intellectual response and to the general impressions of the examiner. The non-risk group presented higher scores regarding emotional and intellectual responses. In this group, a negative correlation was observed between age and verbal communication suggesting improvement in this capacity in older patients (p <0.05, Pearson\'s correlation). In the subsequent analysis considering three groups at risk for (a) ASD only, (b) ID only, and (c) ASD and ID, patients at risk for ASD presented higher scores (indicating higher severity of symptoms of ASD) on the item concerning fear and nervousness, while the group at risk for ASD and ID obtained higher scores on the items regarding auditory response and verbal communication (CARS scale, p <0.05, Kruskal-Wallis test). The group at risk for only ID showed significant lower scores (suggesting lower severity of symptoms) on items related to imitation, emotional response, body use, object use, taste, smell and touch, nonverbal communication and general impressions of the examiner (p <0.05). Analyzes of genetic features showed that patients with mutations downstream from exon 45 presented higher propensity to intellectual deficiency according to the item concerning intellectual response (CARS scale). Executive functions analyses revealed impairments in cognitive flexibility (IED) and planning (SOC) among DMD patients in relation to the control group (p <0.05, Mann-Whitney test). Planning deficits were not attributed to motor difficulties, since there was no significant difference between groups regarding the response times in the SOC tasks. The study showed evidence of the association of Duchenne Muscular Dystrophy with Autistic XI Spectrum Disorder, intellectual disability and impairment in executive functions. The present results support previous findings regarding the association between distal mutations in the dystrophin gene and intellectual impairment. The CARS scale was sensitive to differentiation between autism and intellectual disability in patients with DMD. Future studies are needed to elucidate autism characteristics in DMD patients with and without the diagnosis of this developmental disorder

Este trabalho teve por objetivo analisar o papel da frequência lexical em dois fenômenos fonológicos variáveis que apresentam condicionamentos morfológicos, quais sejam Redução da Nasalidade e Apagamento de /r/ Final, no intuito de verificar: 1) a interação de frequência lexical com variáveis linguísticas; 2) a interação de frequência lexical com variáveis sociais; 3) o melhor modelo, se abstracionista ou exemplarista, para tratar dos resultados aqui obtidos. Para a redução da nasalidade, reanalisamos dados de Schwindt e Bopp da Silva (2010), adicionando a informação de frequência lexical para a análise dos dados; para o fenômeno de apagamento de /r/ final, novos dados foram coletados de entrevistas com informantes do Rio de Janeiro. A informação de frequência lexical dos dados foi obtida do corpus de referência ASPA (Avaliação Sonora do Português Atual). Nossos resultados apontam que frequência lexical apresenta correlação positiva com a aplicação dos fenômenos, estando condicionada à classe gramatical, não à estrutura morfológica interna da palavra. Idade apresenta uma interação bastante significativa em dados de redução da nasalidade, principalmente na classe de não verbos sem gem. A mesma interação com idade não foi encontrada no fenômeno de apagamento de /r/. Uma explicação para isso talvez esteja relacionada com a caracterização de frequência de type e frequência de token. Tendo em vista que, na análise de frequência lexical, regras relativas à morfologia interna à palavra e à fonologia parecem não ser determinantes na aplicação dos fenômenos, defendemos neste trabalho a superioridade das abordagens exemplaristas. / The purpose of this study was to analyze the role of lexical frequency in two variable phonological phenomena which present morphological conditioning, such as Nasality Reduction and Final /r/ Deletion, in order to verify: 1) lexical frequency interaction with linguistic variables ; 2) lexical frequency interaction with social variables; 3) the best model, if abstractionist or exemplarist, to deal with the results obtained here. For nasality reduction, we reanalyzed data from Schwindt and Bopp da Silva (2010), adding lexical frequency information for data analysis; for the final /r/ deletion phenomenon, new data were collected from interviews with informants from Rio de Janeiro. Lexical frequency information of the data was obtained from the reference corpus ASPA (Sound Evaluation of Current Portuguese). Our results point out that lexical frequency has positive correlation with the application of the phenomena, being conditioned by grammatical class, not by the internal morphological structure of the word. Age has a very significant interaction in nasality reduction data, especially in the class of non-verbs without gem. The same interaction with age was not found in the final /r/ deletion phenomenon. An explanation for this may be related to the characterization of type and token frequency. In lexical frequency analysis, considering that rules regarding the internal morphology of words and regarding phonology do not seem to be decisive in the application of both phenomena, we argue for the superiority of the exemplarist approaches.

Many texts we encounter in our everyday lives are lexically and syntactically very complex. This makes them difficult to understand for people with intellectual or reading impairments, and difficult for various natural language processing systems to process. This motivated the need for text simplification (TS) which transforms texts into their simpler variants. Given that this is still a relatively new research area, many challenges are still remaining. The focus of this thesis is on better understanding the current problems in automatic text simplification (ATS) and proposing new data-driven approaches to solving them. We propose methods for learning sentence splitting and deletion decisions, built upon parallel corpora of original and manually simplified Spanish texts, which outperform the existing similar systems. Our experiments in adaptation of those methods to different text genres and target populations report promising results, thus offering one possible solution for dealing with the scarcity of parallel corpora for text simplification aimed at specific target populations, which is currently one of the main issues in ATS. The results of our extensive analysis of the phrase-based statistical machine translation (PB-SMT) approach to ATS reject the widespread assumption that the success of that approach largely depends on the size of the training and development datasets. They indicate more influential factors for the success of the PB-SMT approach to ATS, and reveal some important differences between cross-lingual MT and the monolingual v MT used in ATS. Our event-based system for simplifying news stories in English (EventSimplify) overcomes some of the main problems in ATS. It does not require a large number of handcrafted simplification rules nor parallel data, and it performs significant content reduction. The automatic and human evaluations conducted show that it produces grammatical text and increases readability, preserving and simplifying relevant content and reducing irrelevant content. Finally, this thesis addresses another important issue in TS which is how to automatically evaluate the performance of TS systems given that access to the target users might be difficult. Our experiments indicate that existing readability metrics can successfully be used for this task when enriched with human evaluation of grammaticality and preservation of meaning.

Objetivos: estudar indivíduos com (G1) e sem (G2) sinais da Síndrome Velocardiofacial (SVCF) para verificar diferenças entre eles quanto à extensão e espessura velar, profundidade nasofaríngea, razão entre profundidade nasofaríngea e extensão velar (PNF/EV), tamanho da falha velofaríngea, ângulo velar, movimento do véu palatino e das paredes laterais e posterior da faringe e à presença da tonsila faríngea; diferenças para as medidas de extensão e espessura velar, profundidade nasofaríngea e razão PNF/EV dos grupos estudados com os valores de normalidade propostos por Subtelny (1957); e correlação entre o tamanho da falha velofaríngea e a razão PNF/EV. Material e Método: estudo prospectivo com 60 indivíduos de ambos os sexos sem fissura palatina evidente e com disfunção velofaríngea (DVF), não operados, sendo 30 com sinais clínicos da SVCF (G1) (idade de 5,4 a 51 anos, com média de 15,7±9,5 anos) e 30 sem os sinais da SVCF (G2) (idade de 4,5 a 33 anos, com média de 15±7,6 anos). O exame videofluoroscópico foi realizado nas projeções lateral e frontal para análise da extensão e espessura velar, profundidade nasofaríngea, falha velofaríngea e ângulo velar, movimento do véu palatino, paredes laterais e posterior da faringe e presença da tonsila faríngea. Resultados: quanto às medidas de extensão e espessura velar, profundidade nasofaríngea e razão PNF/EV, não houve diferença entre os grupos quando se comparou os casos maiores de 18 anos, bem como os menores de 18 anos pareados por idade; mas quando a idade não foi pareada nos casos menores de 18 anos, a espessura velar foi menor (p=0,019) e a razão PNF/EV foi maior (p=0,048) no G1 e, ao se analisar independente da faixa etária, a razão PNF/EV foi maior no G1 (p=0,024). Em relação às medidas de normalidade, a extensão velar foi menor no G1 (p=0,007), a espessura velar foi menor no G1 (p=0,000) e G2 no (p=0,000), a profundidade nasofaríngea e a razão PNF/EV foram maiores no G1 (p=0,000) e no G2 (p=0,000), entretanto ao se considerar a variação de 2 desvios-padrão em relação aos valores de normalidade, não houve diferença entre os grupos para todas as medidas. Também não houve diferença entre os grupos quanto ao ângulo de elevação velar (p=0,232) e presença (p=0,698) e tamanho da falha velofaríngea (p=0,293), movimento velar (p=0,085) e das paredes laterais (p=0,763) e posterior (p=0,237) da faringe, além do tamanho da tonsila faríngea (p=0,307). Não houve correlação entre a falha velofaríngea e a razão PNF/EV no G1 (p=0,153) e no G2 (p=0,598). Conclusões: a razão PNF/EV foi maior nos indivíduos com DVF e sinais da SVCF comparados aos indivíduos com DVF sem os sinais da síndrome, sugerindo ser este um indicador velofaríngeo para a SVCF, enquanto os aspectos funcionais da velofaringe não diferiram entre os indivíduos com e sem os sinais da SVCF. Não houve correlação entre o tamanho da falha no fechamento velofaríngeo e a razão PNF/EV nos grupos com e sem sinais da SVCF. / Objective: to study individuals with (G1) and without (G2) signs of velocardiofacial syndrome (VCFS), so as to verify differences in terms of length and thickness of the soft palate, nasopharyngeal depth, ratio of nasopharyngeal depth to velar length (PD/VL), velopharyngeal gap size, velar angle, soft palate movement, lateral and posterior pharyngeal walls movement and the presence of adenoidal tissue; differences for the measurements of velar length and thickness, nasopharyngeal depth and PD/VL ratio for the groups studied with the normality values proposed by Subtelny (1957); and correlation between the size of velopharyngeal gap and the PD/VL ratio. Methods: a prospective study with 60 subjects from both genders, with no evident cleft palate, with velopharyngeal dysfunction (VPD), non operated, being 30 with clinical signs of VCFS (age range 5.4 to 51 yrs, mean 15.7±9.5 yrs), and 30 with no signs of VCFS (age range 4.5 to 33 yrs, mean 15±7.6 yrs). The videofluoroscopy was performed in lateral and frontal views, for the analysis of velar length and thickness, nasopharyngeal depth, velopharyngeal gap, velar angle, soft palate movement, lateral and posterior pharyngeal walls movement and the presence of adenoidal tissue. Results: there was no difference in the measurements of velar length and thickness, nasopharyngeal depth and PD/VL ratio, between the groups, when cases over 18 yrs, as well as those under 18, paired by age, were compared; however, when age was not paired in the cases under 18 yrs, the velar thickness was smaller (p=0.019) and the PD/VL ratio was greater (p=0.048) in G1 and, by analyzing regardless the age range, the PD/VL ratio was greater in G1 (p=0.024). In relation to normality measurements, the velar length was smaller in G1 (p=0.007), the velar thickness was smaller in G1 (p=0.000) and G2 (p=0.000), the nasopharyngeal depth and the PD/VL ratio were greater in G1 (p=0.000) and G2 (p=0.000), nevertheless, when considering the variation of 2 standard deviations in relation to the normality values, there was no difference between the groups for all measurements. No difference was seen between the groups, as to the velar angle (p=0.232) and presence (p=0.698) and size of velopharyngeal gap (p=0.293), velar movement (p=0.085) and lateral (p=0.763) and posterior (p=0.237) pharyngeal walls movement, besides the size of adenoidal tissue (p=0.307). No correlation was seen between the velopharyngeal gap and the PD/VL ratio in G1 (p=0.153) and G2 (p=0.598). Conclusions: the PD/VL ratio was greater in individuals with VPD and signs of VCFS, as compared to individuals with VPD with no signs of the syndrome, suggesting that this is a velopharyngeal indicator for VCFS, whereas velopharyngeal functional aspects did not differ between individuals with and without signs of VCFS. There was no correlation between the size of the velopharyngeal gap and the PD/VL ratio, in the groups with and without signs of VCFS.

Objetivo: Descrever as manifestações clínicas de indivíduos com hipótese diagnostica da Sindrome de deleção 22q11 (SD22q11) confirmados por testes genéticos, na primeira avaliação e durante o acompanhamento dos mesmos em avaliações subsequentes para uma melhor definição do curso natural da doença. Local: Laboratório de Genética e Citogenética Humana do HRAC-USP Bauru/SP. Casuística e metodologia: O presente trabalho é retrospectivo e analisou os dados de prontuários de 72 indivíduos cadastrados no HRAC-USP, os quais receberam hipótese diagnóstica da SD22q11 e foram confirmadas por teste genético (MLPA ou FISH). A avaliação envolveu a analise dos dados relatados por todos os setores do HRAC-USP. Resultados e discussão: Foramanalisados 72prontuários deindivíduos com a SD22q11. Constatamos que a idade media dos indivíduos quando do cadastro no HRAC-USP foi de seis anos. Também constatamos que houve um longo período de tempo entre os retornos ao hospital e que, nesses retornos, nem todas as especialidades foram contempladas. Esses fatos prejudicaram a analise da historia natural da anomalia em questão. Com relação às características fenotipicas, observamos a presença de sinais clínicos típicos, como por exemplo: face alongada, lábios finos, hipoplasia alar, anormalidades menores na orelha, dígitos longos e fendas palpebrais, fissuras labiopalatinas, cardiopatias congenitas, dificuldade de aprendizagem, atraso de linguagem e distúrbios comportamentais. A fissura oral foi à manifestação otorrinolaringológica mais frequente, presente em 75% dos pacientes, onde as fissuras submucosas foram as mais frequentes (43%). As características cognitivas como, atraso de fala (87%), dificuldades de aprendizagem (95%) e distúrbios comportamentais (81%), tiveram um resultado significativo, descritas em quase todos os indivíduos. As cardiopatias congênitas estavam presentes em 47,2% dos prontuários analisados. De um modo geral, comparando a frequência dos sinais clínicos encontrados neste trabalho com dados da literatura, constatamos que as frequências encontram-se dentro do esperado. Conclusão: A maioria dos indivíduos cadastrados no HRAC-USP, pertencentes ao grupo de estudo, apresentou idade superior a 06 anos. Portanto, a observação do curso natural da historia da SD22q11 para avaliar características fenotípicas que surgissem ao longo da evolução clinica do indivíduo e que pudessem ajudar no diagnóstico, ficou prejudicada. Mesmo nos casos onde o indivíduo foi cadastrado no HRAC-USP com idade inferior a dois anos, o diagnóstico foi tardio devido a falta de uma ação multidisciplinar e interdisciplinar no hospital. Mesmo não sendo possível avaliar as características fenotípicas surgidas durante a historia natural da doença, constatamos que as manifestações clínicas relatadas nos prontuários cursam com as características da SD22q11 e em frequências que corroboram com as da literatura / To describe clinical manifestations observed in medical records of individuals registered in the hospital with a diagnostic hypothesis of 22q11.2DS confirmed by genetic tests (MLPA OR FISH), since the first assessment in the HRAC-USP and during the follow up of these individuals in subsequent assessments, in order to achieve a better definition to the natural courses of the disease. Local: Laboratory of Human Genetics and Cytogenetics (HRAC-USP Bauru/SP). Methods: This retrospective study analyzed 72 medical records of individuals registered at the HRAC-USP, who were diagnosed with 22q11DS and who had this diagnosis confirmed by a genetic test (MLPA OR FISH). The assessment concerned the analysis of reported data in all sectors of the HRAC-USP. Results and Discussion: 72 medical records of individuals with 22q11DS were analyzed. It was verified that the average age of individuals when registering at the HRAC-USP was six years old. It was also verified that it took a long period of time for these individuals to return to the hospital and, when they did, not all specialties were contemplated. These facts harmed the analysis of the natural history of the anomaly. About the phenotypic characteristics, some typical clinical signs were observed, such as: long face, thin lips, hypoplasia nasal alar, minor abnormalities in the ear, long digits and narrow palpebral fissures, palatal abnormalities, congenital heart defects, learning disabilities, delay speech and behavioral disorders. An oral cleft was the most frequent otorhinolaryngology manifestation, present in 75% of the patients; among which submucous cleft palate were the most frequent (43%). Cognitive features such as, delay speech (87%), learning disabilities (95%) and behavioral disorders (81%) had a significant result, described in almost all individuals. Congenital heart defects were observed at 4% to 48% of individuals with 22q11.2DS, in this study it was observed in 47.2%. In general, comparing the frequency of some clinical signs observed in this study with the literature data, it was verified that the frequencies were within expectations. Conclusion: Most of the individuals registered at the HRAC belonging to the study group were over 6 years old. Therefore, the observation of natural course of the history of 22q11DS to evaluate the phenotypic characteristics that would arise during the clinical evolution of the individual and that could help in the diagnosis was harmed. Even in cases when the individual was registered at the HRAC-USPunder the age of two, the diagnosis was delayed due to lack of a multidisciplinary and interdisciplinary action in the hospital. Even not being possible to measure the phenotypic characteristics that emerged during the natural history of the disease, it was verified that the clinical manifestations reported in the records occur with the 22q11DS characteristics and in frequencies that corroborate with the literature

Introdução: A Holoprosencefalia (HPE) é uma malformação craniofacial decorrente de falhas no processo de formação do sistema nervoso durante o desenvolvimento embrionário. Sua etiologia é heterogênea e complexa, envolvendo fatores ambientais e genéticos. Estudos recentes sugerem que alterações subteloméricas possam ser um dos fatores etiológicos para o aparecimento da HPE. Objetivos: Investigar possíveis alterações (microdeleções/duplicações) nas regiões subteloméricas em indivíduos com diagnóstico de HPE. Metodologia: Avaliação genética de 25 amostras de DNA de indivíduos matriculados no HRAC-USP, com diagnóstico de HPE através da técnica de MLPA (Multiplex ligation-dependent probe amplification), segundo protocolo proposto por Schouten et al. (2002). Estes indivíduos já foram previamente triados para mutações/deleções nos genes candidatos à HPE (SHH, ZIC2, TGIF, GLI2 e PTCH1) através do sequenciamento direto de Sanger e da técnica de MLPA, sem resultado positivo para qualquer alteração. Resultados: Dentre os 25 indivíduos analisados, o fenótipo predominante foi a microforma da doença. Os principais achados clínicos da amostra estudada foram: hipotelorismo, microcefalia e fissura de lábio e palato (100%), nariz achatado (84%), presença de um incisivo central único (24%) e ponte nasal baixa (64%). Quatro pacientes apresentaram comprometimento no SNC (16%). Nenhum indivíduo apresentou quaisquer alterações, como microdeleções e/ou duplicações nos genes contidos nas regiões subteloméricas, através da análise pela técnica de MLPA. Dessa forma, estes permanecem sem diagnóstico genético definido. Discussão: A não detecção de alterações subteloméricas sugere que o fenótipo predominante na amostra, microforma, possa não apresentar alterações subteloméricas relacionadas ao aparecimento da doença. Entretanto, deve-se salientar que o universo amostral é relativamente pequeno, de forma que este possa não ser um exemplo fiel dos casos de microforma da HPE. Reforça-se, ainda, a grande variedade de fatores envolvidos no surgimento desta patologia, bem como o envolvimento de outros genes ainda não estabelecidos, além das causas ambientais, ainda não completamente elucidadas. Conclusões: Não foram encontradas alterações subteloméricas nos pacientes com diagnóstico de HPE estudados. A técnica de MLPA consiste em uma metodologia rápida, sensível, eficaz e de baixo custo, quando comparada a outras técnicas, sendo indicada para o uso em laboratórios de diagnóstico genético, uma vez que diversos estudos já mostraram que consiste em um método confiável de diagnóstico. Devido ao tamanho relativamente pequeno da amostra utilizada neste estudo, e os dados ainda inconsistentes da literatura atual, estudos adicionais são necessários para que seja possível a realização de um diagnóstico diferencial, explicando o amplo espectro fenotípico observado nesta doença, conforme sugerido pela hipótese dos múltiplos fatores. / Introduction: Holoprosencephaly (HPE), a craniofacial malformation, results from flaws in formation process of the nervous system during embryonic development. The etiology is heterogeneous and complex, involving environmental and genetic factors. Recent studies suggest that subtelomeric aberrations could be an etiological factor to the onset of HPE. Objectives: Investigate possible changes (microdeletions/duplications) in subtelomeric regions in individuals diagnosed with HPE. Methodology: Genetic evaluation of 25 DNA samples from individuals enrolled at HRAC-USP, diagnosed with HPE (performed by Syndromology Division HRAC/USP) by MLPA technique, as proposed by Schouten et al (2002). Patients were previously screened for mutations/deletions in HPE candidate genes (SHH ZIC2, TGIF, GLI2 and PTCH1) by direct Sanger sequencing and MLPA technique, without any match. Analyses were performed at the Laboratory of Molecular Genetics, Hospital for Rehabilitation of Craniofacial Anomalies, HRAC-USP. Results: Among the 25 individuals analyzed, the predominant phenotype was HPE microform. The main clinical findings of the study sample were: hypotelorism, microcephaly, and cleft lip/palate (100%); flat nose (84%); presence of a single central incisor (24%) and low nasal bridge (64%). Four patients had CNS commitment (16%). No subtelomeric mutations were found in our sample, such as microdeletions/duplications of genes analyzed by MLPA technique. Thus, they remain without defined genetic diagnosis. Discussion: Subtelomeric changes were not found, suggesting that the predominant sample phenotype, microform HPE, could not be related with subtelomeric changes associated to the disease outbreak. However, it should be noted that the sample universe is relatively small, so this may not be a true example of HPE microform cases. It should also be reinforced the wide variety of factors involved in the onset of this pathology, as well as the involvement of other genes not yet established and environmental causes, not completely understood. Conclusions: No subtelomeric mutations were found in the HPE individuals studied. MLPA technique consists in a rapid, sensitive and cost effective methodology, when compared to other techniques being suitable for use in genetic diagnostic laboratories, since several studies have shown that consists of a reliable method of diagnosis. Due to the relatively small sample size used in this study, and even inconsistent data in literature, further studies are needed to make it possible to perform a differential diagnosis, explaining the wide phenotypic spectrum observed in this disease, as suggested by the multiple hit hypothesis.

Les translocations chromosomiques qui consistent en l’échange de morceaux de chromosomes sont une des caractéristiques génétiques de nombreux cancers. Les séquences des jonctions des chromosomes transloqués chez les patients correspondent à une réparation par NHEJ. Nous avons étudié le rôle du complexe de ligation XRCC4/LigaseIV du C-NHEJ dans la formation de ces réarrangements chromosomiques dans les cellules humaines. Nous avons utilisé différentes nucléases artificielles (ZFN, TALEN, et CRISPR/Cas9) afin d'introduire deux CDB sur deux chromosomes et nous avons ainsi réussi à générer différentes translocations. Des lignées sauvages et mutantes pour ce complexe de ligation ont été utilisées et la fréquence formation de translocations a été quantifiée par PCR. Nous avons pu observer que celle-ci est souvent diminuée dans les différentes lignées mutantes. Les jonctions des translocations obtenues par séquençage sont modifiées dans des cellules déficientes pour ce complexe. En effet, elles présentent de longues délétions et un biais d’utilisation de microhomologies, indiquant l’utilisation d’un mécanisme alt-NHEJ. Une altération de cette voie dans les cellules humaines n’affecte d’ailleurs pas la formation de ces réarrangements chromosomiques. Ainsi, contrairement aux cellules de souris, les translocations dans les cellules humaines sont générées par le C-NHEJ. / Chromosomal translocations involve the exchange of chromosome pieces and are often associated with oncogenesis. It has been shown that breakpoint junctions of translocated chromosomes found in patients are typical of a repair by NHEJ. Here we investigated the specific role of XRCC4/LigaseIV, the ligation complex of C-NHEJ, on chromosomal translocation formation in human cells. Using different nucleases (ZFN, TALEN, et CRISPR/Cas9) targeting two chromosomes, we studied the induction of translocation in wt and KO human cells, expressing or not the XRCC4/LigaseIV complex. We found that translocation frequency was mostly reduced in XRCC4/LigaseIV deficient cells when we quantified the induction of translocation by PCR. In addition, we analyzed the breakpoint junctions by sequencing. Strikingly, we found that junctions of translocations show large deletions, and a bias towards the use of longer microhomologies only in XRCC4/LigaseIV KO cells, signature of the alt-NHEJ activity. In contrast, translocation formation was not affected in alt-NHEJ deficient cells. Thus conflicting with results obtained in rodent cells where alt-NHEJ promotes translocation formation, translocations in human cells are generated by the C-NHEJ.

As mutações no gene PROP1 são a causa genética mais comum da deficiência combinada de hormônios hipofisários. Até o momento, diversas mutações missense e pequenas deleções foram descritas sendo a mutação 301-302 delAG a mais freqüente. Nosso objetivo foi estudar as mutações em DNA de pacientes com hipopituitarismo e padronizar a extração de DNA de células de swab oral, usando um método com NaCl e comparar com um kit comercial (Purigene, Minneapolis, EUA). Amplificamos os 3 exons do gene PROP1 do DNA obtido de células orais e de sangue periférico. Identificamos a mutação 301-302delAG em 6 pacientes, 4 em homozigose (33%) e 2 em heterozigose (16%) e a mutação G51A em heterozigose em um único paciente. Em dois irmãos, filhos de pais consangüíneos, não foi possível amplificar os 3 exons do gene PROP1 enquanto que os os genes LHX3 e LHX4 foram amplificados com sucesso. Para confirmar a hipótese de deleção do PROP1, o Southern blotting foi realizado usando como sonda o produto de PCR do exon 2 do gene PROP1 e um fragmento do gene CYP21A2 como sonda controle. A banda referente ao CYP21A2 estava presente nos pacientes e nos controles enquanto a banda referente ao PROP1 estava ausente nos irmãos e presente na mãe e nos controles. Para definir a extensão da deleção usamos um mapa de STS próximos ao gene e o STS GDB:314805 localizado a 6,0 kb a montante do PROP1 não foi amplificado nos pacientes. Entretanto, o gene Q8N6H0 a 18 kb a juzante e o STS WI-16216 a 59 kb a montante do PROP1 foram amplificados com sucesso nos pacientes e controles indicando que a deleção está localizada dentro de 81 Kb. Para determinar os limites da deleção, várias reações de PCR foram realizadas com primers desenhados progressivamente distantes de gene PROP1, cobrindo toda a região. Isto nos permitiu determinar a região deletada de 9,6 kb a juzante e 11 kb a montante do gene PROP1, com o tamanho máximo deletado de 18,4 kb. Por ambos os métodos de extração obtivemos um DNA de boa qualidade, permitindo o amplificação dos 3 exons do gene PROP1. A extração com NaCl foi mais rápida e mais barata resultando em maior quantidade de DNA quando comparada com o kit comercial. Em conclusão, descrevemos a deleção completa do gene PROP1 em dois irmãos com o fenótipo clássico de hipopituitarismo associado à hipófise hipoplásica ou aumentada e padronizamos a extração de DNA de células de mucosa oral com NaCl, que apresentou custo mais baixo e resultado mais rápido quando comparado a extração por um kit comercial, indicando que o swab oral é uma fonte prática de obtenção de DNA para estudos genéticos. / PROP1 gene mutations are the most common cause of genetic combined pituitary hormone deficiency. To date, several missense mutations and small deletions have been described and the 301-302 del AG is the most frequent. Our objective was to study PROP1 mutations in patients with hypopituitarism and standardize DNA extraction from an oral swab, using the NaCl method, comparing it with a commercial kit (Purigene, Minneapolis, USA). We amplified the 3 exons of PROP1 gene in DNA obtained from oral cells and peripheral blood cells. We identified the delAG301-302 mutation in 6 patients, 4 in homozygous (33%) and 2 in heterozygous (16%) state and G51A mutation in heterozygous state in a single patient. In two siblings, a boy and a girl, born to consanguineous parents we failed to amplify PROP1 gene by PCR whereas LHX3 and LHX4 genes were successfully amplified. To confirm the hypothesis of PROP1 gene deletion, Southern blotting was performed using PROP1 exon 2 gene PCR product as a probe and a fragment of CYP21A2 gene as a control probe. The CYP21A2 band was present in patients and controls whereas PROP1 band was absent in both siblings and present in their mother and in controls. To define the extension of this deletion we used STS mapping approach and no amplification for a STS GDB:314805 6.0 kb downstream of PROP1 was found. However, Q8N6H0 gene located 18 kb upstream and the STS WI-16216 located 59 kb downstream of PROP1 were successfully amplified indicating that the deletion is placed within 81 Kb. To determine the limits of the deletion a number of PCR covering this region were then carried out with primers located progressively distant from PROP1. This allowed us determine the deleted region from 9.6 kb upstream to 11 kb downstream of PROP with a maximum deletion size of 18.4 kb. Both methods yielded good quality DNA, allowing the amplification of 3 exons of PROP1 gene. The NaCl method showed to be faster and less expensive, resulting in a larger amount of DNA when compared with the commercial kit. In conclusion, we describe a complete deletion of PROP1 gene in two siblings with classical hypopituitarism phenotype associated with hypoplastic or enlarged pituitary gland and standardized the DNA extraction of oral cells with NaCl, which presented lower costs and faster results, when compared with the extraction by a commercial kit indicating that oral swabs are a reliable DNA source for genetic studies.

<p>Produktnedläggning är ett sätt för företag att få bort produkter som inte presterar tillfredsställande. Tidigare studier har visat att företag med ordentliga rutiner kring produktavveckling har kunnat tillgodogöra sig flera fördelar i form av bland annat bättre lönsamhet och försäljning. Ändå har forskning visat att de flesta företag inte har några formella rutiner för nedläggning av produkter och att produktnedläggning inte ses som lika viktigt som andra produktaktiviteter. Av den anledningen studeras i denna uppsats hur väl utvecklade rutiner två utvalda företag har för avveckling av produkter.</p><p>Syftet med uppsatsen är att undersöka hur två utvalda företags rutiner för produktnedläggning ser ut. Resultaten jämförs sedan med den teoretiska referensramen samt mellan företagen i fråga för att urskilja likheter och olikheter.</p><p>Till denna uppsats har en kvalitativ metod använts för att klargöra hur företag går tillväga vid nedläggning av produkter. Genom intervjuer med AB Karl Hedin och Spendrups Bryggeri AB har empirisk data samlats in för att möjliggöra jämförelser företagen emellan samt med den teori som sammanställts.    </p><p>De två studerade företagen har helt skilda förfaranden kring produktavvecklingar. Spendrups har en högre grad av formalitet i sina nedläggningsprocesser där hela kedjan från uppkomsten av en produkt till en eventuell avveckling sker i samråd mellan projektledare, företagsledning och produktansvariga. AB Karl Hedin har inga utformade rutiner kring produktavveckling utan hanterar frågan på respektive ort.  Vad som orsakar produktnedläggning hos respektive företag är dålig lönsamhet och försäljning tillsammans med flera andra individuella faktorer.    </p> / <p>Product deletion is a method that companies can use to abandon products that are not performing satisfying. Studies have shown that companies with routines for product elimination have achieved several benefits in terms of higher sales volume and profitability. Nonetheless, research has proved that most companies do not have formal procedures for product deletion and that it is not considered quite as important as other product activities. For that reason, this essay will examine how highly developed the routines for product deletion are in two selected companies.</p><p>The purpose of the study is to observe the procedures for product deletion in two selected companies. The findings will then be compared between companies in the survey and related to the theory to define similarities and differences.</p><p>A qualitative method has been used to clarify how companies proceed when to abandon products. Empirical data has been collected by interviewing AB Karl Hedin and Spendrups Bryggeri AB for comparison to the theory and between the companies in our study in order to distinguish similarities and differences.</p><p>The companies in the study have different routines when it comes to deleting products. Spendrups has a higher level of formality in their elimination procedures where the top management together with project managers and product managers collaborate all the way from the birth of the product to a possible deletion. AB Karl Hedin has no formal routines for product abandonment. These questions are dealt with at each separate district. What cause the companies to eliminate a product are mainly low profitability and sales volume among several individual factors.  </p>