Thesis (MSc (Genetics))--University of Stellenbosch, 2005. / Breast cancer is a major cause of morbidity and mortality as it is the most common invasive cancer in women worldwide. The lifetime risk for South African women to develop breast cancer is one in 31. A family history of the disease is a well-established risk factor and germline mutations in the BRCA1 (breast cancer one) and BRCA2 (breast cancer two) tumour suppressor genes markedly increase the risk of developing breast cancer. A few hundred mutations spanning the entire coding sequences of both genes have already been reported. Numerous other breast cancer susceptibility loci have been identified, but results from association studies are discrepant. The checkpoint kinase gene, CHEK2, and specifically the CHEK2*1100delC variant has, however, consistently been implicated as a candidate low-penetrance breast cancer allele. To date, few comprehensive molecular-genetic studies have been completed for the various South African breast cancer populations. The aim of this study was to determine the BRCA1 and BRCA2 mutation spectrum and prevalence in two South African populations, namely Mixed Ancestry and Caucasian. The frequency of the CHEK2*1100delC mutation was also investigated. The patient group comprised 101 unrelated patients (98 women and 3 men), presenting with invasive breast cancer. Patients with a moderate family history of breast cancer (n=48) were screened for the CHEK2*1100delC allele and the coding sequences of the BRCA1 (partly completed in a previous study) and BRCA2 genes. Patients without a family history of the disease (n=53) were only screened for the CHEK2*1100delC allele. Mutation detection was done using combined single-strand conformation polymorphism and heteroduplex analysis (SSCP/HA), followed by DNA sequencing of the identified variants. Due to its size (~5kb), exon 11 of BRCA2 was sequenced directly after amplification, in seven overlapping fragments. Three deleterious BRCA1 mutations, 1623_1627delTTAAA, E881X and 5313delC have previously been identified in three patients from the study population. No additional pathogenic mutations have been detected in this gene during this study. Two deleterious BRCA2 mutations, 6677_6678insTA and 8162delG, were identified in two and three patients respectively. Overall, BRCA1 and BRCA2 mutations have been identified in 17% of the Mixed Ancestry patients and in 15.8% of the Caucasian patients. Together BRCA1 and BRCA2 mutations account for 16.7% of breast cancer in the study population. In addition, a number of silent polymorphisms as well as variants of unknown functional significance, both known and novel, were identified. The E881X variant, which has been reported as an Afrikaner founder mutation (Reeves et al. 2004), was identified in one patient of Mixed Ancestry, but none of the published European founder mutations have been detected in our patient group. This suggests a unique mutation spectrum for South African breast cancer patients. The prevalence of the BRCA2 mutations, 8162delG and 6677_6678insTA, has to be elucidated within a larger study group. Haplotype analysis will reveal whether these patients have a common ancestor. Our findings do not suggest the presence of the CHEK2 variant in South African breast cancer patients, but a larger study population has to be analysed to confirm this. The results of this study are in agreement with those from other populations, indicating that less than 20% of breast cancers that occur in individuals with a moderate-risk for developing breast cancer are due to BRCA1 and BRCA2 mutations. By determining the contribution of BRCA1 and BRCA2 mutations to breast cancer in this group of patients, one can assess the appropriateness of predictive or diagnostic DNA testing in the clinical setting.

Made available in DSpace on 2014-06-11T19:28:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-07-27Bitstream added on 2014-06-13T20:37:59Z : No. of bitstreams: 1 faco_vmg_me_bauru.pdf: 553811 bytes, checksum: 424ea74506d016c2115f73d3386e4bb0 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / No Brasil, estudar família é um desafio devido à grande diversidade cultural existente e a variedade de arranjos familiares. Dentro dessa perspectiva, pode-se falar em “famílias brasileiras” formadas por padrões econômicos, sociais e culturais diversos. Partindo do pressuposto de que o conceito de família deve considerar a subjetividade dos indivíduos, esse estudo teve como objetivo caracterizar e conceituar famílias de zona rural e urbana de uma cidade do interior de São Paulo, segundo a perspectiva de adolescentes. Os participantes foram 48 adolescentes de 13 a 18 anos, sendo 16 da zona rural e 32 da urbana. Para atingir o objetivo proposto buscou-se uma abordagem metodológica capaz de permitir uma ampla coleta de informações, sendo utilizados dois instrumentos, um Questionário de Caracterização do Sistema Familiar e um Roteiro de Entrevista de Conceituação Familiar. Os resultados indicam que, nessa amostra, o percentual de famílias nucleares ainda é alto. O nível de escolaridade dos pais e a renda familiar são maiores na cidade do que no campo. A ocupação dos pais na área rural é mais ligada ao setor agropecuário e na urbana predomina os setores administrativos e gerenciais. Aproximadamente metade das mães rurais não exerce atividade remunerada e na cidade esse índice é de 9%. Nas duas localidades, a maioria tem casa própria (cerca de 77%). Com relação à rede social de apoio, a pessoa da família mais procurada pelos adolescentes é a mãe, seguida do pai que está assumindo várias funções, além do suporte financeiro tradicionalmente esperado; fora da família, os amigos são os mais procurados e, algumas vezes, fornecem mais apoio que os próprios membros familiares. A principal representação de família, para os adolescentes das duas localidades, é a de suporte emocional/afetivo. Quando abordam a concepção da própria família... / In Brazil, due to the big cultural diversity and the variety of family arrangements, studying family is a challenge. In this perspective, it is possible to talk about Brazilian families composed according to different economic, social and cultural standards. Presuming that the concept of family should consider individual subjectivity, this paper sought to characterize and conceptualize families from rural and urban areas of a city in São Paulo state, according to the perspective of teenagers. The participants were 48 teenagers between 13 and 18 years old, 16 from the rural area and 32 from the urban area. To reach this goal, we sought a methodological approach which could permit a broad information collection utilizing two tools, a questionnaire for familial system. Characterization and instructions for family conceptualization interview. The results indicate that the percentage of nuclear families in still high in that pattern. The parents' education level and the family's income are higher in the urban area than in the country side. The parents' jobs are more linked to agro business in the rural area and to the management and administration sectors in the urban area. About half the rural mothers do not have a paid job, while in the city they are 9%. In both places, the majority owns their houses (around 77). When it comes to social support net, mothers are the most wanted by teenagers, followed by fathers, who are taking several functins, besides the tradicional and expected financial support; apart from the family, friends are the most wanted and sometimes give even more support than family members. For the adolescents from both places, the main representation of family is that of emotional/affective support. When approaching the concept of their own families, this category continues to predominate, with inferior percentages... (Complete abstract click electronic access below)

La transmission des entreprises familiales d’une génération à l’autre est la principale difficulté à laquelle se heurtent les familles qui cherchent à assurer la pérennité de leur patrimoine. Il s’agit pourtant, pour ces organisations, d’un moment stratégique. La plupart des travaux consacrés à ce processus de succession intrafamiliale, se sont intéressés à des entreprises installées dans les pays anglo-saxons ou en Europe, et, dans une moindre mesure, en Asie. Les PME marocaines, familiales dans leur immense majorité, sont confrontées aujourd’hui à un phénomène de succession de grande ampleur, qui se transforme en enjeu national. Pourtant, les modèles de succession, élaborés à partir de la littérature existante, ne sont pas nécessairement en adéquation avec la culture nationale et le système familial dominant au Maroc, un système communautaire endogame (Todd, 1999), dont le fonctionnement est très différent de celui qui prévaut dans les sociétés anglo-saxonnes ou européennes. Cette recherche cherche à combler ce vide. A partir de neuf études de cas, construites sur la base de récits de vie de successeurs, elle élabore un ensemble de propositions, destiné à tenir compte des caractéristiques de ce système familial sur les dynamiques de succession, qu’il s’agisse du processus lui-même, de la gouvernance, de la gestion des émotions ou de la transmission de la propriété psychologique. Elle montre que l’autorité patriarcale, l’égalité entre frères et sœurs, et l’endogamie sont des dimensions critiques de la réussite du processus de succession. Cette recherche cherche à combler ce vide. A partir de neuf études de cas, construites sur la base de récits de vie de successeurs, elle élabore un ensemble de propositions, destiné à tenir compte des caractéristiques de ce système familial sur les dynamiques de succession, qu’il s’agisse du processus lui-même, de la gouvernance, de la gestion des émotions ou de la transmission de la propriété psychologique. Elle montre que l’autorité patriarcale, l’égalité entre frères et sœurs, et l’endogamie sont des dimensions critiques de la réussite du processus de succession. / Family firms’ transmission from one generation to the next is the main challenge families are facing as they intend to ensure the sustainability of their heritage. For such organisations, it is however a strategic turning point. Most of the research dedicated to successions within the family, deal with Anglo-Saxon or European companies, and to a lesser extend to Asian ones. Moroccan SMEs, most of which are family firms, are going through a large-scale succession phenomenon. Over there, this is becoming a national challenge. Nevertheless, the existing succession models, derived from mainstream literature, do not fit with the national culture and the Moroccan family system. This family system is communitarian and endogamous (Todd, 1999), and its dynamics are very different from the ones visible in Europe or in Anglo-Saxon countries. This research aims at filling this gap. Built upon nine case studies analysing successors’ life stories, it manages to define a number of proposals allowing to deal with the specific impacts of this family system during the succession process. The different steps of the succession process, the governance issues, the management of emotions and the transfer of psychological ownership are taken into account. It concludes that patriarchal authority, equality among siblings and endogamy are critical determinants of the succession process’ success.

Schizophrenia is often a debilitating psychiatric disorder, characterised by both positive and negative symptoms, and cognitive and psychosocial impairments. The established disorder has been associated with a number of brain abnormalities, however it is at present unknown whether these brain changes occur prior to onset of schizophrenia, or in unaffected relatives with a familial vulnerability to develop the disorder, or only in those at high risk that go on to develop the disorder. Furthermore, most studies have been conducted cross-sectionally, which may have obscured subtle longitudinal changes in familial high risk individuals, and these studies tend to have focused on localized cortical gray matter , and thus it is unclear whether they affect different cortical parameters differentially. Prospective familial high risk studies utilizing surface based MRI programmes provide a good method to investigate this. In the Edinburgh High Risk Study, structural magnetic resonance imaging (MRI) scans of 150 young individuals at familial high risk of schizophrenia, 34 patients with first-episode schizophrenia and 36 matched controls were obtained. Of the high risk participants with scans suitable for analysis, 17 developed schizophrenia after the scans were taken, whilst 57 experienced isolated or sub-clinical psychotic symptoms, and 70 remained well. We used Freesurfer to extract volumetric and surface-based measurements of several cortical and localized sub-cortical regions with the aim of assessing whether any alterations found were present in all those at high risk, or selectively in the high risk cohort based on future clinical outcome, or only in those experiencing their first-episode of psychosis. It was found that those experiencing their first episode of schizophrenia exhibited significantly more widespread brain alterations compared to those at high risk or controls, both on a more global cortical level and in more localized regions of the cortex, with cortical thickness being generally thinner than in the other groups, and cortical surface area and gyrification increased compared to the other groups. An increased global surface area was also shared with the HR[ill] group, suggesting that this could be a marker that is predictive of future transition to psychosis. Within the high risk cohort, some brain alterations seemed to present as general vulnerability markers, specifically in the temporal lobe at baseline, whilst longitudinally both localized and global cortical alterations distinguished the high risk cohort from the control group, and a different developmental trajectory of the hippocampus was also found. These findings show that some brain alterations may be more accurately characterized as general vulnerability markers of the disorder, whilst some are specifically present in patients who have experienced their first episode of schizophrenia, whilst some also occur before disorder onset in those at high risk that go on to develop schizophrenia. The findings have some clinical implications, as they suggest that it is possible to assess who at high risk will go on to develop schizophrenia based on brain structural alterations. This may provide clinicians with an early window of opportunity for intervention, as it has been found that early intervention may improve patient's prognosis. The findings also have important implications for the understanding of the underlying eitology of schizophrenia, as they suggest that some of these alterations are present before illness onset, and not associated with medication effects, thus potentially lying on the causal path of developing schizophrenia.

Il est communément admis par les cliniciens tenant au modèle psychodynamique, qui ont entrepris des démarches d’exploration intrapsychique, que l’adolescent engagé dans des agirs sexuels violents présente un réaménagement psychique paradoxal des liens à ses figures parentales et un blocage de son processus de séparation-individuation.Il n’existe pas de recherches qui se soient spécifiquement intéressées à l’étude de facteurs facilitateurs/où empêcheurs du processus de séparation-individuation chez ces adolescents, alors que l’agir sexuel violent est pensé inscrit dans un continuum développemental spécifique à la période de l’adolescence. Il y aurait ainsi une nécessité à élargir l’angle de vue de la problématique. Un facteur, qui pourtant présente une grande influence sur le processus de séparation-individuation de l’adolescent à cette période, a pour l’instant peu été investigué : il s’agit de la capacité psychique inconsciente du groupe familial à offrir un espace de séparation-individuation durant cette phase de développement.L’hypothèse de cette recherche est que le blocage constaté chez ces adolescents, pourrait être en lien avec une difficulté plus globale du groupe familial auquel il appartient, à effectuer de son côté, ce travail psychique de séparation, auquel il est convoqué en parallèle à cette phase de développement. En appui sur les concepts psychodynamiques et psychanalytiques familiaux de « position dépressive familiale » (Roman, 1999), de « miroir familial » (Cuynet, 2001, sur notre proposition pour l’adolescence, Bernard, 2016) et « d’image inconsciente du corps familial » (Cuynet, 2005, 2010), nous avons élaboré un dispositif clinique d’évaluation pour évaluer cette dimension spécifique de la séparation au sein du groupe familial. Ce dispositif comprend des entretiens semi-dirigés avec le recours à des épreuves projectives familiales telles que « l’épreuve de génographie projective familiale » (Cuynet, 2001) et « l’épreuve du dessin de la maison de rêve » (Cuynet, 2005). A partir de la rencontre de sujets adolescents présentant cette problématique avec leur famille, le dispositif d’évaluation a été expérimenté, et la dimension de séparation évaluée. Les résultats de cette recherche, par la constitution d’études de cas approfondies, dans un esprit de recherche initiale qualitatif, ont mis en évidence l’intérêt de la réalisation d’un tel dispositif pour l’étude de la dimension de la séparation mais aussi pour la constitution d’hypothèses élargies de la compréhension de la problématique et du passage à l’acte des jeunes suivis. Les analyses de l’ensemble des études de cas montrent la difficulté pour le groupe familial à offrir un espace de différenciation pourtant nécessaire à cette période pour l’adolescent et répondent en faveur de la validation de l’hypothèse de cette recherche. La prise en compte de l’état de la position dépressive familiale du groupe, et du miroir familial ainsi constitué et de leur mise en travail, pourraient être des facteurs considérés comme leviers thérapeutiques pour favoriser l’évolution psychique de ces adolescents. Ils pourraient constituer également une piste de travail nécessaire dans la lutte contre la transmission transgénérationnelle fréquemment repérée dans cette problématique. / It is commonly recognized by psychodynamic model clinicians, who undertook intrapsychic exploration approaches, that a teenager responsible of violent sexual acts experiences a paradoxal psyche redevelopment of his links towards his parent figures and a blockade of his separation-individuation process.There is no research focusing on easing or inhibitor factors of the separation-individuation process of these teenagers, whereas the violent sexual acting is meant to belong to a development continuum, specific to the puberty period. There might be then a need to lead the problematic further. One factor that has a great influence on the separation-individuation process during puberty has not been much investigated until now: it is the unconscious psyche capacity of the family group to offer a space of separation-individuation during this development phase.The hypothesis of this research is that the blockade these teenagers suffer from is related to a greater difficulty of the family group those teenagers belong to, to achieve on its part this separation psyche work which it is responsible for, during this development phase.Thanks to the familial psychoanalytic and psychodynamic concepts of “Familial Depressive Position” (Roman, 1999), “Familial Mirror” (Cuynet, 2001, after our proposition for puberty, Bernard, 2016) and “Unconscious Image of the Familial Body” (Cuynet, 2005, 2010) we have created a clinical device to evaluate this particular dimension of the separation inside the family group. This device includes semi-structured interviews with family projective tests such as “The Familial Projective Genography” (Cuynet, 2001) and “The Dreamhouse Drawing Test” (Cuynet, 2005).This evaluation device has been tested and the separation dimension has been evaluated thanks to interviews with teenagers concerned by this situation in their family. The result of this research through in-depth case studies brought to light the interest of such a device for the study of the separation dimension but also to set up extended hypotheses about the comprehension of the problematic and the acting of the teenagers taking part to the device.The analyses of all the case studies show the difficulty the family group has to offer a differentiation space that is however necessary for the teenager during this period. Those analyses are then in favor of the validation of the hypothesis. Taking into account the state of the familial depressive position of the group and the familial mirror thus constituted and applying them, could be factors considered as therapeutic solutions to enable the psyche evolution of those teenagers. They could also represent a useful working basis in the fight against the transgenerational transmission that is frequently observed with this problematic.

Submitted by NADIA FAGUNDES GARCIA null (nadia.garcia@usp.br) on 2016-04-26T14:05:01Z No. of bitstreams: 1 Dissertação_Nádia_final.pdf: 1655190 bytes, checksum: 3ec5b191317646b34cbfef983fcdf111 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-04-29T15:08:25Z (GMT) No. of bitstreams: 1 garcia_nf_me_rcla.pdf: 1655190 bytes, checksum: 3ec5b191317646b34cbfef983fcdf111 (MD5) / Made available in DSpace on 2016-04-29T15:08:25Z (GMT). No. of bitstreams: 1 garcia_nf_me_rcla.pdf: 1655190 bytes, checksum: 3ec5b191317646b34cbfef983fcdf111 (MD5) Previous issue date: 2016-03-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A inatividade física e as dislipidemias são considerados fatores de risco para a gênese das doenças cardiovasculares. Estudos em animais mostram que o consumo de dieta contendo alto teor de lipídios leva à diminuição da resposta relaxante dependente do endotélio, o que pode ser prevenido pela realização de exercício físico. Entretanto, a maioria dos estudos investigou os efeitos do exercício físico em artérias de maior calibre e em modelos de dislipidemia induzida por dieta. Há escassez de estudos que avaliam os efeitos do exercício físico em artéria de menor calibre e, principalmente, em modelo que mimetiza a hipercolesterolemia familiar (HF). Portanto, o objetivo do presente estudo foi avaliar o efeito do treinamento físico aeróbio de moderada intensidade na reatividade vascular da artéria ilíaca em camundongos knockout para o receptor de LDL alimentados com dieta hiperlipídica. Foram utilizados camundongos wild type e knockout para o receptor de LDL (LDLR-/-) divididos em quatro grupos experimentais: wild type sedentário (WT), wild type treinado (WT/Ex), knockout sedentário (KO) e knockout treinado (KO/Ex). Os grupos WT foram alimentados com ração balanceada e os grupos KO com dieta hiperlipídica (38% lipídios). Os grupos WT/Ex e KO/Ex realizaram corrida em esteira (60-70%Vmax, 5 dias/semana, 60 min) durante oito semanas. A reatividade vascular em artéria ilíaca foi verificada através de curvas concentração-resposta a acetilcolina (ACh), nitroprussiato de sódio (SNP), fenilefrina (PHE) e ao análogo do tromboxano A2 (U46619). A determinação da produção de óxido nítrico (NO) foi realizada pela análise de fluorescência ao 4,5-diaminofluoresceína (DAF-2) e a produção de ânion superóxido pela análise da fluorescência derivada da oxidação da dihidroetidina (DHE). Foi quantificada a glicose, o colesterol total e os triglicerídeos sanguíneos. O grupo KO aumentou em 640% o ganho de peso corporal, 510% a gordura epididimal, 35% a glicose, 180% o colesterol total e 99% os triglicerídeos comparado ao grupo WT e o treinamento físico aeróbio foi eficaz em prevenir o ganho de peso e a gordura epididimal no grupo KO/Ex (167% e 121%, respectivamente). Nenhuma alteração foi verificada na glicose, colesterol total e triglicerídeos no soro. O relaxamento máximo induzido por ACh foi reduzido em 24% no grupo KO comparado ao grupo WT, sendo esta resposta normalizada no grupo KO/Ex, sem alteração na potência. A resposta máxima a PHE foi 65% maior, e resposta da potência foi 3 vezes maior na artéria ilíaca de animais do grupo KO comparado ao grupo WT. Nenhuma alteração na resposta foi encontrada aos agentes SNP e U46619. A produção de NO foi 47% menor no grupo KO comparado ao WT e a produção de espécies reativas de oxigênio (ERO) foi 48% maior no grupo KO comparado ao grupo WT. O treinamento físico preveniu o aumento na produção de ERO no grupo KO/Ex. Em conclusão, o exercício físico aeróbio, realizado por oito semanas, preveniu a disfunção endotelial na artéria ilíaca de camundongos LDLR-/- alimentados com dieta hiperlipídica. Este achado pode estar relacionado à menor produção de ERO, o que aumentaria a biodisponibilidade do NO. / Physical inactivity and dyslipidemia are considered risk factors for cardiovascular disease. A decrease in endothelium-dependent relaxation response, which can be prevented by physical exercise, had been showing in animals fed with high fat diet. However, most studies investigated the effects of physical exercise on large-caliber arteries using models of diet induced dyslipidemia. There are few studies that evaluate the effects of physical exercise in small-caliber artery and, mainly, in a model that mimics familial hypercholesterolemia (FH). Therefore, the aim of this study was to evaluate the effect of moderate intensity exercise training on vascular reactivity of iliac artery in FH model using mice lacking LDL receptor. Wild type and knockout mice (LDLR-/-) were divided into four groups: sedentary control (WT), trained control (WT/Ex), sedentary knockout (KO) and trained knockout (KO/Ex). Control groups were fed with standard chow and knockout groups with high fat diet. Trained groups ran on a treadmill (60-70% Vmax, 60 min, 5 days/week for 8 weeks). Concentration-response curves to acetylcholine (ACh), sodium nitroprusside (SNP), phenylephrine (PHE) and thromboxane A2 analogue (U46619) were done in iliac artery rings. Arterial production of nitric oxide and oxygen reactive species formation were assessed using fluorescence analysis (DAF-2 and DHE). Serum concentration of glucose, total cholesterol and triglyceride were determined using commercial kits. After eight weeks, the KO group had higher body weight gain (around 640%), epididymal fat (510%), glucose (35%), total cholesterol (180%) and triglycerides (99%) compared with WT group. Exercise training was effective to prevent body weight and epididymal fat gain in KO/Ex group (less 167% and 121%, respectively). No changes were observed in glucose, total cholesterol and triglycerides concentration. KO animals had lower maximal response evoked by ACh (about 24%) and higher maximal response to PHE (about 65%) compared with WT group. Exercise training prevented these alterations since KO/Ex group had vascular response similar to WT and WT/Ex groups. Endothelial dysfunction observed in KO group could be related to the reduced production of NO (about 47%) and the increased formation of oxygen reactive species (about 48%). These features were partially prevented by exercise training, KO/Ex group had lower formation of oxygen species and a slight higher NO prodution compared with KO group. In conclusion, aerobic exercise training carried out for eight weeks, prevented endothelial dysfunction in iliac artery from LDLR-/- mice fed with high fat diet. This finding could be related to the lower formation of reactive oxygen species in situ that increases the NO bioavailability.

Pneumonia intersticial familiar (PIF) é definida como a ocorrência de pelo menos dois casos de pneumonia intersticial fibrosante em uma mesma família biológica. Apesar do avanço sobre o tema em anos recentes, ainda não há estudos brasileiros nessa área. O objetivo deste estudo foi caracterizar uma amostra de pacientes brasileiros com PIF quanto aos aspectos clínicos, radiológicos, anatomopatológicos e genéticos, bem como analisar os respectivos comprimentos teloméricos (CT). Entre março de 2014 e novembro de 2017 foi realizada uma busca ativa por casos de PIF. Os pacientes identificados foram submetidos a testes de função pulmonar; tomografia computadorizada de alta resolução (TCAR) de tórax e a coleta de amostras de sangue. Ainda foram realizadas avaliações empregando ficha clínica padronizada. Amostras de tecido pulmonar foram obtidas e revisadas em seis casos. Empregando-se um kit apropriado, DNA genômico foi extraído de células brancas do sangue periférico. A avaliação do CT foi feita pelo método de Southern blot. Um painel envolvendo 154 genes de interesse foi desenvolvido pelo grupo de pesquisa e construído pela empresa Agilent Technologies. A pesquisa desses genes foi realizada empregando-se técnicas de sequenciamento de nova geração (NGS-Illumina). Foram selecionados 35 pacientes, com idade mediana de 66 (35,5-89,3) anos. Tabagismo e outras exposições ambientais estiveram presentes em 45,7% e 80% dos casos, respectivamente. Estertores finos foram identificados em 91,4% dos pacientes e baqueteamento digital em 20%. Os testes de função pulmonar foram classificados como restritivos em 20 (57,1%), normais em 10 (28,6%), inespecíficos em 4 (11,45) e obstrutivo leve em 1 (2,9%). A DCO esteve reduzida nos 30 pacientes em que foi possível pesquisá-la. Padrão típico de PIU na TCAR foi detectado em 6 (17,1%) pacientes e padrão indeterminado para PIU em outros 4 (11,4%). A maioria dos casos, 25 (71,4%), exibiu padrão tomográficoinconsistente com PIU. A revisão do material anatomopatológico revelou pneumonite intersticial com acentuação bronquiolocêntrica em quatro indivíduos. A grande maioria (85,7%) mostrou CTs inferiores ao percentil 50%. Quatro indivíduos exibiram CTs curtos e um muito curto. Foram identificadas variantes genéticas comuns no promotor do gene MUC5B (rs35705950), nos genes TOLLIP (rs111521887, rs5743894 e rs5743890) ou TERT (rs2736100) em 90% dos casos. Detectaram-se ainda sete variantes raras distintas. As alterações c.2594G>A e c.2146G>A do gene TERT, e c.394C>T do gene RTEL1, previamente descritas e associadas a telomeropatias. Uma anormalidade de TERT (c.1730G>A) e outra de RTEL1 (c.2299C>T) inéditas. Duas outras variantes raras encontradas já conhecidas, ainda não haviam sido associadas a doenças pulmonares: gene SHQ1 (c.828_831del) e gene WRAP53 (c.1558dupG). Em conclusão, pacientes brasileiros com PIF demonstram acentuada heterogeneidade fenotípica e genotípica. Este estudo identificou ainda duas novas variantes genéticas raras associadas a PIF e dois possíveis novos genes implicados na patogênese dessa doença. / Familial interstitial pneumonia (FIP) is defined as the occurrence of at least two cases of interstitial fibrosing pneumonias in the same biological family. Despite advances in the field in recent years, there are no Brazilian studies in this area. The aim of this study was to characterize a sample of Brazilian patients with PIF regarding clinical, radiological, histological and genetic aspects, as well as to analyze the respective telomeric lengths (TL). Between March 2014 and November 2017 an active search was conducted for FIP cases. The patients identified were submitted to pulmonary function tests, high resolution computed tomography (HRCT) of the chest and the collection of blood samples. In addition, a standardized clinical file was also fulfilled. Pulmonary tissue samples were obtained and reviewed in six cases. Using a suitable kit, genomic DNA was extracted from white peripheral blood cells. The TL measurements were done by Southern blot method. A panel of 154 genes of interest was developed by the research group and built by Agilent Technologies. Research on these genes was carried out using next generation sequencing techniques (NGSIllumina). Thirty-five patients were selected, with a median age of 66 (35.5-89.3) years. Smoking and other environmental exposures were present in, respectively, 45.7% and 80% of the cases. Fine crackles were identified in 91.4% of patients and digital clubbing in 20%. Pulmonary function tests were classified as restrictive in 20 (57.1%), normal in 10 (28.6%), non-specific in 4 (11.45) and mild obstructive in 1 (2.9%). The DLCO was reduced in the 30 patients in whom it was possible to investigate it. Typical pattern of UIP in HRCT was detected in 6 (17.1%) patients and undetermined pattern for UIP in another 4 (11.4%). The majority of cases, 25 (71.4%), showed a tomographic pattern inconsistent with UIP. The review of histological material revealed interstitial pneumonitis with bronchiolocentric accentuation in four individuals. The vast majority (85.7%) showed TL lower than the 50th percentile. Four individuals had short TL and one a very short TL. Commongenetic variants were identified in the MUC5B gene promoter (rs35705950), in the TOLLIP genes (rs111521887, rs5743894 and rs5743890) or TERT (rs2736100) in 90% of the cases. Seven different rare variants were also detected: the changes c.2594G> A and c.2146G> A of the TERT gene, and c.394C> T of the RTEL1 gene, previously described and associated with telomeropathy; one previously unknown abnormality of TERT (c.1730G> A) and another of RTEL1 (c.2299C> T); two additional rare genetic variants, had not yet been associated with pulmonary diseases: SHQ1 gene (c.828_831del) and WRAP53 gene (c.1558dupG). In conclusion, FIP Brazilian patients demonstrate marked phenotypic and genotypic heterogeneity. This study also identified two new rare genetic variants associated with FIP and two other possible new genes implicated in the pathogenesis of this disease.

A hipercolesterolemia familiar (HF) é uma doença autossômica dominante causada por mutações no gene do receptor de LDL (LDLR) (cromossomo 19p13.1 - p13.3), que alteram parcialmente ou totalmente a função do LDLR. A HF é também uma das doenças genéticas mais comuns com freqüências estimadas de heterozigotos e homozigotos de 1/500 e 1/1.000.000, respectivamente. Manifesta-se com altos níveis de LDL colesterol, arco corneal, xantomas tendíneos e sintomas prematuros de doença coronariana.. A grande heterogeneidade observada na manifestação clínica desta doença pode ser explicada, ao menos parcialmente, pelo amplo espectro de mutações no gene do LDLR. O presente estudo teve por objetivo a caracterização molecular do gene LDLR em pacientes com HF do Rio Grande do Sul (RS), Brasil. Para isso, foram obtidas amostras de DNA de 40 indivíduos provenientes de cinco macrorregiões do Estado, representando seis diferentes populações de ascendência européia, para a realização do seqüenciamento direto do gene do LDLR, com posterior análise por meio das ferramentas de bioinformática. Quinze mutações pontuais foram identificadas no gene do LDLR, a saber: c.408C>T (D115D), c.1616C>T (P518L), c.1773C>T (N570N) e c.2243A>G (D727G) na região codificadora, IVS6+36G>A, IVS6+171G>A, IVS11+56C>T, IVS11- 69G>T, IVS11-55A>C, IVS15-136A>G, IVS16+46C>T e IVS17-42A>G na região intrônica, e *52G>A, *105T>G e *141G>A na região 3\'-UTR. Destas, oito ainda não foram descritas na literatura (três situadas nos exons, quatro nos introns e uma na região 3\'-UTR). A mutação*52G>A foi previamente identificada em pacientes com HF da região Sudeste do Brasil, sugerindo que possa exercer um importante efeito na patogênese da HF em pacientes brasileiros. Em relação às macrorregiões do RS, os portugueses, italianos e espanhóis apresentaram o maior número de mutações dentre os grupos étnicos analisados. Assim, os resultados obtidos confirmam que existe um amplo de espectro de mutações no gene do LDLR. As mutações nas regiões intrônicas precisam ser investigadas sobre seu efeito potencial no desenvolvimento de HF. Considerando que este é o primeiro estudo que teve por objetivo a caracterização molecular de pacientes com HF no RS, novos estudos que visem a elucidação das bases moleculares da HF devem ser realizados, a fim de obter uma melhor caracterização genética desta doença no Brasil. / Familial hypercholesterolemia (FH) is an autosomal dominant disorder caused by mutations in the low-density lipoprotein receptor (LDLR) gene (chromosome 19p13.1 - p13.3), which alter partially or totally the LDLR function. FH is also one of the most common inherited disorders with frequencies of heterozygotes and homozygotes estimated to be 1/500 and 1/1.000.000, respectively. Affected individuals display high levels of LDL cholesterol, arcus corneae, tendon xanthomas and premature symptomatic coronary heart disease. The extensive heterogeneity observed in the clinical manifestation of this disorder may be explained, at least partially, by the broad spectrum of mutations identified in the LDLR gene. The present study had as the main goal the molecular characterization of the LDLR gene in patients with FH from Rio Grande do Sul (RS) State, Brazil. For this, DNA samples were obtained from 40 individuals living in five macroregions of RS, representing six different isolated populations of European ascendancy. The LDLR gene was subjected to the direct sequencing with further analysis through bioinformatics tools. Fifteen punctual mutations were identified in the LDLR gene, namely: c.408C>T (D115D), c.1616C>T (P518L), c.1773C>T (N570N) and c.2243A>G (D727G) in the coding region, IVS6+36G>A, IVS6+171G>A, IVS11+56C>T, IVS11-69G>T, IVS11-55A>C, IVS15-136A>G, IVS16+46C>T and IVS17-42A>G in the intronic region, and *52G>A, *105T>G and *141G>A in the 3\'-UTR region. Of these, eight were not yet described in the literature (three situated in exons, four in introns and one in 3\'- UTR region). The *52G>A mutation was previously identified in FH patients from Southeast Brazil, suggesting that it can exert an important effect in the pathogenesis of FH in Brazilian patients. In relation to the macroregions of Rio Grande do Sul, Portuguese, Italian and Spanish subjects carried the highest number of mutations among the ethnic groups analyzed. Thus, the results obtained confirm the existence of a broad spectrum of mutations in the LDLR gene. The mutations in intronic regions need to be investigated in relation to its potential effect in the development of FH. Taking into account that this is the first study that had as the goal the molecular characterization of FH patients in RS, further studies aimed at elucidating the molecular bases of FH should be performed, in order to obtain the better characterization of this disease in Brazil.

Hipercolesterolemia familiar (HF) é uma doença autossômica dominante, caracterizada por elevados níveis plasmáticos da lipoproteína de baixa densidade (LDL), desenvolvimento de xantoma tendíneo e arco corneal, além do aumento do risco de doença coronariana e acidente vascular cerebral prematuros. Frequentemente subdiagnosticada, estima-se que apenas 10% dos 400.000 indivíduos com HF no Brasil têm conhecimento da própria doença; afetando, desta forma, a qualidade e a expetativa de vida dos pacientes. Mutações no gene do receptor da LDL (LDLR) são consideradas as alterações genéticas mais frequentes para o desenvolvimento da hipercolesterolemia familiar, pois comprometem a capacidade de remoção das partículas de LDL circulantes, promovendo seu aumento em níveis plasmáticos. Já foram descritas mais de 1600 mutações diferentes no gene LDLR associadas ao fenótipo da HF; entretanto, ainda é difícil determinar em muitas delas o efeito deletério na atividade do receptor. O objetivo desse estudo foi identificar e caracterizar funcionalmente mutações no gene LDLR não descritas na literatura para determinar sua patogenicidade na hipercolesterolemia familiar. Foi avaliada a atividade residual de LDLR através da captação de LDL marcado com fluoróforo lipofílico em cultura de linfócitos T dos pacientes portadores das mutações analisadas após estimulação dos linfócitos T por mitógenos específicos. As mutações Cys82Ser, Thr404Ser, Gly529Arg e His285Tyr foram consideradas patogênicas por acarretarem diminuição da atividade residual do receptor de LDL. As mutações Glu 602X e His388ProfsX53 confirmaram sua patogenicidade e podem ser considerados como controle positivo para futuros ensaios funcionais. Estudos que esclareçam os mecanismos moleculares da HF e da relação genótipo/fenótipo abrem perspectivas para o desenvolvimento de terapias mais específicas na redução dos níveis de colesterol e, consequentemente, da morbidade e mortalidade associadas às doenças cardiovasculares. / Familial hypercholesterolemia (FH) is an autosomal dominant disorder characterized by elevated plasma levels of low-density lipoprotein (LDL), and development of corneal arcus tendinous xanthoma, and increased risk of coronary heart disease and premature stroke. Often misdiagnosed, it is estimated that only 10% of the 400.000 patients with FH in Brazil has knowledge of the disease itself, affecting in this way the quality and life expectancy of patients. Mutations in the LDL receptor (LDLR) are considered the most frequent genetic alterations for the development of familial hypercholesterolemia because compromise the ability of removal of circulating LDL particles, promoting its increase in plasma levels. Have been described over 1600 different mutations in the LDLR gene associated with the phenotype of FH, however, it is still difficult to determine in many of the deleterious effects on receptor activity. The aim of this study was to identify mutations in the LDLR gene and functionally characterize mutations not described in the literature to determine its pathogenicity in familial hypercholesterolemia. The residual activity of LDLR was evaluated by raising LDL labeled with lipophilic fluorophore in cultured T lymphocytes of patients with the analyzed mutations after stimulation of T lymphocytes by specific mitogen. The substitution mutations Cys82Ser, Thr404Ser, Gly529Arg e His285Tyr were considered pathogenic because it causes decrease of the residual activity of the LDL receptor in T lymphocytes. The His388ProfsX53 and Glu602X mutations confirmed their pathogenicity and can be considered as positive control for future functional assays. Studies to clarify the molecular mechanisms of HF and genotype/ phenotype open perspectives for the development of more specific therapies for reducing cholesterol levels, and therefore the morbidity and mortality associated with cardiovascular diseases.

L’élevage familial gaucho, une catégorie spéciale dans le monde de l’agriculture familiale, a été longuement ignoré par le milieu académique et par les décideurs. Le scénario où se déroule l’élevage familial gaucho – la Pampa – est un biome en profonde transformation qui, même riche, a le plus petit pourcentage de surface légalement protégée, en plus des passifs environnementaux à réversibilité difficile. Cette dévalorisation naît du manque de politiques d’appui au secteur et à l’exode massif des éleveurs familiaux qui, marginalisés ou forcés à choisir un système d’élevage en confinement des animaux [feedlot], finissent par louer ou vendre leurs terres à des entreprises multinationales dédiées aux monocultures de soja ou eucalyptus. Dans ce travail, nous analyserons les perceptions et les stratégies d’adaptation des éleveurs familiaux de viande bovine du biome Pampa de l’Argentine, de l’Uruguay et du Brésil face à la concurrence entre l’élevage traditionnel et l’agriculture à grande échelle. On utilise les concepts de vulnérabilité, résilience, adaptation et de double exposition aux risques climatiques et socio-économiques comme l’explication théorique plus consistante pour la réalité trouvée. Comme données primaires, nous avons interviewé 75 familles d’éleveurs [25 par pays] avec des formulaires semi-structurés qui cherchaient à identifier, à travers la méthode de l’analyse thématique, leurs modes de vie. C’est à partir de ces données que nous avons conclu que l’élevage familial gaúcho constitue un lien important entre emploi, production, durabilité et contention sociale. / Gaucho family livestock producers are a special social category in the family agriculure world. They have been traditionally ignored by researchers and decision-makers. Like their livelihoods, the context in which gaúcho family farming takes place – the Pampa biome – is also experiencing deep transformations. In spite of its rich biodiversity, the grasslands of the Pampas have one of the smallest areas of legally protected land in the world and is subject to environmental passives of difficult reversibility. This lack of interest in the Pampas and its family livestock producers has led to the absence of public policies tailored to that sector and the massive exodus of family farmers to the cities. Thus marginalized, and forced to intensify their production through systems such as feedlots, many of them choose to rent or directly sell their land to multinational soybean or eucalypt companies. We have conducted a thematic analysis of 75 semi-structured interviews to family livestock producers [25 per country]. The semi-structured questionnaires aimed at discovering their livelihoods and adaptation strategies. We use the concepts of adaptation, resilience, vulnerability and the double exposure framework as theoretical tools to explain this complex reality. We conclude that gaucho family livestock production is a positive link between employement, production, sustainability and social contention.