Inaug.-Diss.--Kiel. / At head of title: Aus der psychiatrischen und nerven-klinik in Kiel. Lebenslauf. "Literatur": p. 19.

Orientador: Pedro Henrique Saldanha / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-07-16T14:52:57Z (GMT). No. of bitstreams: 1 CastilloSalgado_MiguelAngel_M.pdf: 2635627 bytes, checksum: 7c48c3afdb3ef9ba2d2a0ba78a25bb18 (MD5) Previous issue date: 1986 / Resumo: Não informado / Abstract: Dermatoglyphics of the 183 patients with congenital heart disease (CHD), their parents and brothers and a sample of 170 subjects of the normal populations of são Paulo were studied. The propositi were patients of the Clinical Hospital of the Faculty of Medicine of the University of são Paulo, with diagnoses established through surgical treatment and clinical criteria such as electrocardiographic, roentgenologic and haemodinamic studies; no case with doubtful diagnosis was included. Informations concerning to sex, race, type of CHD, age of the parents by the time of birth, presence of CHD or another congenital malformations in the relatives, birth order of the propositi, number of abórtion and/or fetal deaths, expositionto the X-rays, infeccious diseases and use of drugs by their mothers during the first, three months of pregnancy. The following dermatoglyphic features were analised: total ridge count (TRC) in the fingers, a-b ridge count, atd angle, ulnar index, number of digital,sub-digital and axial triradii, and ridge count of the halux in the solar area. The statistical treatment consisted in the elaboration of frequency tables and distribution curves, means and their standard errors, analysis of variance between groups and intra-individual correlation coefficients between the different traits. The results allow the following eonelusions: 1. The reeurrenee of the CHD in the relatives suggest the existenee of hereditary faetorspredisposing the development of those anomalies. 2. The high eonsanguinity rate between the propositios parents suggest the existenee of recessive genes participating in the heredity of CHD. 3. The total ridge count on fingers and on the halux are a of the propositi are not significantly different from that of normal population. 4. The a-b ridge count is significantly higher in the propositi and their relatives. 5. The maximum atd anglereveals a statistically significant distal displacement of the axial palmar triradius' in the propositi and in their brothers. 6. The ulnar index reveals a highly significant radial displacement of the axial palmar triradius in the propositi and in their relatives, characterizing them as a group differentiated from the normal population. 7. The correlations between atd angle and ulnar index were highly significant in the relatives of the propositi but not significant in the normal population. This situation suggests that the genic constellation present in the related groups determine pleiotropically a distortion in the displacement of the axial. palmar triradius to the radial side. 8. The large number of significant and highly significant correlations in the propositi and in their normal relatives reveals that these groups share specific genic constellation which, besides determining a larger number of dermatoglyphic associations, it should predispose to cardiac teratogenesis during the critical period of embryogenic development common to the both structures / Mestrado / Mestre em Biologia e Patologia Buco-Dental

Keratoderma – pathological hyperkeratosis of palms and soles - is a cause of disability in many clinical situations, including the rare and heterogeneous group of inherited palmoplantar keratodermas (PPKs). The aim of this study was to work towards better understanding of molecular mechanisms active in the pathogenesis of PPK by the creation of a cell and tissue culture resource and its initial application to laboratory studies. My study was based on a diverse group of autosomal dominant disorders, previously ascertained in families from Scotland, in whom the precise genetic aetiology was known. I established a tissue and cell culture resource of inherited keratodermas of known single-gene aetiology from patients with proven keratin 1, 9, 17, loricrin and mitochondrial mutations. An additional pedigree with striate keratoderma with an unknown mutation was recruited, and the causative mutation identified as a novel heterozygous A-to-T transversion in exon 5 (c.430A>T) of the desmoglein 1 gene, converting an arginine residue to a premature termination codon (p. Arg144stop). The keratinocyte culture resource was established from patients with keratin 1, 9, 17 and loricrin mutations, as well as controls. Due to the pain associated with direct infiltration of plantar skin, biopsies were obtained using peripheral nerve block for plantar biopsy. The effectiveness of this approach, which may be useful for future administration of treatment, was made the subject of an open clinical trial. Histological and immunocytochemical studies were carried out on affected plantar skin obtained from PPK patients and compared to control tissue, in an attempt to identify common and distinct pathways resulting in hyperkeratosis. Histological changes, e.g. hypergranulosis, extent of hyperkeratosis, acanthosis or acantholysis, were not uniform across different subtypes of inherited PPK and varied even between individuals within subtypes. Prominent eosin staining of spinous cells was a common feature in inherited PPK due to underlying K1 and K17 mutations. Electron microscopy showed abnormal keratin filaments in PPK with underlying keratin mutations only but was not a uniform finding within subtypes, and other electron microscopic features also varied between individuals. Immunocytochemical study did not demonstrate significant differences in expression of a selection of markers of differentiation (keratins 1, 9, 14 and 17), and cornified envelope protein filaggrin. Abnormal involucrin expression was observed, with premature expression in basal and lower spinous layers in all PPK subtypes raising the possibility of a common underlying mechanism in the development of hyperkeratosis. Prominent loricrin staining was noted in areas of acantholysis in K1 and K9 subtypes, but was uniform across other subtypes. Markers of proliferation and apoptosis demonstrated no overt change in epidermal turnover, although it is possible that only small changes in proliferative index are required to produce plantar hyperkeratosis. Overall, using morphological criteria, plantar hyperkeratosis was not readily distinguishable between inherited PPK of different underlying genetic causes. This raises the possibility that many of the reported structural features of inherited PPK are secondary phenomena as opposed to critical steps in the pathogenesis of hyperkeratosis. Initial attempts at RNA extraction using laser and manual microdissection have to date been unsuccessful in generating RNA of the quality and concentration to run a pilot microarray experiment, using standard RNA extraction kits. Plans for future projects include the further development of a possible microarray experiment in the Pachyonychia Congenita type 2 pedigree with the McLean laboratory in Dundee. The tissue resource has been made available for collaborative study via the GENESKIN project, as well as through the McLean and Lane laboratories, Dundee for both functional studies and immortalisation of cell lines.

Dyskeratosis congenita (DC) is a multi system disorder that exhibits considerable clinical and genetic heterogeneity. It is characterised by mucocutaneous features, bone marrow failure and a predisposition to cancer. Research has identified mutations affecting several telomerase components and patients often have short telomeres, implicating defective telomere maintenance in this disease. Affected components include dyskerin, NOP10 and NHP2, which together with GAR1 form a protein core common to telomerase and all other H/ACA ribonucleoprotein complexes (H/ACA RNPs). Initially characterised as H/ACA RNP components important for pseudouridylation and rRNA processing, their role in the functionally distinct telomerase complex and telomere maintenance is less defined. In order to better understand their implications in DC, this study investigated the importance of these core proteins for the integrity and function of telomerase in human cells. RNAi knockdown studies demonstrated that dyskerin, NOP10 and NHP2 are necessary for the accumulation of TERC (telomerase RNA component); dyskerin and NOP10 for telomerase activity. Moreover, dyskerin was found to be important for maintaining telomere length over time. The impact of NOP10 and NHP2 missense mutations was also analysed in vitro, which indicated that they impair TERC accumulation. The potential effect on pseudouridylation was also considered in this study; the analysis of other H/ACA RNA levels in these knockdown experiments and in a cohort of patients with DKC1 mutations revealed an irregular and inconsistent impact compared to that observed on TERC. Finally, defective telomere maintenance is heavily implicated as the primary cause of DC and very short telomeres have been proposed as a diagnostic marker. This study investigated telomere length in a patient cohort of unprecedented size. It demonstrated the prevalence of the telomere length defect, but telomere length was not found to correlate with either genetic subtype or disease severity, implicating the rate of telomere shortening as the correlating factor instead.

Malformações congênitas ocorrem em aproximadamente 2% a 3% dos nascidos vivos, podendo compreender desde malformações discretas até graves defeitos que comprometem a sobrevida. Estima-se que cerca de 6% dos casos de defeitos congênitos ocorram devido à presença de anomalias cromossômicas. Essa prevalência é, provavelmente, subestimada, uma vez que malformações mais graves podem levar a perdas fetais antes que um diagnóstico seja possível e, algumas vezes, as alterações cromossômicas ou desequilíbrios genômicos associados às malformações congênitas não são identificados. Este estudo teve como objetivo geral identificar rearranjos cromossômicos e variação do número de cópias (CNVs) do genoma por meio da investigação de microdeleções e microduplicações em recém-nascidos com malformações congênitas de etiologia desconhecida. Este trabalho fez parte de um projeto de desenvolvimento tecnológico para o estabelecimento do método de análise comparativa de genomas em um hospital universitário público. Foi realizado um estudo retrospectivo em 35 amostras de DNA estocadas em biorrepositório por meio da análise total do genoma pela técnica de Hibridação Genômica Comparativa baseada em microarranjos (array-CGH). Todas as CNVs detectadas foram comparadas com as descritas em bancos públicos de dados genômicos, e sua significância clínica foi estimada. Treze alterações genômicas foram detectadas em 12/35 (34,3 %) das amostras. Em 4/35 (11.4%) dessas, os desequilíbrios genômicos puderam ser definidos como patogênicos e causalmente relacionados às anomalias congênitas; em 5/35 (14.3%) das amostras, CNVs de significado clínico incerto foram identificadas; e, em 4/35 (11.4%), variantes normais foram detectadas. Entre os 4 casos cujos resultados foram considerados causalmente relacionados com os achados clínicos, 2/4 (50%) tinham alterações patogênicas que estão reconhecidamente associados à síndromes de microdeleção definidas. Em 2/4 amostras (50%), os desequilíbrios cromossômicos encontrados, ainda que preditos como patogênicos, não haviam sido previamente associados à entidades clínicas reconhecidas. O uso de array-CGH indicou a presença na amostra estudada de rearranjos cromossômicos não identificados previamente, permitindo a identificação de regiões cromossômicas relacionadas à algumas anomalias congênitas. Além disso, ainda que a interpretação dos resultados deva ser refinada, os dados sugerem que a análise comparativa de genomas por array-CGH seja considerada como investigação de primeira linha no rastreamento seletivo para análise prospectiva/retrospectiva de amostras de DNA em programas de monitoramento de defeitos congênitos. / Congenital malformations occur in approximately 2% to 3% of live births and may comprise from mild to severe malformations defects that compromise survival. It is estimated that about 6% of the cases of birth defects occur due to the presence of chromosomal abnormalities. This prevalence is probably underestimated, since more severe malformations may lead to fetal loss before a diagnosis is possible, and sometimes, the chromosomal aberrations or genomic imbalances associated with congenital malformations are not identified. This study had as main objective to identify chromosomal rearrangements and copy number variations (CNVs) in the genome through the investigation of microdeletions and microduplications in newborns with congenital malformations of unknown etiology. This work was part of a technological development project for the establishment of the method of comparative analysis of genomes in a public university hospital. A retrospective study was performed on 35 DNA samples stored in a biorepository through whole-genome based microarrays Comparative Genomic Hybridization (array-CGH). All CNVs detected were compared with those described in public genome databases, and their clinical significance was estimated. Thirteen genomic alterations were detected in 12/35 (34.3%) of the samples. On 4/35 (11.4%) of these, the genomic imbalances were defined as pathogenic and causally related to congenital anomalies; in 5/35 (14.3%) samples CNVs of uncertain clinical significance were identified and in 4/35 (11.4%), normal variants were detected. Among the 4 cases whose results were considered causally related to clinical findings, 2/4 (50%) had pathogenic changes that are associated with well-known microdeletion syndromes. In 2/4 samples (50%), chromosomal imbalances found, although predicted as pathogenic, have not been previously associated with the recognized clinical entities. The use of array-CGH indicated the presence of chromosomal rearrangements not previously identified, allowing the identification of chromosomal regions related to some congenital anomalies. Moreover, although the interpretation of the results should be refined, the data suggest that comparative genome analysis by array-CGH must be considered as first-line investigation in selective screening for prospective/retrospective analysis of DNA samples in birth defects monitoring programs.

Telomeres are regions of repetitive DNA bound with a set of specialized proteins required to protect chromosomes from fusing with each other and from eliciting DNA damage response. Dysfunctional telomere maintenance can lead to premature cellular senescence, premature organismal aging and cancer predisposition. In the last few years the evidence has emerged indicating a link between dysfunctional maintenance of telomeres and defective DNA damage response. The objective of this project was to explore further this link by examining effects of some DNA damage response proteins on telomeres that have not been examined before and examining DNA damage response in cells in which telomeres are dysfunctional as a result of alterations in genes not directly involved in DNA damage response. We have developed a method, termed IQ-FISH, for accurate identification of average telomere length in interphase cells from individuals with defective DNA damage response. By applying IQ-FISH we could successfully measure telomere lengths in cell lines from patients that are heterozygous (+/-) and cell lines from patients or animals that are homozygous (-/-) with respect to mutations in these genes. We then analysed telomere length and function, as well as DNA damage response, in lymphoblastoid cell lines originating from BRCA1 and BRCA2 carriers (+/-) and also a single fibroblast cell line from a patient with bi-allelic mutations in BRCA2 (-/-). In addition we have analysed a mouse embryonic stem cell line in which Brca1 was deleted (Brca1-/-) by gene targeting. Our results show lack of correlation between DNA damage response and telomere maintenance in heterozygous cell lines (with the exception of one BRCA1+/- cell line) but a clear positive correlation in the case of cell lines with homozygous mutations. Finally, as a model for telomere dysfunction we have chosen cell lines from Dyskeratosis Congenita (DC) patients. DC is a rare progressive congenital disorder which results in premature aging. DC is primarily a disorder of dysfunctional telomere maintenance and we used cell lines from patients with mutations in DKC1, a gene encoding a protein termed Dyskerin which forms a part of the telomerase enzyme complex. Our results indicate that DC cells with dysfunctional DKC1 may have a dysfunctional DNA damage response.

Thesis (MAppSc)--University of South Australia, 1996

Thesis (MAppSc)--University of South Australia, 1996

Malformações congênitas ocorrem em aproximadamente 2% a 3% dos nascidos vivos, podendo compreender desde malformações discretas até graves defeitos que comprometem a sobrevida. Estima-se que cerca de 6% dos casos de defeitos congênitos ocorram devido à presença de anomalias cromossômicas. Essa prevalência é, provavelmente, subestimada, uma vez que malformações mais graves podem levar a perdas fetais antes que um diagnóstico seja possível e, algumas vezes, as alterações cromossômicas ou desequilíbrios genômicos associados às malformações congênitas não são identificados. Este estudo teve como objetivo geral identificar rearranjos cromossômicos e variação do número de cópias (CNVs) do genoma por meio da investigação de microdeleções e microduplicações em recém-nascidos com malformações congênitas de etiologia desconhecida. Este trabalho fez parte de um projeto de desenvolvimento tecnológico para o estabelecimento do método de análise comparativa de genomas em um hospital universitário público. Foi realizado um estudo retrospectivo em 35 amostras de DNA estocadas em biorrepositório por meio da análise total do genoma pela técnica de Hibridação Genômica Comparativa baseada em microarranjos (array-CGH). Todas as CNVs detectadas foram comparadas com as descritas em bancos públicos de dados genômicos, e sua significância clínica foi estimada. Treze alterações genômicas foram detectadas em 12/35 (34,3 %) das amostras. Em 4/35 (11.4%) dessas, os desequilíbrios genômicos puderam ser definidos como patogênicos e causalmente relacionados às anomalias congênitas; em 5/35 (14.3%) das amostras, CNVs de significado clínico incerto foram identificadas; e, em 4/35 (11.4%), variantes normais foram detectadas. Entre os 4 casos cujos resultados foram considerados causalmente relacionados com os achados clínicos, 2/4 (50%) tinham alterações patogênicas que estão reconhecidamente associados à síndromes de microdeleção definidas. Em 2/4 amostras (50%), os desequilíbrios cromossômicos encontrados, ainda que preditos como patogênicos, não haviam sido previamente associados à entidades clínicas reconhecidas. O uso de array-CGH indicou a presença na amostra estudada de rearranjos cromossômicos não identificados previamente, permitindo a identificação de regiões cromossômicas relacionadas à algumas anomalias congênitas. Além disso, ainda que a interpretação dos resultados deva ser refinada, os dados sugerem que a análise comparativa de genomas por array-CGH seja considerada como investigação de primeira linha no rastreamento seletivo para análise prospectiva/retrospectiva de amostras de DNA em programas de monitoramento de defeitos congênitos. / Congenital malformations occur in approximately 2% to 3% of live births and may comprise from mild to severe malformations defects that compromise survival. It is estimated that about 6% of the cases of birth defects occur due to the presence of chromosomal abnormalities. This prevalence is probably underestimated, since more severe malformations may lead to fetal loss before a diagnosis is possible, and sometimes, the chromosomal aberrations or genomic imbalances associated with congenital malformations are not identified. This study had as main objective to identify chromosomal rearrangements and copy number variations (CNVs) in the genome through the investigation of microdeletions and microduplications in newborns with congenital malformations of unknown etiology. This work was part of a technological development project for the establishment of the method of comparative analysis of genomes in a public university hospital. A retrospective study was performed on 35 DNA samples stored in a biorepository through whole-genome based microarrays Comparative Genomic Hybridization (array-CGH). All CNVs detected were compared with those described in public genome databases, and their clinical significance was estimated. Thirteen genomic alterations were detected in 12/35 (34.3%) of the samples. On 4/35 (11.4%) of these, the genomic imbalances were defined as pathogenic and causally related to congenital anomalies; in 5/35 (14.3%) samples CNVs of uncertain clinical significance were identified and in 4/35 (11.4%), normal variants were detected. Among the 4 cases whose results were considered causally related to clinical findings, 2/4 (50%) had pathogenic changes that are associated with well-known microdeletion syndromes. In 2/4 samples (50%), chromosomal imbalances found, although predicted as pathogenic, have not been previously associated with the recognized clinical entities. The use of array-CGH indicated the presence of chromosomal rearrangements not previously identified, allowing the identification of chromosomal regions related to some congenital anomalies. Moreover, although the interpretation of the results should be refined, the data suggest that comparative genome analysis by array-CGH must be considered as first-line investigation in selective screening for prospective/retrospective analysis of DNA samples in birth defects monitoring programs.

Malformações congênitas ocorrem em aproximadamente 2% a 3% dos nascidos vivos, podendo compreender desde malformações discretas até graves defeitos que comprometem a sobrevida. Estima-se que cerca de 6% dos casos de defeitos congênitos ocorram devido à presença de anomalias cromossômicas. Essa prevalência é, provavelmente, subestimada, uma vez que malformações mais graves podem levar a perdas fetais antes que um diagnóstico seja possível e, algumas vezes, as alterações cromossômicas ou desequilíbrios genômicos associados às malformações congênitas não são identificados. Este estudo teve como objetivo geral identificar rearranjos cromossômicos e variação do número de cópias (CNVs) do genoma por meio da investigação de microdeleções e microduplicações em recém-nascidos com malformações congênitas de etiologia desconhecida. Este trabalho fez parte de um projeto de desenvolvimento tecnológico para o estabelecimento do método de análise comparativa de genomas em um hospital universitário público. Foi realizado um estudo retrospectivo em 35 amostras de DNA estocadas em biorrepositório por meio da análise total do genoma pela técnica de Hibridação Genômica Comparativa baseada em microarranjos (array-CGH). Todas as CNVs detectadas foram comparadas com as descritas em bancos públicos de dados genômicos, e sua significância clínica foi estimada. Treze alterações genômicas foram detectadas em 12/35 (34,3 %) das amostras. Em 4/35 (11.4%) dessas, os desequilíbrios genômicos puderam ser definidos como patogênicos e causalmente relacionados às anomalias congênitas; em 5/35 (14.3%) das amostras, CNVs de significado clínico incerto foram identificadas; e, em 4/35 (11.4%), variantes normais foram detectadas. Entre os 4 casos cujos resultados foram considerados causalmente relacionados com os achados clínicos, 2/4 (50%) tinham alterações patogênicas que estão reconhecidamente associados à síndromes de microdeleção definidas. Em 2/4 amostras (50%), os desequilíbrios cromossômicos encontrados, ainda que preditos como patogênicos, não haviam sido previamente associados à entidades clínicas reconhecidas. O uso de array-CGH indicou a presença na amostra estudada de rearranjos cromossômicos não identificados previamente, permitindo a identificação de regiões cromossômicas relacionadas à algumas anomalias congênitas. Além disso, ainda que a interpretação dos resultados deva ser refinada, os dados sugerem que a análise comparativa de genomas por array-CGH seja considerada como investigação de primeira linha no rastreamento seletivo para análise prospectiva/retrospectiva de amostras de DNA em programas de monitoramento de defeitos congênitos. / Congenital malformations occur in approximately 2% to 3% of live births and may comprise from mild to severe malformations defects that compromise survival. It is estimated that about 6% of the cases of birth defects occur due to the presence of chromosomal abnormalities. This prevalence is probably underestimated, since more severe malformations may lead to fetal loss before a diagnosis is possible, and sometimes, the chromosomal aberrations or genomic imbalances associated with congenital malformations are not identified. This study had as main objective to identify chromosomal rearrangements and copy number variations (CNVs) in the genome through the investigation of microdeletions and microduplications in newborns with congenital malformations of unknown etiology. This work was part of a technological development project for the establishment of the method of comparative analysis of genomes in a public university hospital. A retrospective study was performed on 35 DNA samples stored in a biorepository through whole-genome based microarrays Comparative Genomic Hybridization (array-CGH). All CNVs detected were compared with those described in public genome databases, and their clinical significance was estimated. Thirteen genomic alterations were detected in 12/35 (34.3%) of the samples. On 4/35 (11.4%) of these, the genomic imbalances were defined as pathogenic and causally related to congenital anomalies; in 5/35 (14.3%) samples CNVs of uncertain clinical significance were identified and in 4/35 (11.4%), normal variants were detected. Among the 4 cases whose results were considered causally related to clinical findings, 2/4 (50%) had pathogenic changes that are associated with well-known microdeletion syndromes. In 2/4 samples (50%), chromosomal imbalances found, although predicted as pathogenic, have not been previously associated with the recognized clinical entities. The use of array-CGH indicated the presence of chromosomal rearrangements not previously identified, allowing the identification of chromosomal regions related to some congenital anomalies. Moreover, although the interpretation of the results should be refined, the data suggest that comparative genome analysis by array-CGH must be considered as first-line investigation in selective screening for prospective/retrospective analysis of DNA samples in birth defects monitoring programs.