Current interventions used to reduce skeletal fragility are insufficient at enhancing bone across multiple hierarchical levels. Bisphosphonates, such as Zoledronate (ZOL), treat a variety of bone disorders by increasing bone mass and bone mineral density to decrease fracture risk. Despite the mass-based improvements, bisphosphonate use has been shown to compromise bone quality. Alternatively, Raloxifene (RAL) has recently been demonstrated to improve tissue quality and overall mechanical properties by binding to collagen and increasing tissue hydration in a cell-independent manner. We hypothesized that a combination of RAL and ZOL would improve mechanical and material properties of bone more than either monotherapy alone by enhancing both quantity and quality of bone. In this study, wildtype (WT) and heterozygous (OIM+/-) male mice from the Osteogenesis Imperfecta (OI) murine model were treated with either RAL, ZOL, or RAL and ZOL from 8 weeks to 16 weeks of age. Combination treatment resulted in higher trabecular architecture, cortical mechanical properties, and cortical fracture toughness in diseased mouse bone. Two fracture toughness properties, direct measures of the tissue’s ability to resist the initiation and propagation of a crack, were significantly improved with combination treatment in OIM+/- compared to control. There was no significant effect on fracture toughness with either monotherapy alone in either genotype. Following the mass-based effects of ZOL, bone volume fraction was significantly higher with combination treatment in both genotypes. Similar results were seen in trabecular number. Combination treatment resulted in higher ultimate stress in both genotypes, with RAL additionally increasing ultimate stress in OIM+/-. RAL and combination treatment in OIM+/- also produced a higher resilience compared to the control. Given no significant changes in cortical geometry, these mechanical alterations were likely driven by the quality-based effects of RAL. In conclusion, this study demonstrates the beneficial effects of using combination therapy to increase bone mass while simultaneously improving tissue quality, especially to enhance the mechanical integrity of diseased bone. Combination therapies could be a future mechanism to improve bone health and combat skeletal fragility on multiple hierarchical levels.

Submitted by Luis Guilherme Macena (guilhermelg2004@gmail.com) on 2013-05-07T15:40:21Z No. of bitstreams: 1 Tese - Lia Martins Moreira.pdf: 3329572 bytes, checksum: fc270c2266a2f89e6d2f0fbfece0fa1e (MD5) / Made available in DSpace on 2013-05-07T15:40:21Z (GMT). No. of bitstreams: 1 Tese - Lia Martins Moreira.pdf: 3329572 bytes, checksum: fc270c2266a2f89e6d2f0fbfece0fa1e (MD5) Previous issue date: 2012 / Fundação Oswaldo Cruz. Instituto Fernandes Figueira. Departamento de Ensino. Programa de Pós-Graduação em Saúde da Criança e da Mulher. Rio de Janeiro, RJ, Brasil / Investigamos a atuação da clínica fisioterapêutica, no tocante à funcionalidade dos pacientes com osteogênese imperfeita (OI), inscritos no Programa de Tratamento de Osteogênese Imperfeita do Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira da Fundação Oswaldo Cruz (INSMCA/IFF/FIOCRUZ), coordenado pelo Centro de Referência de Osteogênese Imperfeita do INSMCA/FIOCRUZ (CROI/IFF). A atuação da clínica fisioterapêutica considera aspectos como: (a) a funcionalidade; (b) os estímulos táteis e, (c) as ações preventivas como facilitadores do desenvolvimento motor e da reabilitação de indivíduos com OI. O objetivo geral foi analisar a funcionalidade dos indivíduos com OI em tratamento fisioterapêutico no CROI/IFF. Os objetivos específicos foram: (a) analisar o processo de locomoção dos pacientes com OI, mapeando os fatores que o influenciam; (b) avaliar a capacidade funcional mediante a mensuração da amplitude de movimento, e (c) discutir os exercícios, atividades fisioterapêuticas e orientações aos familiares propostas para os pacientes com OI. Lançamos mão de duas abordagens: as análises bioestatísticas e a perspectiva etnográfica, compondo uma pesquisa exploratória e um estudo de caso institucional. O estudo foi desenvolvido com pacientes com OI atendidos no CROI/IFF entre 2004 e 2008, totalizando 92 sujeitos de pesquisa. O Epi-Info versão 3.4 e o SPSS versão 15 foram empregados na construção de banco de dados e para cálculos estatísticos. Em relação à perspectiva etnográfica, tomou-se como fonte as anotações feitas num diário de campo durante as diversas avaliações fisioterapêuticas, analisando-as via uma codificação analítica qualitativa seguida pela análise semiótica. Os resultados são apresentados em três artigos. O primeiro expõe a relação entre a marcha independente na OI e os fatores que a influenciam, ressaltando-se a associação negativa entre hipotonia, número de fraturas e os desfechos de interesse e positiva entre marcha independente e OI tipo I. O segundo aborda a hipermobilidade articular como característica clínica da OI e a hipotonia observada nesta amostra como aspectos contribuintes às limitações funcionais, estabelecendo que a primeira se associa à idade e a segunda ao tipo de OI. O terceiro remete-se aos relatos da experiência de campo marcando as múltiplas dimensões da ação fisioterapêutica assentada num diálogo que envolve os pacientes, seus familiares e o fisioterapeuta, aprofundando a compreensão do movimento humano nesta doença. Salienta-se que o incentivo precoce reduziu as contraturas musculares com melhora do tônus muscular; os manuseios fisioterapêuticos facilitaram a integração da percepção do corpo e ajudou a afastar o medo das fraturas, permitindo a construção de uma nova imagem funcional. Concluímos que como a OI é uma doença rara, necessita uma abordagem multidisciplinar e, no que tange à Fisioterapia, esta deve focar questões relevantes à mobilidade dos pacientes, procurando estabelecer estratégias de ação e caminhos para a maximização e ou recuperação de sua independência funcional. / This study investigates a clinical physical therapy approach concerning functionality of patients with osteogenesis imperfecta (OI) enrolled at the OI Treatment Programme at the Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira - Fundação Oswaldo Cruz (INSMCA/IFF/FIOCRUZ), which is coordinated by the OI Reference Center at the same institution (CROI/IFF). Physical therapy assistance provided by the CROI/IFF takes into consideration aspects such as: a) functionality, b) tactile stimuli, and c) preventive actions regarded as key elements for promoting motor development and rehabilitation of individuals with OI. The objective was to analyse the functionality of patients with OI under physical therapy treatment carried out as part of the cited programme, and, more specifically, a) to analyse the locomotion process in those patients, mapping out factors that influence it; b) to evaluate functional capacity by measuring the amplitude of movement, and c) to discuss with patients and their families physical therapy activities, exercises, and orientations. The analysis followed two different approaches: biostatistical analyses and an ethnographic perspective, comprising an exploratory research and an institutional case study. The research subjects were 92 patients with OI treated at the CROI/IFF between 2004 and 2008. Epi-Info version 3.4 and SPSS version 15 were used to construct the database and for statistical analyses. Ethnographic approach was based on fieldnotes taken during physical therapy assessments. Those fieldnotes were submitted to a two-part codification - open and focused - followed by semiotic techniques of analysis. Results are presented in three articles. The first one investigates the relationship between the locomotion process in patients with OI and connected factors, highlighting that a negative association was observed between hypotonia, number of fractures and the outcomes of interest, and a positive association was observed between independent walk and OI type I. The second article focuses on articular hypermobility, a clinical feature of OI, and the presence of hypotonia in those patients as major issues that tend to worsen functional limitations. It discusses the association between hypermobillity and age and hypotonia and type of OI. The third article examines accounts of physical therapy assistance and emphasizes its multiple dimensions. Moreover, the paper argues that this kind of assistance should be based on a dialogue between patients/families and physical therapists, contributing to deepen the understanding of human movement in relation to OI. We argue that early encouragement to perform active movements within a safe environment reduced articular contratures and enhanced muscular tonus; physiotherapy manipulation facilitated the integration of body perception, contributed to reduce fear of fractures and allowed the construction of a new functional image. Finally, we highlight the significance of a multidisciplinary approach to rare diseases such as OI and, in the case of physical therapy, the importance of establishing strategic actions in order to recover or enhance functional independence of patients with OI.

Made available in DSpace on 2014-07-22T13:16:49Z (GMT). No. of bitstreams: 2 Antilia Januário Martins.pdf: 1137257 bytes, checksum: 159a0e5bfe6dfad2a82e1e8948f1b580 (MD5) license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) Previous issue date: 2011 / Fundação Oswaldo Cruz. Instituto Fernandes Figueira. Departamento de Ensino. Programa de Pós-Graduação em Saúde da Criança e da Mulher. Rio de Janeiro, RJ, Brasil. / A Osteogênese Imperfeita (OI) é uma doença genética causada por mutações no colágeno tipo I, caracterizada pela fragilidade óssea e fraturas recorrentes, que podem evoluir com graves deformidades e limitações funcionais. Atualmente, o tratamento baseia-se em procedimentos cirúrgico-ortopédicos, na reabilitação fisioterápica e no uso de medicamentos, como os bisfosfonatos, que buscam reduzir a fragilidade óssea. Em 2001, com a aprovação da Portaria 2.305 do Ministério da Saúde, foi implantado o programa de tratamento com o pamidronato dissódico para crianças, adolescentes e jovens com OI na rede pública de saúde no Brasil. No entanto, o tratamento com bisfosfonatos na OI ainda é inovador e os estudos que existem na literatura, em sua maioria, baseiam-se no resultado positivo do uso dos medicamentos e intervenções em relação ao aumento na densidade mineral óssea, diminuição do número de fraturas esqueléticas, diminuição da dor e melhora no crescimento. Poucos estudos buscam avaliar a qualidade de vida, considerado hoje um desfecho importante na área da saúde. Neste sentido, buscou-se avaliar a qualidade de vida dos adolescentes inseridos no programa de tratamento com bisfosfonatos (pamidronato e alendronato) do Instituto Fernandes Figueira/Fiocruz, através de uma abordagem quantitativa e qualitativa. Participaram deste estudo 37 adolescentes, que na primeira etapa responderam ao instrumento de avaliação de qualidade de vida da Organização Mundial da Saúde (WHOQOL-100) e, seus responsáveis, ao questionário sociodemográfico. Na segunda etapa, os adolescentes participaram do grupo focal. A maioria dos adolescentes tinha idade maior ou igual a 15 anos (62,2%), OI do tipo I (51,4%) e encontrava-se em tratamento ambulatorial fazendo uso de alendronato (59,4%). Não se confirmou a associação entre as características sociodemográficas e o tipo de tratamento e, também, o tipo de OI. Os domínios do WHOQOL-100 que obtiveram os melhores escores de avaliação foram religiosidade/espiritualidade/crenças pessoais (77,6%) e relações sociais (73,7%) e o pior escore foi o do meio ambiente (64,9%). O único domínio que apresentou diferença estatisticamente significativa foi o meio-ambiente entre os tipos de OI (p=0,091). Como não se obteve diferença estatisticamente significante nos domínios, com exceção do meio ambiente, os resultados do WHOQOL-100 sugerem que a percepção que os adolescentes deste estudo têm de sua qualidade de vida independe do tipo de OI (I, III e IV) ou do tipo de tratamento (ambulatório/alendronato e internação/pamidronato). Estes resultados são semelhantes aos do grupo focal, quando se percebeu as relações sociais como a dimensão mais valorizada pelos participantes, o que é comum nesta fase. Estes destacaram as vivências familiares e sociais, muitas vezes, como experiências estigmatizantes em função das marcas que a OI proporciona como a fragilidade, a baixa estatura, as cicatrizes corporais, dentre outras. Estas marcas também afetam o desenvolvimento da sexualidade nestes jovens. Mas a vivência dos adolescentes com OI é muito semelhante e independe da gravidade da doença ou do tipo de tratamento a que estão submetidos. Desta forma, a qualidade de vida é indiscutivelmente uma questão subjetiva, que se estabelece dentro de um contexto social, mas cuja construção não se dá comparativamente nem de forma vertical em relação às marcas da doença, pois um adolescente que possui um marca muito grande pode considerar a sua qualidade de vida melhor do que outro que possui uma marca menor. A qualidade de vida, portanto, não está necessariamente vinculada à marca que o adolescente tem, mas principalmente à forma como ele lida com a marca e enfrenta as barreiras criadas por ela. / Osteogenesis Imperfecta (OI) is a genetic illness caused by mutations in the type I collagen, characterized by the bone fragility and recurring fractures, that can result in severe deformities and functional limitations. Currently, treatment encompasses surgical-orthopedic procedures, physiotherapeutic rehabilitation and the administration of bisphosphonates to increase bone mass and reduce the incidence of fractures. In 2001, with the approval of a federal law, a treatment program using pamidronate disodium for children, adolescents and young adults with OI was implemented in the Brazilian public health system. However, bisphosphonate therapy is still innovative and the current studies found in the literature are in their majority focused on the positive results of drugs and interventions regarding the increase in the bone mineral density, reduction of skeletal fractures rates, pain and improvements in the growth patterns. Few studies aim to evaluate the quality of life of these patients, considered today an important outcome in health discussions. The present study had as its objective the evaluation of the quality of life of adolescents enrolled in the bisphosphonate (pamidronate and alendronate) therapeutic program of the Instituto Fernandes Figueira/Fiocruz, through a qualitative and quantitative approach. The first phase of the study consisted on the administration of the World Health Organization Quality of Life Assessment Instrument (WHOQOL-100) to 37 adolescents and of the sociodemographic questionnaire to parents. The next phase of the study consisted of a focus group. The majority of the adolescents were 15 years of age or older (62,2%), had OI Type I (51,4%) and were on ambulatory treatment using alendronate (59,4%). The association between the socio-demographic characteristics and the type of treatment was not confirmed in addition to the Type of OI. The domains of the WHOQOL-100 that attained the higher scores were spirituality/religion/personal beliefs (77,6%) and social relationships (73,7%) and the worst scores were observed for the environment domain (64,9%). The only domain that presented statistically significant difference when it came to type of OI was environment (p=0,091). The lack of statistical significance within the domains, with the exception of environment, in the WHOQOL-100 suggests that adolescents` perception of their quality of life is independent from type of OI (I, III and IV) and treatment (ambulatory/alendronate and hospitalization/pamidronate). The results are similar those obtained on the focus group, for this technique the social relationships were perceived as the more valuable dimension by the participants, common in adolescence. The subjects point out social and family experiences as frequently stigmatized due to marks associated with OI and such as fragility, decreased stature, bodily scars, amongst others. These marks and blemishes of character also affect the development of sexuality in these adolescents. But the experience of the adolescents with OI is very similar and is independent of the severity of the illness or of the treatment plan. Thus, quality of life is indisputably a subjective construct established in a social context, although its construction can neither be made comparatively nor vertically in relation to the marks present in the disease, thus an adolescent with a very significant mark may have the perception of a better quality of life than another that possesses a smaller mark. Therefore, quality of life is not necessarily associated to the mark the adolescent holds as is the manner in which he/she faces the barriers produced by it.

Made available in DSpace on 2016-12-23T13:49:05Z (GMT). No. of bitstreams: 1 Geise de Aguiar Quirino.pdf: 839323 bytes, checksum: 5ff1a2735aa2271c126e6d624a6760bd (MD5) Previous issue date: 2012-02-17 / Osteogenesis Imperfecta (OI) is a genetic desease characterized by patient s bone fragility and deformity, in which severity ranges from a barely detectable connective tissue disorder to lethality in the perinatal period. The diversity of clinical variability in patients is caused by the different location or type of mutations in one of the ten genes related with the disease. This wide clinical variability difficults the perfect clinical diagnoses, due to that the use of molecular biology techniques becomes necessary to obtain a correct diagnoses and for genotype: phenotype correlation. One of the relevant genes associated with recessive forms of OI is the LEPRE-1 gene, responsible for encoding the prolyl 3 hidroxylase 1 protein. This protein and two others are components of the complex responsible for pro-collagen alfa 1 chains 3 prolyl hydroxylation. The target of this research was to analyze the LEPRE-1 gene in eight non consanguineous patients clinically diagnosed as severe Osteogenesis Imperfecta suggestive of autossomic recessive heritage by DNA sequencing of exons 1, 3, 5, 6 and 14 of the gene. In addition, the data obtained was used to analyze the efficiency of the SSCP technique by comparing the results between screening for mutations methodologies and gene sequencing methodologies. On exon 6, for instance, a mutation in one patient was found: a heterozygose base change (c.1087A>G / p.Lys363Glu), consequently, lysine was produced instead of glutamic acid. On the other exons, there was no mutation found on the patients chosen. All the results obtained in this research were compatible with datas generated by SSCP and suggest high efficient of SSCP technique for LEPRE-1 gene to recessive cases of Osteogenesis Imperfecta / A Osteogênese Imperfeita é uma doença genética caracterizada por fragilidade e deformidade esquelética, onde o quadro clínico pode variar desde simples deformidades ósseas à forma letal perinatal. A alta variabilidade clínica apresentada pelos indivíduos afetados ocorre devido ao tipo e à localização da mutação em um dos dez genes relacionados a doença. A grande heterogeneidade genética existente exige a utilização de técnicas da biologia molecular para o diagnóstico e compreensão das correlações genótipo: fenótipo da doença. Um dos genes relevantes associados com as formas recessivas da Osteogênese Imperfeita é o gene LEPRE-1 codificador da proteína prolil 3 hidroxilase 1. Esta é uma das três proteínas componentes do complexo responsável pela prolil 3 - hidroxilação das cadeias de pró-colágeno alfa 1 formadoras da molécula do colágeno tipo I, expresso, predominantemente em ossos, tendões e pele. Este projeto de pesquisa teve como objetivo analizar o gene LEPRE-1 em oito pacientes não consanguineos com Osteogênese Imperfeita tipo grave sugestivos de herança autossômica recessiva por meio do sequenciamento direto dos exons 1, 3, 5, 6 e 14 do gene. Além disso, o resultado gerado foi utilizado para avaliar a eficiência da técnica de triagem de mutações por Polimorfismo Conformacional de Fita Simples (SSCP) por meio da comparação de resultados entre as metodologias de triagem de mutações e sequenciamento direto do gene. Foi identificada, no exon 6, uma mutação de troca de nucleotídeos em heterozigose, c.1087A>G / p.Lys363Glu, levando a produção do aminoácido ácido glutâmico ao invés da lisina em um dos pacientes avaliados. Não foram encontradas mutações em nenhum dos pacientes para os demais exons analisados. Estes resultados corroboram dados gerados por meio de SSCP, e sugerem grande eficiência da técnica de triagem para o gene LEPRE-1 para formas recessivas de Osteogênese Imperfeita

A osteogênese imperfeita (OI) é caracterizada como uma desordem genética na qual uma osteopenia generalizada leva a baixa estatura, fragilidade óssea excessiva e deformidades ósseas graves. As células mesenquimais estromais multipotentes (CTMs) são precursores presentes na medula óssea adulta capazes de se diferenciar em osteoblastos, adipócitos e mioblastos que passaram a ter grande importância como fonte terapia celular. O objetivo do presente estudo foi analisar o perfil de expressão gênica durante a diferenciação osteogênica a partir de células mesenquimais estromais multipotentes da medula óssea obtidas de pacientes diagnosticados com Osteogênese Imperfeita e de indivíduos controle. Foram coletadas amostras de três indivíduos normais e cinco amostras de pacientes portadores de Osteogênese Imperfeita. As células mononucleares (CMN) foram isoladas para a obtenção de células mesenquimais que foram expandidas até a terceira passagem quando iniciou-se o estímulo para diferenciação osteogênica. Também foram realizadas análises para contagem de CFU-F e para quatro das cinco amostras de pacientes portadores de OI, o número de CFU-F observado foi inferior ao geralmente encontrado para amostras de doadores normais. Foram coletadas células para análises de imunofenotipagem celular por citometria de fluxo e o RNA foi extraído originando a amostra denominada T0. As garrafas restantes tiveram suas células estimuladas para diferenciação osteogênica. Após um dia em cultura com estímulo, mais uma garrafa teve o RNA de suas células extraído (T1), e o mesmo procedimento foi realizado nos dias 2 (T2), 7 (T7), 12 (T12), 17 (T17) e 21 (T21). Todas as amostras demonstraram possuir potencial de diferenciação in vitro em osteoblastos e adipócitos. A imunofenotipagem de células mesenquimais foi realizada e as amostras de todos os pacientes apresentaram perfil imunofenotípico compatível com trabalhos anteriores. Foram identificadas mutações nos genes COL1A1 e/ou COL1A2 responsáveis pelo desenvolvimento da doença para quatro dos cinco pacientes avaliados. Para o paciente portador de Osteogênese Imperfeita e Síndrome de Bruck a região codificadora do gene PLOD2 também foi seqüenciada, porém não foram encontradas mutações. A análise da expressão gênica foi realizada pela técnica de microarranjos e foram identificados vários genes com expressão diferencial. Alguns genes com importância fundamental na diferenciação osteoblástica apresentaram menor expressão nas amostras dos pacientes portadores de OI, sugerindo um menor comprometimento das CTMs desses pacientes com a linhagem osteogênica. Outros genes também tiveram sua expressão diferencial confirmada por PCR em Tempo Real. Foi observado um aumento na expressão de genes relacionados a adipócitos, sugerindo um aumento da diferenciação adipogênica em detrimento à diferenciação osteogênica. A expressão das variantes do gene PLOD2 mostrou-se diferencial entre amostras normais, de OI e do paciente portador de Síndrome de Bruck. Também foi evidenciada uma expressão diferencial do microRNA 29b, um microRNA com papel estabelecido durante a diferenciação osteogênica, sugerindo um mecanismo de regulação dependente da quantidade de RNAm do seu gene alvo, o COL1A1. / Osteogenesis imperfecta (OI) is characterized as a genetic disorder in which a generalized osteopenia leads to short stature, bone fragility and serious skeletal deformities. Mesenchymal stem cells (MSCs) are precursors present in adult bone marrow that can differentiate into osteoblasts, adipocytes and myoblasts that have been given great importance as a source cell therapy. The aim of this study was to analyze the gene expression profile during osteogenic differentiation from mesenchymal stem cells from bone marrow taken from patients diagnosed with Osteogenesis Imperfecta and control subjects. Samples were collected from three normal individuals and five samples from patients with Osteogenesis Imperfecta. Mononuclear cells (MON) were isolated to obtain mesenchymal cells that were expanded until third passage when the stimulus for osteogenic differentiation was induced. Analyses were also conducted to count the CFU-F and for four of the five samples from patients with OI, the number of CFU-F observed was lower than generally found for normal samples. Cells were collected for analysis of cell immunophenotyping by flow cytometry and RNA was extracted from the resulting sample called T0. Remaining cells were stimulated for osteogenic differentiation. After a day in culture with stimulation, cells from another bottle had their RNA extracted (T1), and the same procedure was performed on days 2 (T2), 7 (T7), 12 (T12), 17 (T17) and 21 (T21). All samples have shown potential of in vitro differentiation into osteoblasts and adipocytes. Immunophenotyping of mesenchymal cells was performed and samples of all patients had immunophenotypic profile consistent with previous works. We identified mutations in COL1A1 and / or COL1A2 responsible for developing the disease for four of five patients. For the patient with Osteogenesis Imperfecta and Bruck Syndrome, coding region of the gene PLOD2 was also sequenced, but no mutations were found. The gene expression analysis was performed by microarray and identified several genes with differential expression. Some genes of fundamental importance in osteoblast differentiation showed lower expression in samples from patients with OI, suggesting a minor involvement of MSCs of patients with osteogenic lineage. Other genes also confirmed their differential expression by Real Time PCR. We observed an increased expression of genes related to adipocytes, suggesting an increased adipogenic differentiation at the expense of osteogenic differentiation. The expression of PLOD2 gene variants proved to be different between normal samples, OI and the patient with Bruck Syndrome. There was also evidence of differential expression of 29b microRNA, with established role during osteogenic differentiation, suggesting a mechanism dependent regulation of mRNA abundance of its gene target, COL1A1.

<p dir="ltr">Skeletal fragility can be caused by a wide array of diseases and disorders, but the most difficult etiologies to clinically circumvent are those in which the body loses not just bone mass but the ability to create healthy bone tissue. While in conditions such as osteoporosis (the most prevalent cause of age-related skeletal fragility in which elevated resorption without compensatory elevated formation leads to bone loss), interventions can target bone remodeling pathways to protect and increase bone mass, many other diseases are characterized by genetic and metabolic crippling of the remodeling process, rendering those same mass-based interventions less effective at reducing fracture risk. Osteogenesis imperfecta (OI) is a class of genetic disorders in which gene mutations affect the formation of collagen, a crucial building block of bone tissue that makes up 90% of its organic matrix, leading to lost bone mass and quality. As the main genetic causes of OI cannot currently be directly treated, therapeutic OI treatments are needed that improve tissue-level material properties. Similarly, metabolic conditions such as diabetes, a disorder in which the body cannot properly regulate blood sugar due to loss of insulin production and/or efficacy, can have multi-organ impacts including increased risk of developing chronic kidney disease and skeletal fragility. Type 2 diabetes is especially notorious for increasing fracture risk despite maintained or even increased apparent bone mass, which is strong evidence that intrinsic bone material properties are impaired by the disease state. A possible solution to the bone quality problem may be treatments that increase bone water content, as amplifying the water content of bone can improve multi-scale material properties such as collagen fibril elasticity and whole-bone toughness. Therefore, increasing bone hydration could be a way of improving tissue-level material properties, despite being unable to eradicate the genetic or metabolic disorders that alter how collagen is produced and incorporated into the bone matrix. To that end, this dissertation presents several studies that characterize models of osteogenesis imperfecta and diabetic kidney disease in mice and investigate methods of rescuing skeletal fragility in these animals through treatments that target both bone mass and bone quality with ties to tissue hydration.</p>

Osteogenesis imperfecta, innate brittle bone disease, is a very serious disease. It is inheritable disease of connective tissue, which shows by abnormal fragility of bones. The occurrence of this disease is one case in 10 000 30 000 births. The theoretical part of the thesis deals with the disease itself, also the psychical impact on children suffering from Osteogenesis imperfecta and the impact on their families as well. At the beginning of the research, three goals of this thesis were set: map out (on the basis of theoretical and practical backgrounds) the pitfalls of life of children with the disease Osteogenesis imperfecta, find out what are the most common difficulties by children with the disease Osteogenesis imperfecta and also find out the experiences of nurses with the care for children with disease Osteogenesis imperfecta. The empirical part of the thesis was processed by means of qualitative research conducted by the technique of semi-structured interview and narrative biographical interview. The research set were nurses working at the child departments in hospitals, parents of ill children and also an adult woman with the diagnosis of Osteogenesis imperfecta and two doctors. From the research emerged that among the most common difficulties of children is pain, which decreases the quality of their life. Small children can't engage in typical activities of children, such as going to a playground, older children can't attend for example music festivals. Children feel fear from fractures and are therefore limited in sports. Because of injuries and their treatments, the children have more absences at schools and therefore are isolated from peers. Nevertheless, the children with this disease can live a happy life. From the results of the research also emerges, that nurses working at the child departments of the hospitals attended by children with this illness have a good experiences with their treatment. They are able to give parents important information and know the specifics of application of the treatment. The results of the diploma thesis were presented at a national student conference and will be further published.

本論文是以曾引起臺灣社會輿論熱烈關注的玻璃娃娃案為研究起點，進而探究法律語言、法律論證與日常語言之間的關聯性。首先，分析玻璃娃娃案的四件民事判決，並聚焦在這些判決文內容的法律語言、法律論證與日常語言。其次，從玻璃娃娃案判決的相關新聞報導中，探討日常語言與法律語言。再者 ，檢視與評論民法學者對玻璃娃娃案的第一、第二審判決的個案分析，藉此探究法律語言、法律論證與日常語言之間的關聯性，緊接著運用民法請求權基礎的架構來重新檢視玻璃娃娃案之當事人間的法律關係，例如契約、無因管理和侵權行為等。最後，藉由Robert Alexy的法律論證理論，檢驗玻璃娃娃案的民事第一、二、三審判決和民法學者對此案的案例分析之中的法律論證。 / This thesis investigate the relationships between legal language, legal argumentation and daily language in the fatal accident case of osteogenesis imperfecta in Taiwan. First, we analyze legal language, legal argumentation and daily language used in the verdicts of four civil cases. We focus on the contents of verdicts. Second, from the newspaper reports of this case, we investigate the relationships between legal language and daily language. Third, the case studies by civil law researchers are investigated by the verdicts of trial of first and second instances. From the investigation of cases studies, we can obtain the connection between legal language, legal argumentation and daily language. Using the foundations of claims of civil law, we re-study the legal relationships like contracts, torts and voluntary service among clients in the case of osteogenesis imperfecta. Finally, we re-investigate the legal argumentations of the every verdicts of civil court and the case studies with Theory of Legal Argumentation proposed by Robert Alexy.