Para a execução da etapa inicial da identificação médico-legal de restos humanos (antropometria e exame dos arcos dentários), faz-se necessária uma limpeza prévia da ossada, para a remoção de tecidos moles putrefeitos. Os casos não identificados por esses métodos tradicionais, poderão ser submetidos ao exame de DNA. No entanto, apesar do grande avanço da biologia molecular, utilizando a amplificação de DNA pela PCR, algumas limitações que afetam a habilidade de se obter DNA em restos humanos, permanecem. Dessa forma, o objetivo deste trabalho foi fornecer subsídios morfológicos para os analistas forenses, com ênfase na prática médico-legal, visando uma utilização mais eficiente do DNA obtido de osso compacto de restos humanos em decomposição ou já esqueletizados, sem tecidos moles aderidos. Foi realizado o estudo da microestrutura do tecido ósseo compacto femoral, de restos humanos em decomposição, ainda com tecidos moles, que foram limpos pela fervura em água (n = 7) e ossadas já esqueletizadas pela decomposição natural, que não foram fervidas (n = 8). Destes, seis ossadas foram provenientes de cemitério público regular, após 3 anos de inumação, 1 ossada proveniente da região amazônica, e 1 ossada de origem desconhecida. Estas duas ultimas, apresentado-se porosas ou quebradiças. As análises morfológicas de cortes histológicos foram coradas com hematoxilina e eosina e o DNA amplificado pela PCR para os loci CSF1PO, TPOX, TH01, F13A0, FESFPS, vWA, D16S539, D7S820, D13S317 e amelogenina. Os resultados da análise desses dois grupos foram comparados com os de cadáveres frescos (n = 5) do Serviço de Verificação de Óbitos da Capital. A fervura dos ossos, do modo como é realizada no Instituto Médico Legal de São Paulo, pode aumentar a eosinofilia da matriz óssea e, em alguns casos, pode promover a desagregação dos ósteons. Tal procedimento pode remover células, mas pode também remover possíveis inibidores da PCR, favorecendo a análise do DNA obtido destas amostras. O fator limitante para a obtenção e análise de DNA, em amostras de ossos limpos por fervura, é a quantidade exígua de células. Ossos não submetidos à fervura, após inumação por três anos ou há mais tempo em contato com a terra, podem apresentar alterações da microestrutura. No entanto, a presença de hemácias preservadas e núcleos de osteócitos nestas amostras, indica melhor preservação de células em relação às amostras de ossos fervidos. O fator limitante para a análise de DNA nestas amostras é a presença sugestiva de inibidores da reação de amplificação pela PCR. Restos humanos, sem tecidos moles, macroscópicamente não preservados (porosos e quebradiços), e não submetidos à fervura, apresentam alterações de perda de matriz mineralizada; no entanto, nestas amostras ainda é possível encontrar células preservadas. Os resultados obtidos no neste trabalho permitem traçar algumas estratégias para uma melhor utilização nos protocolos de extração e análise do DNA em osso compacto de restos humanos. / To the first essential step to forensic identification of human remains (anthropological study of race, sex, age, etc) it is necessary a previous cleaning of the bones, to remove decomposing soft tissues. Medico-legal inconclusive or non identified cases, by using these traditional methods, could be subjected to DNA analysis. However, in spite of advances in human identification techniques, specially by PCR amplified DNA, some limitations that affect the ability to obtain DNA in human remains still persist. Therefore, the aim of this study was to provide additional support from morphological analysis, to help forensic analysts personnel to utilise more efficiently the DNA, extracted from compact bones of human remains in decomposition or already skeletonized corpse, it means without soft tissues, with special emphasis in the legal-medicine practice. Femoral compact bones were obtained from: 7 human remains found on the ground, in different degree of decomposition which were cleaned by boiling to remove soft tissues; also studied were collections of bones from 8 corpses having undergone natural decomposition: 6 human remains exhumed after 3 years from a common public cemetery in São Paulo City; 1 case from amazon region and 1 case with no information, both cases remained from long time (more than 3 years) in contact with soil. All eight cases, were not boiled as no soft tissue were adhered. As a control, five cadavers 12 to 16 hours post mortem were also used. The compact bones histological sections were stained by haematoxilin and eosin and the loci CSF1PO, TPOX, TH01, F13A01,FESFPS, vWA, D16S539, D7S820, D13S317 and amelogenin were amplified by PCR.The procedure for boiling the human remains utilised in the Legal Medicine Institute of São Paulo would have increased the eosinophily of bone matrix and, in some cases, promoted the desaggregation of the osteons. In addition these procedures would have removed the cells, but in some cases would have removed possible inhibitors of the PCR, favouring in this way the analysis of DNA obtained from these samples. The limiting factor to obtain successful analysis in bones submitted to boiling seem to be the low quantity of nuclei present in these samples. For the other hand, in bones not cleaned by boiling, the presence of preserved red cells and oscteocyte nuclei inside the lacunae indicates better preservation of cells in relation to those bones cleaned by boiling. The limiting factor to obtain successful DNA analysis in bones exhumed or in contact of soil, is the suggestive presence of inhibitors of PCR. Porous and brittle bones from human remains, without soft tissues that are not processed by boiling, present alterations through loss of mineralised matrix, although it is still possible to found preserved cells in these samples. The results presented in this work clarify concerns about viability of DNA for identification analysis. They also help to establish better strategies for optimisation of DNA extraction and analysis in compact bones of human remains.

INTRODUÇÃO: A doença de Legg-Calvé-Perthes (DLCP) é caracterizada pela necrose idiopática da epífise femoral proximal na criança. A interrupção do suprimento arterial para a cabeça femoral é o fator etiopatogênico mais aceito. Embora as angiografias sejam a base para as investigações das causas vasculares, elas não avançaram na DLCP devido a limitações éticas, por serem procedimentos invasivos com exposição à radiação ionizante e pela potencial toxicidade do contraste iodado. O objetivo primário foi avaliar a rede arterial da epífise proximal do fêmur em crianças com DLCP unilateral, particularmente a artéria circunflexa femoral medial (ACFM) que nutre a referida epífise, comparando o quadril afetado com o quadril contralateral não afetado. A técnica usada foi a angiografia por ressonância magnética (angioRM) que permite o estudo vascular sem as limitações e os riscos inerentes à angiografia convencional. A sequência angiográfica usada foi TRICKS (Time Resolved Imaging of Contrast KinecticS) que utiliza técnica de aquisição de imagens com foco na resolução temporal do contraste (identificação da passagem do contraste pelos vasos ao longo do tempo). MÉTODOS: Os dados foram obtidos a partir de uma amostra consecutiva composta por 24 pacientes com DLCP unilateral admitidos no ambulatório de ortopedia pediátrica de um hospital terciário de maio de 2013 a dezembro de 2016. Os exames de angioRM-TRICKS foram avaliados por dois radiologistas e o consenso foi usado para a análise. RESULTADOS: Foram realizados 24 exames. A idade média foi de 7,9 +/- 2,1 anos. O trajeto da ACFM foi analisado no plano coronal dos exames de angioRM-TRICKS. No segmento 1 (da origem até a borda medial do colo femoral), havia 100% de visibilidade no lado da DLCP, bem como no lado contralateral. No segmento 2 (da borda medial do colo femoral até a borda lateral do colo femoral), houve 63% de visibilidade no lado DLCP e 71% de visibilidade no lado contralateral. No segmento 3 (ponto de entrada no sulco intertrocantérico), houve 25% de visibilidade no lado com DLCP e 17% de visibilidade no lado contralateral. O segmento 4 (ramo terminal e ascendente na face lateral do colo do fêmur) foi 100% não visibilizada no quadril com DLCP nem no lado sem a doença. CONCLUSÕES: A angioRM-TRICKS fornece imagens bem definidas do suprimento arterial para a epífise femoral proximal em crianças com a DLCP. Não foram encontradas diferenças significativas nos trajetos analisados da ACFM entre o lado acometido e o lado sem a DLCP. Com os exames de angioRM-TRICKS não foi possível visibilizar o trecho ascendente epifisário lateral, ramo terminal da ACFM, tanto nos quadris afetados quanto nos quadris sem a DLCP / INTRODUCTION: Legg-Calvé-Perthes disease (LCPD) is characterized by idiopathic necrosis of the proximal femoral epiphysis in the child. Interruption of the arterial supply to the femoral head is the most accepted etiopathogenic factor. Although angiograms are the basis for investigations of vascular causes, they have not progressed in LCPD due to ethical limitations, because they are invasive procedures with exposure to ionizing radiation and the potential toxicity of iodinated contrast. The primary objective was to evaluate the arterial network of the proximal femoral epiphysis in children with unilateral LCPD, particularly the medial femoral circumflex artery (MCFA) that supplies the epiphysis, comparing the affected hip with the unaffected contralateral hip. The technique used was magnetic resonance angiography (MRA) that allows vascular study without the limitations and risks inherent in conventional angiography. The angiographic sequence used was TRICKS (Time Resolved Imaging of Contrast KinecticS), which uses an image acquisition technique focused on temporal resolution of contrast (identification of the passage of contrast by vessels over time). METHODS: Data were obtained from a consecutive sample composed of 24 patients with unilateral LCPD who were admitted to the pediatric orthopedic outpatient clinic of a tertiary hospital from May 2013 to December 2016. Two radiologists evaluated the MRA-TRICKS exams and the consensus was used for the analysis. RESULTS: Twenty-four examinations were performed. The mean age was 7.9 +/- 2.1 years. The MCFA trajectory was analyzed in the coronal plane of the MRA-TRICKS exams. In segment 1 (from the origin to the medial border of the femoral neck), there was 100% visibility on the LCPD side, as well as on the contralateral side. In segment 2 (from the medial border of the femoral neck to the lateral border of the femoral neck), there was 63% visibility on the LCPD side and 71% visibility on the contralateral side. In segment 3 (entry point in the intertrochanteric groove), there was 25% visibility on the LCPD side and 17% visibility on the contralateral side. Segment 4 (terminal and ascending limb on the lateral aspect of the femoral neck) was 100% not seen in the hip with LCPD or on the side without the disease. CONCLUSIONS: MRA-TRICKS provides well-defined images of the arterial supply to the proximal femoral epiphysis in children with LCPD. No significant differences were found in the analyzed ACFM pathways between the affected side and the non- LCPD side. With the MRA-TRICKS exams, it was not possible to visualize the ascending lateral epiphyseal segment, terminal branch of the MCFA, both in the affected hips and in the hips without the LCPD

Submitted by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-28T12:43:48Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO MONIQUE BABINSKI_Pós Defesa 2016 SEM APENDICES (1).pdf: 1899605 bytes, checksum: 5667910f275aadea7cc77dcc4a1ea0c1 (MD5) / Approved for entry into archive by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-28T12:44:09Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO MONIQUE BABINSKI_Pós Defesa 2016 SEM APENDICES (1).pdf: 1899605 bytes, checksum: 5667910f275aadea7cc77dcc4a1ea0c1 (MD5) / Made available in DSpace on 2017-09-28T12:44:09Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO MONIQUE BABINSKI_Pós Defesa 2016 SEM APENDICES (1).pdf: 1899605 bytes, checksum: 5667910f275aadea7cc77dcc4a1ea0c1 (MD5) Previous issue date: 2016 / Complexo Hospitalar de Niterói / O objetivo deste estudo foi avaliar os efeitos da restrição proteica e calórica materna durante a lactação, analisando se haveria recuperação, o conhecido “catch-up” das dimensões morfológicas femorais após a restauração da dieta nutricional. Ao nascimento da ninhada, as progenitoras foram divididas nos seguintes grupos: Controle (C) com dieta a 23% de proteína ad libitum; Restrição Proteico-Calórica (RPC) com dieta a 8% de proteína e Restrição Calórica (RC) com dieta a 23% de proteína, em quantidades restritas de acordo com a ingestão do grupo RPC. Após o desmame, todos os filhotes foram separados das mães e receberam livre acesso a uma dieta padrão de laboratório contendo 23% de proteína, até 180 dias, quando os ratos foram eutanasiados. O fêmur foi excisado e as dimensões femorais macroscópicas foram medidas usando pontos anatômicos pré-estabelecidos. Após a morfometria macroscópica, os fêmures foram fixados em formol 10% (24h) e descalcificados por EDTA. Foram realizadas secções transversais de 10μm de espessura no terço médio diafisário dos espécimes, corados com HE e levados ao microscópio para análise qualitativa e histomorfometria da cortical óssea. Os resultados morfométricos macroscópicos mostraram que a maioria das medidas nos grupos de RC e RPC foram significativamente menores do que no grupo controle. As maiores reduções ocorrendo no grupo de RPC e com várias anormalidades histoarquiteturais, eg., diminuição da área cortical e aumento da área medular. Cortical com lacunas aumentadas significativamente nos grupos RC e RPC. Nossos resultados mostram que a restrição proteico-calórica durante a lactação leva a uma recuperação incompleta na idade adulta. O fêmur mostrou redução significativa na maior parte dos parâmetros macro e microscópicos nos dois grupos tratados, em particular o grupo RPC, quando comparado com o grupo C / The aim of this study was to evaluate the effects of maternal protein and energy restriction during lactation, analyzing on morphological dimensions whether there was catch-up growth through normative nutrition restored, as well as on mechanical axis of femur of the offspring at adulthood. At parturition, Wistar rat dams were randomly assigned to the following groups: 1) control group (C) - free access to a standard laboratory diet containing 23 % protein, 2) protein-energy restricted group (PER) - free access to an isoenergetic, protein-restricted diet containing 8 % protein, and 3) energy-restricted group (ER) – fed with restricted amounts of a standard laboratory diet. At weaning, all pups were separated of dams and received free access to a standard laboratory diet containing 23% protein until 180 days, when the rats were anesthetized and sacrificed. The dimensions of excised pup femur were measured directly using pre-established anatomical points. After macroscopic morphometry, the femurs were fixed in 10% formalin (24h) and decalcified by EDTA. Cross sections of 10μm thickness were made in the middle third (diaphyseal) of the specimens stained with HE and taken to the microscope for qualitative analysis and histomorphometry of cortical bone. Morphometric analysis of the femur (macroscopic) showed that most of the measurements in the ER and PER groups were significantly lower than in the control group, with the greatest reductions occurring in the PER group and several histoarchitectural abnormalities, e.g., decrease in cortical area and increased medullary area. Cortical bone with gaps increased significantly in ER and PER groups. Our results show that protein and energy restriction during lactation leads to an incomplete catch-up growth in adulthood. The femur showed significant reduction in most of the parameters in the two treated groups, particularly the PER group, when compared to the control group

INTRODUÇÃO: A doença de Legg-Calvé-Perthes (DLCP) é caracterizada pela necrose idiopática da epífise femoral proximal na criança. A interrupção do suprimento arterial para a cabeça femoral é o fator etiopatogênico mais aceito. Embora as angiografias sejam a base para as investigações das causas vasculares, elas não avançaram na DLCP devido a limitações éticas, por serem procedimentos invasivos com exposição à radiação ionizante e pela potencial toxicidade do contraste iodado. O objetivo primário foi avaliar a rede arterial da epífise proximal do fêmur em crianças com DLCP unilateral, particularmente a artéria circunflexa femoral medial (ACFM) que nutre a referida epífise, comparando o quadril afetado com o quadril contralateral não afetado. A técnica usada foi a angiografia por ressonância magnética (angioRM) que permite o estudo vascular sem as limitações e os riscos inerentes à angiografia convencional. A sequência angiográfica usada foi TRICKS (Time Resolved Imaging of Contrast KinecticS) que utiliza técnica de aquisição de imagens com foco na resolução temporal do contraste (identificação da passagem do contraste pelos vasos ao longo do tempo). MÉTODOS: Os dados foram obtidos a partir de uma amostra consecutiva composta por 24 pacientes com DLCP unilateral admitidos no ambulatório de ortopedia pediátrica de um hospital terciário de maio de 2013 a dezembro de 2016. Os exames de angioRM-TRICKS foram avaliados por dois radiologistas e o consenso foi usado para a análise. RESULTADOS: Foram realizados 24 exames. A idade média foi de 7,9 +/- 2,1 anos. O trajeto da ACFM foi analisado no plano coronal dos exames de angioRM-TRICKS. No segmento 1 (da origem até a borda medial do colo femoral), havia 100% de visibilidade no lado da DLCP, bem como no lado contralateral. No segmento 2 (da borda medial do colo femoral até a borda lateral do colo femoral), houve 63% de visibilidade no lado DLCP e 71% de visibilidade no lado contralateral. No segmento 3 (ponto de entrada no sulco intertrocantérico), houve 25% de visibilidade no lado com DLCP e 17% de visibilidade no lado contralateral. O segmento 4 (ramo terminal e ascendente na face lateral do colo do fêmur) foi 100% não visibilizada no quadril com DLCP nem no lado sem a doença. CONCLUSÕES: A angioRM-TRICKS fornece imagens bem definidas do suprimento arterial para a epífise femoral proximal em crianças com a DLCP. Não foram encontradas diferenças significativas nos trajetos analisados da ACFM entre o lado acometido e o lado sem a DLCP. Com os exames de angioRM-TRICKS não foi possível visibilizar o trecho ascendente epifisário lateral, ramo terminal da ACFM, tanto nos quadris afetados quanto nos quadris sem a DLCP / INTRODUCTION: Legg-Calvé-Perthes disease (LCPD) is characterized by idiopathic necrosis of the proximal femoral epiphysis in the child. Interruption of the arterial supply to the femoral head is the most accepted etiopathogenic factor. Although angiograms are the basis for investigations of vascular causes, they have not progressed in LCPD due to ethical limitations, because they are invasive procedures with exposure to ionizing radiation and the potential toxicity of iodinated contrast. The primary objective was to evaluate the arterial network of the proximal femoral epiphysis in children with unilateral LCPD, particularly the medial femoral circumflex artery (MCFA) that supplies the epiphysis, comparing the affected hip with the unaffected contralateral hip. The technique used was magnetic resonance angiography (MRA) that allows vascular study without the limitations and risks inherent in conventional angiography. The angiographic sequence used was TRICKS (Time Resolved Imaging of Contrast KinecticS), which uses an image acquisition technique focused on temporal resolution of contrast (identification of the passage of contrast by vessels over time). METHODS: Data were obtained from a consecutive sample composed of 24 patients with unilateral LCPD who were admitted to the pediatric orthopedic outpatient clinic of a tertiary hospital from May 2013 to December 2016. Two radiologists evaluated the MRA-TRICKS exams and the consensus was used for the analysis. RESULTS: Twenty-four examinations were performed. The mean age was 7.9 +/- 2.1 years. The MCFA trajectory was analyzed in the coronal plane of the MRA-TRICKS exams. In segment 1 (from the origin to the medial border of the femoral neck), there was 100% visibility on the LCPD side, as well as on the contralateral side. In segment 2 (from the medial border of the femoral neck to the lateral border of the femoral neck), there was 63% visibility on the LCPD side and 71% visibility on the contralateral side. In segment 3 (entry point in the intertrochanteric groove), there was 25% visibility on the LCPD side and 17% visibility on the contralateral side. Segment 4 (terminal and ascending limb on the lateral aspect of the femoral neck) was 100% not seen in the hip with LCPD or on the side without the disease. CONCLUSIONS: MRA-TRICKS provides well-defined images of the arterial supply to the proximal femoral epiphysis in children with LCPD. No significant differences were found in the analyzed ACFM pathways between the affected side and the non- LCPD side. With the MRA-TRICKS exams, it was not possible to visualize the ascending lateral epiphyseal segment, terminal branch of the MCFA, both in the affected hips and in the hips without the LCPD

Αναδρομική μελέτη των 13 ασθενών με σηπτική ψευδάρθρωση κάτω πέρατος μηριαίου που αντιμετωπίστηκαν με ευρύ χειρουργικό καθαρισμό και με τη μέθοδο Ilizarov. Κατά την έναρξη της τελικής αντιμετώπισης όλοι οι ασθενείς είχαν σημαντικό περιορισμό της κίνησης της άρθρωσης του γόνατος. Ο μέσος όρος προηγούμενων χειρουργικών επεμβάσεων ήταν τρεις. Ο μέσος όρος οστικού ελλείμματος ήταν 8.3 εκ. Ο μέσος χρόνος εξωτερικής οστεοσύνθεσης ήταν 309.8 ημέρες. Σύμφωνα με τα κριτήρια Paley σε οκτώ ασθενείς είχαμε άριστο οστικό αποτέλεσμα, ενώ το λειτουργικό αποτέλεσμα ήταν σε τρεις περιπτώσεις άριστο, σε τέσσερις καλό. Πώρωση του κατάγματος, εκρίζωση της λοίμωξης και αποκατάσταση της στηρικτικής ικανότητας του σκέλους επιτεύχθηκε σε όλους τους ασθενείς. Η αύξηση του χρόνου εξωτερικής οστεοσύνθεσης παρατηρήθηκε: 1) η οριστική αντιμετώπιση εφαρμόστηκε 6 μήνες μετά από τον αρχικό τραυματισμό. 2) ο ασθενής υποβλήθηκε σε 4 τουλάχιστον προηγούμενες χειρουργικές επεμβάσεις 3) η αρχική αντιμετώπιση συμπεριλάμβανε ανοικτή ανάταξη και εσωτερική οστεοσύνθεση. Με την μέθοδο Ilizarov επιτυγχάνεται πλήρη εκρίζωση της οστικής λοίμωξης, υψηλό ποσοστό πώρωσης και αποκατάσταση της στηρικτικής ικανότητας του σκέλους. Ωστόσο συχνά η δυσκαμψία του γόνατος και η χωλότητα αποτελούν χρόνιο πρόβλημα για αρκετούς ασθενείς. / We retrospectively reviewed 13 patients with infected nonunion of the distal femur, which had been treated by radical surgical debridement and Ilizarov method. All had severely restricted movement of the knee and a mean of 3.1 previous operations. The mean bone defect was 8.3 cm and no patient was able to bear weight. The mean external fixation time was 309.8 days. According to the Paley’s grading system, eight patients had an excellent bone result and seven excellent and good functional results. Bone union, the ability to bear weight fully, and eradication of infection were achieved in all the patients. The external fixation time was increased when the definitive treatment started six months or more after the initial trauma, the patient had been subjected to more than four previous operations and the initial operation had been ORIF. The treatment of infected defect pseudarthrosis of the distal femur using the Ilizarov device is a salvage procedure, as it offers complete eradication of infection, high union rate and ability for full weight bearing. Nevertheless problems such as, impaired knee joint motion and limping bother the patients permanently.

Pour comprendre et diagnostiquer des pathologies telles que l'ostéoporose, qui est un problème majeur de santé publique et, est un facteur de risque important de fractures notamment la Fracture de l'Extrémité Supérieure du Fémur (FESF), il est essentiel d’aborder ces problématiques en trois dimensions (3D) pour fournir au praticien un outil de diagnostic et de dépistage du risque fracturaire. De plus, la visualisation en 3D de l'anatomie joue un rôle important dans le domaine de la chirurgie orthopédique guidée (assistée par ordinateur). En préopératoire, pour la planification chirurgicale ou la conception de prothèses sur mesure, en per opératoire, pour assister le chirurgien durant l'acte chirurgical et enfin en postopératoire pour le suivi. La reconstruction de modèles anatomiques en 3D peut être réalisée par l'utilisation de techniques d'imagerie 3D directes telles que la tomodensitométrie. Cependant, l'utilisation d'une telle imagerie est limitée à des procédures complexes en raison des contraintes imposées par le coût, la disponibilité et les risques de radiation pour le patient. Ainsi, l'alternative à ce type d'imagerie 3D est de développer des méthodes de reconstruction 3D qui s'appuient uniquement sur quelques radiographies 2D. L'objectif principal de cette thèse est de proposer une technique permettant de reconstruire de façon automatique une surface 3D du fémur proximal à partir d'un nombre restreint de radiographies. Les études précédentes sur la reconstruction de surface ont généralement besoin de connaissances supplémentaires comme l'utilisation d'un modèle générique ou statistique 3D de la forme à reconstruire. La méthode décrite dans cette thèse nécessite seulement les coordonnées 3D de points calculées à partir de quelques paires de clichés radiographiques. Deux approches sont proposées. La première méthode repose sur la mise en correspondance de contours extraits de paires de radiographies et sur un modèle mathématique basé sur le principe de la stéréovision pour le calcul d'un nuage de points 3D. La deuxième technique utilise les résultats de l’approche précédente ainsi que des points extraits d’un autre fémur pour améliorer la précision au niveau de certaines régions sensibles choisies par l’opérateur. La reconstruction du modèle surfacique à partir des nuages de points obtenus par les deux techniques est obtenue par un maillage basé sur l'équation de Poisson. Un recalage 3D/3D est effectué entre le nuage de points calculé et le nuage de points extrait du modèle générique connu ("Gold Standard" obtenu avec des coupes CT-Scan) du même fémur proximal afin de pouvoir comparer la surface reconstruite à un modèle "vérité terrain" et ainsi estimer la précision de la méthode. / To understand and diagnose pathologies such as osteoporosis, which is considered as a major public health issue and, an important risk factor of fractures in particular the proximal femur fracture. The classical tools of diagnosis are mainly based on the analysis of X-rays photographs. These techniques have shown many limitations to carry out the key information for the physician. Through the last decade the 3D visualization of anatomy demonstrated the effectiveness for analysis and diagnosis, particularly for the guided orthopedic surgery (computer aided). In preoperative, for the surgical planning or the design of prostheses, in per operative, to assist the surgeon during the surgical act and finally in postoperative for the monitoring. It is also essential to provide the practitioner with a 3D tool for the diagnosis and analysis of the osteoporosis and the fracture risk. Reconstruction of 3D anatomical models can be achieved by the use of direct 3D imaging modalities such as Computed Tomography. However, such technique is limited to complex procedures because of the constraints imposed by cost, availability and risk of radiation to the patient. Thus, the alternative to this kind of 3D imaging is to develop methods for 3D reconstruction which are based only on few 2D radiographs. The main objective of this work is to propose a tool able to reconstruct automatically a 3D surface of the proximal femur from a limited number of X-ray images. Previous studies on the reconstruction of surfaces usually need additional knowledge such as the use of a 3D generic or statistical model of the shape to be reconstructed. The method described in this thesis requires only the 3D coordinates of points calculated from a few pairs of radiographs. Two approaches are proposed. The first method is based on the matching of extracted contours from pairs of radiographs and on a mathematical model based on the principle of the stereovision for the calculation of a 3D point cloud. The second technique uses the results of the previous method as well as new points, chosen by an operator from another proximal femur to improve accuracy at sensitive areas. The reconstruction of the surface model from this cloud of points is obtained by a meshing based on the Poisson's equation. A 3D/3D registration is made between the cloud of the calculated points and the cloud of the extracted points from the generic model ("Gold Standard" obtained with CT-Scan) of the same proximal femur in order to compare the reconstructed surface with a model "ground truth" and thus estimate the accuracy of the method.

Para a execução da etapa inicial da identificação médico-legal de restos humanos (antropometria e exame dos arcos dentários), faz-se necessária uma limpeza prévia da ossada, para a remoção de tecidos moles putrefeitos. Os casos não identificados por esses métodos tradicionais, poderão ser submetidos ao exame de DNA. No entanto, apesar do grande avanço da biologia molecular, utilizando a amplificação de DNA pela PCR, algumas limitações que afetam a habilidade de se obter DNA em restos humanos, permanecem. Dessa forma, o objetivo deste trabalho foi fornecer subsídios morfológicos para os analistas forenses, com ênfase na prática médico-legal, visando uma utilização mais eficiente do DNA obtido de osso compacto de restos humanos em decomposição ou já esqueletizados, sem tecidos moles aderidos. Foi realizado o estudo da microestrutura do tecido ósseo compacto femoral, de restos humanos em decomposição, ainda com tecidos moles, que foram limpos pela fervura em água (n = 7) e ossadas já esqueletizadas pela decomposição natural, que não foram fervidas (n = 8). Destes, seis ossadas foram provenientes de cemitério público regular, após 3 anos de inumação, 1 ossada proveniente da região amazônica, e 1 ossada de origem desconhecida. Estas duas ultimas, apresentado-se porosas ou quebradiças. As análises morfológicas de cortes histológicos foram coradas com hematoxilina e eosina e o DNA amplificado pela PCR para os loci CSF1PO, TPOX, TH01, F13A0, FESFPS, vWA, D16S539, D7S820, D13S317 e amelogenina. Os resultados da análise desses dois grupos foram comparados com os de cadáveres frescos (n = 5) do Serviço de Verificação de Óbitos da Capital. A fervura dos ossos, do modo como é realizada no Instituto Médico Legal de São Paulo, pode aumentar a eosinofilia da matriz óssea e, em alguns casos, pode promover a desagregação dos ósteons. Tal procedimento pode remover células, mas pode também remover possíveis inibidores da PCR, favorecendo a análise do DNA obtido destas amostras. O fator limitante para a obtenção e análise de DNA, em amostras de ossos limpos por fervura, é a quantidade exígua de células. Ossos não submetidos à fervura, após inumação por três anos ou há mais tempo em contato com a terra, podem apresentar alterações da microestrutura. No entanto, a presença de hemácias preservadas e núcleos de osteócitos nestas amostras, indica melhor preservação de células em relação às amostras de ossos fervidos. O fator limitante para a análise de DNA nestas amostras é a presença sugestiva de inibidores da reação de amplificação pela PCR. Restos humanos, sem tecidos moles, macroscópicamente não preservados (porosos e quebradiços), e não submetidos à fervura, apresentam alterações de perda de matriz mineralizada; no entanto, nestas amostras ainda é possível encontrar células preservadas. Os resultados obtidos no neste trabalho permitem traçar algumas estratégias para uma melhor utilização nos protocolos de extração e análise do DNA em osso compacto de restos humanos. / To the first essential step to forensic identification of human remains (anthropological study of race, sex, age, etc) it is necessary a previous cleaning of the bones, to remove decomposing soft tissues. Medico-legal inconclusive or non identified cases, by using these traditional methods, could be subjected to DNA analysis. However, in spite of advances in human identification techniques, specially by PCR amplified DNA, some limitations that affect the ability to obtain DNA in human remains still persist. Therefore, the aim of this study was to provide additional support from morphological analysis, to help forensic analysts personnel to utilise more efficiently the DNA, extracted from compact bones of human remains in decomposition or already skeletonized corpse, it means without soft tissues, with special emphasis in the legal-medicine practice. Femoral compact bones were obtained from: 7 human remains found on the ground, in different degree of decomposition which were cleaned by boiling to remove soft tissues; also studied were collections of bones from 8 corpses having undergone natural decomposition: 6 human remains exhumed after 3 years from a common public cemetery in São Paulo City; 1 case from amazon region and 1 case with no information, both cases remained from long time (more than 3 years) in contact with soil. All eight cases, were not boiled as no soft tissue were adhered. As a control, five cadavers 12 to 16 hours post mortem were also used. The compact bones histological sections were stained by haematoxilin and eosin and the loci CSF1PO, TPOX, TH01, F13A01,FESFPS, vWA, D16S539, D7S820, D13S317 and amelogenin were amplified by PCR.The procedure for boiling the human remains utilised in the Legal Medicine Institute of São Paulo would have increased the eosinophily of bone matrix and, in some cases, promoted the desaggregation of the osteons. In addition these procedures would have removed the cells, but in some cases would have removed possible inhibitors of the PCR, favouring in this way the analysis of DNA obtained from these samples. The limiting factor to obtain successful analysis in bones submitted to boiling seem to be the low quantity of nuclei present in these samples. For the other hand, in bones not cleaned by boiling, the presence of preserved red cells and oscteocyte nuclei inside the lacunae indicates better preservation of cells in relation to those bones cleaned by boiling. The limiting factor to obtain successful DNA analysis in bones exhumed or in contact of soil, is the suggestive presence of inhibitors of PCR. Porous and brittle bones from human remains, without soft tissues that are not processed by boiling, present alterations through loss of mineralised matrix, although it is still possible to found preserved cells in these samples. The results presented in this work clarify concerns about viability of DNA for identification analysis. They also help to establish better strategies for optimisation of DNA extraction and analysis in compact bones of human remains.

Osteoporose und gestörte Heilungsverläufe von Knochenbrüchen verursachen immer noch beachtliche klinische Komplikationen. Ein vielversprechender Ansatz für die nichtinvasive und nichtionisierende Abschätzung des Frakturrisikos und der Bildgebung von Frakturheilung ist quantitativer Ultraschall (QUS). Dennoch liegt die derzeitige Akzeptanz für die Knochenqualitätsabschätzung noch weit hinter herkömmlichen röntgenbasierten Anwendungen. Es wurden akustische Mikroskopie und Synchrotronstrahlen-Mikrotomographie für die Anatomie und altersabhängige Erfassung von strukturellen und elastischen Variationen auf der mikroskopischen Ebene von humanen Femora verwendet. Die gewonnenen Daten dienten als Grundlage für die Erstellung mikromechanischer Modelle von Knochen für numerische Simulationen der Schallausbreitung im humanen Femurhals. Dabei wurde der Aufbau eines US-basierten Femur-Scanners in transversaler Transmission (TT) nachempfunden. Im letzten Abschnitt der Arbeit wurde QUS in TT in in vitro Experimenten am Rattenfrakturmodell auf eine Anwendung für die Bildgebung der Frakturheilung getestet. Die Studien konnten zeigen, dass ein Großteil der adaptiven Fähigkeiten von Knochen auf mikroskopischer Ebene auf eine Kombination von extrazellulärer Matrixelastizität und Gewebeporosität zurückzuführen ist. Die Simulationen des zweiten Teils konnten die Existenz von geführten Wellen im humanen Femurhals bestätigen. Die sensitive Abhängigkeit von US-parametern von frakturrelevanten Knocheneigenschaften zeigt das hohe Potential von QUS für die Frakturrisikoabschätzung. Der zweite Teil der Arbeit konnte erfolgreich die Möglichkeit von QUS in TT zur Diskriminierung von zeitigen Heilungsstadien demonstrieren. Zusammenfassend bestätigt die Studie das hohe Potential von QUS für die Frakturrisikoabschätzung und die Bildgebung der Frakturheilung. / Osteoporosis and impaired bone healing are of high relevance. A promising non-invasive, non-ionizing candidate for fracture risk prediction and monitoring fracture healing is quantitative ultrasound (QUS). However, the acceptance of QUS for bone quality assessment is still not comparable to X-ray based methods. Scanning acoustic microscopy (SAM) and Synchrotron Radiation micro-computer tomography (SRµCT) has been used to investigate anatomical and age dependent variations of micro elastic, structural and mineralization parameters at the tissue level of human femoral bone. Femoral neck models were created based on these data for numerical sound propagation simulations emulating a transverse transmission (TT) setup of an in vivo QUS prototype. In the last part of the project the TT approach has been tested in ex vivo experiments in a rat healing model. The power of QUS, to discriminate two early healing stages has been compared to µCT measurements at the same specimens. It was found that the major contributor to bone adaptation is related to a combination of extracellular matrix elasticity and tissue porosity. It is hypothesized that these parameters are likely to have a considerable impact on the reliability of in silico models. The simulations of the second part confirmed the existence of guided wave propagation in the cortical shell and a high dependency of US parameters on fracture relevant bone properties. The results demonstrate the high potential for bone fracture risk prediction at the femoral neck using QUS. Finally, it was successfully demonstrated that early healing stage discrimination of QUS in TT was superior compared to µCT. In summary these investigations not only show the importance for a precise estimation of micro mechanical properties for numerical modelling but also demonstrate the feasibility and high potential of QUS for bone quality assessment and monitoring of fracture healing.

CAPES / Desde a década de 80, a obesidade mundial aumentou cerca de 36%. O cicloergômetro é um recurso para combater a obesidade e também pode ser utilizado como instrumento para manter, reabilitar e avaliar o condicionamento físico. Pedalar promove uma série de benefícios ao organismo como a melhora do condicionamento cardiorrespiratório, do sistema cardiovascular e musculoesquelético, das conexões neurais e da autoestima. A termografia é um recurso utilizado para medir a temperatura emitida pela superfície da pele. A eletromiografia (EMG) e a mecanomiografia (MMG) quantificam e qualificam a contração muscular. O objetivo desta pesquisa é investigar a viabilidade da análise térmica superficial da pele na identificação das alterações metabólicas e vasculares geradas pela contração muscular durante a pedalada, em indivíduos hígidos, por meio da correlação com os sinais eletromiográficos e mecanomiográficos. Onze voluntários do sexo masculino foram separados em dois grupos: sedentários (GS) e dos ativos fisicamente (GA). Eles participaram do protocolo de pedalada com acréscimo de carga até a fadiga muscular do músculo reto femoral. Dois testes (1 e 2), com intervalo de 1h entre ambos, foram realizados. A coleta dos dados antropométricos, cardíacos, térmicos e dos sinais neuromusculares resultou em tabelas exportadas ao software IBM SPSS Statistics para análise da distribuição normal, percentagens, testes de Wilcoxon, Mann-Whitney e correlação da temperatura com os sinais neuromusculares pelo teste de Spearman. As medianas antropométricas dos participantes mostram para o GS a idade de 33±4,5 anos, massa de 82±12 kg, altura de 1,85±0,09 m, IMC de 23,95±2,65 kg/m2, adipometria da coxa de 11,73±4,98 mm e perimetria de 0,55±0,04 cm. Já para o GA, a idade de 28±12 anos, massa de 66,5±8,58 kg, altura de 1,76±0,04 m, IMC de 21,75±2,78 kg/m2 , adipometria da coxa de 10±2,09 mm e perimetria de 0,57±0,02 cm. A frequência cardíaca diminuiu 25% do primeiro para o segundo teste, assim como a pressão arterial, sistólica e diastólica, em cerca de 3 e 15%, respectivamente, no GS. A temperatura diminuiu de 1 a 3 °C em ambos os grupos; entretanto, o teste de Wilcoxon comprovou o resultado significativo apenas para o GS no primeiro teste, com p= 0,018, e no segundo, com p=0,043. Houve aumento da amplitude dos sinais da MMG e EMG, entretanto, os testes de Wilcoxon e Mann-Whitney não apresentaram p-valor significativo. Na correlação da temperatura com a contração muscular, por meio do teste de Spearman, somente o GA apresentou um p-valor significativo e forte na MMG (p=0,01 e r=1,000) e na EMG (p=0,01 e r= -1,000). Com esses achados, concluiu-se que a variação térmica se comporta de modo inversamente proporcional à variação de amplitude dos sinais de EMG e MMG e a temperatura representa um parâmetro viável para análise das alterações metabólicas e vasculares geradas pela contração durante as atividades aeróbicas com acréscimo de carga até a exaustão. / From the 80s on, global obesity increased around 36%. Cycle-ergometer is a resource to oppose obesity and it is also used as a tool to maintain, rehabilitate and evaluate physical conditioning. Cycling promotes a number of benefits to the organism as the improvement of cardiorespiratory condition, cardiovascular and musculoskeletal system, neural connections and self-esteem. Thermography is a resource used to measure body skin temperature. Electromyography (EMG) and mechanomyography (MMG) allows quantifying and qualifying muscle contraction. The goal of this research is to investigate the feasibility of surface skin thermal analysis in the identification of metabolic and vascular disorders created by muscle contraction during cycling in healthy individuals, through correlation with electromyographical and mechanomiographical signals. Eleven male volunteers were separate on two groups: sedentary (GS) and physically active (GA). Both participated on cycling protocol with load increase until muscular fatigue of the rectus femoris occurs. There were two tests (1 and 2) with 1h interval in between. After collection of anthropometric data, cardiac, thermal and neuromuscular signals, tables were compiled and their values exported to IBM's SPSS Statistics in order to check the normal distribution, percentages, Wilcoxon’s and Mann-Whitney’s test and the correlation of temperature with neuromuscular signals using the Spearman's test. The median anthropometric values of the GS showed average age of 33±4,5 years, weight of 82±12 kg, height of 1,85±0,09 m, BMI of 23,95±2,65 kg/m2, measurement of thigh’s skinfold thickness of 11,73±4,98 mm and perimeter of 0,55±0,04 cm. The average values for GA were: age of 28±12 years, weight of 66,5±8,58 kg, height of 1,76±0,04 m, BMI of 21,75±2,78 kg/m2, measurement of thigh’s skinfold thickness of 10±2,09 mm and perimeter of 0,57±0,02 cm. The heart rate decreased 25% from de first to second test, likewise systolic and diastolic arterial pressure around 3% and 15%, respectively, for GS. Temperature decreased from 1 to 3 °C in both groups, however, Wilcoxon’s test proved a significant result only to GS first test p=0,018 and second test p=0,043. The amplitude of MMG and EMG signal increased, however Wilcoxon’s and MannWhitney’s tests did not show a significant p-value. Concerning temperature correlation to muscle contraction by Spearman’s test, only GA showed a significant p-value and strong correlation for MMG (p=0,01 and r=1,000) and EMG (p=0,01 e r= -1000) signals. From these findings, it was concluded that thermal variation behaviors inversely proportional to EMG and MMG amplitude variation, and temperature represents a feasible parameter to analyze the metabolic and vascular disorders created by muscle contraction during aerobic activities with load increase until exhaustion.

CAPES / Desde a década de 80, a obesidade mundial aumentou cerca de 36%. O cicloergômetro é um recurso para combater a obesidade e também pode ser utilizado como instrumento para manter, reabilitar e avaliar o condicionamento físico. Pedalar promove uma série de benefícios ao organismo como a melhora do condicionamento cardiorrespiratório, do sistema cardiovascular e musculoesquelético, das conexões neurais e da autoestima. A termografia é um recurso utilizado para medir a temperatura emitida pela superfície da pele. A eletromiografia (EMG) e a mecanomiografia (MMG) quantificam e qualificam a contração muscular. O objetivo desta pesquisa é investigar a viabilidade da análise térmica superficial da pele na identificação das alterações metabólicas e vasculares geradas pela contração muscular durante a pedalada, em indivíduos hígidos, por meio da correlação com os sinais eletromiográficos e mecanomiográficos. Onze voluntários do sexo masculino foram separados em dois grupos: sedentários (GS) e dos ativos fisicamente (GA). Eles participaram do protocolo de pedalada com acréscimo de carga até a fadiga muscular do músculo reto femoral. Dois testes (1 e 2), com intervalo de 1h entre ambos, foram realizados. A coleta dos dados antropométricos, cardíacos, térmicos e dos sinais neuromusculares resultou em tabelas exportadas ao software IBM SPSS Statistics para análise da distribuição normal, percentagens, testes de Wilcoxon, Mann-Whitney e correlação da temperatura com os sinais neuromusculares pelo teste de Spearman. As medianas antropométricas dos participantes mostram para o GS a idade de 33±4,5 anos, massa de 82±12 kg, altura de 1,85±0,09 m, IMC de 23,95±2,65 kg/m2, adipometria da coxa de 11,73±4,98 mm e perimetria de 0,55±0,04 cm. Já para o GA, a idade de 28±12 anos, massa de 66,5±8,58 kg, altura de 1,76±0,04 m, IMC de 21,75±2,78 kg/m2 , adipometria da coxa de 10±2,09 mm e perimetria de 0,57±0,02 cm. A frequência cardíaca diminuiu 25% do primeiro para o segundo teste, assim como a pressão arterial, sistólica e diastólica, em cerca de 3 e 15%, respectivamente, no GS. A temperatura diminuiu de 1 a 3 °C em ambos os grupos; entretanto, o teste de Wilcoxon comprovou o resultado significativo apenas para o GS no primeiro teste, com p= 0,018, e no segundo, com p=0,043. Houve aumento da amplitude dos sinais da MMG e EMG, entretanto, os testes de Wilcoxon e Mann-Whitney não apresentaram p-valor significativo. Na correlação da temperatura com a contração muscular, por meio do teste de Spearman, somente o GA apresentou um p-valor significativo e forte na MMG (p=0,01 e r=1,000) e na EMG (p=0,01 e r= -1,000). Com esses achados, concluiu-se que a variação térmica se comporta de modo inversamente proporcional à variação de amplitude dos sinais de EMG e MMG e a temperatura representa um parâmetro viável para análise das alterações metabólicas e vasculares geradas pela contração durante as atividades aeróbicas com acréscimo de carga até a exaustão. / From the 80s on, global obesity increased around 36%. Cycle-ergometer is a resource to oppose obesity and it is also used as a tool to maintain, rehabilitate and evaluate physical conditioning. Cycling promotes a number of benefits to the organism as the improvement of cardiorespiratory condition, cardiovascular and musculoskeletal system, neural connections and self-esteem. Thermography is a resource used to measure body skin temperature. Electromyography (EMG) and mechanomyography (MMG) allows quantifying and qualifying muscle contraction. The goal of this research is to investigate the feasibility of surface skin thermal analysis in the identification of metabolic and vascular disorders created by muscle contraction during cycling in healthy individuals, through correlation with electromyographical and mechanomiographical signals. Eleven male volunteers were separate on two groups: sedentary (GS) and physically active (GA). Both participated on cycling protocol with load increase until muscular fatigue of the rectus femoris occurs. There were two tests (1 and 2) with 1h interval in between. After collection of anthropometric data, cardiac, thermal and neuromuscular signals, tables were compiled and their values exported to IBM's SPSS Statistics in order to check the normal distribution, percentages, Wilcoxon’s and Mann-Whitney’s test and the correlation of temperature with neuromuscular signals using the Spearman's test. The median anthropometric values of the GS showed average age of 33±4,5 years, weight of 82±12 kg, height of 1,85±0,09 m, BMI of 23,95±2,65 kg/m2, measurement of thigh’s skinfold thickness of 11,73±4,98 mm and perimeter of 0,55±0,04 cm. The average values for GA were: age of 28±12 years, weight of 66,5±8,58 kg, height of 1,76±0,04 m, BMI of 21,75±2,78 kg/m2, measurement of thigh’s skinfold thickness of 10±2,09 mm and perimeter of 0,57±0,02 cm. The heart rate decreased 25% from de first to second test, likewise systolic and diastolic arterial pressure around 3% and 15%, respectively, for GS. Temperature decreased from 1 to 3 °C in both groups, however, Wilcoxon’s test proved a significant result only to GS first test p=0,018 and second test p=0,043. The amplitude of MMG and EMG signal increased, however Wilcoxon’s and MannWhitney’s tests did not show a significant p-value. Concerning temperature correlation to muscle contraction by Spearman’s test, only GA showed a significant p-value and strong correlation for MMG (p=0,01 and r=1,000) and EMG (p=0,01 e r= -1000) signals. From these findings, it was concluded that thermal variation behaviors inversely proportional to EMG and MMG amplitude variation, and temperature represents a feasible parameter to analyze the metabolic and vascular disorders created by muscle contraction during aerobic activities with load increase until exhaustion.