Amphibians possess a great diversity of skin glands, among which sexually dimorphic skin glands (SDSGs) are present in males and can form macroglands usually referred to as \"swollen\" regions. Dendrobatoidea is the only clade in which adult males present a swollen in finger IV supposedly associated with the cephalic amplexus, also exclusive to the superfamily. However, few studies of this character have been carried out and the histology of the structure has never been examined. We characterized the different morphologies of the swelling, assessed the anatomical distribution of its glands and tested its homology by taxonomic analysis. We studied the external and internal morphologies and the histology of the hands of adult males and females and juveniles of 27 species to compare all fingers and its gland types. To compare and describe the gland types found at the fingers we ran histochemical studies. We also analyzed the skin of the dorsum of dendobatoid frogs to determine if the gland types found at the swollen region were in fact SDSGs. Our findings showed typical serous and mucous glands at the dorsum and fingers of all individuals and we discovered specialized mucous glands of types I, II and III only at males with the swollen fourth finger. Type I glands are present in all males with the swelling, type II are restricted to Dendrobatidae and type III occur only in Anomaloglossus stepheni. We have also observed that males whose fourth finger was previously coded as \"without the swelling\" had in fact the swelling present, a fact corroborated by the presence of the specialized gland types. According to our samplings, the presence of the swelling in finger IV and of type I glands is a synapomorphy of Dendrobatoidea, with secondary absence at species without the swelling. The presence of type II glands is a synapomorphy of Dendrobatidae and the presence of type III glands is an autapomorphy of An. Stepheni. Our findings will guide studies in the most diverse fields such as dendrobatoid\'s reproductive behavior, biochemistry of glandular secretion and taxonomy / A pele dos anfíbios possui uma grande diversidade de glândulas responsáveis por diversas funções. Glândulas cutâneas sexualmente dimórficas (GCSDs) são geralmente encontradas em machos e podem formar macroglândulas conhecidas como regiões \"inchadas\". Dendrobatoidea é o único clado de anfíbios em que machos adultos possuem um inchaço no dedo IV, supostamente associado ao amplexo cefálico que também é exclusivo da superfamília. No entanto, poucos estudos referentes a este caráter foram realizados e sua histologia nunca foi examinada. Buscamos caracterizar as morfologias do inchaço, avaliar a distribuição anatômica de suas glândulas e realizar sua análise taxonômica. Observamos as morfologias externa e interna das mãos de machos e fêmeas adultos e juvenis de 27 espécies da superfamília e fizemos a histologia das mãos de todos os indivíduos para comparar os dedos e os tipos glandulares presentes neles. Realizamos análises histoquímicas para diferenciar e descrever os tipos glandulares encontrados e para determinar se encontramos GCSDs na região do inchaço, analisamos também a pele do dorso de alguns dendrobatóideos. Encontramos glândulas granulosas e mucosas comuns no dorso e nos dedos de todos os indivíduos. Exclusivamente nos machos adultos com o inchaço, encontramos glândulas mucosas especializadas de tipos I, II e III no dedo IV. Glândulas de tipo I estão presentes em todos os machos que apresentam o inchaço do dedo IV, de tipo II estão restritas aos Dendrobatidae e de tipo III ocorrem apenas em Anomaloglossus stepheni. Observamos ainda que machos de espécies anteriormente codificadas sem o inchaço possuem o inchaço no dedo IV e corroboramos a existência do caráter pela presença dos tipos glandulares descritos. Segundo nossa amostragem, a presença do inchaço e de glândulas de tipo I no inchaço é uma sinapomorfia de Dendrobatoidea, com ausência secundária nas espécies que não possuem o caráter. A presença de glândulas de tipo II é uma sinapomorfia de Dendrobatidae, com perdas independentes em duas espécies e a presença de glândulas de tipo III é uma autapomorfia de An. Stepheni. Trazemos, portanto, informações que poderão guiar estudos em diversos campos como: comportamento reprodutivo, bioquímica das secreções glandulares e taxonomia dos dendrobatóideos

La dextérité manuelle est au centre de notre interaction physique avec le monde. La sophistication des mouvements des doigts chez l’homme nécessite le contrôle de plusieurs composants clés comme la force, l’indépendance, le timing et le séquençage des mouvements des doigts. La dextérité est souvent affectée dans de nombreuses pathologies impactant l’indépendance et la vie quotidienne des patients. Le but global de cette thèse est d’améliorer la rééducation de la dextérité chez ces patients par une meilleure mesure et compréhension de la dextérité et de ses composants. Nous avons développé le Finger Force Manipulandum (FFM), un outil permettant de quantifier les principaux composants de la dextérité chez des sujets sains et des patients. Afin de valider cet appareil, nous avons testé la faisabilité de son utilisation chez des patients souffrant d’un important déficit de dextérité après un accident vasculaire cérébral. Le FFM permet, chez ces patients (N= 10 vs N= 10 sujets sains), de quantifier les différents composants de la dextérité et d’identifier des déficits dans chacun d’entre eux (exemple : les patients font trois fois plus d’erreur que les témoins pour le contrôle de force ; P=0,0002). Les mesures sont plus sensibles que certains tests cliniques comme l’ARAT : elles permettent de détecter des déficits de la dextérité même chez des patients atteignant le score maximum de l’ARAT. Le FFM permet également de créer un profil de dextérité affectée chez chaque patient permettant ainsi de détecter quel composant est significativement affecté et permet aussi de suivre la récupération. Dans une maladie affectant légèrement la dextérité, comme la schizophrénie, lorsque l’on compare les scores FFM des patients stabilisés (N= 35) avec ceux des témoins (N= 20) on constate que les patients ont une performance diminuée de façon significative dans chacun des quatre composants de la dextérité. Certaines des mesures du FFM corrèlent avec des échelles cliniques comme la PANSS (R=0,53, P=0,0019) mais aussi avec des échelles neuropsychologiques. Ces mesures FFM sont également assez sensibles pour détecter une évolution au cours du temps : certains composants restent stables après une remédiation cognitive alors que d’autres s’améliorent. En conclusion, le FFM est un nouvel outil qui permet de quantifier les différents composants de la dextérité. Il est utilisable même chez des patients avec un important déficit manuel et permet d’identifier des profils individuels de dextérité affectée. Il est également assez sensible pour détecter de faibles diminutions de performances motrices comme celles retrouvées chez des patients schizophrènes et pourrait permettre d’identifier certains marqueurs moteurs ayant trait au background neuro-développemental des patients schizophrènes (détection précoce) et à l’évolution de la maladie. / Manual dexterity is essential for our physical interaction with the world. The high degree of dexterity in humans requires sophisticated control of several key components such as the control of force, of independence, timing and sequencing of finger movements. Manual dexterity is affected in various pathologies, impacting activities of daily living and leading to loss of independence. The main purpose of this thesis is to improve rehabilitation of dexterity in these patients by a better behavioral quantification and a clearer understanding of manual dexterity and its components of control. We developed the Finger Force Manipulandum (FFM), a new tool allowing for the quantification of the main components of the dexterity in healthy subjects and in patients. To validate this device, we tested the feasibility of its use with stroke patients suffering from moderate-to-severe deficits of dexterity. In these patients, the FFM allowed for quantification of four components of dexterity and for identification of deficits in each of them (example: patients (N=10) made three times more error than controls (N=10) in force control; P=0.0002). These measures (components) are more sensitive than clinical tests, such as the ARAT: patients reaching the maximum ARAT score still showed deficits of dexterity with the FFM. Based on the four FFM scores, individual profiles of affected dexterity were calculated, highlighting the individual deficit of each patient. This allowed for quantitative longitudinal follow up during recovery. In a disease affecting dexterity mildly, such as schizophrenia, the FFM scores of stabilized patients (N = 35) indicated a significantly lower performance compared to control subjects (N = 20) in each of the four dexterity components. Some of the FFM measures correlated with clinical scales, such as the PANSS (R=0.53, P=0.0019), and also with some neuropsychological scales. These FFM measures also provide indicators for the evolution of dexterity over time: certain components remained stable after cognitive remediation, while others improved. In conclusion, the FFM is a new tool, which allows for quantification of manual dexterity (by measuring various underlying components). It is suitable for patients with moderate-to-sever manual deficits and allows for identification of individual profiles of affected dexterity. It also detects minor manual deficits in schizophrenic patients, and may allow for identification of potential behavioral markers related to the neurodevelopmental background of schizophrenic patients (early detection) and to the evolution of the disease.

Die einzellige Grünalge Chlamydomonas reinhardtii ist ein vielseitiger Modellorganismus sowohl in der Grundlagenforschung als auch für biotechnologische Anwendung. Für die genetische Veränderung wurden verschiedene Methoden entwickelt, jedoch ist die gezielte Modifikation kerncodierter Gene immernoch sehr schwierig. In dieser Arbeit wird eine Strategie vorgestellt, die es ermöglicht, kerncodierte Gene in Chlamydomonas gezielt mit sequenzspezifischen Zinkfinger-Nukleasen zu verändern. Das COP3-Gen, welches den lichtaktivierbaren Ionenkanal Kanalrhodopsin-1 codiert, diente hierbei als Zielsequenz der für die Deletion hergestellten Zinkfinger-Nukleasen. Um eine Charakterisierung der ZFNs zu ermöglichen, wurde ein Modelstamm generiert, der ein inaktiviertes Markergen enthält. Die Inaktiverung erfolgte hierbei durch Insertion der COP3-ZFN Zielsequenz. Die Transformation dieses Modellstamms mit ZFN codierender Plasmid-DNA und einem Reparatur-Template ermöglichte die Wiederherstellung der Markeraktivität und eine Selektion Antibiotika-resistenter Kolonien. Wenn in diesen Experimenten zusätzlich ein COP3 veränderndes Template benutzt wurde, enthielt 1% der analysierten Klone ein mutiertes COP3-Gen. Der Chlamydomonas Augenfleck ist ein lichtsensitives Organell mit entscheidender Funktion für die phototaktische Orientierung der Alge. Eine Deletionsmutante des Blaulicht-Photorezeptors Phototropin zeigte in Experimenten eine veränderte Regulation der lichtabhängigen Augenfleckgröße. Durch Komplementierung der Phototropin-Dysfunktion konnte der lichtabhängige Regulationsprozess wiederhergestellt werden. Die Expression der Phototropin-Kinasedomäne führte zu einer lichtunabhängigen Reduktion der Augenfleckfläche. Interessanterweise führte auch die Expression der N-terminalen LOV-Domänen zu einer geänderten Regulation des Augenflecks und der Phototaxis. Dies deutet, zusätzlich zur Lichtregulation der Kinasedomäne, auf eine zelluläre Signalfunktion der LOV-Domänen hin. / The unicellular green alga Chlamydomonas reinhardtii is a versatile model for fundamental and biotechnological research. A wide toolset for genetic manipulation has been developed for this alga, but specific modification of nuclear genes is still not routinely possible. Here we present a nuclear gene targeting strategy for Chlamydomonas that is based on the application of zinc-finger nucleases (ZFNs). Initially, we designed a set of ZFNs for targeting the COP3 gene that encodes the light-activated ion channel channelrhodopsin-1. To evaluate the designed ZFNs, we constructed a model strain by inserting a non-functional selection marker interspaced with a short COP3 target sequence into the nuclear genome. Upon co-transformation of this recipient strain with the engineered ZFNs and a DNA repair template, we were able to restore marker activity and select antibiotic resistant clones with active nucleases. In cases where cells were co-transformed with a modified COP3 template, 1% of these clones contained a modified COP3 locus as well. The eyespot of Chlamydomonas is a light-sensitive organelle important for phototactic orientation of the alga. Here we found that eyespot size is downregulated in light. In a strain in which the blue light photoreceptor phototropin was deleted, the light regulation of the eyespot size was affected. We restored this dysfunction in different phototropin complementation experiments. Complementation with the phototropin kinase fragment reduced the eyespot size, independent of light. Interestingly, overexpression of the N-terminal LOV-domains alone also affected eyespot size and phototaxis, suggesting that aside from activation of the kinase domain, they fulfill an independent signaling function in the cell. We propose that phototropin is a light regulator of phototaxis that desensitizes the eyespot when blue light intensities increase.

<div>Stability of motor performance is important for voluntary movement control, but it should not be maximized to the exclusion of all else. To transition to a new task, the current task must be destabilized. When expecting to switch tasks, people are known to reduce their stability prior to initiating the change. Here, we determine if the observed stability modulation is influenced by the expectation of future movement, is a relic of the movements performed in the recent past, or is a consequence of both those processes. Furthermore, this work explores the relation between stability modulation observed in isometric finger force production tasks to cognitive flexibility and clinical measures of manual dexterity. Stability modulation can be viewed as a motor response to the recognition of altered environmental demands or internally generated desires to change body movements or postures. Therefore, it is hypothesized that cognitive flexibility – the efficacy of cognitive processing – will relate to stability modulation. Finally, it is hypothesized that the motor adjustments in response to changing task/environment demands will correlate with clinical tests of manual dexterity that involve placing pegs into holes.</div><div>Twenty-two young-adult participants (age 21.05 +/- 0.44 years) completed tasks in the three domains. The Grooved Pegboard and NIH 9-Hole tests of manual dexterity measured their manual function by time to complete the tests. Cognitive flexibility was measured by a task-switching task which required adjusting to a changing set of rules, and the reaction time and accuracy costs of task-switching were recorded. Lastly, participants’ stability of performance in an isometric finger-pressing task was assessed using the uncontrolled manifold analysis and root-mean-square error (RMSE) in the performance. Participants produced pressing forces with four fingers to match a single total force targets presented as feedback on a computer screen. In the ‘Steady’ task, target remained motionless. In the ‘Future Effects’ task, the target remained motionless for several seconds and then began moving. The ‘Past Effects’ task comprised of a dynamic initial portion followed by a stationary target. Lastly, the ‘Combined’ task had a constant force section flanked on either side by epochs of target movement. </div><div>The RMSE results confirmed the existence of stability modulation and established that this is driven by the expectation of future movement, and not by the history of previous movements. The Steady and Past Effects tasks exhibited higher stability than the Future Effects and Combined tasks. The stability estimates obtained from the uncontrolled manifold analysis showed similar trends. Cognitive flexibility (quantified as global accuracy cost) correlated with stability modulation indicating that individuals who show greater cognitive flexibility tend to demonstrate greater stability modulation. However, an association between stability modulation and clinical pegboard tests of manual function were not observed. This may possibly be due to the homogeneity of the test sample, or because the finger-force-production task and pegboard task measure disparate aspects of manual function. </div><div><br></div>

Neste trabalho propõe-se contribuir para o desenvolvimento de tecnologia nacional investigandoa aplicação de uma nova geração de adesivos para a confecção de elementos estruturais de madeira laminada colada (MLC), viabilizando um menor custo final do produto e tornando-o competitivo no mercado interno e externo. Nesse contexto, avaliou-se a resistência à tração paralela em emendas dentadas com dimensão estrutural, em condições de temperatura ambiente e umidade padrão, usando as espécies Pinus e Eucalipto e vários tipos de adesivos, dentre eles o adesivo poliuretano à base de óleo de mamona, foco principal de estudo deste trabalho. Outro aspecto abordado é o da durabilidade, em que foi avaliado o desempenho dos adesivos Cascophen e poliuretano à base de óleo de mamona por meio de ensaios de resistência ao cisalhamento na lâmina de cola e de resistência à tração de emendas dentadas, ambos em corpos-de-prova isentos de defeitos. Também foi avaliada a qualidade da colagem das lâminas, por meio de ensaios de flexão buscando-se a ruptura por cisalhamento, em vigas de MLC com dimensões estruturais, fabricadas em laboratório com os adesivos Cascophen, poliuretano à base de óleo de mamona e Purbond. Os resultados obtidos permitem concluir o bom desempenho do adesivo poliuretano à base de óleo de mamona, demonstrando a sua adequação para aplicação na produção de madeira laminada colada. / The aim of this work is the development of national technology investigating the application of a new adhesive generation to fabrication of glued laminated timber (Glulam) structural members, making possible a lesser final cost of the product and becoming it competitive in the domestic and external market. In this context, the parallel tensile to the fibers tests in joint pieces with structural dimension, in conditions of ambient temperature and standard relative humidity, was evaluated using the species Pinus and Eucalyptus and some types of adhesives, among them the polyurethane adhesive base on castor oil, that is the main focus of study of this work. Another approached aspect is the durability, in which was evaluated the performance of the Cascophen and the polyurethane adhesive base on castor oil adhesive, by means of shear strength tests in the lamina of glue and tensile strength tests of joint laminas, both made in specimens exempt of defects. Also the quality of the collage of lamina, by means of bending and shearing tests, in glulam structural beams was evaluated, which were manufactured in laboratory with the adhesives Cascophen, polyurethane adhesive base on castor oil and Purbond. The results allow concluding the well-behaved performance of polyurethane adhesive base on castor oil, demonstrating its adequacy for application in glued laminated timber fabrication.

No início dos anos 90, os primeiros indícios do tráfico ilícito de materiais nucleares e radioativos foram observados principalmente na região européia. Uma década marcada por inúmeros casos de apreensão desse tipo de material. Como resultado, esses atos passaram a ser alvo de investigações criminais forenses, desenvolvendo-se a partir daí, a ciência forense nuclear. No Brasil não há registros oficiais do tráfico ilícito de material nuclear, entretanto, é amplamente conhecida a extração e o transporte ilegal de materiais geológicos radioativos, assim como a apreensão de fragmentos de materiais utilizados como blindagem de fontes radioativas. Uma das principais ferramentas utilizadas na ciência forense nuclear consiste no estabelecimento de bancos de dados de materiais nucleares. Esses documentos devem conter o maior número possível de informações sobre as características físicas, químicas e nucleares do material apreendido, permitindo a identificação de sua origem, processo de fabricação ou mesmo a idade (age). Assim, se estabelecem padrões de composição característicos de cada material, denominados assinaturas químicas (chemical finger print). Nesse trabalho foi adotado o protocolo forense nuclear seguindo as três etapas de avaliação sugeridas pela Agência Internacional de Energia Atômica (AIEA) na identificação da origem de Siliceto de urânio (U3Si2). Realizaram-se ensaios de caracterização física, química e isotópica dos materiais em estudo e compararam-se os dados com aqueles obtidos para outros compostos de urânio (Tetrafluoreto de urânio, UF4; óxidos de urânio, UO2 e U3O8; tricarbonato de amônio e uranila, TCAU) estabelecendo-se uma assinatura característica para cada um. A partir dos resultados foi possível classificar os compostos por grupos de origem, uma vez que são provenientes de diferentes processos de fabricação e/ ou origem. Demonstrou-se também a importância da criação e manutenção de um banco de dados nuclear na investigação de um evento forense nuclear. / In the early 1990´s, the first illicit trafficking of nuclear and radioactive materials was observed mainly in Europe. A decade marked by numerous cases of seizures of these materials. As a result, these events have become the subject of criminal forensic investigations and develop from there, nuclear forensics. In Brazil there are no illicit trafficking official records of nuclear material, however, is widely known the extraction and illegal transportation of radioactive geological materials, and the materials pieces attachment used as shielding for radioactive sources. One of the main tools used in nuclear forensics is the nuclear materials databases establishment. These documents must contain the most information as possible about the physical, chemical and nuclear material seized, allowing the identification of their origin, manufacturing process or age. Thus, it sets characteristic composition standards of each material, called \"chemical signatures (chemical finger print). In this work nuclear forensic protocol was adopted as well as the three stages of assessment suggested by International Atomic Energy Agency (IAEA) in identifying the origin of uranium silicide (U3Si2). Assays were performed in order to make physical, chemical and isotopic characterization of the studied materials and compared the data with those obtained for other uranium compounds (Uranium tetrafluoride, UF4; uranium oxide, UO2 and U3O8; Yellow cake) by establishing a characteristic signature for each one. Through the assays the uranium compounds were classify by origin groups, as far as they are from different manufactured process and/ or origin. It was also possible to show the importance of a nuclear forensic database during an investigation of a nuclear forensic event.

Le sujet porte sur les bagues et les anneaux découverts dans les contextes archéologiques datés du Ve au IXe siècle, dans une zone géographique allant de la Loire aux Pyrénées, et de la façade atlantique à l’ouest du Rhône. La période chronologique, assez large, permet d’aborder la question de l’héritage antique du Ve siècle, particulièrement important dans des régions romanisées. Le IXe siècle correspond quant à lui à une forte diminution des dépôts funéraires qui constituent les principaux contextes de découverte des objets de parure. Les études portant sur les bagues et les anneaux ont jusqu’alors essentiellement consisté en l’élaboration d’une typochronologie, que ce soit pour la Gaule romaine ou pour les royaumes mérovingiens septentrionaux. Pour ce travail doctoral un catalogue a été élaboré, il comprend plus de 300 artefacts pour lesquels un classement a été proposé. La singularité de ce travail repose sur l’analyse des pratiques sociales menée à partir des artefacts du Sud-Ouest. En tant qu’objets précieux, le port des bagues et des anneaux reste l’apanage des élites ; elles constituent un marqueur important des identités sociales, de la mémoire familiale et des échanges économiques. Afin d’aborder pleinement cette thématique, l’étude est articulée en quatre parties. Les deux premières sont dévolues à la mise en place de la méthodologie, en portant une attention particulière à l’héritage historiographique dans laquelle l’étude se place. Les deux dernières parties sont consacrées à l’inscription de ces bijoux dans l’espace social du haut Moyen Âge. De leur fabrication à leur réception et à leur transmission, il s’agit de documenter les manipulations dont ils ont fait l’objet et qui leur ont conféré leur valeur, jusqu’à leur dépôt en sépulture. / The subject-matter is the finger ornaments discovered in archaeological contexts, dating from the 5th to the 9th century. The geographical area ranges from the Loire to the Pyrenees, and from the Atlantic coast to the west of the Rhône. The chronological period, which is quite broad, makes it possible to approach the question of the ancient heritage of the fifth century, which is particularly important in romanised regions. The 9th century corresponds to a sharp decrease in funeral deposits, which are the main contexts for the discovery of ornaments. Studies on rings have so far essentially consisted in the development of a typochronology, whether for Roman Gaul or for the northern Merovingian kingdoms. For this doctoral study a catalogue has been elaborated, it includes more than 300 artifacts for which a classification has been proposed. The uniqueness of this work is based on the analysis of social practices using artifacts from the Southwest. As precious objects, the wearing of rings remains the prerogative of the elite; they are an important marker of social identities, family memory and economic exchanges. In order to fully address this theme, the study is divided into four parts. The first two are devoted to the implementation of the methodology, paying particular attention to the historiographical heritage in which the study is placed. The last two parts are devoted to the inscription of these jewels in the social space of the early Middle Ages. From their manufacture to their reception and transmission, it is a question of documenting the manipulations to which they have been subjected and which have given them their value, up to their burial.

No início dos anos 90, os primeiros indícios do tráfico ilícito de materiais nucleares e radioativos foram observados principalmente na região européia. Uma década marcada por inúmeros casos de apreensão desse tipo de material. Como resultado, esses atos passaram a ser alvo de investigações criminais forenses, desenvolvendo-se a partir daí, a ciência forense nuclear. No Brasil não há registros oficiais do tráfico ilícito de material nuclear, entretanto, é amplamente conhecida a extração e o transporte ilegal de materiais geológicos radioativos, assim como a apreensão de fragmentos de materiais utilizados como blindagem de fontes radioativas. Uma das principais ferramentas utilizadas na ciência forense nuclear consiste no estabelecimento de bancos de dados de materiais nucleares. Esses documentos devem conter o maior número possível de informações sobre as características físicas, químicas e nucleares do material apreendido, permitindo a identificação de sua origem, processo de fabricação ou mesmo a idade (age). Assim, se estabelecem padrões de composição característicos de cada material, denominados assinaturas químicas (chemical finger print). Nesse trabalho foi adotado o protocolo forense nuclear seguindo as três etapas de avaliação sugeridas pela Agência Internacional de Energia Atômica (AIEA) na identificação da origem de Siliceto de urânio (U3Si2). Realizaram-se ensaios de caracterização física, química e isotópica dos materiais em estudo e compararam-se os dados com aqueles obtidos para outros compostos de urânio (Tetrafluoreto de urânio, UF4; óxidos de urânio, UO2 e U3O8; tricarbonato de amônio e uranila, TCAU) estabelecendo-se uma assinatura característica para cada um. A partir dos resultados foi possível classificar os compostos por grupos de origem, uma vez que são provenientes de diferentes processos de fabricação e/ ou origem. Demonstrou-se também a importância da criação e manutenção de um banco de dados nuclear na investigação de um evento forense nuclear. / In the early 1990´s, the first illicit trafficking of nuclear and radioactive materials was observed mainly in Europe. A decade marked by numerous cases of seizures of these materials. As a result, these events have become the subject of criminal forensic investigations and develop from there, nuclear forensics. In Brazil there are no illicit trafficking official records of nuclear material, however, is widely known the extraction and illegal transportation of radioactive geological materials, and the materials pieces attachment used as shielding for radioactive sources. One of the main tools used in nuclear forensics is the nuclear materials databases establishment. These documents must contain the most information as possible about the physical, chemical and nuclear material seized, allowing the identification of their origin, manufacturing process or age. Thus, it sets characteristic composition standards of each material, called \"chemical signatures (chemical finger print). In this work nuclear forensic protocol was adopted as well as the three stages of assessment suggested by International Atomic Energy Agency (IAEA) in identifying the origin of uranium silicide (U3Si2). Assays were performed in order to make physical, chemical and isotopic characterization of the studied materials and compared the data with those obtained for other uranium compounds (Uranium tetrafluoride, UF4; uranium oxide, UO2 and U3O8; Yellow cake) by establishing a characteristic signature for each one. Through the assays the uranium compounds were classify by origin groups, as far as they are from different manufactured process and/ or origin. It was also possible to show the importance of a nuclear forensic database during an investigation of a nuclear forensic event.

Ho Kar Fai, William. / Thesis submitted in: July 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 121-129). / Abstracts in English and Chinese. / Abstract --- p.I / Acknowledgement --- p.V / Table of Contents --- p.VI / Abbreviations --- p.VIII / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- The recent situation of hepatitis B infection and HBV-induced HCC in Hong Kong / Chapter 1.2 --- Natural history of HBV infection in human / Chapter 1.3 --- The genomic organization of HBV / Chapter 1.4 --- Potential oncogenic mechanism of HBV-induced hepatocarcinogenesis / Chapter 1.5 --- Aim of the present study / Chapter Chapter 2 --- Materials and methods --- p.16 / Chapter 2.1 --- Transformation in E.coli for subtracted normal-counterpart library / Chapter 2.2 --- PCR amplification of subtracted clones / Chapter 2.3 --- Sequencing of subtracted clones with dye-terminator cycle sequencing technology / Chapter 2.4 --- Sequence analysis and database construction / Chapter 2.5 --- Molecular cloning and characterization of novel gene / Chapter 2.6 --- In silico structural and functional analysis of Z313 / Chapter 2.7 --- Cloning and sequencing analysis of zinc finger protein 313 (Z313) / Chapter 2.7.1 --- PCR amplification of target gene -Z313 / Chapter 2.7.2 --- Mini-preparation of plasmid DNA / Chapter 2.7.3 --- Cycle sequencing of cloned cDNA -Z313 with dye-primer technology / Chapter 2.8 --- Multiple Tissue Northern (MTN) blot hybridisation / Chapter 2.9 --- RT-PCR analysis of Z313 / Chapter 2.10 --- Subcellular localization study of Z313 by Green Fluorescent Protein (GFP) / Chapter 2.10.1 --- Directional cloning of Z313 into pEGFP-Cl / Chapter 2.10.2 --- Mini-preparation of plasmid DNA / Chapter 2.10.3 --- Transient transfection of plasmids in different cell lines / Chapter 2.10.4 --- Microscope observation of GFP transfected cells / Chapter Chapter 3 --- Results --- p.49 / Chapter 3.1 --- PCR selection of subtracted clones for sequencing analysis / Chapter 3.2 --- Partial sequencing of selected subtracted clones / Chapter 3.3 --- DNA homology searching using program - BLASTN / Chapter 3.4 --- Catalogue of the 467 ESTs from the subtracted normal-counterpart library / Chapter 3.5 --- Classification and frequency of the subtracted normal-counterpart cDNA clones / Chapter 3.6 --- Identification of putative differentially expressed genes in HCC surrounding normal liver / Chapter 3.7 --- Categorization of ESTs exclusively appeared in the subtracted normal- counterpart library / Chapter 3.8 --- In silico structural and functional analysis of zinc finger protein313 (Z313) / Chapter 3.9 --- Molecular cloning of zinc finger protein 313 (Z313) / Chapter 3.10 --- Northern analysis of zinc finger protein 313 (Z313) / Chapter 3.11 --- RT-PCR analysis of zinc finger protein 313 (Z313) / Chapter 3.12 --- Subcellular localization study of zinc finger protein 313 (Z313) / Chapter Chapter 4 --- Discussion --- p.104 / Chapter 4.1 --- EST analysis on the subtracted normal-counterpart cDNA clones / Chapter 4.1.1 --- Characterization of ESTs generated from the subtracted normal-counterpart library / Chapter 4.1.2 --- Putative differentially expressed genes in HCC surrounding normal liver related to hepatocellular carcinoma / Chapter 4.2 --- Molecular cloning and characterization of zinc finger protein313 (Z313) / Chapter 4.3 --- Future aspects / References --- p.121

By means of reverse-transcription polymerase chain reaction, overexpression of ZNF146 was detected in two human HCC cell lines HepG2 and Hep3B and a clear relationship between HCC and overexpression of ZNF146 has been established. Subcellular localization of ZNF146 protein in liver cells was studied by generation and expression of a green fluorescent protein (GFP) fusion protein. The nuclear localization and the reported DNA binding ability of ZNF146 protein provided a hint that ZNF146 may carry out its function in the cell system by interacting with specific genomic DNA sequences. Recombinant ZNF146 protein was expressed using bacterial and yeast system for the genomic DNA pull down assay in the identification of potential interacting genomic DNA sequences. Several potential genomic DNA sequences that interact with ZNF146 were identified and the gene MDM2 is the one of the candidates that is directly related to human carcinogenesis. MDM2 is a negative regulator of the tumor suppresser protein p53. Deregulation of MDM2 will impair the cell's ability in cell cycle arrest, DNA repair and apoptosis upon induced DNA damage. / Hepatocellular carcinoma (HCC) is a type of primary malignant liver tumor. And is one of the most frequent malignancies worldwide. The focus of this research project is the characterization of a Kruppel zinc finger protein, zinc Finger Protein 146 (ZNF146) using HCC as a disease model. The aim of this project is to understand the functional role ZNF146 and try to explore the mechanism of how ZNF146 might be involved in the carcinogenesis of HCC. / In order to have a better understanding with the protein ZNF146, SUMOylation properties of this protein has been studied. SUMO1 modification on ZNF146 has already been reported. And in our study, experimental result demonstrated that ZNF146 is also modified by SUMO2 and SUMO3 in liver cells. Other than the SUMOylation sites for SUMO1 protein which has been reported, modification sites for SUMO2 at the K247 and K275 positions were mapped, while K191R, K219R, K247R, K256R and K275R, five positions were mapped for SUMO3 modification. A more complete picture of the SUMOylation properties of ZNF146 has been revealed. Since we hypothesized that ZNF146 is related to the p53 tumor suppressor, cell cycle control and DNA repair pathway, a cell cycle study using flow cytometry was performed for the investigation of the effect on cell cycle regulation by ZNF146 overexpression. In our study, ZNF146 overexpression promoted the G1/S transition in the cell division cycle, which indicated that liver cells were more active for the progression of cell cycle. / On the other hand, using cDNA microarray technology expression profiles of ZNF146 overexpressing and non-overexpressing liver cell lines were compared and with real-time polymerase chain reaction, six candidate genes CRLF1, IFI44, ST6GAL1, LOC441601, IL18 and RAD17 were confirmed with their deregulation induced by the overexpression of ZNF146. Four of the candidates, IFI44, LOC441601, IL18 and RAD17 were found to be related to the p53 tumor suppressor activity or DNA damage, repair response and control. This observation, together with the result of genomic DNA pull down assay, gives us a hint that ZNF146 is possibly involved in liver carcinogenesis by affecting DNA repair and cell cycle control upon induced DNA damage. / The gene ZNF146 codes for a member of the Kruppel zinc finger proteins, however ZNF146 protein is different from most members of the Kruppel zinc finger proteins subfamily. It encodes a 33 kDa protein solely composed of 10 zinc finger motifs and is devoid of any non-zinc finger regulatory domain for interactions with other proteins. ZNF146 overexpression has been reported in a number of cancers including colon cancer and pancreatic carcinoma. However, the functional role of ZNF146 overexpression in tumorigenesis is yet to be solved and not much research on how ZNF146 might be invovled in the establishment of HCC was published. / To conclude, the experimental results of this study support the hypothesis that ZNF146 overexpression may deregulating the cell division cycle and some genes differentially regulated upon over-expression of ZNF146 are related to the regulations of DNA damage response. Future research on ZNF146 can be focused on the detail regulatory pathway of ZNF146 overexpression and its interaction between the p53 tumor suppressor, DNA damage response and cell cycle regulation, and a fuller picture of how ZNF146 overexpression might induce hepatocarcinogenesis can be revealed. / Yeung, Tsz Lun. / Adviser: Miu Yee (Mary) Waye. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3329. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 287-304). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.