A estimulação epidural do córtex motor (ECM) é eficaz no tratamento da dor neuropática refratária, porém seus mecanismos de ação ainda são incertos. Sabendo que a ECM ativa a via analgésica descendente em ratos, fomos investigar o efeito da ECM sobre os núcleos serotonérgicos descendentes, dorsal da rafe (NDR) e magno da rafe (NMR) e sobre os neurônios da coluna posterior da medula espinhal (CPME). Ratos Wistar, submetidos à ECM, foram avaliados no teste de pressão da pata e seus tecidos foram avaliados frente à imunorreatividade (IR) para Egr-1 (marcador de ativação neuronal), serotonina (5HT) e substância P (SP). A ECM induziu antinocicepção em 62% nos animais, não alterou a ativação do NDR, entretanto ativou o NMR (67%), quando comparado com ratos não estimulados. A ECM aumentou a IR-5HT em 75% no NDR e em 92% no NMR. Na CPME, a ECM inibiu os neurônios nociceptivos (48%), porém não interferiu com a IR-SP. Estes resultados sugerem que a ECM induz analgesia, em parte, via ativação do sistema serotonérgico descendente. / Motor cortex stimulation (MCS) is effective in the treatment of refractory neuropathic pain; however, its mechanisms of action remain unclear. Since the MCS activates the descending pain pathway in rats, we investigated the MCS effect on the descending serotonergic nuclei, dorsal raphe nucleus (DRN) and the magnus raphe nucleus (MRN) and also on the neurons of the dorsal horn of the spinal cord (DHSC). Wistar rats, submitted to MCS, were evaluated by paw pressure test and its tissues were evaluated by immunoreactivity (IR) to Egr-1 (neuronal activation marker), serotonin (5HT) and substance P (SP). MCS induced antinociception by 62% in animals, although did not modify the NDR activation; however, activated the MRN (67%), when compared to control groups. MCS increased the IR-5HT by 75% in the NDR and 92% in the MRN. In the DHSC, MCS inhibited the nociceptive neurons (48%), however did not change the IR-SP. These results suggest that MCS induce antinociception, partly, by the descending serotonergic pathway activation.

O Tronco encefálico é uma estrutura singular do sistema nervoso central, pois nele passam tratos sensoriais ascendentes da medula espinal, tratos sensoriais da cabeça e do pescoço, os tratos descendentes motores originados no prosencéfalo, as vias ligadas a centros de movimento dos olhos, contemos núcleos dos nervos cranianos, e também está envolvido na regulação do nível de consciência através de projeções ao prosencéfalo oriundas da formação reticular. Tudo isto compactado em um espaço muito exíguo o que faz deste um local particularmente sensível às alterações patológicas, mesmo que pequenas, que acabam cursando com uma riqueza de sinais neurológicos devido a presença muito próxima das estruturas já citadas. Compreender a anatomia interna do tronco encefálico é essencial para o diagnóstico neurológico e a prática da medicina clínica. Por tudo que foi exposto, o tronco encefálico é um terreno fértil para o estudo através do diagnóstico por imagem, principalmente quando realizado por novas tecnologias, como exames em aparelho de ressonância magnética de alto campo (3 teslas). No entanto, pouco se sabe sobre as correlações existentes entre a microscopia e as imagens de ressonância magnética do tronco encefálico. Sendo assim, o objetivo deste estudo foi analisar e correlacionar as diversas estruturas encontradas no tronco encefálico, visualizadas em peças microscópicas de encéfalos humanos post mortem, com as imagens de ressonância magnética dos mesmos, antes da dissecção, mapeandoas e discernindo-as, contribuindo assim para diagnósticos mais precisos e topográficos das patologias que acometem o tronco encefálico, justificando o presente estudo. O estudo foi de caráter observacional exploratório e descritivo, adotando as seguintes técnicas para coleta da informações: os encéfalos humanos (n=3) foram submersos em recipiente contendo água e então lacrados de forma que o ar ambiente não entrasse no recipiente. As imagens de RM foram adquiridas em sequência gradiente echo (FFE) 2D em equipamento de campo 3T (PHILIPS ACHIEVA), com bobina de 8 canais de encéfalo. Foi utilizado TE=9,0 ms, TR=1000 ms e o ângulo de flip 90°; número de médias igual 10 e BW por pixel igual a 72 Hz/pixel. O Fator EPI utilizado foi igual a um e a resolução espacial de 0,219x0,219x2,0 mm3 com FOV de 210x210x90mm3. O tempo total de aquisição foi de 3 horas e 01 minuto e 96 segundos. Imagens histológicas utilizadas no presente estudo são do banco de dados do departamento de patologia da UNICAMP e foram comparadas às imagens obtidas na ressonância magnética. Demonstramos que foi possível a identificação das estruturas visíveis, histologicamente, nas imagens obtidas, com definição e resolução suficiente para a geração de um atlas de imagens de ressonância magnética de cortes do tronco encefálico / The Brainstem is a unique structure of the central nervous system, because in it pass ascending sensory tracts of the spinal cord, sensory tracts of head and neck, descending tracts originated in the forebrain, the pathways linked to eye movement centers, contains nuclei of cranial nerves, and is also involved in regulating the level of consciousness through projections to the forebrain that arise from the reticular formation. All these estructures are packed into a very small space which makes the brainstem a particularly sensitive place to pathological changes, that bring up a large amount of neurological signs due to very close packing of the aforementioned structures.Understanding the internal anatomy of the brainstem is essential for the neurological diagnosis and the clinical medicine practice. Thus, the brainstem is fertile ground for the study through diagnostic imaging, especially when performed by new technologies such as high-field (3 tesla) MRI machines. However, little is known about the correlation between the microscopy and magnetic resonance imaging of the brainstem. The aim of this study was to analyze and correlate the various structures found in the brainstem, viewed in microscopic slides of human brains post mortem, with the magnetic resonance imaging thereof, prior to dissection, mapping them and defining them, thus contributing to more accurate diagnoses and surveying of pathologies that affect the brainstem. Human brains (n = 3) were submerged in a container containing water, and then sealed so that the ambient air does not enter the container. MRI images were acquired in gradient echo sequence (FFE) 2D 3T field equipment (PHILIPS ACHIEVA) with coil 8- channel brain. It was used TE = 9.0 ms, TR = 1000 ms and flip angle 90°; number of averages equal to 10 and BW per pixel equal to 72 Hz/pixel. Factor PPE used was equal to one and the spatial resolution of FOV with 0,219x0,219x2,0 mm3 210x210x90mm3. The total acquisition time was 3 hours, 01 minute and 96 seconds. Histological images used in this study are from the pathology department of State University of Campinas (UNICAMP) database and compared to images obtained in MRI. We demonstrated that it was possible to identify histologically visible structures in images acquired with sufficient resolution and definition to generate an magnetic resonance imaging atlas of the brainstem sections

Orientador: Jorg Kobarg / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-21T03:42:39Z (GMT). No. of bitstreams: 1 Migueleti_DeividLucasdosSantos_M.pdf: 8057948 bytes, checksum: ef239a4533686a90e511e97afd0f97ba (MD5) Previous issue date: 2012 / Resumo: A proteína UNC-76 foi identificada como necessária para a fasciculação e elongação de axônios do verme Caenorhabditis elegans durante o desenvolvimento do sistema nervoso. A homóloga de mamíferos FEZ1 apresenta altos níveis de expressão em tecidos neuronais e camundongos knockout para o gene FEZ1 apresentam desvios de comportamento que remetem a desordens neurológicas. O papel de FEZ1 no desenvolvimento do sistema nervoso parece residir na sua associação com elementos do citoesqueleto e vias de sinalização (e.g., PKC?, E4B, DISC1) que conduzem o crescimento axonal e a polarização celular. Trabalhos do grupo mostram que FEZ1 é uma proteína multifuncional (hub), capaz de interagir com mais de 50 parceiros através de seus domínios coiled-coil. Além disso, a superexpressão de FEZ1 em células HEK293 provoca o aparecimento de núcleos multilobulados, um fenótipo comum em alguns tipos de leucemia. Nesse trabalho foi investigado o papel de FEZ1 nos mecanismos causadores dos núcleos multilobulados e as consequências funcionais de sua superexpressão na viabilidade celular, tentando extrapolar esse modelo para leucemias. Análises in silico de diversas leucemias mostraram que FEZ1 está superexpressa em LMAs e que isso pode se relacionar à ocorrência da fusão 11q23/MLL. A expressão de FEZ1 na linhagem leucêmica THP-1 foi detectada por Western blotting, mas, a expressão em PBMCs de pacientes ainda permanece sem provas empíricas. Para avaliar as consequências funcionais da superexpressão, uma linhagem com expressão estável e indutível foi obtida e utilizada em ensaios de proliferação e resistência a quimioterápicos. Porém, não foram observadas diferenças entre as linhagens expressando a fusão FLAG-FEZ1 e as que expressavam o FLAG tag apenas. Em um ensaio de IP-MS utilizando tais linhagens, foram identificadas proteínas cuja interação com FEZ1 pode ser modulada pela atividade de PKCs. Finalmente, a cotransfecção de FEZ1 inteira com coiled-coils C-terminais diminui a formação de núcleos multilobulados em quase 40%. A transfecção com o mutante FEZ1 nocys contendo 5 cisteínas mutadas não teve o mesmo efeito, mas, novos experimentos são necessários para determinar o potencial de sinergismo que esses dois componentes podem ter sobre a ocorrência desse fenômeno / Abstract: The protein UNC-76 was identified as necessary for fasciculation and elongation of axons of the worm Caenorhabditis elegans during development of the nervous system. The mammalian homologue FEZ1 is mostly expressed in neuronal tissues and FEZ1 knockout mice present behavior abnormalities that resemble neurological disorders. The role of FEZ1 in the development of the nervous system seems to lie in its association with cytoskeletal elements and signaling pathways (e.g., PKC?, E4B, DISC1) regulating axon outgrowth and cell polarization. The studies of our group have shown that FEZ1 is a hub, able to interact with more than 50 partners through its coiled-coil domains. Furthermore, overexpression of FEZ1 in HEK293 cells causes the appearance of flower-like nuclei, a common phenotype to certain types of leukemia. In this work the role of FEZ1 in the mechanisms of flower-like nuclei formation and functional consequences of its overexpression on cell viability were investigated, attempting to extrapolate this model for leukemias. In silico analysis of several leukemias showed that FEZ1 is overexpressed in AML patients and that this may relate to the occurrence of 11q23/MLL genetic fusion. FEZ1 expression in leukemic THP-1 cells was detected by Western blotting, but the expression in PBMCs of leukemic patients still lacks empirical evidence. To assess the functional consequences of overexpression, cell lineage with stable and inducible expression of FEZ1 was obtained and used in proliferative assays. However, it was not observed any differences between lineages expressing FLAG-FEZ1 fusion protein or FLAG tag alone. IP-MS assay using these lineages identified proteins whose interaction with FEZ1 could be modulated by the activity of PKCs. Finally, cotransfection of C-terminal coiled-coils and FEZ1 full-length decreases flower-like nuclei formation to nearly 40%. Transfection with FEZ1nocys mutant containing five substituted cysteines did not play the same, but further experiments are needed to determine the potential synergism these two components may have on this phenomenon / Mestrado / Genetica Animal e Evolução / Mestre em Genética e Biologia Molecular

The environment inside the atomic nucleus is one of the most fascinating arenas for the study of quantum chromodynamics (QCD). The strongly-interacting nature of the nuclear medium affects the nature of both QCD processes and the quark-gluon structure of hadrons, allowing several unique aspects of the strong nuclear force to be investigated in reactions involving nuclear targets. The research presented in this dissertation explores two aspects of nuclear QCD: firstly, the partonic structure of the nucleus itself; and secondly, the use of the nucleus as a micro-laboratory in which QCD processes can be studied. The partonic structure of the nucleus is calculated in this work by deriving and utilizing a convolution formula. The hadronic structure of the nucleus and the quark-gluon structure of its constituent nucleons are taken together to determine the nuclear partonic structure. Light cone descriptions of short range correlations, in terms of both hadronic and partonic structure, are derived and taken into account. Medium modifications of the bound nucleons are accounted for using the color screening model, and QCD evolution is used to connect nuclear partonic structure at vastly different energy scales. The formalism developed for calculating nuclear partonic structure is applied to inclusive dijet production from proton-nucleus collisions at LHC kinematics, and novel predictions are calculated and presented for the dijet cross section. The nucleus is investigated as a micro-laboratory in vector meson photoproduction reactions. In particular, the deuteron is studied in the break-up reaction γd → Vpn, for both the ϕ(1020) and J/ψ vector mesons. The generalized eikonal approximation is utilized, allowing unambiguous separation of the impulse approximation and final state interactions (FSIs). Two peaks or valleys are seen in the angular distribution of the reaction cross section, each of which is due to an FSI between either the proton and neutron, or the produced vector meson and the spectator nucleon. The presence and size of the latter FSI valley/peak contains information about the meson-nucleon interaction, and it is shown that several models of this interaction can be distinguished by measuring the angular distribution for the deuteron breakup reaction.

Solid-state nuclear magnetic resonance (SSNMR) has proven to be a useful tool in the characterization of non-covalent interactions such as hydrogen bonding and cation-π interactions. In recent years, the scientific community has manifested a renewed interest towards an important class of non-covalent interaction, halogen bonding (XB), as it has applications in various fields such as crystal engineering and biological processes. This dissertation demonstrates that NMR parameters measured in the solid state are sensitive to changes in electronic structure, which are caused by halogen bonds. A series of halogen bonded compounds exhibiting interactions between different diiodoperfluorobenzenes (p- C6F4I2, o- C6F4I2, sym- C6F3I3, p- C6H4I2) and various halogen bond acceptors have been synthesized as part of this work. These new halogen bonded compounds were characterized with a combined theoretical and experimental SSNMR, X-ray diffraction (XRD) methods. The complete multinuclear magnetic resonance spectroscopy of the nuclei involved directly in the halogen bond (13C, 14/15N, 31P, 77Se, 35/37Cl and 79/81Br) were recorded at multiple magnetic fields (4.7, 9.4, 11.75 and 21.1 T). The specialized SSNMR experiments provided high-resolution spectra of quadrupolar nuclei, which were WURST-QCPMG or solid-echo type experiments combined with the variable offset cumulative spectral (VOCS) method, as for spin 1/2 nuclei cross polarization magic angle spinning (CPMAS) experiments were usually appropriate. This dissertation will discuss successful applications of SSNMR spectroscopy to characterize halogen bonds, it will demonstrate the significant changes in NMR observables in the presence of XB interaction and thus establish that NMR parameters are very sensitive to halogen bonding interaction. Furthermore, this work explains why the NMR parameters are correlated with the halogen bonding interaction. The different trends observed between the NMR observables and the halogen bonding were further understood with a ZORA-DFT natural localized molecular orbital (NLMO) study.

L’étude des noyaux à la drip-line proton est un outil récent et puissant pour sonder la structure nucléaire loin de la vallée de stabilité. En particulier, le phénomène de radioactivité deux protons prédit théoriquement en 1960 a été découvert expérimentalement en 2002. Ce travail de thèse concerne une expérience réalisée au GANIL dans le but d’étudier la radioactivité 2pde 54Zn avec une chambre à projection temporelle, développée pour la détection individuelle de chaque proton et la reconstruction de leur trajectoire en trois dimensions. L’analyse des données a permis de déterminer les corrélations en énergie et en angle entre les deux protons.Celles-ci ont été comparées à un modèle théorique qui prend en compte la dynamique de la décroissance, permettant d’obtenir des informations sur la structure de l’émetteur. La statistique obtenue étant très faible, l’interprétation des résultats reste encore limitée mais ces résultats ouvrent de très belles perspectives sur les études futures des noyaux aux limites d’existence. / The study of nuclei at the proton drip-line is a recent and efficient tool to prove the nuclearstructure far from stability. In particular, the two proton radioactivity phenomenonpredicted in 1960 has been discovered in 2002. This PhD thesis concerns an experiment realisedat GANIL, in order to study the 2p radioactivity of 54Zn with a time projection chamber,developed for the individual detection of each proton and the reconstruction of their tracksin three dimensions. The data analysis allowed to determine the correlations in energy andangle between the two protons. They have been compared to a theoretical model, which takesinto account the dynamics of the emission, giving information about the structure of theemitter. However, due to the very low statistics, the interpretation of the results is limitedbut these results open very interesting prospects for further studies of nuclei at the limits ofthe existence.

Since the construction of the IceCube Neutrino Observatory was completed in 2010, many amazing discoveries have been made in the field of neutrino physics. Recently a neutrino event has been linked to an blazar-type active galactic nucleus source, bringing us one step closer to understanding the production of high-energy extragalactic neutrinos and ushering in a new era of multimessenger astronomy. This was found by linking the neutrino event to one of the Fermi Collaboration’s gamma ray sources which had a blazar counterpart. The quest to link other neutrino events to AGN (active galactic nuclei) sources through collaboration with the Fermi Large Area Telescope has turned up some interesting candidates. The fact that some of these potential sources are not blazars is curious and, although unconfirmed as neutrino sources, these objects merit further investigation due to their unusual nature.

La physique à la drip-line offre l'opportunité d'étudier les noyaux à la limite du paysage couvert par la physique nucléaire, où l'addition de nouveaux protons et neutrons ne peut plus être supportée. A l'approche de la drip-line, la structure même des noyaux se trouve modifiée par rapport aux plus stables. Les fermetures de couches classiques disparaissent et de nouveaux phénomènes surgissent, tels que la clusterisation et les halos. Dans ce travail, nous avons revisité la spectroscopie du noyau non-lié 10He par l'intermédiaire d'une réaction de transfert d'un proton 11Li(d,3He) à 50 AMeV étudiée par la méthode de masse manquante. Pour ce faire, nous avons utilisé le détecteur MUST2 sur la ligne RIPS du RIKEN Nishina center. La partie expérimentale de ce travail repose sur un dispositif novateur utilisant des détecteurs de silicium très fins. Pour la première fois, la mise en oeuvre de ces derniers en conjonction avec des détecteurs sensibles en position de haute granularité a été démontrée. L'approche par réaction de transfert conduit à des résultats très clairs sur la position du premier état résonant du 10He, trouvé dans ce travail à 1.4(3) MeV, en accord avec la plupart des précédents travaux expérimentaux. L'obtention pour la première fois d'un facteur spectroscopique permet de nous renseigner sur la nature du recouvrement <10He|11Li>, trouvé égale à 0.13(6). / Physics at the drip-line offer the opportunity to study nuclei at the limits of the nuclear landscape, where additional protons or neutrons can no longer be kept in the nucleus. In the vicinity of the drip-lines, the structural features of the nuclei change with respect to the more stable nuclei, the normal shell closure of particles disappear and new phenomena appear, such as clusterization and halo. In this work we have revisited the spectroscopy of the unbound 10He by means of the 11 Li(d,3 He) reaction at 50 AMeV studied in missing mass method with the MUST2 array at the RIPS beam line of RIKEN. The experimental part of this study rely on new detection set-up using very thin silicon detectors. For the first time, the effective operability of these detectors, in conjunction with high granularity position sensitive detectors, was proved. The transfer reaction approach led to clear results on the 10He first resonant state position, found in this work at 1.4(3) MeV, in agreement with most of the previous experimental works. The extraction of a spectroscopic factor for the first time shed light on the <10He|11Li> overlap, found to be 0.13(6).

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第21570号 / 理博第4477号 / 新制||理||1642(附属図書館) / 京都大学大学院理学研究科物理学・宇宙物理学専攻 / (主査)准教授 延與 佳子, 講師 村上 哲也, 教授 田中 貴浩 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DFAM

Afin de sonder la persistance de la fermeture de couches à N=50 pour les isotopes de nickel, une expérience a été réalisée au RIBF (RIKEN, Japon) pour produire des noyaux autour du ⁷⁸Ni par fission en vol d'un faisceau de ²³⁸U. Les spectromètres BigRIPS et ZeroDegree ont été utilisés pour la purification et la sélection des noyaux d'intérêt. La décroissance de ces noyaux a été observée avec le dispositif EURICA pour la détection des rayonnements gamma, couplé avec le détecteur à particules chargées WAS3aBi.L'analyse des corrélations entre l'implantation des ions ⁷⁹′⁸⁰Cu et leur décroissance, ainsi que les spectres de décroissance gamma en simple et double coïncidence, nous ont permis de proposer un schéma de niveaux excités pour le ⁷⁹Zn, avec assignation de spin et parité pour la plupart des états. Nous avons aussi mis une contrainte entre 3hbar et 5hbar sur la valeur du spin de l'état fondamental du ⁸⁰Cu. La comparaison avec un modèle schématique de couplage cœur-particule nous a permis d’interpréter le schéma de niveau et de proposer des hypothèses sur la nature des états observés. Les énergies des états de particule individuelle extraites du calcul théorique indiquent la persistance du gap N=50 à Z=28. / In order to investigate the persistence of the N=50 gap for the Nickel isotopes, an experiment has been performed at RIBF (RIKEN, Japan) to produce exotic nuclei around ⁷⁸Ni by in-flight fission of a ²³⁸U beam. The BigRIPS and ZeroDegree spectrometers were used to purify and select the nuclei of interest. Their decay was detected with the EURICA detector for γ-ray detection, coupled with the charged particle WAS3aBi detector. The analysis of the correlations between the ion implantation and their decay, as well as the γ decay spectra in simple and double coincidence allowed us to propose a level scheme of the excited states for the ⁷⁹Zn, and we assigned the spin and parity for most of the states. We also constrained the spin value for the fundamental state of the ⁸⁰Cu, between 3hbar and 5hbar. Comparison with a core-particle coupling model allowed us to interpret the level scheme and propose hypothesis about the nature of the observed states. The single particle state energies extracted from the theoretical calculation indicate the persistence of the N=50 gap at Z=28.