Traumatic brain injury is a major health problem with much of its morbidity associated with traumatic axonal injury (TAI). To date, significant insight has been gained into the initiating pathogenesis of TAI. However, the specific anterograde and retrograde sequelae of TAI are poorly understood because the diffuse nature of TAI complicates data analysis. To overcome this limitation, we subjected transgenic mice expressing yellow fluorescent protein (YFP) within the visual system to central fluid percussion injury, and consistently generated diffuse TAI within the optic nerve that could easily be followed in the organized YFP positive fibers. We demonstrated progressive axonal swelling, disconnection and proximal and distal axonal dieback, with regression and reorganization of the proximal swellings, and the persistence of the distal disconnected and degenerating swellings. Antibodies targeting the C-terminus of amyloid precursor protein, a marker of TAI, mapped to the proximal axonal segments without distal targeting. Antibodies targeting microglia/macrophages, revealed activated microglia/ macrophages closely encompassing the distal disconnected, degenerating axonal segments at 7 - 28 days post injury, suggesting their role in the delayed axonal degeneration. In contrast, in the proximal reorganizing axonal segments, microglia/macrophages appeared less reactive with their processes paralleling preserved axonal profiles. Concomitant with these events, YFP fluorescence quenching also occurred, complicating data analysis. This quenching mapped to Texas-Red-conjugated-IgG immunoreactive loci, suggesting that blood–brain barrier disruption and its attendant edema participated in fluorescence quenching. This was confirmed through antibodies targeting endogenous YFP, which identified the retention of intact axons despite YFP fluorescent loss. Paralleling these events, TAI was not accompanied by retrograde retinal ganglion cell (RGC) death. Specifically, no TUNEL+ or cleaved caspase-3 immunoreactive RGCs were observed from 2 days to 3 months post-TBI. Further, Brn3a immunoreactive RGC quantification revealed no significant RGC loss. This RGC preservation was accompanied by the persistent phospho-c-Jun expression for up to 3 months post-TBI, a finding linked to neuronal survival and potential axonal repair. Parallel ultrastructural study again failed to identify RGC death. Collectively, this study provides unprecedented insight into the evolving pathobiology associated with TAI, and offers advantages for future studies focusing on its therapeutic management and neuronal reorganization.

Introdução: O sucesso da cateterização da via biliar é de importância para o diagnóstico e terapêutica nas afecções biliopancreáticas nos procedimentos de colangiopancreatografia retrógrada endoscópica (CPRE) e está associado a complicações graves e mortalidade. O objetivo do estudo foi comparar o sucesso, perfil laboratorial e as complicações da técnica de fistulotomia papilar direta com o acesso cateter e fio-guia. Métodos: No período de julho de 2010 a maio de 2017 foram selecionados e randomizados para CPRE em dois grupos: cateterização com cateter e fioguia (Grupo I) e a fistulotomia papilar (Grupo II). As curvas de amilase, lipase e proteína C reativa (T0, 12 e 24 horas) e as complicações (pancreatite, sangramento e perfuração) foram avaliadas após CPRE. Resultados: Foram incluídos 102 pacientes (66 do sexo feminino e 36 do masculino, com idade média de 59,11±18,7 anos) e divididos em 51 pacientes para Grupo I e 51 no Grupo II. Os sucessos das cateterizações dos Grupos I e II foram de 76,47% e 100%, respectivamente (p=0,0002). Doze pacientes (23,53%) do Grupo I foram considerados pacientes de cateterização difícil e submetidos à fistulotomia papilar com sucesso no acesso biliar. Foram observadas 13,7% (2 perfurações e 5 pancreatites leves) e 2 % (1 paciente com perfuração e pancreatite) complicações nos Grupos I e II, respectivamente (p=0,062). Conclusão: A fistulotomia papilar demonstrou maior eficácia na cateterização da via biliar e com menor índice de amilasemia e lipasemia em comparação a cateterização com papilótomo e fio guia. As complicações foram semelhantes entre as duas técnicas / Background: The success of biliary tract cannulation is important for the diagnosis and treatment of biliopancreatic diseases in endoscopic retrograde cholangiopancreatography (ERCP) procedures. ERCP is associated with severe complications and mortality. The aim of the study was to compare the success, laboratory profile and complications of the direct papillary fistulotomy technique with standard catheter and guidewire access. Methods: In the period from July 2010 to May 2017, two groups were selected and randomized for ERCP: cannulation with catheter and guidewire (Group I) and papillary fistulotomy (Group II). The curves of amylase, lipase and C-reactive protein (T0, 12 and 24 hours) and complications (pancreatitis, bleeding and perforation) were evaluated after ERCP. Results: A total of 102 patients (66 females and 36 males, mean age 59.11 ± 18.7 years) were divided into 51 patients for Group I and 51 for Group II. The success of cannulation was 76.47% and 100%, in Groups I and II, respectively (p = 0.0002). Twelve patients (23.53%) of Group I were considered to have difficult cannulation and were submitted to fistulotomy with successful biliary access. There were 13.73% (2 perforations and 5 mild pancreatitis) and 2% (1 patient with perforation and pancreatitis) complications in Groups I and II, respectively (p=0,062). Conclusion: Papillary fistulotomy demonstrated greater efficacy in the bile duct cannulation and presented lower serum amylase and lipase compared with standard catheter and guidewire cannulation. Complications were similar in the two techniques

Network dynamics are thought to be the substrate of brain information processing and of mental representations. Moreover, network-wide dysfunctions are recognized to be at the core of several psychiatric and neurodegenerative disorders. Yet, our ability to target specific networks for functional or genetic manipulations remains limited. The development of monosynaptically-restricted Rabies virus, G-deleted Rabies virus (ΔG-Rabies), has greatly facilitated the anatomical investigation of neural circuits, revealing the network synaptic structure upstream of defined neuronal populations. However, the inherent cytotoxicity of the Rabies virus largely restrains its use to the mere structural characterisation of neural networks. To overcome this limitation, I generated novel tools that allow the manipulation of neural networks for the entire life of the animal, without affecting neuronal and circuit properties. I first developed a viral system obtained by engineering the Rabies virus genome to eliminate its cytotoxicity. This led to the generation of a Self-inactivating Rabies virus (SiR) that transcriptionally disappears from the infected neurons while leaving permanent genetic access to the traced network. I showed that SiR provides a virtually unlimited temporal window for the functional manipulation of neural circuits in vivo without adverse effects on neuronal physiology. To further expand our ways of intervening on neural networks function I then developed a completely virus-free system, named Genetically-Encoded TransSynaptic Shuttle (GETSS), which is the only specific genetically-encoded transsynaptic tracer to date. In this thesis, I established novel approaches that provide, for the first time, the functional and genetic access to traced network elements in vivo for the lifetime of the animal, with no cytotoxic effects, no changes in the electrophysiological properties of the traced neurons and no adverse effects on network function. This opens new horizons in the functional investigation of neural circuits and potentially represent the first approaches to experimentally monitor neural circuit remodelling in vivo.

Challenges with integrating high-κ gate dielectric,retrograde Si1-xGexchannel and silicided contacts in future CMOStechnologies are investigated experimentally and theoreticallyin this thesis. ρMOSFETs with either Si or strained Si1-xGex surface-channel and different high-κgate dielectric are examined. Si1-xGex ρMOSFETs with an Al2O3/HfAlOx/Al2O3nano-laminate gate dielectric prepared by means ofAtomic Layer Deposition (ALD) exhibit a great-than-30% increasein current drive and peak transconductance compared toreference Si ρMOSFETs with the same gate dielectric. Apoor high-κ/Si interface leading to carrier mobilitydegradation has often been reported in the literature, but thisdoes not seem to be the case for our Si ρMOSFETs whoseeffective mobility coincides with the universal hole mobilitycurve for Si. For the Si1-xGexρMOSFETs, however, a high density ofinterface states giving riseto reduced carrier mobility isobserved. A method to extract the correct mobility in thepresence of high-density traps is presented. Coulomb scatteringfrom the charged traps or trapped charges at the interface isfound to play a dominant role in the observed mobilitydegradation in the Si1-xGexρMOSFETs. Studying contacts with metal silicides constitutes a majorpart of this thesis. With the conventional device fabrication,the Si1-xGexincorporated for channel applications inevitablyextends to the source-drain areas. Measurement and modelingshow that the presence of Ge in the source/drain areaspositively affects the contact resistivity in such a way thatit is decreased by an order of magnitude for the contact of TiWto p-type Si1-xGex/Si when the Ge content is increased from 0 to 30at. %. Modeling and extraction of contact resistivity are firstcarried out for the traditional TiSi2-Si contact but with an emphasis on the influenceof a Nb interlayer for the silicide formation. Atwo-dimensional numerical model is employed to account foreffects due to current crowding. For more advanced contacts toultra-shallow junctions, Ni-based metallization scheme is used.NiSi1-xGex is found to form on selectively grown p-typeSi1-xGexused as low-resistivity source/drain. Since theformed NiSi1-xGex with a specific resistivity of 20 mWcmreplaces a significant fraction of the shallow junction, athree-dimensional numerical model is employed in order to takethe complex interface geometry and morphology into account. Thelowest contact resistivity obtained for our NiSi1-xGex/p-type Si1-xGexcontacts is 5´10-8Ωcm2, which satisfies the requirement for the 45-nmtechnology node in 2010. When the Si1-xGexchannel is incorporated in a MOSFET, it usuallyforms a retrograde channel with an undoped surface region on amoderately doped substrate. Charge sheet models are used tostudy the effects of a Si retrograde channel on surfacepotential, drain current, intrinsic charges and intrinsiccapacitances. Closed-form solutions are found for an abruptretrograde channel and results implicative for circuitdesigners are obtained. The model can be extended to include aSi1-xGexretrograde channel. Although the analytical modeldeveloped in this thesis is one-dimensional for long-channeltransistors with the retrograde channel profile varying alongthe depth of the transistor, it should also be applicable forshort-channel transistors provided that the short channeleffects are perfectly controlled. Key Words:MOSFET, SiGe, high-k dielectric, metal gate,mobility, charge sheet model, retrograde channel structure,intrinsic charge, intrinsic capacitance, contactresistivity.

ABA insensitive mutants (abi1 - abi5) have been isolated in germination screens that use high concentrations of exogenous ABA, and this method is believed to be saturated. To overcome this problem, a sensitized screen that used much lower concentration of exogenous ABA was performed to isolate new ABA insensitive mutants. Some of the isolated mutants had defects in light or retrograde signalling. One particular mutant (18-11) developed long hypocotyls under normal light condition. Based on its response to blue, red and far-red light conditions, this mutant is likely a novel hy mutant. Genetic analysis revealed that while ABA insensitivity in this mutant is recessive, the long hypocotyl phenotype is dominant. Positional cloning is currently being carried out to identify the gene. Findings from this study supports that ABA signalling interacts with light signalling networks.

ABA insensitive mutants (abi1 - abi5) have been isolated in germination screens that use high concentrations of exogenous ABA, and this method is believed to be saturated. To overcome this problem, a sensitized screen that used much lower concentration of exogenous ABA was performed to isolate new ABA insensitive mutants. Some of the isolated mutants had defects in light or retrograde signalling. One particular mutant (18-11) developed long hypocotyls under normal light condition. Based on its response to blue, red and far-red light conditions, this mutant is likely a novel hy mutant. Genetic analysis revealed that while ABA insensitivity in this mutant is recessive, the long hypocotyl phenotype is dominant. Positional cloning is currently being carried out to identify the gene. Findings from this study supports that ABA signalling interacts with light signalling networks.

<p>Challenges with integrating high-κ gate dielectric,retrograde Si_1-xGe_xchannel and silicided contacts in future CMOStechnologies are investigated experimentally and theoreticallyin this thesis. ρMOSFETs with either Si or strained Si_1-xGex surface-channel and different high-κgate dielectric are examined. Si_1-xGex ρMOSFETs with an Al₂O₃/HfAlO_x/Al₂O₃nano-laminate gate dielectric prepared by means ofAtomic Layer Deposition (ALD) exhibit a great-than-30% increasein current drive and peak transconductance compared toreference Si ρMOSFETs with the same gate dielectric. Apoor high-κ/Si interface leading to carrier mobilitydegradation has often been reported in the literature, but thisdoes not seem to be the case for our Si ρMOSFETs whoseeffective mobility coincides with the universal hole mobilitycurve for Si. For the Si_1-xGe_xρMOSFETs, however, a high density ofinterface states giving riseto reduced carrier mobility isobserved. A method to extract the correct mobility in thepresence of high-density traps is presented. Coulomb scatteringfrom the charged traps or trapped charges at the interface isfound to play a dominant role in the observed mobilitydegradation in the Si_1-xGe_xρMOSFETs.</p><p>Studying contacts with metal silicides constitutes a majorpart of this thesis. With the conventional device fabrication,the Si_1-xGe_xincorporated for channel applications inevitablyextends to the source-drain areas. Measurement and modelingshow that the presence of Ge in the source/drain areaspositively affects the contact resistivity in such a way thatit is decreased by an order of magnitude for the contact of TiWto p-type Si_1-xGe_x/Si when the Ge content is increased from 0 to 30at. %. Modeling and extraction of contact resistivity are firstcarried out for the traditional TiSi₂-Si contact but with an emphasis on the influenceof a Nb interlayer for the silicide formation. Atwo-dimensional numerical model is employed to account foreffects due to current crowding. For more advanced contacts toultra-shallow junctions, Ni-based metallization scheme is used.NiSi_1-xGex is found to form on selectively grown p-typeSi_1-xGe_xused as low-resistivity source/drain. Since theformed NiSi1-xGex with a specific resistivity of 20 mWcmreplaces a significant fraction of the shallow junction, athree-dimensional numerical model is employed in order to takethe complex interface geometry and morphology into account. Thelowest contact resistivity obtained for our NiSi_1-xGe_x/p-type Si_1-xGe_xcontacts is 5´10^-8Ωcm², which satisfies the requirement for the 45-nmtechnology node in 2010.</p><p>When the Si_1-xGe_xchannel is incorporated in a MOSFET, it usuallyforms a retrograde channel with an undoped surface region on amoderately doped substrate. Charge sheet models are used tostudy the effects of a Si retrograde channel on surfacepotential, drain current, intrinsic charges and intrinsiccapacitances. Closed-form solutions are found for an abruptretrograde channel and results implicative for circuitdesigners are obtained. The model can be extended to include aSi_1-xGe_xretrograde channel. Although the analytical modeldeveloped in this thesis is one-dimensional for long-channeltransistors with the retrograde channel profile varying alongthe depth of the transistor, it should also be applicable forshort-channel transistors provided that the short channeleffects are perfectly controlled.</p><p><b>Key Words:</b>MOSFET, SiGe, high-k dielectric, metal gate,mobility, charge sheet model, retrograde channel structure,intrinsic charge, intrinsic capacitance, contactresistivity.</p>

Introdução: O sucesso da cateterização da via biliar é de importância para o diagnóstico e terapêutica nas afecções biliopancreáticas nos procedimentos de colangiopancreatografia retrógrada endoscópica (CPRE) e está associado a complicações graves e mortalidade. O objetivo do estudo foi comparar o sucesso, perfil laboratorial e as complicações da técnica de fistulotomia papilar direta com o acesso cateter e fio-guia. Métodos: No período de julho de 2010 a maio de 2017 foram selecionados e randomizados para CPRE em dois grupos: cateterização com cateter e fioguia (Grupo I) e a fistulotomia papilar (Grupo II). As curvas de amilase, lipase e proteína C reativa (T0, 12 e 24 horas) e as complicações (pancreatite, sangramento e perfuração) foram avaliadas após CPRE. Resultados: Foram incluídos 102 pacientes (66 do sexo feminino e 36 do masculino, com idade média de 59,11±18,7 anos) e divididos em 51 pacientes para Grupo I e 51 no Grupo II. Os sucessos das cateterizações dos Grupos I e II foram de 76,47% e 100%, respectivamente (p=0,0002). Doze pacientes (23,53%) do Grupo I foram considerados pacientes de cateterização difícil e submetidos à fistulotomia papilar com sucesso no acesso biliar. Foram observadas 13,7% (2 perfurações e 5 pancreatites leves) e 2 % (1 paciente com perfuração e pancreatite) complicações nos Grupos I e II, respectivamente (p=0,062). Conclusão: A fistulotomia papilar demonstrou maior eficácia na cateterização da via biliar e com menor índice de amilasemia e lipasemia em comparação a cateterização com papilótomo e fio guia. As complicações foram semelhantes entre as duas técnicas / Background: The success of biliary tract cannulation is important for the diagnosis and treatment of biliopancreatic diseases in endoscopic retrograde cholangiopancreatography (ERCP) procedures. ERCP is associated with severe complications and mortality. The aim of the study was to compare the success, laboratory profile and complications of the direct papillary fistulotomy technique with standard catheter and guidewire access. Methods: In the period from July 2010 to May 2017, two groups were selected and randomized for ERCP: cannulation with catheter and guidewire (Group I) and papillary fistulotomy (Group II). The curves of amylase, lipase and C-reactive protein (T0, 12 and 24 hours) and complications (pancreatitis, bleeding and perforation) were evaluated after ERCP. Results: A total of 102 patients (66 females and 36 males, mean age 59.11 ± 18.7 years) were divided into 51 patients for Group I and 51 for Group II. The success of cannulation was 76.47% and 100%, in Groups I and II, respectively (p = 0.0002). Twelve patients (23.53%) of Group I were considered to have difficult cannulation and were submitted to fistulotomy with successful biliary access. There were 13.73% (2 perforations and 5 mild pancreatitis) and 2% (1 patient with perforation and pancreatitis) complications in Groups I and II, respectively (p=0,062). Conclusion: Papillary fistulotomy demonstrated greater efficacy in the bile duct cannulation and presented lower serum amylase and lipase compared with standard catheter and guidewire cannulation. Complications were similar in the two techniques

The proposal of this work was to determine the contribution of functional MR imaging techniques, i.e. secretin-enhanced MRCP (S-MRCP) and dynamic contrast-enhanced magnetic resonance (DCE-MR) imaging in the quantitative assessment of exocrine pancreatic function and perfusion.<p><p>The pancreas is both an exocrine and endocrine organ, though the exocrine tissue accounts for more than 90%. The exocrine pancreas is specialized in the synthesis and storage of digestive enzymes and in bicarbonate and water secretion in response respectively to various secretagogues (CCK, ach, GRP, VIP,…) and to secretin. <p>The arterial supply of the pancreas derives from branches of the celiac trunk and of the superior mesenteric artery. The microvascularity of the exocrine and the endocrine parts of the gland are anatomically and functionally separated, with differentially regulated blood perfusion. <p>Based on the knowledge of a close relationship between the activity of the gland and its blood supply, in normal conditions pancreatic perfusion responds to the functional state of the exocrine parenchyma: increased demands for exocrine secretions are associated with increased pancreatic blood flow. <p>The pancreatic gland can be involved at different degrees of severity in acute and chronic inflammatory processes due to various causes. In both processes microcirculatory changes occur and the pancreatic exocrine function can be impaired. Moreover, an exiguous microvascular component characterizes pancreatic ductal adenocarcinoma (PDA) related to a prominent stroma.<p><p><p>In the first section of this thesis, quantitative assessment of the pancreatic exocrine secretions was performed with S-MRCP in physiologic and non-physiologic conditions. The stimulating effect of secretin as well as the inhibitory effect of somatostatin on normal pancreas, both administered at different dose-regimens, were tested. The results of these investigations showed that quantitative S-MRCP is able to detect changes in pancreatic exocrine secretions correlated to the degree of stimulation or inhibition. <p>In pathologic settings, pancreatic exocrine secretions were assessed in chronic pancreatitis patients showing different degrees of severity, before and after endoscopic pancreatic duct drainage procedures (PDDP). In the group of patients presenting a reduced pancreatic exocrine reserve before treatment, quantitative S-MRCP showed a short-term improvement after PDDP. <p><p>In the second section, the feasibility and the reproducibility of DCE-MR imaging to quantify regional pancreatic perfusion was firstly investigated. DCE-MR imaging was performed in normal volunteers. Reference values for regional pancreatic perfusion were achieved with an intra-individual variability of 21%.<p>DCE-MR investigations were repeated during secretin stimulation and disclosed a significant increase of regional pancreatic perfusion in all individuals. <p>Secondly, DCE-MR imaging investigated benign and malignant focal pancreatic solid lesions and non tumoral tissue in patients undergoing pancreatic surgical resection. The purpose was to correlate DCE-MR quantitative parameters, (reflecting perfusion and/or permeability and the distribution volume fraction) with histologic features such as the degree of fibrosis and the microvascular density (MVD) in the corresponding tissues. A significant correlation was found between DCE-MR and histologic parameters: Ktrans was negatively correlated with the degree of fibrosis (high fibrosis was correlated with low perfusion), while the distribution volume fraction was positively correlated with the degree of fibrosis and with MVD (larger EES was correlated with high fibrosis and higher MVD). <p> / Doctorat en Sciences médicales / info:eu-repo/semantics/nonPublished

:.In our work we wanted to confirm our clinical experience with therapy of biliary and pancreatic duct injuries from the endoscopic retrograde cholangiopancreatography (ERCP) which was done in 267. Children and infants with a variety of biliary tract disorders and traumatic injuries in the area of biliary and pancreatic duct.. Pressure of the bile plays the key role in the therapy of biliary tract injuries Therefore we have measured the pressure in biliary tract and duodenum before and after the sphincterotomy of Oddi sphincter. Thea aim of our study was to confirm the insertion of drainage into the biliary and pancreatic duct in children with injury in this area. Our results showed significant differences between biliary duct pressure and duodenal pressure in the patients before and after sphincterotomy of Oddi sphincter. This results on theoretical basis confirmed, that it is necessary in children after traumatic rupture of biliary duct to provide ERCP and insert a biliary drainage after sphincterotomy. With this procedure the biliary tract injury is healed ad integrum without surgical liver resection. To provide only papilosphincterotomy without biliary drainage is not sufficient. This new miniinvasive procedure plays a fundamental role in the therapy of blunt abdominal injuries in a children and infants...