Dissertations / Theses on the topic 'Stabilità mediale'

Thrombosis, the major cause of heart attack and strokes,1 is triggered by localized clotting of the blood as the result of deregulated platelet aggregation. During the repair of vascular injury, clotting usually occurs when platelets adhere to each other at the site of vascular injury in order to stop bleeding.2 Distinct protein receptors and adhesive ligands together with the blood flow conditions govern this process. One of the negative regulators in platelet aggregation is Disabled-2 (Dab2), a modular protein that is released upon platelet activation to the extracellular platelet surface.3 Dab2 inhibits platelet aggregation through its phosphotyrosine-binding (PTB) domain by competing with fibrinogen for ï ¡IIï ¢3 integrin binding on the activated platelet surface.3 Sulfatides are also found on the platelet surface,4 interacting with adhesive and coagulation proteins5-7 and, thus, they are thought to play a major role in haemostasis and thrombogenesis. Here, we show that the Dab2 PTB domain specifically interacts with sulfatides through two conserved basic motifs. The sulfatide-binding site overlaps with that of phosphatidylinositol 4,5-biphosphate (PtdIns(4,5)P2) in the PTB domain. Whereas sulfatides recruit the Dab2 PTB domain to the platelet surface, thus sequestering the protein from thrombin-mediated platelet aggregation, the phosphoinositide mediates its internalization. Experimental data support the hypothesis that two pools of Dab2 co-exist at the platelet surface and that the balance between them controls the extent of the clotting response.
Master of Science

Biomedical Sciences
Ph.D.
This work identifies the Fragile X-related protein (FXR1) as a reciprocal regulator of HuR target transcripts in vascular smooth muscle cells (VSMC). FXR1 was identified as an HuR interacting protein by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The-HuR-FXR1 interaction is abrogated in RNase-treated extracts, indicating that their association is tethered by mRNAs. FXR1 expression is induced in diseased, but not normal arteries. SiRNA knock down of FXR1 increases abundance and stability of inflammatory mRNAs, while overexpression of FXR1 reduces their abundance and stability. RNA-EMSA and RIP demonstrate that FXR1 directly interacts with an ARE and a previously uncharacterized element in the 3’UTR of TNFa. FXR1 expression is increased in VSMC challenged with the anti-inflammatory cytokine IL-19, and FXR1 is required for IL-19 reduction of HuR. This suggests FXR1 is an anti-inflammation responsive, HuR counter-regulatory protein that reduces abundance of pro-inflammatory transcripts. Additionally, we observed significantly increased poly-A-Binding protein (PABP) expression localizing to discrete punctate structures in both vascular smooth muscle (VSMC) and endothelial cells (EC) of the aortic arch of Ldlr-/- mice, as compared to WT controls. EIF2α phosphorylation, requisite for SG formation, was also induced by clotrimazole and oxLDL in these cells. Interestingly, VSMCs pre-treated with anti-inflammatory cytokine IL-19 followed by clotrimazole significantly reduced the formation of SGs and eIF2a phosphorylation, suggesting a relationship between inflammation and SG formation in vascular cells. Reduction of SG component G3BP1 by siRNA knockdown significantly reduced stress granule formation and inflammatory gene abundance in hVSMC. Microtubule inhibitors reduced SG formation in hVSMC. These results support the hypothesis that SG formation in atherosclerosis is driven by inflammation, SG may mediate the cellular response to inflammation, and that anti-inflammatory treatment may lessen atherosclerosis progression and plaque formation by reduction of SGs.
Temple University--Theses

Understanding how memory is formed requires looking beyond the genes involved to the neural circuitry and temporal aspects of memory. In this dissertation I have focused my investigation on Dorsal Paired Medial (DPM) neurons, two modulatory neurons essential for memory in Drosophila. DPM neurons highly express the amnesiac (amn) gene, which encodes for a putative pre-pro-neuropeptide. amn function in DPM neurons is required for memory. Here I provide evidence that DPM neurons are cholinergic and that acetylcholine (ACh) and AMN act as co-transmitters essential for DPM function. In order to investigate the temporal requirements of DPM output I blocked transmitter release during discrete intervals in the memory process using shibirets1 and tested flies for shock and sugar-reinforced memory. These experiments demonstrated that stable memory requires persistent transmitter release from DPM neurons. Furthermore these results suggest AMN and DPM neurons act as general stabilizers of mushroom body dependent memory. To further investigate the neural circuitry underlying DPM function I disrupted DPM projections onto the mushroom body lobes by ectopically expressing DScam17-2::GFP in DPM neurons. Flies with DPM neurons that predominantly project to the mushroom body α´/β´ lobes exhibit normal memory, and blocking transmitter release from the mushroom body prime lobes neurons themselves abolishes memory indicating DPM neuron-mushroom body α´/β´ neuron interaction that are critical for memory. Taken together, the experimental evidence presented here are used to provide a rudimentary model of the neural circuitry involved in memory stability, where DPM neurons form a recurrent feedback loop with the mushroom body α´/β´ lobe neurons and act to stabilize odorspecific conditioned memories at Kenyon cell synapses.

La recherche de nouvelles molécules pour la médecine nucléaire est un domaine en expansion croissante. Ces dernières années, le développement de nouveaux radiopharmaceutiques à visée thérapeutique a relancé l'intérêt pour la chimie du rhénium. En effet, les deux isotopes 186Re et 188Re, du fait de leurs propriétés adéquates et de leur analogie avec le 99mTc, largement utilisé pour les examens cliniques, semblent très prometteurs pour la préparation de radiopharmaceutiques.
Dans la première partie de ce manuscrit, la synthèse de complexes du rhénium et du technétium-99, [M(RPhCS3)2(RPhCS2)] (M = Re, Tc), est décrite. La préparation de radiopharmaceutiques à base de technétium-99m, analogues des composés obtenus à l'échelle pondérale, est également décrite. La stabilité/réactivité de ces complexes a été étudiée, au moyen de réactions d'échange avec d'autres ligands potentiels, et notamment par des dithiocarbamates, ainsi que par spectrophotométrie d'absorption UV-visible et analyse thermogravimétrique.
La réactivité des complexes vis-à-vis des dithiocarbamates conduit à la possibilité du greffage de biomolécules sur le cœur métallique, via le fragment dithiocarbamate. Cette méthode constitue une alternative potentielle aux procédures actuelles utilisant l'approche bifonctionnelle.

Dans la seconde partie de ce manuscrit, la mise au point d'un kit pour le marquage du lipiodol par le rhénium-188 est décrite, à partir du complexe analogue des complexes décrits dans la première partie. L'huile radiomarquée ainsi obtenue est potentiellement utilisable pour le traitement de l'hépatocarcinome. La stabilité in vitro et in vivo du complexe rhénié 188Re-SSS lipiodol et de son analogue technétié 99mTc-SSS lipiodol a été étudiée, ainsi que leur comportement in vivo sur un modèle de porc sain.
Cette étude a permis de montrer la fixation quasi-exclusive du radiopharmaceutique au niveau du foie, ainsi que la stabilité de ce composé. Sa sélectivité pour les tumeurs reste à démontrer avant de passer aux premiers essais chez l'homme.

Foam flow in porous media is important for many industrial operations such as enhanced oil recovery, remediation of contaminated aquifers and CO2 sequestration. The application of foam in these processes is due to its unique ability to reduce gas mobility and to divert gas to low permeability zones in porous media which otherwise would not be reached. To achieve optimum success with foam as a displacing fluid in oil recovery and remediation operations, it is essential to understand how different parameters influence foam flow in porous media. In this thesis, a variety of experimental techniques were used to study foam stability, foam rheology as well as the dynamics and patterns of oil displacement by foam under different boundary conditions such as surfactant formulation, oil type, foam quality (gas fraction) and porous media geometry. Bulk scale studies showed that foam stability was surfactant and oil dependant such that decreasing oil carbon number and viscosity decreased the stability of foam. However, no meaningful correlation was found between foam stability at bulk scale and the efficiency of oil displacement in porous media for the various surfactants studied in this work. Additionally, our results show that foams consisting of smaller bubbles do not necessarily correspond to higher apparent viscosity as the foam quality is also crucial. For the same foam quality decreasing bubble size resulted in higher apparent viscosity. Although in theory a higher apparent viscosity (i.e. higher foam quality) would be ideal for displacement purposes, increasing foam quality resulted in less stable foam in porous media due to formation of thin films which were less stable in the presence of oil. The effect of pore geometry on foam generation and oil displacement has also been investigated. Our findings provide new insights about the physics and complex dynamics of foam flow in porous media.

Despite significant efforts have been directed toward reducing waste generation and encouraging alternative waste management strategies, landfills still remain the main option for Municipal Solid Waste (MSW) disposal in many countries. Hence, landfills and related impacts on the surroundings are still current issues throughout the world. Actually, the major concerns are related to the potential emissions of leachate and landfill gas into the environment, that pose a threat to public health, surface and groundwater pollution, soil contamination and global warming effects. To ensure environmental protection and enhance landfill sustainability, modern sanitary landfills are equipped with several engineered systems with different functions. For instance, the installation of containment systems, such as bottom liner and multi-layers capping systems, is aimed at reducing leachate seepage and water infiltration into the landfill body as well as gas migration, while eventually mitigating methane emissions through the placement of active oxidation layers (biocovers). Leachate collection and removal systems are designed to minimize water head forming on the bottom section of the landfill and consequent seepages through the liner system. Finally, gas extraction and utilization systems, allow to recover energy from landfill gas while reducing explosion and fire risks associated with methane accumulation, even though much depends on gas collection efficiency achieved in the field (range: 60-90% Spokas et al., 2006; Huitric and Kong, 2006). Hence, impacts on the surrounding environment caused by the polluting substances released from the deposited waste through liquid and gas emissions can be potentially mitigated by a proper design of technical barriers and collection/extraction systems at the landfill site. Nevertheless, the long-term performance of containment systems to limit the landfill emissions is highly uncertain and is strongly dependent on site-specific conditions such as climate, vegetative covers, containment systems, leachate quality and applied stress. Furthermore, the design and operation of leachate collection and treatment systems, of landfill gas extraction and utilization projects, as well as the assessment of appropriate methane reduction strategies (biocovers), require reliable emission forecasts for the assessment of system feasibility and to ensure environmental compliance. To this end, landfill simulation models can represent an useful supporting tool for a better design of leachate/gas collection and treatment systems and can provide valuable information for the evaluation of best options for containment systems depending on their performances under the site-specific conditions. The capability in predicting future emissions levels at a landfill site can also be improved by combining simulation models with field observations at full-scale landfills and/or with experimental studies resembling landfill conditions. Indeed, this kind of data may allow to identify the main parameters and processes governing leachate and gas generation and can provide useful information for model refinement. In view of such need, the present research study was initially addressed to develop a new landfill screening model that, based on simplified mathematical and empirical equations, provides quantitative estimation of leachate and gas production over time, taking into account for site-specific conditions, waste properties and main landfill characteristics and processes. In order to evaluate the applicability of the developed model and the accuracy of emissions forecast, several simulations on four full-scale landfills, currently in operative management stage, were carried out. The results of these case studies showed a good correspondence of leachate estimations with monthly trend observed in the field and revealed that the reliability of model predictions is strongly influenced by the quality of input data. In particular, the initial waste moisture content and the waste compression index, which are usually data not available from a standard characterisation, were identified as the key unknown parameters affecting leachate production. Furthermore, the applicability of the model to closed landfills was evaluated by simulating different alternative capping systems and by comparing the results with those returned by the Hydrological Evaluation of Landfill Performance (HELP), which is the most worldwide used model for comparative analysis of composite liner systems. Despite the simplified approach of the developed model, simulated values of infiltration and leakage rates through the analysed cover systems were in line with those of HELP. However, it should be highlighted that the developed model provides an assessment of leachate and biogas production only from a quantitative point of view. The leachate and biogas composition was indeed not included in the forecast model, as strongly linked to the type of waste that makes the prediction in a screening phase poorly representative of what could be expected in the field. Hence, for a qualitative analysis of leachate and gas emissions over time, a laboratory methodology including different type of lab-scale tests was applied to a particular waste material. Specifically, the research was focused on mechanically biologically treated (MBT) wastes which, after the introduction of the European Landfill Directive 1999/31/EC (European Commission, 1999) that imposes member states to dispose of in landfills only wastes that have been preliminary subjected to treatment, are becoming the main flow waste landfilled in new Italian facilities. However, due to the relatively recent introduction of the MBT plants within the waste management system, very few data on leachate and gas emissions from MBT waste in landfills are available and, hence, the current knowledge mainly results from laboratory studies. Nevertheless, the assessment of the leaching characteristics of MBT materials and the evaluation of how the environmental conditions may affect the heavy metals mobility are still poorly investigated in literature. To gain deeper insight on the fundamental mechanisms governing the constituents release from MBT wastes, several leaching experiments were performed on MBT samples collected from an Italian MBT plant and the experimental results were modelled to obtain information on the long-term leachate emissions. Namely, a combination of experimental leaching tests were performed on fully-characterized MBT waste samples and the effect of different parameters, mainly pH and liquid to solid ratio (L/S,) on the compounds release was investigated by combining pH static-batch test, pH dependent tests and dynamic up-flow column percolation experiments. The obtained results showed that, even though MBT wastes were characterized by relatively high heavy metals content, only a limited amount was actually soluble and thus bioavailable. Furthermore, the information provided by the different tests highlighted the existence of a strong linear correlation between the release pattern of dissolved organic carbon (DOC) and several metals (Co, Cr, Cu, Ni, V, Zn), suggesting that complexation to DOC is the leaching controlling mechanism of these elements. Thus, combining the results of batch and up-flow column percolation tests, partition coefficients between DOC and metals concentration were derived. These data, coupled with a simplified screening model for DOC release, allowed to get a very good prediction of metal release during the experiments and may provide useful indications for the evaluation of long-term emissions from this type of waste in a landfill disposal scenario. In order to complete the study on the MBT waste environmental behaviour, gas emissions from MBT waste were examined by performing different anaerobic tests. The main purpose of this study was to evaluate the potential gas generation capacity of wastes and to assess possible implications on gas generation resulting from the different environmental conditions expected in the field. To this end, anaerobic batch tests were performed at a wide range of water contents (26-43 %w/w up to 75 %w/w on wet weight) and temperatures (from 20-25 °C up to 55 °C) in order to simulate different landfill management options (dry tomb or bioreactor landfills). In nearly all test conditions, a quite long lag-phase was observed (several months) due to the inhibition effects resulting from high concentrations of volatile fatty acids (VFAs) and ammonia that highlighted a poor stability degree of the analysed material. Furthermore, experimental results showed that the initial waste water content is the key factor limiting the anaerobic biological process. Indeed, when the waste moisture was lower than 32 %w/w the methanogenic microbial activity was completely inhibited. Overall, the obtained results indicated that the operative conditions drastically affect the gas generation from MBT waste, in terms of both gas yield and generation rate. This suggests that particular caution should be paid when using the results of lab-scale tests for the evaluation of long-term behaviour expected in the field, where the boundary conditions change continuously and vary significantly depending on the climate, the landfill operative management strategies in place (e.g. leachate recirculation, waste disposal methods), the hydraulic characteristics of buried waste, the presence and type of temporary and final cover systems.

Mms1 and Mms22 are important mediators of genome stability in Saccharomyces cerevisiae. Deletion mutants of both genes display decreased viability in the presence of a number of known DNA damaging agents as well as agents that perturb DNA replication. Here I present a detailed analysis of Mms1 and Mms22 function in stabilizing DNA replication forks. I find that mms1∆ and mms22∆ strains accumulate spontaneous DNA damage and have an increased rate of mutation during a normal cell cycle. Additionally, treatment with genotoxic agents causes defects in induced recombination as well as inhibiting the ability of cells to efficiently recover from DNA damage. Genetic interaction data support a model where Mms1 and Mms22 function with components of the replication fork. Accordingly I observed that the enrichment of key replication fork proteins at regions proximal to replication origins is decreased in mms1∆ and mms22∆ cells during replication stress. Furthermore, I monitored DNA replication in wild-type and mms1∆ strains and found that mms1∆ strains exhibit irregular fork progression under conditions of replication stress. I therefore concluded that Mms1 and Mms22 function at replication forks. Lastly, I uncovered that Mms1 is regulated by Rtt101-dependent ubiquitin-mediated proteolysis. Mms1 degradation is likely necessary for its function as overexpression of Mms1 results in decreased cell viability both during a normal cell cycle, and during treatment with DNA damaging agents. I found that Mms1 and Mms22 operate at sites of DNA replication to promote the stability of the genome, especially under conditions of DNA damage and replication fork stress.

碩士
國立陽明大學
物理治療暨輔助科技學系
103
Background: Joint instability is a risk factor for knee osteoarthritis (KOA). The presence of self-reported knee instability is commonly associated with impaired proprioceptive accuracy, poor sensorimotor control, muscle weakness and high laxity that frequently results in biomechanical aberrations during functional activities. Recent studies found that visualized varus thrust which occurred suddenly during the early stance phase of gait in KOA suffers would be a suitable index for knee instability, which may further result in increased joint translation during movement, especially in the mediolateral direction. Exercise is the first-line treatment for KOA. However, the effect of exercise intervention for knee joint arthro-kinematics in individuals with KOA remains unclear. Purpose: To investigate 1) whether varus thrust would be associated with arthrokinematic alterations, as detected by a 3-dimension (3D) motion tracking system; and 2) whether neuromuscular training could improve knee stability and other clinical outcomes in individuals with medial KOA and varus thrust. Methods: We recruited 24 subjects with medial KOA and visualized varus thrust and 12 healthy volunteers matched with age, gender and body mass index (BMI). The KOA group received 6-week neuromuscular training emphasizing specific muscle strengthening and functional re-training. The VICON motion system was used to capture arthrokinematic data including translation, velocity and acceleration of the tibiofemoral joint during level walking. Clinical outcomes including muscle strength, functional performance and pain were also assessed. Multivariate analyses of variance (MANOVA) were used to analyze the differences between subjects with the KOA and healthy control volunteers. Repeated measures MANOVA were used to determine the statistically significant differences between pre and post-test of the KOA group for arthrokinematic data and muscle strength. Improvement in functional performance and pain were analyzed with Paired t test. Level of significance was set at p < 0.05. Results: Subjects with KOA and varus thrust walked with significantly greater and faster mediolateral translation and had weaker hip/knee strength as compared with the matched controls. After 6-week neuromuscular training, these KOA subjects demonstrated significantly decreased mediolateral/posterioanterior translation, better hip/knee strength, and other clinical outcomes. Discussion and conclusion: Through 3D motion tracking, varus thrust in knee OA subjects was found to be associated with increased mediolateral translation and velocity of tibiofemoral joint. Moreover, 6-week neuromuscular training could decrease translations of the tibiofemoral joint to enhance knee stability that was parallel to other clinical outcomes. Clinical relevance: 3D motion tracking was a feasible tool with discriminant validity and clinical responsiveness to detect knee stability. 6-week neuromuscular exercise was beneficial on knee stability and other clinical performance in individuals with medial KOA.

The ulnar collateral ligament (UCL) is the primary restraint against valgus forces at the elbow. This structure cannot support the entire load placed upon the medial elbow during overhand throwing motions such as pitching a baseball. In this study we measured the contributions that different muscle groups make to the stability of the medial elbow, under conditions intended to reproduce the loads during pitching by varying the forearm position and loading conditions. We also evaluated the strength of the elbow musculature for the possibility of a training effect in the dominant arm of 11 male high school baseball pitchers. We collected surface EMG data in the two forearm positions to determine if the different positions used in various pitches have an effect on muscular action. We also tested an isometric and dissipative loading condition to determine if the muscles activity was load reactive. The four muscle groups tested were the flexor-pronator group (FP), the extensor-supinator group (ES), the tricep brachii (TB), and the pectoralis major (PM). We found significantly (p-value=0.001) higher peak activity levels of the flexor-pronator group in the neutral forearm position (79.4% MVIC �� 27.0% MVIC full trial peak, 30.8% MVIC �� 20.8% MVIC initial l50ms peak following activation) when compared to the supinated position (55.5% MVIC �� 29.6% MVIC full trial peak, 16.9% MVIC �� 14.8% MVIC initial l50ms peak following activation), which may explain the link between breaking pitches and medial elbow injury. We found an increase (p-value=0.001) in force output of the dominant arm (49.3 N/kg �� 12.5 N/kg) over the non-dominant arm (38.1 N/kg �� 11.0 N/kg). This finding is attributed to a training effect, which assists in protecting the elbow. These findings help provide baseline muscle activity information on protection of the medial soft tissue structures of the elbow.
Graduation date: 2004