Les oligonucléotides antisens possèdent un immense potentiel thérapeutique. Cependant, la faible efficacité avec laquelle ils traversent les membranes biologiques limite leur utilisation. De nombeuses stratégies de délivrances ont été proposées pour contourner ce problème mais la plupart restent peu adaptées à une utilisation in vivo. Durant cette dernière décennie, plusieurs peptides capables de traverser la membrane plasmique ont été caractérisés. Regroupés sour le nom de Cell Penetrating Peptide, ces peptides sont polycationiques et parfois amphipatiques. Nos travaux d'évaluation de ces CPPs dans le modèle cellulaire de correction d'épissage indiquent que ces vecteurs, couplé à des PNA ou PMO, restent bloqués dans les vésicules d'endocytose. L'utilisation d'agents endosomolytiques comme la chloroquine, libère ces conjugués améliorant ainsi l'efficacité de la correction d'épissage. D'une manière générale il est admis que le développement de nouveaux peptides vecteurs présentant une propriété endosomolytique intrinsèque constituerait une avancée majeure dans le domaine de la délivrance. Deux conjugués (R-Ahx-R)4-PMO et R6Pen-PNA corrigent efficacement l'épissage sans addition de chloroquine. Ces conjugués sont internalisés dans les cellules par un mécanisme endocytotique. Les études de structure activité ont indiqué une corrélation entre l'affinité des conjugués aux héparanes sulfates ainsi que de leur hydrophobicité et l'efficacité de correction. Les travaux sur les modèles animaux ont montré une large biodisponibilité du conjugé (R-Ahx-R)4-PMO. Nos collaborations continuent pour améliorer ces deux peptides de délivrance.

Submitted by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-05-11T14:30:59Z No. of bitstreams: 1 DIS_RUIZ_RITTER_COMPLETO.pdf: 1478533 bytes, checksum: 401f51d6785b69fc2255986a7e168081 (MD5) / Made available in DSpace on 2017-05-11T14:30:59Z (GMT). No. of bitstreams: 1 DIS_RUIZ_RITTER_COMPLETO.pdf: 1478533 bytes, checksum: 401f51d6785b69fc2255986a7e168081 (MD5) Previous issue date: 2016-12-01 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / La presente investigaci?n objetiva analizar la inefectividad del principio de la imparcialidad en el derecho procesal penal brasile?o, desde los estudios de la psicolog?a social, m?s propiamente de la teor?a de la disonancia cognitiva. Se desarrolla con esa intenci?n, en tres cap?tulos, que tratan respectivamente sobre: 1) la imparcialidad en el ?mbito de los sistemas procesuales penales, su debida conceptuaci?n, dimensi?n e importancia para el Estado de Derecho; 2) las bases de la teor?a mencionada y sus principales contribuciones para el tema de la imparcialidad jurisdiccional - que pueden ser traducidas en la revelaci?n de los procesos cognitivo-comportamentales desarrollados en los sujetos (contextualizados en la figura de los jueces) despu?s de la toma de las decisiones y el conocimiento de nuevas informaciones sobre alguien, que pongan en jaque la primera impresi?n fijada sobre esa persona; y, 3) los obst?culos a esa imparcialidad previstos en el C?digo de Proceso Penal vigente, revelados por el marco te?rico profundizado en el cap?tulo anterior (de que son ejemplos la prevenci?n como criterio fijador de competencia, la presencia f?sica de los autos de la investigaci?n preliminar en el proceso y la permisibilidad de los poderes instructores del juzgador), proponi?ndose, entonces, cuatro medidas reductoras de da?os para la concretizaci?n de un debido proceso penal, de cariz acusatorio-constitucional e jurisdicci?n imparcial, con especial atenci?n para la Reforma del C?digo de Proceso Penal, que est? en tramitaci?n en la C?mara de los Diputados (PL 8045/2010). / A presente pesquisa objetiva analisar a (in)efetividade do princ?pio da imparcialidade no direito processual penal brasileiro, a partir dos estudos da psicologia social, mais especificamente da teoria da disson?ncia cognitiva. Desenvolve-se, com esse intuito, em tr?s cap?tulos, que versam, respectivamente, sobre: 1) a imparcialidade no ?mbito dos sistemas processuais penais, sua devida conceitua??o, dimens?o e import?ncia para o Estado de Direito; 2) as bases da teoria referida e suas principais contribui??es para o tema da imparcialidade jurisdicional - que podem ser traduzidas no desvelamento dos processos cognitivo-comportamentais desencadeados nos indiv?duos (contextualizados na figura dos ju?zes) ap?s a tomada de decis?es e o conhecimento de novas informa??es sobre algu?m, que coloquem em xeque a primeira impress?o fixada sobre essa pessoa; e, 3) os obst?culos ? essa imparcialidade previstos no C?digo de Processo Penal vigente, desvelados pelo arcabou?o te?rico aprofundado no cap?tulo anterior (do que s?o exemplos a preven??o como crit?rio fixador de compet?ncia, a presen?a f?sica dos autos do inqu?rito policial no processo e a permissibilidade de se atribu?rem poderes instrut?rios ao julgador), propondo-se, ent?o, quatro medidas redutoras de danos para a concretiza??o de um devido processo penal, de cariz acusat?rio-constitucional e jurisdi??o imparcial, com especial aten??o para a Reforma do C?digo de Processo Penal, que est? em tramita??o na C?mara dos Deputados (PL 8045/2010).

Clinical studies reported side effects of muscular spasms and muscle stiffness following the administration of clofibrate, a drug once used to treat hyperlipidaemia in patients. Experiments with clofibrate and its analogues in animal models showed it produced these myotonic symptoms in muscle by reducing the chloride conductance of the muscle membrane. The effects of 2-(4-chlorophenoxy)propionic acid, an analogue of clofibric acid, was assessed on the rat ClC-1 channel (rClC-1). Racemic 2-(4-chlorophenoxy)propionic acid shifted the voltage dependence of rClC-1 activation to more depolarising potentials, a mechanism accounting for myotonic symptoms previously reported. Experiments with resolved enantiomers revealed that the effects recorded were due exclusively to S-(–) 2-(4- chlorophenoxy)propionic acid. The R-(+) enantiomer was ineffective at the concentrations tested. Further experiments with the compound at differing Cl- concentrations in the extracellular solution suggested that S-(–) 2-(4-chlorophenoxy)propionic acid altered the gating of ClC-1 by decreasing the affinity of the binding site where Cl- normally acts to ‘gate’ the channel. Similarities in the effects reported for most dominant mutations in the CLCN1 gene that lead to myotonia congenita and 2-(4-chlorophenoxy)propionic acid prompted experiments that introduced these point mutations in the human ClC-1 (hClC-1) gene to compare their mode of action to that of the drug. These mutations, F307S and A313T, predominantly altered the slow, or common, gate of the channel. Conversely, the effect of 2-(4-chlorophenoxy)propionic acid was predominantly on the fast gating process of hClC-1. A macroscopically similar effect therefore, can be produced by two different modes of action. Results suggested that both drug and mutations exert their action by affecting the transition of the channel from its closed to open state subsequent to Cl- binding. Investigation of the interaction between rClC-1 gating and a further 25 compounds structurally related to clofibric acid identified a number of compounds effective at shifting the open probability of fast gating to depolarising potentials. Fewer were identified that influence slow gating. Some compounds affected both gating processes, however, none were identified which influenced slow gating alone. Ability to displace the voltage dependent activation of the fast gate appeared to depend largely on the lipophilicity of the molecules tested, indicating the importance of hydrophobic interactions between drug and channel protein. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1474724 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 2009

Today, the healthcare IT vendors develop interfaces for their products to comply with the IHE Profiles and test them with other vendors in annual events, called IHE Connactathons which are one week events organized to perform interoperability testing. Network configuration including hostnames, IP addresses,port numbers for healthcare services and URLs for the various web services used in IHE profiles are stored on online databases accessible from the IHE website. When all companies register for participation to the connectathon, a default configuration is created in the database, assigning default values for healthcare service specific parameters. Afterwards, a company is expected to use the configuration parameters from that database during the connectathon. Except from the connectathons, the vendors could not test their products with other vendors&rsquo / . In this thesis, research on IHE Configuration Management Tool based on IHE profiles, ebXML ebBP, CPP and CPA specifications is presented. The intention behind developing such a tool is to automate the creation and exchange of configuration information between IHE Actors in a Virtual Connectathon Portal. The results of this research enable the IHE product vendors to test their products in interactions with other vendors through the web anytime/anywhere by automatic business service interface configuration. The research enables the healthcare systems to engage with many trading partners at the same time automatically. The work presented in this thesis is realized as a part of IST-027065 RIDE project funded by European Commission.

The hydrophobic plasma membrane constitutes an indispensable barrier for cells in living animals. Albeit being pivotal for the maintenance of cells, the inability to cross the plasma membrane is still one of the major obstacles to overcome in order to progress current drug development. A group of substances, with restricted access to the interior of cells, which has shown great promise for future clinical use is oligonucleotides that are exploited to interfere with gene expression. Short interfering RNAs that are utilized to confer gene silencing and splice correcting oligonucleotides, applied for the manipulation of splicing patterns, are two classes of oligonucleotides that have been explored in this thesis. Cell-penetrating peptides (CPPs) are a class of peptides that has gained increasing focus in last years. This ensues as a result of their remarkable ability to convey various, otherwise impermeable, macromolecules across the plasma membrane of cells in a relatively non-toxic fashion. This thesis aims at further characterizing well-established, and newly designed, CPPs in terms of toxicity, delivery efficacy, and internalization mechanism. Our results demonstrate that different CPPs display different toxic profiles and that cargo conjugation alters the toxicity and uptake levels. Furthermore, we confirm the involvement of endocytosis in translocation of CPPs, and in particular the importance of macropinocytosis. All tested peptides facilitate the delivery of splice correcting oligonucleotides with varying efficacy, the newly designed CPP, M918, being the most potent. Finally we conclude that by promoting endosomolysis, by exploring new CPPs with improved endosomolytic properties, the biological response increases significantly. In conclusion, we believe that these results will facilitate the development of new CPPs with improved delivery properties that could be used for transportation of oligonucleotides in clinical settings.

There is an increasing awareness in clinical dentistry of the need to better understand the aetiology and management of tooth wear, as increasing numbers of elderly patients are retaining their natural teeth to a stage when they present with extensive wear. In addition, more younger patients are presenting with wear of both primary and permanent teeth. In order to comply with the philosophy of minimal intervention dentistry, clinical management of tooth wear should focus on early detection and prevention before a restorative approach is considered. Fluoride is a commonly used agent in the management of tooth wear, with previous studies showing that it can protect teeth against erosion (at around pH 2.0 and 3.0) and toothbrush abrasion in an acidic environment (at around pH 3.0). However, it does not reduce attritional wear between opposing tooth specimens. Nightguards are commonly used to prevent attritional wear but there is still a need to improve preventive strategies to manage the risk of erosive tooth wear. Previous in vitro studies have shown that a remineralizing agent in the form of CPP-ACP (casein phosphopeptide – amorphous calcium phosphate) can reduce erosion of both enamel and dentine by white wine. A recent study has also highlighted the potential lubricating and remineralizing properties of a CPP-ACP containing paste (Tooth Mousse®, G C Asia Pty Ltd) in reducing dentine wear in both an acidic environment (pH 3.0) and a near neutral environment (pH 6.1). In this context, it is desirable to investigate the effect of CPP-ACP on enamel wear under conditions simulating various clinical situations and to better understand the nature of third-body components at the wear interface. The aims of the present study were: (i) to determine the effect of CPP-ACP on enamel wear under conditions simulating three clinical situations: heavy attrition with gastric regurgitation (at pH 1.2); toothbrush abrasion after an erosive episode (at pH 3.2); and erosion from gastric regurgitation (at pH 1.2), and (ii) to clarify its mode of action by characterizing the enamel surface treated with CPP-ACP. Electro-mechanical tooth wear machines were used to simulate attrition and toothbrush abrasion in combination with erosion, but no machine was used in experiments investigation erosion alone. The effect of paste containing CPP-ACP in reducing erosive tooth wear was also compared with that without CPP-ACP. Enamel surfaces with and without treatment with CPP-ACP were then characterized by using Time of Flight – Secondary Ion Mass Spectrometry (ToF-SIMS). CPP-ACP was found to reduce enamel wear under conditions simulating heavy attrition combined with gastric regurgitation, and toothbrush abrasion after an erosive episode. However, it did not protect enamel from erosion under conditions simulating gastric regurgitation alone, indicating that its lubricating effect was more pronounced than its remineralizing effect. Silicone and ethyl siloxane were present in trace amounts on the enamel surface treated with Tooth Mousse®, thus they were found to be included in its formulation. These findings imply that CPP-ACP nanocomplexes and silicon can act as third body components to lubricate the wear interface. The findings presented in this thesis have implications for clinical management of tooth wear and may lead to new strategies of tooth wear prevention. Although some caution is needed in extrapolating these findings to the in vivo situation, it should be noted that in vitro studies provide valuable insights into separate components of the tooth wear process that are often difficult to distinguish in clinical situations. Overall, this thesis provides new information about the role of CPP-ACP in reducing erosive enamel wear and provides a basis for future experimental and clinical studies focussing on tooth wear prevention. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1379897 / Thesis (Ph.D.) -- University of Adelaide, School of Dentistry, 2009

The effects of dopamine D1 and D2 antagonists on conditioned place preference (CPP) and locomotor activity were assessed. A total of three experiments were conducted. In each experiment there were two conditioning days followed by a test day. The results indicate that DA D1 and D2 receptors have distictly different roles in the mediation of behavior.

Novel psychoactive substances (NPS) are a novel problem of the drug scene. NPS mimic effects of the "classic" illicit drugs, but since they have a different chemical structure, they are usually not covered by legislative control. The exact nature of the aforementioned effects depends merely on the description by users, with a proper scientific assessment still absent. Aim of this study is to evaluate effects and addictive potential of naphyrone (a derivative of cathinone) in Wistar rats. High concentration of naphyrone in the brain tissue discovered by pharmacokinetic analysis proved its high blood-brain barrier permeability. Brain level of naphyrone peaked at approximately 30 min after the treatment, nearly at the same time as in the serum. Since naphyrone administration significantly rises body temperature and increases overall locomotion, its stimulant effect is prominently apparent. Our study failed to prove any effect of naphyrone on sensorimotor gating. Tendency to produce conditioned place preference was observed but was not significant. My thesis reports on initial and novel findings about impact of naphyrone administration on physiological parameters of the animal model.

No description available.

Autonomous lawn mowers and floor cleaning robots are today easily accessible and areutilizing well-studied Coverage Path Planning algorithms. They operate in single-floorenvironments that are small with simple geometry compared to general urban environments such as city parking garages, highway bridges or city crossings. A next step for autonomous cleaning is road sweeping of these complex urban environments. In this work,a new Coverage Path Planning approach, Sampled BA* & Inward Spiral , handling this taskwas compared with existing well-performing algorithms BA* and Inward Spiral. The proposed approach combines the strengths of existing algorithms and demonstrates state-of-the-art performance on three large-scale 3D environments. It generated paths with lessrotation, while keeping the length of the path on the same level. For a given starting point,the new approach had consistently lower cost (length + rotation) for all environments. Forrandom starting points, randomness in the new approach caused less robustness, givingsignificantly higher cost. To improve the performance of the algorithms and remove biasfrom manual tuning, the parameters were automatically tuned using Bayesian Optimization. This makes the evaluation more robust and the results stronger.