No description available.

Post-translational modifications (PTMs) of proteins are now known to increase the complexity of biological signaling networks. The ability to selectively control the activity of individual proteins and enzymes involved in PTMs enables studies of biological systems and in designing therapeutic agents. The phosphorylation of serine, threonine, and tyrosine catalyzed by protein kinases play an important role in signaling and when deregulated are involved in different types of cancer amongst other diseases. Therefore, selective regulation of kinase activity by inhibition could provide a means to understand signaling and also aid in developing therapies. The design of specific kinase inhibitors is challenging because most inhibitors target the common ATP-binding site, leading to several off-target hits with adverse effects. With this in mind, a fragment-based bivalent strategy was developed to potentially increase affinity and selectivity. Our approach seeks to target the ATP-binding site with a small molecule while simultaneously targeting any available site on the kinase with a cyclic peptide that potentially increases selectivity and affinity. The bivalent approach has produced effective and selective inhibitors for cAMP-dependent protein kinase (PKA). Herein, we extended this approach to Aurora kinase (Aurora A), RSK1 and MSK1. Using our optimized selection conditions, we have successfully selected several cyclic peptide ligands to later generate bivalent ligands for these kinases. This strategy may prove a robust method to discover new allosteric sites on kinases as well as other proteins, furthermore it also has the potential to provide insight towards the design of new ATP targeted approaches. A second area of research focuses on PTMs involved in the acetylation of lysine residues by enzymes dubbed histone acetyl transferases (HATs) or lysine acetyl transferases (KATs). Recent studies have shown that a large fraction of the proteome may be modifies by KATs but it is challenging to develop uniquely selective inhibitors as the KATs like Kinases have similar active sites. Towards the goal of developing a method to control a single KAT in the presence of other KATs, we designed ligand-gated split-proteins based on a sequence dissimilarity based approach. Potential fragmentation sites were identified by insertion of a 25-residue loop insertion. Successful loop insertion mutants provided guidance for the dissection of KATs into fragments tethered to FKBP and FRB that cannot spontaneously assemble, but that do upon the addition of a ligand, rapamycin to generate catalytically active. The method was successfully applied to design the very first split-KAT, GCN5,which only showed activity in the presence of an added small molecule. The method was shown to be potentially general by application to a second KAT, PCAF. The above studies provide potentially new approaches for selectively targeting and studying PTMs catalyzed by kinases and lysine acetyl transferase.

No description available.

As tecnologias utilizadas em estações de tratamento de água e efluentes não são eficientes para a remoção total de resíduos farmacêuticos e os efeitos dessas substâncias sobre o meio ambiente e a saúde humana ainda não são bem conhecidos. No presente trabalho, estudou-se a degradação do antibiótico enrofloxacina (ENRO) por fotólise e pelo processo H2O2/UV na presença de compostos bio-orgânicos (BOS), que têm se apresentado como interessantes promotores da oxidação de poluentes. Os experimentos foram realizados em um reator fotoquímico tubular de imersão com fonte radiante (lâmpada de xenônio) concêntrica, operado em batelada com recirculação. Utilizaram-se concentrações iniciais de ENRO e de BOS iguais a 50 mg L-1 e 20 mg L-1, respectivamente. Para todos os pH mantidos constantes (3, 5, 7 ou 9), a solução foi irradiada por 240 minutos. Os resultados indicam que o antibiótico não sofreu hidrólise em qualquer dos pH estudados em um período de 24 horas. Por sua vez, a fotólise da enrofloxacina mostrou-se eficiente somente na presença do composto bioorgânico CVT 230 (BOS C), com remoção de ENRO de quase 90% em meio neutro (pH 7). Resultados da literatura, associados a experimento realizado em meio anóxico, sugerem a participação de oxigênio singlete como principal espécie oxidante da enrofloxacina. Por outro lado, a degradação da ENRO pelo processo H2O2/UV apresentou remoção máxima do fármaco de 48% em pH 7, o que sugere que a ação de oxigênio singlete e/ou radicais hidroxila não foi eficaz na presença de peróxido de hidrogênio. Portanto, o BOS C pode ser empregado como promotor no tratamento de águas e efluentes aquosos contaminados com enrofloxacina sob baixas potências radiantes ou em sistemas irradiados por luz solar. / The technologies used in water and wastewater treatment plants are not efficient for the total removal of pharmaceutical compounds whose effect to the environment and to human health are still not well known. In this work, the degradation of the antibiotic enrofloxacin (ENRO) was studied by photolysis and by the H2O2/UV process in the presence of bio-organic substances (BOS), which have been identified as interesting promoters of pollutant oxidation. The experiments were carried out in a tubular immersion photochemical reactor equipped with a concentric radiant source (xenon lamp), and operated in batch mode with recirculation. Initial ENRO and BOS concentrations of 50 mg L-1 and 20 mg L-1 were used, respectively. The solution was irradiated for 240 minutes for all pH studied at constant values (3, 5, 7, and 9). The results show that the antibiotic did not undergo hydrolysis at any pH after 24 hours. The photolysis of enrofloxacin showed to be efficient only in the presence of the bio-organic substance CVT 230 (BOS C), with almost 90% ENRO removal in neutral solution (pH 7). Results from the literature, associated with an experiment carried out in anoxic conditions, suggest singlet oxygen as the main species responsible for enrofloxacin oxidation. On the other hand, ENRO degradation by the H2O2/UV process showed a maximum removal of 48% at pH 7, suggesting that the action of singlet oxygen and/or hydroxyl radicals was not effective in the presence of hydrogen peroxide. BOS C can therefore be used as an efficient promoter for the treatment of enrofloxacin-containing water and wastewater under low irradiant power or in solar-irradiated systems.

Depuis plusieurs années, les quadruplexes de guanine ou G4, structures particulières de l’ADN, suscitent un intérêt grandissant. Ces structures, largement étudiées in vitro, sont encore peu connues in vivo mais semblent jouer un rôle important dans la régulation de l’expression génétique. Elles ont rapidement été considérées comme des cibles thérapeutiques potentielles pour certaines maladies telles que le cancer. Le premier indice de leur existence dans les cellules n’a été obtenu qu’en 2013 par immunodétection sur des cellules fixées. Les recherches sont actuellement tournées vers le développement de nouveaux outils moléculaires qui permettraient la visualisation des G4 dans des cellules vivantes.C’est dans ce cadre que nous avons conçu et développé une série de complexes polyazaaromatiques de ruthéniumII à base de ligands plans étendus (heptacycle dpqp et octacycle dppqp). La combinaison des propriétés photophysiques des complexes de RuII associée à la présence d’un large plan étendu devant permettre l’interaction avec les G4, positionne ces molécules comme outils potentiels pour l’étude des G4 in vivo.La première partie de ce projet porte sur la synthèse de ces nouveaux complexes de ruthénium. Une méthode originale de "chimie sur complexe" a permis d'obtenir, entre autres, un complexe possédant le ligand dpqp, fonctionnalisé par une triple liaison. Il a également été possible, « par chimie sur complexe », de construire un cycle supplémentaire sur le ligand heptacyclique (dpqp) chélaté pour obtenir les complexes [Ru(L)2dppqp]2+. Les propriétés photophysiques des différents complexes ont été étudiées. Seuls deux complexes, [Ru(phen)2dpqp-Cl]2+ et [Ru(TAP)2dpqp-Cl]2+, présentent un comportement s’approchant de celui des complexes de référence; c’est à dire des rendements quantiques comparables à [Ru(bpy)3]2+ et des durées de vie de l’état excité de l’ordre de la centaine de nanosecondes. Les autres complexes sont non luminescents et l’hypothèse d’un quenching par transfert de proton à l’état excité a été avancée pour expliquer ce comportement.Les complexes ont aussi été évalués vis à vis de différentes structures oligonucléotidiques G4 et duplexes. Tous les complexes possèdent une affinité correcte envers les G4. Comme nous l'espérions, le complexe porteur du ligand octacyclique semble être particulièrement sélectif envers les G4 par rapport à l'ADN double brin. Il a aussi été montré que deux des complexes testés peuvent être utilisés comme sondes moléculaires "light-switch ON" pour les structures G4 en milieu cellulaire. Certains des complexes synthétisés sont donc d’excellents candidats en tant qu’outils moléculaires pour l’étude des G4 in vivo. / The interest for some unusual structures of DNA, the G4 quadruplexes was growing these past few years. Those structures were extensively studied in vitro, but a lot remains unkwown about their existence in vivo. Intererstingly, G4 structures seem to play key regulatory roles in gene expression and are therefore potential therapeutic targets for diseases such as cancer. Consequently, visualisation of those structures in cells is the present challenge today, and the use of small molecules with luminescent properties is promising in this context.In this framework, we designed a series of new rutheniumII complexes chelating a large planar aromatic ligand, (heptacycle dpqp or octacycle dppqp). The combination of the photophysical properties of RuII complexes with proposed favorable stacking properties of the ligand with G4, would make these complexes promising molecular tools for G4 visualization.The first part of this project focusses on the synthesis of ruthenium complexes and their specific ligands. One particular ligand is formed by the condensation between acridine and phenanthroline moieties. A chemistry on the complex approach enabled us 1) to functionnalize the chelated ligand by a triple bond that will be used for the formation of an heterodimer by click chemistry, and 2) to build an additional cycle on the chelated ligand. Photophysical properties were evaluated and only two complexes, [Ru(phen)2dpqp-Cl]2+ and [Ru(TAP)2dpqp-Cl]2+, exhibit the same photophysical behaviour than the reference complexes. Indeed, the quantum yields are in the same order of magnitude than for [Ru(bpy)3]2+ and the excited state lifetime are of a few hundred nanosecondes. The other complexes are non-luminescent and a quenching by excited state proton transfer was hypothesised to explain this unusual behaviour. Interaction properties with G4 DNA were evaluated. All complexes display good affinity towards G4 and as expected, the complexe bearing the octacyclic ligand shows a strong selectivity towards G4 compared to dsDNA. Two of the complexes were proven to be potential luminescent light-switch ON probes for G4 detection in cells. In conclusion, this work highlights the possibility to use some of the newly synthetized complexes as efficient molecular tools for G4 visualization in cells.

As tecnologias utilizadas em estações de tratamento de água e efluentes não são eficientes para a remoção total de resíduos farmacêuticos e os efeitos dessas substâncias sobre o meio ambiente e a saúde humana ainda não são bem conhecidos. No presente trabalho, estudou-se a degradação do antibiótico enrofloxacina (ENRO) por fotólise e pelo processo H2O2/UV na presença de compostos bio-orgânicos (BOS), que têm se apresentado como interessantes promotores da oxidação de poluentes. Os experimentos foram realizados em um reator fotoquímico tubular de imersão com fonte radiante (lâmpada de xenônio) concêntrica, operado em batelada com recirculação. Utilizaram-se concentrações iniciais de ENRO e de BOS iguais a 50 mg L-1 e 20 mg L-1, respectivamente. Para todos os pH mantidos constantes (3, 5, 7 ou 9), a solução foi irradiada por 240 minutos. Os resultados indicam que o antibiótico não sofreu hidrólise em qualquer dos pH estudados em um período de 24 horas. Por sua vez, a fotólise da enrofloxacina mostrou-se eficiente somente na presença do composto bioorgânico CVT 230 (BOS C), com remoção de ENRO de quase 90% em meio neutro (pH 7). Resultados da literatura, associados a experimento realizado em meio anóxico, sugerem a participação de oxigênio singlete como principal espécie oxidante da enrofloxacina. Por outro lado, a degradação da ENRO pelo processo H2O2/UV apresentou remoção máxima do fármaco de 48% em pH 7, o que sugere que a ação de oxigênio singlete e/ou radicais hidroxila não foi eficaz na presença de peróxido de hidrogênio. Portanto, o BOS C pode ser empregado como promotor no tratamento de águas e efluentes aquosos contaminados com enrofloxacina sob baixas potências radiantes ou em sistemas irradiados por luz solar. / The technologies used in water and wastewater treatment plants are not efficient for the total removal of pharmaceutical compounds whose effect to the environment and to human health are still not well known. In this work, the degradation of the antibiotic enrofloxacin (ENRO) was studied by photolysis and by the H2O2/UV process in the presence of bio-organic substances (BOS), which have been identified as interesting promoters of pollutant oxidation. The experiments were carried out in a tubular immersion photochemical reactor equipped with a concentric radiant source (xenon lamp), and operated in batch mode with recirculation. Initial ENRO and BOS concentrations of 50 mg L-1 and 20 mg L-1 were used, respectively. The solution was irradiated for 240 minutes for all pH studied at constant values (3, 5, 7, and 9). The results show that the antibiotic did not undergo hydrolysis at any pH after 24 hours. The photolysis of enrofloxacin showed to be efficient only in the presence of the bio-organic substance CVT 230 (BOS C), with almost 90% ENRO removal in neutral solution (pH 7). Results from the literature, associated with an experiment carried out in anoxic conditions, suggest singlet oxygen as the main species responsible for enrofloxacin oxidation. On the other hand, ENRO degradation by the H2O2/UV process showed a maximum removal of 48% at pH 7, suggesting that the action of singlet oxygen and/or hydroxyl radicals was not effective in the presence of hydrogen peroxide. BOS C can therefore be used as an efficient promoter for the treatment of enrofloxacin-containing water and wastewater under low irradiant power or in solar-irradiated systems.

Thesis deals with analysis of two consumer eco-labels, specifically BIO - organic farming product and Environmentally friendly product. In the practical part is made. The result of the marketing research is the identification of knowledge and awareness of consumers of selected eco-labels.

The thesis defines a concept of organic food and a closer look is given to an organic agriculture - a place of origin for organic food. The second part discusses benefits of organic food for consumers and looks for consumer purchase motives. Third and fourth part is devoted to rules of labeling and opportunities for improvement in this area. Other consumer brands and a current state on organic food market are presented here as well. Variety of products offered and availability of organic food at the particular points of sale are analyzed in the final part.

The project is about piezoelectric energy harvesters and piezoelectric bio-organic materials.Nowadays, various kinds of energy harvesters based on micro or nano materials are appliedinmanyelectronic applications, such as wearable devices and electricity generators. Amongthem, thepiezoelectric effect-based energy harvesters are more attractive in research and industryfields. Inrecent years, piezoelectric biomaterials become a popular option because they are availabletocouple electrical and mechanical energy in a biological, ecofriendly systemto generate electricityinreal time. Among them, γ- glycine crystals have been recently synthesized in wafer scale throughasimple polyvinyl alcohol (PVA)-assisted evaporation process exhibiting good piezoelectricperformance. However, so far there are no metal-free energy-harvesting devices basedonPVA-glycine film to enable green manufacturing. In this project, we proposed the direct inkwritingorganic PEDOT:PSS electrodes and PVA-glycine-PVA piezoelectric crystals to fabricate metal-freeenergy harvesters. The output voltage reaches 1.5 V at a load resistance of 500 MΩandunderaforce of 10 N. The performance is comparable to other glycine-based devices in recent literature.Our scalable, sustainable and low-cost printing process is expected to greatly contribute tothefieldof biomaterials-based piezoelectric energy harvesting. / Projektet handlar om piezoelektriska energiskördare och piezoelektriska bioorganiska material. Nuförtiden används olika typer av energiskördare baserade på mikro- eller nanomaterial i mångaelektroniska applikationer, såsom bärbara enheter och elgeneratorer. Bland dem är de piezoelektriskaeffektbaserade energiskördarna mer attraktiva inom forsknings- och industriområden. På senare år harpiezoelektriska biomaterial blivit ett populärt alternativ eftersom de är tillgängliga för att koppla elektrisk och mekanisk energi i ett biologiskt, miljövänligt system för att generera elektricitet i realtid. Bland dem har γ-glycinkristaller nyligen syntetiserats i waferskala genom en enkel polyvinylalkohol (PVA)-assisterad förångningsprocess som uppvisar god piezoelektrisk prestanda. Än så länge finnsdet dock inga metallfria energiskördande enheter baserade på PVA-glycinfilm för att möjliggöra gröntillverkning. I detta projekt föreslog vi direkt bläckskrivande organiska PEDOT:PSS-elektroder ochPVA-glycin-PVA piezoelektriska kristaller för att tillverka metallfria energiskördare. Utspänningennår1,5 V vid en belastningsresistans på 500 MΩ och under en kraft på 10 N. Prestandan är jämförbar medandra glycinbaserade enheter i nyare litteratur. Vår skalbara, hållbara och billiga utskriftsprocess förväntas i hög grad bidra till området för biomaterialbaserad piezoelektrisk energiskörd.

This diploma theses deals with the construction of the neoplasm Biomother in the Czech media between 2010-2015. The 'bio' concept, label borrowed from organic agriculture, resonates in public for quite long. The research component of this theses brings an explanation, why the notion of bio was connected with motherhood, and also why media has changed its meaning and brought new defining topics to it. There are 3 main questions to be answered in the theses. How did the picture of Biomothers change between the years 2010-2015? What other characteristics have been included in the topic? What role does media have in this case: Do they serve as a booster of alternative values or, do they tend to promote conservative values? Media portrayal of Biomothers showed considerable dynamism during the examined period and it altered to greater complexity and controversy. Some features, that accompanied the notion biomatka from the start, remained unchanged and are key for its interpretation. This includes the tendency to search and buy organic products, support an all natural lifestyle, and searching for forms of alternative treatments. During the analysed period (2010- 2015) the relatively uncomplicated and positive image of Biomothers, as a worshiper of organic farming, natural lifestyle and organic products,...