A large number of anti-cancer chemotherapeutics target DNA topoisomerases. Etoposide is a specific topoisomerase II poison which causes reversible double strand DNA breaks. The focus of this project is to analyze the repair of DNA damage induced by etoposide.. Double strand DNA break repair is mediated by through either non-homologous end joining (NHEJ) or homologous recombination. NHEJ repairs through direct ligation of a double stranded break while homologous recombination utilizes a homologous template to recover the wild type sequence. A reporter cassette, RYDR-GFP, has been stably integrated into HeLa cells. This reporter contains an ultra-high affinity topoisomerase II cleavage site (RY) placed in the middle of a mutant GFP sequence. Flanking this sequence is a corresponding stretch of wild type GFP that is used as template to repair the break and restore gene function yielding GFP positive cells. Titrations with etoposide have shown that a logarithmic increase in drug concentration yields a corresponding increase in repair through homologous recombination (HR). This result demonstrates that topoisomerase II mediated damage is efficiently repaired by the process of HR. To examine NHEJ repair, a doxycycline inducible, stably integrated NHEJ HeLa cell reporter cassette was also evaluated. The data indicates that repair of topoisomerase II mediated DNA damage occurs more efficiently through the HR pathway. Collectively, the data suggests that tumor cells proficient in HR repair may effectively elude treatment by topoisomerase II targeting drugs.

No description available.

Le contexte environnemental (réchauffement climatique) et politique (augmentation du parc EnR) entraîne une mutation du paysage énergétique français. Le méthane de synthèse est présenté comme un vecteur énergétique permettant le stockage et le transport de l’électricité renouvelable en surproduction tout en recyclant le CO2 (Power-to-Methane). Un des objectifs de la thèse est de développer un catalyseur actif en co-méthanation d’un mélange post co-électrolyse H2/CO/CO2/H2O/CH4 (projet ANR CHOCHCO). Le catalyseur synthétisé est de type Ni/CZP. L’optimisation de la synthèse coprécipitation par l’utilisation combinée du sel précipitant (NH4)2CO3 et du tensioactif CTAB a mené à un catalyseur performant (absence de poison alcalin et augmentation de la surface spécifique) capable de produire du CH4 à basse température (250 °C) avec un rendement élevé. Le catalyseur a montré une résistance satisfaisante à la désactivation par dépôt de carbone et par frittage, indispensable pour le fonctionnement intermittent du procédé. Un modèle cinétique, de type Langmuir-Hinshelwood, a pour la première fois été développé sur un catalyseur de type Ni/CZP. / The environmental (global warming) and political (increase of the renewable electric farm) context leads to a mutation of the French energy landscape. Synthetic methane is presented as a energy carrier for storing and transporting renewable electricity in overproduction while recycling CO2, a process called Power-to-Methane. One of the objectives of the thesis is to develop an active catalyst for the co-methanation of a post-co-electrolysis mixture H2/CO/CO2/H2O/CH4 (CHOCHCO ANR project). A Ni/CZP type catalyst was synthesized in this purpose. The optimization of the coprecipitation synthesis by the combination of the precipitating salt (NH4)2CO3 and the surfactant CTAB has led to a high performance catalyst (absence of alkaline poison and increase of the specific surface area) able to produce CH4 at low temperature (250 °C) with a high yield. The catalyst showed a satisfactory resistance to carbon deposition and sintering deactivation, which is a key point for the intermittent operating conditions of the process. A kinetic model, of the Langmuir-Hinshelwood type, was developed for the first time on a Ni/CZP type catalyst.

L’excès de réactivité neutronique dans les centrales nucléaires est compensé par des sys-tèmes actifs de contrôle du réacteur : acide borique et barres de contrôle. L’apport d’antiréactivité peut se faire passivement avec des poisons consommables, c'est-à-dire des absorbants de neutrons, en particulier du gadolinium (Gd).Dans le cadre d’une augmentation de l’enrichissement en U²³⁵ et de réduction de l’utilisation d’acide borique, cette thèse a pour objectif d’optimiser la distribution du ga-dolinium dans des céramiques d’UO₂ afin d’obtenir un apport optimisé d’antiréactivité dans un Réacteur à Eau sous Pression.Dans ce sens, le travail est orienté à trouver des nouveaux matériaux riches en gadolinium. Le diagramme de phase U-O-Gd a donc été exploré dans le domaine des fortes teneurs en Gd. Deux phases cubiques ont été trouvées et caractérisées : les phases C1 et C2. En vue d’une application industrielle, la phase C1 a été retenue comme candidate pour l’ajout du Gd dans les pastilles d’UO₂.La distribution optimale de cette phase C1 dans les assemblages de combustible nucléaire a été étudiée avec le code de calcul neutronique APOLLO2.8. Des études paramétriques ont été réalisées. Ces études neutroniques ont aboutit à un concept performant de pastille empoisonnée. Finalement, des pastilles prototype ont été fabriquées en laboratoire suivant ce concept. L’ensemble des résultats obtenus montre qu’un concept de pastille avec un dépôt superficiel neutrophage de phase C1 est une manière d’apporter de l’antiréactivité de manière optimisée dans le cadre de cycles longs. Ceci pourrait potentiellement être appliquée à l‘échelle industrielle. Un brevet a été déposé en ce sens. / Reactivity excess in Nuclear Power Plants is controlled by reactor’s active systems: boric acid dilution and control rods. Alternatively, negative reactivity insertion can be made in a passive way using burnable poisons, i.e. neutron absorbers, this is the case of gadolinium (Gd).In the industrial framework of U²³⁵ enrichment increase and boric acid restraint, the goal of this thesis is to optimize the distribution of gadolinium in UO₂ ceramics to obtain a high-performance provision of negative reactivity in Pressurized Water Reactors.In this sense, the work is focus on new gadolinium-rich materials. Thus, U-Gd-O phase diagram was explored in the field of high Gd contents. Two cubic phases were found and characterized: the C1 and C2 phases. With the aim of an industrial application, C1 phase was selected as candidate for Gd addition into UO₂ pellets.The optimal distribution of C1 phase within a nuclear fuel assembly was studied using APOLLO 2.8 neutron transport code. Parametrical calculations were performed. These neutronic studies have ends in a successful “concept of poisoned pellet”.Finally, some prototype pellets following this concept were made in laboratory to proof it feasibility.All the obtained results shows that the proposed concept of a neutrophage C1-phase coating on UO₂ pellets is a convenient way to reduce reactivity excess within the framework of long irradiation cycles. This concept could be potentially applied in industrial scale. Consequently a patent application process was initiated.

Neste trabalho avaliamos se as Natterinas (NTR), proteínas majoritárias presentes no veneno do peixe peçonhento T. nattereri, são as responsáveis pela a ação inibitória do recrutamento de leucócitos durante a inflamação e os mecanismos moleculares envolvidos neste processo. Utilizando análises de microscopia intravital tratamos os animais com as NTR e 6 h depois induzimos o rolamento de leucócitos com KC, LPS ou agonista de PAR-4 sobre a microcirculação. Os dados mostraram que o tratamento com as NTR inibem o rolamento e a migração de leucócitos induzido pela KC e pelo LPS. Além disso, o efeito inibitório das NTR depende parcialmente da sua atividade proteásica e não está relacionado com a clivagem proteolítica da quimiocina KC ou com a indução de reguladores endógenos como IL-10, corticóides, a enzima HO-1 ou IL-1Ra. O efeito inibitório parece ser dependente da ativação da sinalização TLR/MyD88 e PI3K e das proteínas serina/treonina fosfatases, contribuindo de forma significativa na sobrevida de animais induzidos a endotoxemia por LPS. / In this work we evaluated whether Natterins (NTR) the majority proteins present in the venom of Thalassophryne nattereri venomous fish are responsible for the inhibitory action of leukocyte recruitment during inflammation and the molecular mechanisms involved in this process. Using intravital microscopy analysis we treated the animals with NTR and 6 h after we induced the leukocyte rolling by topical of KC, LPS or agonist of PAR-4 on the microcirculation. The data showed that treatment with NTR inhibited the leukocyte rolling and migration induced by LPS and KC. Moreover, the inhibitory effect of NTR depends partly on its proteinase activity and is not related to the proteolytic cleavage of the chemokine or KC nor with the induction of endogenous regulators such as IL-10, corticoids, the HO-1 or IL-1Ra. The inhibitory effect is dependent on the activation of PI3K and TLR/MyD88 signaling and protein serine/threonine phosphatases, contributing significantly to the survival of animals endotoxemia induced by LPS.

A Toxicologia tem desempenhado um importante papel na identificação de efeitos nocivos à população, gerando subsídios para a tomada de decisões na prática clínica, auxiliando desta forma no bom prognóstico de pacientes intoxicados. De acordo com a Pan American Health Organization, o sucesso em qualquer intervenção em saúde somente pode ser obtido com a criação e manutenção de um banco de dados confiável, que seja capaz de predizer as diversas particularidades das intoxicações, como a população-alvo e suas suscetibilidades. Assim, recomenda-se que médicos, especialistas, legisladores e administradores em saúde adotem em sua rotina uma coleta de dados sistêmica e integrada para o mapeamento e caracterização das intoxicações. Neste cenário, técnicas de análises multivariadas poderiam ser empregadas para evidenciar possíveis intercorrelações; entretanto, seu uso ainda não é comum na toxicologia clínica. Neste trabalho foram identificados e quantificados agentes exógenos em amostras biológicas (sangue e urina) provenientes do Centro de Controle de Intoxicações da cidade de São Paulo, correlacionando os dados obtidos dessas análises com o perfil clínico e prognóstico dos pacientes. Fármacos benzodiazepínicos, antidepressivos, anticonvulsivantes, paracetamol, drogas de abuso e praguicidas foram selecionados de acordo com a incidência reportada por esse centro no período de 2013 a 2014. Do total de amostras analisadas (n = 320), 192 foram positivas para alguma substância, sendo 101 positivas para etanol e 131 positivas para as demais substâncias. Os dados obtidos foram submetidos à análise de correspondência múltipla e análise hierárquica de cluster. A partir da análise multivariada foi possível agrupar os indivíduos em 3 clusters, o que correspondeu a 66,5% do total de informações. No primeiro eixo houve a separação dos pacientes do gênero feminino, que se intoxicaram ou foram expostos a medicamentos e drogas de abuso, por tentativa de suicídio, dos pacientes do gênero masculino, de idade entre 30 a 39 anos, que se intoxicaram com drogas de abuso. No segundo eixo fatorial foram agrupados os pacientes que se intoxicaram com etanol isoladamente, juntamente com pacientes que se intoxicaram com diazepam. Este trabalho contribuiu para o mapeamento dos casos de intoxicação atendidos pelo CCI-SP e foi um estudo inicial para a criação de um banco de dados que poderá ser alimentado constantemente e assim, oferecer ao sistema de toxicovigilância uma base para políticas educativas. / Toxicology has played an important role in the identification of harmful effects to the population, generating subsidies for decision making in clinical practice, helping in this way the good prognosis of acutely intoxicated patients. According to the Pan American Health Organization, success in any health intervention can only be achieved by creating and maintaining a reliable database that is capable of predicting the various characteristics of intoxications, such as the target population and their susceptibilities. Thus, it is recommended that doctors, specialists, legislators and health administrators adopt in their routine a systemic and integrated data collection for the mapping and characterization of intoxications. In this scenario, multivariate analysis techniques could be used to evidence possible intercorrelations; however, its use is not yet common in clinical toxicology. In this work, were identified and quantified exogenous agents in biological samples (blood and urine) from the Poison Control Center São Paulo city, correlating the data obtained from these analyzes with the clinical and prognostic profile of the patients. Benzodiazepines, antidepressants, anticonvulsants, acetaminophen, drugs of abuse and pesticides were selected according to the incidence reported by this center in the period from 2013 to 2014. Of the total number of samples analyzed (n = 320), 192 samples have shown to be positive for some of the analytes, from these 100 were positive for ethanol and 131 positive for other substances. The data were submitted to multiple correspondence analysis and hierarchical cluster analysis. From the multivariate analysis it was possible to group the individuals into 3 clusters, which corresponded to 66.5% of the total information. In the first axis, the patients were separated from the female gender, who were intoxicated or were exposed to drugs and suicide attempt of the male patients, aged between 30 and 39 years, who became intoxicated with drugs of abuse. In the second factorial axis were grouped the patients who were intoxicated with ethanol, together with patients who became intoxicated with diazepam. This work contributed to the mapping of intoxication cases attended by the CCI-SP and was a initial study for the creation of a database that could be fed constantly and thus provide the toxicovigilance system with a basis for educational policies.

A Toxicologia tem desempenhado um importante papel na identificação de efeitos nocivos à população, gerando subsídios para a tomada de decisões na prática clínica, auxiliando desta forma no bom prognóstico de pacientes intoxicados. De acordo com a Pan American Health Organization, o sucesso em qualquer intervenção em saúde somente pode ser obtido com a criação e manutenção de um banco de dados confiável, que seja capaz de predizer as diversas particularidades das intoxicações, como a população-alvo e suas suscetibilidades. Assim, recomenda-se que médicos, especialistas, legisladores e administradores em saúde adotem em sua rotina uma coleta de dados sistêmica e integrada para o mapeamento e caracterização das intoxicações. Neste cenário, técnicas de análises multivariadas poderiam ser empregadas para evidenciar possíveis intercorrelações; entretanto, seu uso ainda não é comum na toxicologia clínica. Neste trabalho foram identificados e quantificados agentes exógenos em amostras biológicas (sangue e urina) provenientes do Centro de Controle de Intoxicações da cidade de São Paulo, correlacionando os dados obtidos dessas análises com o perfil clínico e prognóstico dos pacientes. Fármacos benzodiazepínicos, antidepressivos, anticonvulsivantes, paracetamol, drogas de abuso e praguicidas foram selecionados de acordo com a incidência reportada por esse centro no período de 2013 a 2014. Do total de amostras analisadas (n = 320), 192 foram positivas para alguma substância, sendo 101 positivas para etanol e 131 positivas para as demais substâncias. Os dados obtidos foram submetidos à análise de correspondência múltipla e análise hierárquica de cluster. A partir da análise multivariada foi possível agrupar os indivíduos em 3 clusters, o que correspondeu a 66,5% do total de informações. No primeiro eixo houve a separação dos pacientes do gênero feminino, que se intoxicaram ou foram expostos a medicamentos e drogas de abuso, por tentativa de suicídio, dos pacientes do gênero masculino, de idade entre 30 a 39 anos, que se intoxicaram com drogas de abuso. No segundo eixo fatorial foram agrupados os pacientes que se intoxicaram com etanol isoladamente, juntamente com pacientes que se intoxicaram com diazepam. Este trabalho contribuiu para o mapeamento dos casos de intoxicação atendidos pelo CCI-SP e foi um estudo inicial para a criação de um banco de dados que poderá ser alimentado constantemente e assim, oferecer ao sistema de toxicovigilância uma base para políticas educativas. / Toxicology has played an important role in the identification of harmful effects to the population, generating subsidies for decision making in clinical practice, helping in this way the good prognosis of acutely intoxicated patients. According to the Pan American Health Organization, success in any health intervention can only be achieved by creating and maintaining a reliable database that is capable of predicting the various characteristics of intoxications, such as the target population and their susceptibilities. Thus, it is recommended that doctors, specialists, legislators and health administrators adopt in their routine a systemic and integrated data collection for the mapping and characterization of intoxications. In this scenario, multivariate analysis techniques could be used to evidence possible intercorrelations; however, its use is not yet common in clinical toxicology. In this work, were identified and quantified exogenous agents in biological samples (blood and urine) from the Poison Control Center São Paulo city, correlating the data obtained from these analyzes with the clinical and prognostic profile of the patients. Benzodiazepines, antidepressants, anticonvulsants, acetaminophen, drugs of abuse and pesticides were selected according to the incidence reported by this center in the period from 2013 to 2014. Of the total number of samples analyzed (n = 320), 192 samples have shown to be positive for some of the analytes, from these 100 were positive for ethanol and 131 positive for other substances. The data were submitted to multiple correspondence analysis and hierarchical cluster analysis. From the multivariate analysis it was possible to group the individuals into 3 clusters, which corresponded to 66.5% of the total information. In the first axis, the patients were separated from the female gender, who were intoxicated or were exposed to drugs and suicide attempt of the male patients, aged between 30 and 39 years, who became intoxicated with drugs of abuse. In the second factorial axis were grouped the patients who were intoxicated with ethanol, together with patients who became intoxicated with diazepam. This work contributed to the mapping of intoxication cases attended by the CCI-SP and was a initial study for the creation of a database that could be fed constantly and thus provide the toxicovigilance system with a basis for educational policies.

Made available in DSpace on 2015-07-22T20:50:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-01-27 / Fundação de Amparo à Pesquisa do Estado do Amazonas (FAPEAM) / Introdução: Centros de Informações Toxicológicas (CIT) prestam papel essencial na assistência de pacientes intoxicados, sem contar com financiamento seguro de suas atividades. Objetivo: verificar a diferença no tempo de internação dos pacientes intoxicados que receberam atenção remota de um CIT em comparação aos que não receberam. Métodos: foi organizada uma coorte retrospectiva incluindo todos pacientes intoxicados hospitalizados em um pronto-socorro de Manaus de 2005 a 2007, dos quais os pacientes selecionados (com agente tóxico conhecido, tempo de exposição inferior a 12 horas e sem comorbidades graves) tiveram a gravidade avaliada por dois revisores independentes e divergências resolvidas por outro revisor. A concordância foi calculada através do índice Kappa. Resultados: Foram incluídos 198 pacientes; aqueles com auxílio remoto do CIT ficaram em média 3,43 dias (- 6,10 a -0,77 IC 95%) a menos internados quando comparados a nenhum auxílio do CIT. Noventa pacientes tiveram gravidade avaliada; não houve diferença estatística na gravidade entre os pacientes com ou sem assistência do CIT (p > 0,5). A concordância entre os revisores foi significativa. Conclusão: Pacientes com assistência remota do CIT tiveram tempo de internação inferior a pacientes sem este auxílio. A análise de gravidade mostrou-se factível de ser incorporada à prática dos CIT brasileiros. / Introduction: Poison Control Centers (PCC) play an essential role in caring for poisoned patients, albeit without secure funding for their activities. Objective: to investigate differences in length of hospital stay among poisoned patients, between those who received remote assistance from a PCC and those who did not. Methods: a retrospective cohort including all poisoned patients hospitalized at an emergency service in Manaus between 2005 and 2007 was set up. Patients presenting a known toxic agent, with less than 12 hours elapsed since exposure and without severe comorbidities, were selected. Their severity of poisoning was evaluated by two independent reviewers and divergence was solved by another reviewer. Agreement was obtained by Kappa index. Results: 198 patients were included. Those who received remote assistance from a PCC stayed in hospital on average for 3.43 days less than those without PCC assistance (95% CI: -6.10 to -0.77). Severity was assessed in the cases of 90 patients: there was no statistical difference in severity between the patient groups (p > 0.5). Agreement between reviewers was substantial. Conclusion: Patients with PCC aid had a lower length of stay then patients without this aid. Severity assessment is likely to be incorporated into Brazilian PCC routine. / TEDE / BV UNIFESP: Teses e dissertações

ABSTRACT Toxicology and Health Aspects of Non-lethal Chemical Weapons. Non-lethal chemical weapons, which belong among the mass destruction weapons, have been one of the most frequently discussed topics recently. These weapons are able to disbar manpower or combat technology and weapons smartly and temporarily with minimum costs. The range of possible application of chemical weapons as non-lethal is probably wider compared to any other type, and there are a lot of means capable of immediate wide application nowadays. The possibility to produce highly efficient psychotropic substances with controllable action time, human immobilizing substances is particularly pointed out. The main aim of my work is to clarify the importance of non-lethal chemical weapons as possible means of suppressing inner turmoil and to find the extent of knowledge of non-lethal chemical weapons among the informed and the general public by means of obtained answers and follow-up research, then to confirm hypotheses that the informed public consider non-lethal chemical weapons to be combat means and weapon systems intended for temporary putting people out of combat or other intentional activity; that the general public perceive non-lethal chemical weapons as health endangering substances and have only limited information about them, by means of selected statistical methods. I chose questionnaire survey to examine awareness of the general and informed public of this matter. The statistic sets in both the groups consist of a hundred respondents from the city of České Budějovice. Stratified selection was performed for both the examined sets upon consultation with a member of the Fire Brigade in České Budějovice. In terms of threat to the population it is important to ensure its safety, namely upon the knowledge among the units of the Integrated Emergency System intervening in the instance of terroristic attack.

Neste trabalho avaliamos se as Natterinas (NTR), proteínas majoritárias presentes no veneno do peixe peçonhento T. nattereri, são as responsáveis pela a ação inibitória do recrutamento de leucócitos durante a inflamação e os mecanismos moleculares envolvidos neste processo. Utilizando análises de microscopia intravital tratamos os animais com as NTR e 6 h depois induzimos o rolamento de leucócitos com KC, LPS ou agonista de PAR-4 sobre a microcirculação. Os dados mostraram que o tratamento com as NTR inibem o rolamento e a migração de leucócitos induzido pela KC e pelo LPS. Além disso, o efeito inibitório das NTR depende parcialmente da sua atividade proteásica e não está relacionado com a clivagem proteolítica da quimiocina KC ou com a indução de reguladores endógenos como IL-10, corticóides, a enzima HO-1 ou IL-1Ra. O efeito inibitório parece ser dependente da ativação da sinalização TLR/MyD88 e PI3K e das proteínas serina/treonina fosfatases, contribuindo de forma significativa na sobrevida de animais induzidos a endotoxemia por LPS. / In this work we evaluated whether Natterins (NTR) the majority proteins present in the venom of Thalassophryne nattereri venomous fish are responsible for the inhibitory action of leukocyte recruitment during inflammation and the molecular mechanisms involved in this process. Using intravital microscopy analysis we treated the animals with NTR and 6 h after we induced the leukocyte rolling by topical of KC, LPS or agonist of PAR-4 on the microcirculation. The data showed that treatment with NTR inhibited the leukocyte rolling and migration induced by LPS and KC. Moreover, the inhibitory effect of NTR depends partly on its proteinase activity and is not related to the proteolytic cleavage of the chemokine or KC nor with the induction of endogenous regulators such as IL-10, corticoids, the HO-1 or IL-1Ra. The inhibitory effect is dependent on the activation of PI3K and TLR/MyD88 signaling and protein serine/threonine phosphatases, contributing significantly to the survival of animals endotoxemia induced by LPS.