This study analyzed the interaction of the sleep aid eszopiclone (ESZ) and antidepressant fluoxetine (FLX) on social defeat stress (SDS) in the mouse. Beta adrenoreceptors, brain-derived neurotrophic factor (BDNF) and cAMP response element binding protein (CREB) expression in the hippocampus and frontal cortex were also analyzed. Subjects were adult male 'intruder' C57/B6 mice that were exposed to a retired 'resident' male breeder ICR mouse in this animal's home cage for a 5 min period for each of 10 consecutive days, and the resident established physical dominance. The following day, all animals were assigned to one of four drug treatment groups, and treatment was given for up to 18 days: vehicle, ESZ only (3mg/kg), FLX (10mg/kg) only, or ESZ+FLX. A social interaction test was given on days 1, 5, 10, and 15 of drug treatment to assess SDS. Results showed that the ESZ+FLX group spent less time in avoidance zones during the interaction test at days 1 and 5, and more time in the interaction zone at day 5 compared to defeated mice given vehicle. All drug treatment groups spent more time in the interaction zone compared to defeated mice given vehicle on day 1 as well as day 10. SDS completely dissipated by the fourth interaction test according to both behavioral measures. Neurochemically, SDS did not produce changes in any marker analyzed. This study shows the combination of ESZ and FLX alleviated SDS, but a neurochemical correlate remains elusive.

The focus of this study was to analyze whether the psychostimulant nicotine would enhance reference and working memory consolidation in rats tested on the 8-arm radial arm maze. Mecamylamine, a nicotine antagonist, was used to attempt to block the enhancement of memory consolidation. All rats were given one training trial/day for 12 consecutive days, and 4 arms were baited. Rats were separated into five groups: the saline-nicotine group received an intraperitoneal (i.p.) injection of saline immediately after each trial followed 15 min later by an subcutaneous (s.c.) injection of nicotine (0.6 mg/kg free base); the nicotine-delay group received an s.c. injection of nicotine 2 h after each training trial, two groups received an i. p. injection of one of two different doses of mecamylamine (2.5 and 6.0 mg/kg) immediately after each trial, which was followed 15 min later by an s.c. nicotine injection, and a control group received an i.p. injection of saline immediately and 15 min after each training trial. Results showed that the saline-nicotine group made fewer reference and working memory errors than the saline- or nicotine-delay groups, but only the effect of nicotine on reference memory was blocked by the higher dose of mecamylamine. It appears from these results that nicotine's effects on reference and working memory may be mediated through different mechanisms.

Reading has long been acknowledged to be a critical skill that is best acquired early in life. According to the most recent National Assessment of Educational Progress (NAEP) reports, American public school children continue to struggle to master the basics of reading. Although federal funding in real dollars has increased consistently over time, reading scores have not followed suit, suggesting that fiscal resources have not been applied successfully to the variables that are directly related to reading acquisition and achievement. The current state of affairs therefore suggests the need for identifying a fiscally-targetable, instructionally-relevant variable with a direct, causal relationship to early-reading achievement. One way to determine whether such a relationship exists between two variables is by means of dose-response methodology. Although this methodology has not been broadly implemented in educational research, it is attractive because it allows for the formal characterization and comparison of cause-effect relationships, and may also inform practice in readily implementable ways. Researchers have noted that time spent learning (TSL), and in particular academic learning time (ALT), is a promising candidate for a dose-response relationship with student achievement in early reading. Although ALT holds promise, there have traditionally been significant difficulties in operationalizing and quantifying it. The growing prevalence of academic software in the American public classroom holds promise for overcoming these challenges and provides an opportunity to test the hypothesis that there is a dose-response relationship between ALT and student achievement in early reading.

Campylobacter enteritis in humans is primarily associated with C. jejuni/coli infection. Other species cause campylobacteriosis relatively infrequently; while this could be attributed to bias in diagnostic methods, the pathogenicity of non-jejuni/coli Campylobacter spp. such as C. upsaliensis and C. helveticus (isolated from dogs and cats) is uncertain. Galleria mellonella larvae are suitable models of the mammalian innate immune system and have been applied to C. jejuni studies. This study compared the pathogenicity of C. jejuni, C. upsaliensis, and C. helveticus isolates. Larvae inoculated with either C. upsaliensis or C. helveticus showed significantly higher survival than those inoculated with C. jejuni. All three Campylobacter species induced indistinguishable histopathological changes in the larvae. C. jejuni could be isolated from inoculated larvae up to eight days post-inoculation whereas C. upsaliensis and C. helveticus could only be isolated in the first two days. There was a significant variation in the hazard rate between batches of larvae, in Campylobacter strains, and in biological replicates as random effects, and in species and bacterial dose as fixed effects. The Galleria model is applicable to other Campylobacter spp. as well as C. jejuni, but may be subject to significant variation with all Campylobacter species. While C. upsaliensis and C. helveticus cannot be considered non-pathogenic, they are significantly less pathogenic than C. jejuni.

The dual-specificity protein tyrosine phosphatases (PTPs) play integral roles in the regulation of cell signaling. There is a need for new tools to study these phosphatases, and the identification of inhibitors potentially affords not only new means for their study, but also possible therapeutics for the treatment of diseases caused by their dysregulation. However, the identification of selective inhibitors of the protein phosphatases has proven somewhat difficult. PTP localized to mitochondrion 1 (PTPMT1) is a recently discovered dual-specificity phosphatase that has been implicated in the regulation of insulin secretion. Screening of a commercially available small-molecule library yielded alexidine dihydrochloride, a dibiguanide compound, as an effective and selective inhibitor of PTPMT1 with an in vitro concentration that inhibits response by 50% of 1.08 microM. A related dibiguanide analog, chlorhexidine dihydrochloride, also significantly inhibited PTPMT1, albeit with lower potency, while a monobiguanide analog showed very weak inhibition. Treatment of isolated rat pancreatic islets with alexidine dihydrochloride resulted in a dose-dependent increase in insulin secretion, whereas treatment of a pancreatic beta-cell line with the drug affected the phosphorylation of mitochondrial proteins in a manner similar to genetic inhibition of PTPMT1. Furthermore, knockdown of PTPMT1 in rat islets rendered them insensitive to alexidine dihydrochloride treatment, providing evidence for mechanism-based activity of the inhibitor. Taken together, these studies establish alexidine dihydrochloride as an effective inhibitor of PTPMT1, both in vitro and in cells, and support the notion that PTPMT1 could serve as a pharmacological target in the treatment of type II diabetes. / Dissertation

A expansão das áreas cultivadas com cana-de-açúcar no Brasil é evidente, e para um eficaz manejo dessas extensas áreas, é necessário um bom planejamento das operações agrícolas. Nesse sentido, a aplicação de herbicidas pode ocorrer até no período seco do ano e em condições de solo bem distintas. Sabe-se que os diferentes ambientes nos quais os herbicidas são aplicados têm grande influencia em sua eficácia, e entender essas diferenças é importante para se obter o melhor rendimento e se fazer o correto uso dessa excelente ferramenta de manejo. Assim, este trabalho teve como objetivo avaliar o comportamento da formulação comercial contendo os herbicidas diurom + hexazinona + sulfometurom-metílico quando aplicado em solos com texturas contrastantes, além de avaliar a lixiviação desse herbicida e seus componentes e também como condições de restrição hídrica em presença de palha de cana-de-açúcar podem afetar o seu efeito residual e de seus componentes. Foram realizados ao todo 5 experimentos sendo 3 com o uso de curvas de dose-resposta, avaliando-se capim-colchão e corda-de-viola; um com lixiviação em tubos de PVC, avaliando-se corda-de-viola, e um com períodos de restrição hídrica, avaliando-se corda-de-viola e capim-colonião. Os resultados obtidos nos experimentos com as curvas de dose-resposta demonstraram a grande influencia da textura na eficácia do herbicida, com quantidades de argila menores que 30% sendo suficientes para gerar respostas consideravelmente distintas. O experimento com lixiviação demonstrou que as três moléculas do herbicida comportam-se diferentemente no solo, o que também é importante para o seu uso em diferentes ambientes. Os resultados obtidos no experimento com períodos de restrição hídrica em palha de cana-de-açúcar permitem concluir que apesar de as três moléculas comportarem-se diferentemente no ambiente, elas se complementam no controle de plantas daninhas mono e dicotiledôneas, e têm o efeito residual pouco influenciado por períodos de restrição hídrica. / The expansion of areas planted with sugar cane in Brazil is clearly noticeable and a good planning of agricultural operations is necessary for an effective management of these extensive areas. Accordingly, the application of herbicides may occur even in the dry season and in distinctive soil conditions. It is known that the different environments in which the herbicides are applied have great influence on their effectiveness and understanding these differences is an important factor in order to obtain best performance and make proper use of this excellent management tool. Thus, this work aimed to evaluate the behavior of the commercial formulation containing the herbicides diuron + hexazinone + sulfometurom-methyl when applied to soils with contrasting textures, and also to evaluate the leaching of this herbicide and its components as well as how conditions of water restriction with presence of sugarcane staw can affect your residual effect and its components. Five experiments were performed, three of those using dose-response curves, to evaluate crabgrass and morning glory; one with leaching using PVC tubes, evaluating morning glory and one with periods of water restriction, evaluating morning glory and guinea grass. Obtained results of the experiments with the dose-response curves demonstrated the great influence of the texture on the effectiveness of the herbicide, clay amounts of less than 30% being sufficient to generate considerably distinct responses. The experiment with leaching showed that the three molecules behave differently in the soil, which is also important to their use in different environments. Obtained results in the experiment with periods of water restriction on straw sugarcane allow the conclusion that although the three molecules behave differently in the environment, they complement each other in control of mono and dicotyledonous weeds, and they have little residual effect influenced by periods of water restriction.

A Doença de Chagas é um dos maiores problemas de saúde pública na América do Sul causando um elevado prejuízo à população. A despeito dos inúmeros esforços para o seu controle, a doença não tem cura e apresenta problemas científicos ainda não esclarecidos. Considerando-se que vários pesquisadores têm usado a radiação ionizante para modificar protozoários ou propriedades imunológicas de biomoléculas, neste trabalho foram estudados aspectos da resposta imunológica induzida em camundongos, resistentes e suscetíveis ao T. cruzi, utilizando formas irradiadas deste parasita. Doses baixas de radiação preservaram a capacidade reprodutiva e de invasão celular. Animais resistentes e suscetíveis, imunizados com os parasitas tratados por radiação, produziram anticorpos específicos. Após o desafio, os animais apresentaram baixa parasitemia, com exceção dos grupos imunizados com parasitas que receberam apenas altas doses de radiação. A seleção de formas tripomastigotas foi obtida irradiando-se os parasitas com baixas doses, o que promoveu aprimoramento da qualidade da resposta imune, a exemplo do que se observa quando da utilização de complemento. Estes dados evidenciam a importância da seleção das formas tripomastigotas para a imunização contra o T. cruzi e apontam a radiação ionizante como alternativa para este fim, uma vez que quando a seleção é feita utilizando-se complemento, depara-se com a dificuldade de sua remoção, colocando em risco o processo de imunização por introduzir substancias estranhas ao organismo. / Chagas\'s disease is one of the major public health problems in South America, promoting high prejudice to the local population. Despite the massive efforts to control it, this disease has no cure and presents puzzling unsolved questions. Considering that many researchers have used ionizing radiation to modify protozoans or biomolecules, we investigated the immunological response aspects of susceptible and resistant mice using irradiated parasites. Low radiation doses preserved the reproductive and invasive capacities of the parasite. Both susceptible and resistant animals, after immunization with irradiated parasites produced specific antibodies. After a challenge, the animals presented low parasitaemia, excepting those immunized with the antigen irradiated with higher doses. Using low radiation doses, we were able to selectively isolate trypomastigotes, leading to an improvement in the quality of the immune response, as previously reported when performing complement system assays. These data highlight the importance of selecting trypomastigote forms for immunization against T. cruz; and point towards ionizing radiation as an alternative to achieve this selection, since when this procedure is performed using complement, the subsequent steps are impaired by the difficulties to remove this component from the system.

O crescimento de investimentos em biotecnologia na agricultura tem sido um elemento primordial para a segurança alimentar global. Isso tem levado a uma reorganização da indústria mundial de sementes na busca por técnicas mais adequadas de cultivo, mecanização, uso de fertilizantes, defensivos agrícolas e a utilização de sementes. O Brasil por ser um dos ambientes mais sólidos nesse contexto tem contribuído com o crescimento e a diversificação da produção de sementes levando as lavouras brasileiras a atingirem um novo patamar de produtividade. Diferentes metodologias estatísticas têm sido utilizadas para analisar o comportamento da germinação de uma população de sementes. Entretanto, usar abordagens estatísticas que analisam os dados de germinação da melhor maneira possível permitirá uma maior confiabilidade dos resultados, bem como, ganho de informações pertinentes. Como em testes de germinação de sementes estuda-se o tempo até a ocorrência do evento, que envolve medidas repetidas feitas no mesmo lote e não se conhece o tempo exato da germinação da semente, propõe-se o uso de modelos de dose-resposta com censura intervalar, que permitem a interpretação biológica dos parâmetros usados para medir o processo germinativo e refletem o desenho experimental dos dados. Neste trabalho foram utilizadas duas metodologias estatísticas usuais na análise de dados de germinação de sementes e seus resultados foram comparados com os da abordagem de modelos dose-resposta com censura intervalar. Foram utilizados os modelos de dose-resposta Weibull 2 e log-logístico para explicar o processo germinativo de sementes de Brachiaria e Citrumelo Swingle respectivamente, com diferentes tempos de observação. Os experimentos foram realizados em delineamento inteiramente aleatorizado e os procedimentos dos testes de germinação de acordo com as Regras para Análise de Sementes (RAS). As conclusões obtidas a partir da análise dos dados por meio da metodologia proposta, em geral, divergiram das conclusões obtidas por abordagens tradicionais (modelos de regressão não-linear considerando a distribuição normal e índices de germinação com o uso da análise de variância) utilizadas para analisar dados de germinação. Os modelos de dose-resposta com censura intervalar apresentaram ajustes satisfatórios e sendo portanto uma análise mais adequada que as abordagens usuais. / The growth of investment in biotechnology in agriculture has been a vital element for global food security. This has led to a reorganization of the world seed industry in the seeking of the most appropriate techniques of cultivation, mechanization, use of fertilizers, pesticides and seeds. The Brazil for being one of the most solid in the world context of the seed industry has contributed to the growth and diversification of seed production, leading Brazilian crops to a new level of productivity. Different statistical methodologies have been used to analyze the germination behavior. However, using statistical approaches that analyze germination data as efficiently as possible will allow a greater reliability of the results, as well as relevant information gain. As in seed germination test studies the time until the occurrence of the event, involves repeated measurements on the same experimental unit and do not know the exact time of germination, we propose the use of dose-response models with interval censured that allow biological interpretation of parameters used to measure the germination process and reflect the experimental design of the germination data. In this work we used two methodologies of usual statistical analysis and their results were compared with those of the approach that uses the dose-response models with interval censured. We used the dose response models Weibull 2 and log-logistic to explain the process of seed germination of Brachiaria and Citrumelo Swingle with different observation times. The experiments were carried out in completely randomized design and testing procedures according to the rules for seed analysis (RAS). The conclusions obtained from the analysis of the data by the proposed methodology in general diverged from the conclusions obtained by traditional approaches (regression models nonlinear considering normal distribution and germination indices using analysis of variance) used to analyze data germination. The dose-response models with interval-censored showed satisfactory adjustments and therefore a more accurate analysis than the usual approaches.

O prognóstico de pacientes portadores de sarcomas de partes moles (SPM) avançados é reservado. Aumentar o benefício de quimioterapia é necessário, sendo, uma das estratégias, quimioterapia em dose densa, a qual demonstrou benefício em câncer de mama. Por outro lado, a busca de um marcador prognóstico é importante para uma melhor seleção de pacientes que se beneficiariam de protocolo de tratamento mais intensivo. A ezrina é uma proteína que liga o citoesqueleto celular a proteínas de membrana, está associada a invasão celular e metástase e sua hiperexpressão tem sido associada a um pior prognóstico em sarcomas de partes moles. O objetivo deste estudo foi avaliar o papel de quimioterapia com dose densa em pacientes portadores de SPM de alto grau, avançados. O desfecho primário foi taxa de resposta e os secundários foram sobrevida global (SG), sobrevida livre de progressão (SLP), perfil de toxicidade, qualidade de vida e controle de dor. Avaliou-se também a expressão de ezrina por imunohistoquímica como marcador de prognóstico, com o intuito de estratificação da população que poderia se beneficiar mais desta abordagem intensificada. Neste estudo de fase II prospectivo, vinte e um pacientes foram incluídos. A idade mediana foi 37 anos (23-60) e extremidades inferiores foram o sítio primário mais comum. Sarcoma sinovial, leiomiossarcoma e sarcoma sem outras especificações foram as histologias mais frequentes. O protocolo consistiu de seis ciclos seqüenciais de doxorubicina 30mg/m2 D1-3 e ifosfamida 2,5g/m2 D1-5 a cada 14 e 21 dias, respectivamente, seguidos por sete dias de suporte hematopoiético. As intensidades de dose medianas de doxorrubicina e ifosfamida foram, respectivamente, 42mg/m2/semana e 3,63g/m2/semana (93% e 87% do planejado, respectivamente) e 15 pacientes (71%) receberam todo o tratamento. Toxicidades graus 3 e 4 foram observadas em 19 pacientes e em 77/105 ciclos, neutropenia febril em 6 ciclos (5 pacientes) e reduções da fração de ejeção de ventrículo esquerdo de pelo menos 10% em três pacientes. Não houve toxicidade renal provavelmente pela adminsitração da ifosfamida em duas horas. A resposta foi avaliada pelos critérios de RECIST, com três respostas parciais, totalizando uma taxa de resposta de 14%. Seis respostas deveriam ser observadas para que o estudo completasse a inclusão de todos os pacientes programados. Como não se atingiu a taxa de resposta prevista, o protocolo foi fechado. Três mortes precoces foram observadas com suspeita de toxicidade. Após seguimento mediano de 11 meses, a SLP e a SG medianas foram 8,1 e 20,1 meses respectivamente. Pacientes com sarcoma sinovial e idade inferior a 45 anos apresentaram maior sobrevida na análise univariada. A expresão de ezrina foi positiva em 10 pacientes (47%) e houve tendência a uma correlação direta entre sua expressão e sobrevida mais longa (p=0,1191). Todos os pacientes com histologia sinovial foram positivos para ezrina (teste de Fischer, p= 0,0325). Este esquema de quimioterapia sequencial com dose densa de doxorubicina e ifosfamida foi tóxico, a taxa de resposta foi baixa em um grupo de pacientes com doença avançada e não pode ser empregado na prática clínica diária fora de protocolo de pesquisa / Advanced soft tissue sarcoma (STS) patients have a dismal prognosis. Efforts to increase benefit from chemotherapy are needed and dose-dense chemotherapy could be an option, since this approach has demonstrated survival benefit in breast cancer. On the other hand, the identification of a prognostic marker is essential to stratify which patients could benefit most from intensified strategies. Ezrin is a member of the ERM (ezrin, radixin, moesin) cytoeskeleton-associated protein family associated with invasion and metastasis, and has been pointed as important prognostic marker in sarcomas. The objective of this study was to explore the role of dose-dense doxorubicin- and ifosfamide-based chemotherapy in advanced high grade STS patients. Primary endpoint was response rate and secondary endpoints were overall survival (OS), progression free survival (PFS), toxicity profile, quality of life and pain control evaluation. Tumor ezrin immunoreactivity was an exploratory endpoint as a predictor of response to chemotherapy and as a prognostic factor in this population, trying to find which patients could benefit most from this intensified strategy. This prospective, single arm, phase II study included 21 advanced STS patients. Median age was 37 years (23-60y) and lower limbs were the most frequent primary site. Synovial, leiomyo and unclassified sarcoma were the most common histologies. Protocol consisted of 6 cycles of sequential dose-dense doxorubicin 30 mg/m2 D1-3 and ifosfamide 2.5 g/m2 D1-5 every 14 and 21 days, respectively, followed by seven days of hematopoietic support. The median doxorubicin and ifosfamide dose-intensities were, respectively, 42 mg/m2/week and 3.63 g/m2/week (93% and 87% of planned, respectively) and 15 patients (71%) received all cycles. Grade 3/4 toxicities occurred in 19 patients and 77/105 cycles, febrile neutropenia in 5 patients (six cycles) and three LVEF drops of at least 10%, one symptomatic. No renal toxicity was observed what could occurred due to the two-hour-schedule of ifosfamide. Responses were evaluated by RECIST criteria and three patients presented partial response (response rate of 14%). Six responses were necessary to the inclusion of the target population, however, this was not observed and the study was closed. Three deaths were probably related to toxicity. After a median follow-up was 11 months, PFS and OS were 8.1 months and 20.1 months, respectively. Patients with synovial sarcoma and those younger than 45y presented better survival at univariate analysis. Ezrin expression was positive in 10 patients (47%) and a trend was observed for a correlation between positive ezrin expression and longer survival (p= 0.1191). There was a statistically significant correlation between positive ezrin expression and synovial hystology (Fishers exact test, p= 0.0325). This sequential dosedense doxorubicin/ifosfamide-based chemotherapy protocol was toxic, response rate was low in advanced STS patients and can not be considered for routine practice outside clinical trials

A expansão das áreas cultivadas com cana-de-açúcar no Brasil é evidente, e para um eficaz manejo dessas extensas áreas, é necessário um bom planejamento das operações agrícolas. Nesse sentido, a aplicação de herbicidas pode ocorrer até no período seco do ano e em condições de solo bem distintas. Sabe-se que os diferentes ambientes nos quais os herbicidas são aplicados têm grande influencia em sua eficácia, e entender essas diferenças é importante para se obter o melhor rendimento e se fazer o correto uso dessa excelente ferramenta de manejo. Assim, este trabalho teve como objetivo avaliar o comportamento da formulação comercial contendo os herbicidas diurom + hexazinona + sulfometurom-metílico quando aplicado em solos com texturas contrastantes, além de avaliar a lixiviação desse herbicida e seus componentes e também como condições de restrição hídrica em presença de palha de cana-de-açúcar podem afetar o seu efeito residual e de seus componentes. Foram realizados ao todo 5 experimentos sendo 3 com o uso de curvas de dose-resposta, avaliando-se capim-colchão e corda-de-viola; um com lixiviação em tubos de PVC, avaliando-se corda-de-viola, e um com períodos de restrição hídrica, avaliando-se corda-de-viola e capim-colonião. Os resultados obtidos nos experimentos com as curvas de dose-resposta demonstraram a grande influencia da textura na eficácia do herbicida, com quantidades de argila menores que 30% sendo suficientes para gerar respostas consideravelmente distintas. O experimento com lixiviação demonstrou que as três moléculas do herbicida comportam-se diferentemente no solo, o que também é importante para o seu uso em diferentes ambientes. Os resultados obtidos no experimento com períodos de restrição hídrica em palha de cana-de-açúcar permitem concluir que apesar de as três moléculas comportarem-se diferentemente no ambiente, elas se complementam no controle de plantas daninhas mono e dicotiledôneas, e têm o efeito residual pouco influenciado por períodos de restrição hídrica. / The expansion of areas planted with sugar cane in Brazil is clearly noticeable and a good planning of agricultural operations is necessary for an effective management of these extensive areas. Accordingly, the application of herbicides may occur even in the dry season and in distinctive soil conditions. It is known that the different environments in which the herbicides are applied have great influence on their effectiveness and understanding these differences is an important factor in order to obtain best performance and make proper use of this excellent management tool. Thus, this work aimed to evaluate the behavior of the commercial formulation containing the herbicides diuron + hexazinone + sulfometurom-methyl when applied to soils with contrasting textures, and also to evaluate the leaching of this herbicide and its components as well as how conditions of water restriction with presence of sugarcane staw can affect your residual effect and its components. Five experiments were performed, three of those using dose-response curves, to evaluate crabgrass and morning glory; one with leaching using PVC tubes, evaluating morning glory and one with periods of water restriction, evaluating morning glory and guinea grass. Obtained results of the experiments with the dose-response curves demonstrated the great influence of the texture on the effectiveness of the herbicide, clay amounts of less than 30% being sufficient to generate considerably distinct responses. The experiment with leaching showed that the three molecules behave differently in the soil, which is also important to their use in different environments. Obtained results in the experiment with periods of water restriction on straw sugarcane allow the conclusion that although the three molecules behave differently in the environment, they complement each other in control of mono and dicotyledonous weeds, and they have little residual effect influenced by periods of water restriction.