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Introducción. En la enseñanza suele utilizarse como herramienta la comparación de dos situaciones que comparten una estructura relacional similar. Este recurso se denomina razonamiento analógico y ha sido ampliamente estudiado, aunque los conocimientos al respecto no se encuentran sistematizados. Objetivo. Realizar una revisión de los artículos empíricos referidos a la enseñanza mediante analogías y, específicamente, indagar los resultados en cuanto a su efectividad. Metodología. Se utilizó el método PRISMA-NMA para realizar una revisión bibliográfica sistemática. Las bases de datos utilizadas fueron Dialnet, DOAJ, Pubmed y ScienceDirect, introduciendo los términos de búsqueda: razonamiento analógico + enseñanza y pensamiento analógico + enseñanza, en español e inglés. Luego de aplicar los criterios de inclusión/exclusión se obtuvieron 19 artículos que contenían un total de 24 elementos muestrales. Resultados. Se encontró que estos trabajos refieren con mayor frecuencia al nivel superior y a las ciencias exactas. Además, el principal uso dado a las analogías fue la enseñanza de un concepto novedoso. Principalmente las analogías utilizadas fueron provistas por las personas investigadoras y se expusieron de manera explícita los análogos y sus puntos de comparación. Discusión. La mayoría de los estudios obtuvieron resultados positivos, lo que indicaría la utilidad de este recurso para la enseñanza. Resulta llamativo que, al contrario de lo sugerido por especialistas, la proveniencia de los análogos y el grado de explicitación de la analogía parecen no afectar su efectividad. Se encuentran como áreas de vacancia otros usos menos estudiados de las analogías y su utilización en otros niveles educativos y áreas disciplinares.

Environmental changes and biodiversity loss have emphasized the need to understand how communities affect ecosystem functioning and services. In riparian ecosystems, integrative, generalizable, broad-scale models of ecosystem functioning are still required to fulfill this need. However, few studies have explored the links between functional traits, ecosystem functions, and the services of riparian vegetation. Here we adapt the response-effect trait framework to link drivers, traits, ecosystem functions, and services in riparian ecosystems and assess ecosystem functioning sensitivity to environmental changes. The response-effect trait framework distinguishes between traits related to responses to the environment (response traits) and effects on ecosystem functioning (effect traits). The framework predicts that if response and effect traits are tightly linked, shifts in environmental drivers may alter communities' traits and ecosystem functioning. We adapted the response-effect trait framework for riparian plant communities and used it to assess the overlap between response and effect traits. We tested for correlation among traits identified in the framework and for community functional responses to climatic, topographic, soil, and land cover factors using riparian plant communities along a Temperate-Mediterranean climate gradient in North Portugal. We found a high overlap between response and effect traits, with seven out of thirteen traits identified as both response and effect. Additionally, we found trait linkages in four groups of positively correlated community mean traits. Precipitation and aridity were the most predictive drivers of community functional structure, and life form and leaf area were the most responsive traits. Overall, our findings suggest riparian plant communities are likely to propagate the effects of environmental changes to ecosystem functioning and services, affecting several regulation ecosystem services. This work highlights the sensitivity of riparian ecosystems to environmental changes and how it can affect ecosystem services. Similar functional approaches can be useful for adaptive ecosystem management to sustain biodiversity and ecosystem services.

O presente trabalho procurou avaliar a influência de dois modelos de estresse e do tratamento neonatal com capsaicina sobre a inflamação pulmonar em ratos Wistar. Através do registro de vocalizações ultrassõnicas (VU\'s), constataram-se os níveis de ansiedade e de estresse. A resposta inflamatória foi induzida através de uma reação anafilática decorrente do desafio inalatório ao antígeno ovoalbumina, em ratos OVA-sensibilizados há 14 dias. Os modelos de estresse aplicados nos animais foram, respectivamente: 1 - Estresse por confrontação social de ratos invasores contra ratos residentes dominantes e 2 - Estresse em esquiva ativa composta por 13 sessões diárias de choques escapáveis, pareados ao som de uma campainha, seguidas de uma sessão de choques inescapáveis, pareados ao mesmo som. Através de testes de função respiratória, avaliaram-se os índices de elastância e resistência do sistema respiratório. Observou-se maior intensidade da resposta inflamatória pulmonar de elastância e resistência nos animais submetidos aos modelos de estresse, em relação aos animais do grupo controle não submetidos aos modelos de estresse descritos; constatou-se que os animais tratados neonatalmente com capsaicina e submetidos ao modelo de estresse apresentaram uma diminuição nos índices de elastância basal do sistema respiratório, em relação aos não estressados. Não foram observadas diferenças significativas nos percentuais de aumento do óxido nítrico encontrado no ar exarado dos animais não tratados com capsaicina. Tomados em conjunto, os presentes resultados sugerem: 1- uma correlação positiva entre os níveis de ansiedade/estresse e a resposta inflamatória pulmonar; 2- um efeito inibitório do estresse, em animais tratados com capsaicina sobre os índices de elastância do sistema respiratório. Estes dados foram interpretados levando-se em conta as relações bidirecionais existentes entre os sistemas nervoso e imunológico, bem como uma ação de neuropeptídeos liberados junto ao tecido peribrõnquico das vias aéreas. The present work was designed to study the influence of two stress models and the capsaicin neonatal treatment on lung inflammation in Wistar rats. Stress and anxiety leveis were measured through ultrasonic vocalizations. The inflammatory response was induced by anaphylatic reaction to an OVA-inhalatory challenge 14 days after a sensitization with OVA antigen. The stress models applied to the animais were respectively: 1 - Social confrontation of intruder rats vs dominant and resident rats; and 2 - 13 daily sessions of escapable shocks discriminated by a sound stimulus, and by one session of inescapable shocks discrimínated by the same stimulus unto a shuttle box chamber. The pulmonary indexes e.g. elastance and resistance of airway were increased in stressed rats. Moreover, stressed animais showed a larger elastance index of airways evaluated by respiratory function in mechanical ventilation when compared to non-stressed control groups. Capsaicin neonatally-treated animais that were submitted to stress exhibited decreased airways basal elastance indexes in relation to animais treated with control solution compared to non-stressed animais. Differences were not observed in the content of nitric oxide in the exhaled air of rats not treated with capsaicin. Taken together, these data suggest: 1 . The existence of a positive correlation between anxiety and stress leveis, and decrement of lung inflammatory response; 2. An inhibitory effect of capsaicin on airways elastance indexes. Data were discussed considering the bidirectional relationships between nervous and immune systems as well as the actions of neuropeptides, released in the proximity of the peribronchial airway tissues.

PISA (2018) demonstrated that 55% of Brazilian students have a low performance in Science, highlighting the need to introduce new methodologies in order to reframe the teaching-learning of the discipline. It was then proposed to build a booklet on Biological Pest Control by students, a theme that starts from the reality of the subjects. The objective was to build knowledge from textual and illustrative production on the topic, considering the development of scientific literacy indicators: writing in science, creating, articulating ideas and problematizing. Were carried out: bibliographic survey; socialization; textual and illustrative production - in pairs; correction; textual and illustrative production (second version); organization and assembly of the booklet for further reading to the Didactic Book. The students demonstrated appropriation of knowledge from the productions, corroborating that writing and illustrative production practices are favorable in the Science discipline, which is intensified by the relationship between scientific knowledge and everyday knowledge., O PISA (2018) demonstrou que 55% dos estudantes brasileiros estão com um baixo rendimento em Ciências, evidenciando a necessidade de introduzir novas metodologias com vistas em ressignificar o ensino-aprendizagem da disciplina. Foi, então, proposta a construção de um livreto sobre o Controle Biológico de pragas pelos estudantes, temática que parte da realidade dos sujeitos. O objetivo foi construir conhecimento a partir da produção textual e ilustrativa sobre o tema, considerando o desenvolvimento dos indicadores de Alfabetização Científica: escrever em ciências, criar, articular ideias e problematizar. Foram realizados: levantamento bibliográfico; socialização; produção textual e ilustrativa em dupla; correção; produção textual e ilustrativa (segunda versão); organização e montagem do livreto para leitura complementar ao Livro Didático. Os estudantes demonstraram apropriação do conhecimento a partir das produções, corroborando que práticas de escrita e produção ilustrativa são favoráveis na disciplina de Ciências, sendo isto intensificado pela relação do conhecimento científico com conhecimento cotidiano.

96 Background: Radium-223 represents an option for symptomatic metastatic castration-resistant prostate cancer (mCRPC) after demonstrated an overall survival improvement in the ALSYMPCA trial. Radium-223 real-life experiences are scarce, particularly after the results of the ERA-223 trial. To describe Radium-223 outcomes in real life practice, and specifically those related with bone health, sequence of treatment, and prognostic factors. Methods: Multicenter retrospective study of a cohort of 143 patients with mCRPC treated with Radium-223 in either the second line or third line of therapy or beyond in the context of routine clinical practice between 11 March 2013 and 26 July 2022, in seven Galician hospitals. Survival estimates were calculated by the Kaplan-Meier method, and groups were compared with the log-rank test. The Cox proportional hazards regression model was used to evaluate factors independently associated with OS. Between March 11, 2013, and July 26, 2022, 143 patients were enrolled from 5 Galician hospitals. With a median follow-up of 9.68 months. All patients signed an informed consent form agreeing to participate in this observational study, that was approved by the Galician Ethics Committee, registration code 2018/282. Results: Baseline patient and disease characteristics are summarized. Median follow-up was 9.68 months. Median overall survival (OS) was 12 months (95% CI 9.7 – 16.0). Best OS outcomes were achieved in second and third line, 18 months (95% CI 12.0 – 26.0) and 9.4 months (95% CI 8.0 – 12.0) months, respectively. Among those baseline clinic-analytical factors analyzed, only alkaline phosphatase level >354 UI/L was correlated with a worse OS (HR 2.56 (95% CI 1.39 – 4.72; p=0.003) and the number of cycles of Radium-223 received (HR 0.63 (95% CI 0.54 – 0.73; p

In Saccharomyces cerevisiae cAMP regulates different cellular processes through PKA. The specificity of the response of the cAMP-PKA pathway is highly regulated. Here we address the mechanism through which the cAMP-PKA pathway mediates its response to heat shock and thermal adaptation in yeast. PKA holoenzyme is composed of a regulatory subunit dimer (Bcy1) and two catalytic subunits (Tpk1, Tpk2, or Tpk3). PKA subunits are differentially expressed under certain growth conditions. Here we demonstrate the increased abundance and half-life of TPK1 mRNA and the assembly of this mRNA in cytoplasmic foci during heat shock at 37 °C. The resistance of the foci to cycloheximide-induced disassembly along with the polysome profiling analysis suggest that TPK1 mRNA is impaired for entry into translation. TPK1 expression was also evaluated during a recurrent heat shock and thermal adaptation. Tpk1 protein level is significantly increased during the recovery periods. The crosstalk of cAMP-PKA pathway and CWI signalling was also studied. Wsc3 sensor and some components of the CWI pathway are necessary for the TPK1 expression upon heat shock. The assembly in foci upon thermal stress depends on Wsc3. Tpk1 expression is lower in a wsc3∆ mutant than in WT strain during thermal adaptation and thus the PKA levels are also lower. An increase in Tpk1 abundance in the PKA holoenzyme in response to heat shock is presented, suggesting that a recurrent stress enhanced the fitness for the coming favourable conditions. Therefore, the regulation of TPK1 expression by thermal stress contributes to the specificity of cAMP-PKA signalling.

In Saccharomyces cerevisiae, cAMP regulates a number of different cellular processes, such as cell growth, metabolism, stress resistance and gene transcription. The intracellular target for this second messenger in yeast cells is the cAMP-dependent protein kinase (PKA). The way in which a broad specificity protein kinase mediates one right physiological response after cAMP increase indicates that specificity is highly regulated in the cAMP / PKA system. Here we address the mechanism through which cAMP-PKA signalling mediates its response to heat shock thermotolerance in Saccharomyces cerevisiae. Yeast PKA is a tetrameric holoenzyme composed of a regulatory (Bcy1) subunit dimer and two catalytic subunits (Tpk1, Tpk2 and Tpk3). PKA subunits are differentially expressed under certain stress conditions. In the present study we show that, although the mRNA levels of TPK1 are upregulated upon heat shock at 37°C, no change is detected in Tpk1 protein levels. The half-life of TPK1 mRNA increases and this mRNA condensates in cytoplasmic foci upon thermal stress. The resistance of TPK1 mRNA foci to cycloheximide-induced disassembly, together with the polysome profiling analysis suggest that TPK1 mRNA is impaired for entry into translation. TPK1 mRNA foci and TPK1 expression were also evaluated during thermotolerance. The crosstalk of cAMP-PKA pathway and cell wall integrity (CWI) signalling was also studied. Wsc3 sensor and other components of the CWI pathway are necessary for the upregulation of TPK1 mRNA upon heat shock conditions. The assembly in cytoplasmic foci upon thermal stress shows to be dependent of Wsc3. Finally, evidence of an increase in the abundance of Tpk1 in the PKA holoenzyme in response to heat shock is presented, suggesting that a recurrent stress enhanced the fitness for the coming favorable conditions The results indicate the existence of a mechanism that exclusively regulates Tpk1 subunit expression and therefore contributing to the specificity of cAMP-PKA.SUMMARY STATEMENTPKA subunits are differentially expressed under heat-shock conditions. The mRNA of the TPK1 subunit is upregulated upon heat-shock at 37°C and thermotolerance, the half-life increases upon heat-stress and also this transcript condensates in cytoplasmic foci upon thermal stress and thermotolerance. The resistance to cycloheximide treatment of TPK1 mRNA foci together with the analysis by polysome profiling suggest that TPK1 mRNA is impaired for entry into translation upon thermal stress. An increase in Tpk1 protein and PKA activity was detected after the heat stress treatments.Cell Wall Integrity pathway, through Wsc3 sensor, is involved in TPK1 expression. Heat-stress regulates TPK1 expression through this pathway from an intermediate step of the cascade and independently of the upstream elements of the CWI pathway activation. These results demonstrate a new crosstalk between the two signalling pathways.The increment in Tpk1-dependent PKA activity during cell adaptation to heat stress might contribute to the overall cellular fitness when more favorable environmental conditions are restored.The results indicate the existence of a mechanism that exclusively regulates TPK1 subunit expression and therefore contributing to the specificity of cAMP-PKA.

PISA (2018) demonstrated that 55% of Brazilian students have a low performance in Science, highlighting the need to introduce new methodologies in order to reframe the teaching-learning of the discipline. It was then proposed to build a booklet on Biological Pest Control by students, a theme that starts from the reality of the subjects. The objective was to build knowledge from textual and illustrative production on the topic, considering the development of scientific literacy indicators: writing in science, creating, articulating ideas and problematizing. Were carried out: bibliographic survey; socialization; textual and illustrative production - in pairs; correction; textual and illustrative production (second version); organization and assembly of the booklet for further reading to the Didactic Book. The students demonstrated appropriation of knowledge from the productions, corroborating that writing and illustrative production practices are favorable in the Science discipline, which is intensified by the relationship between scientific knowledge and everyday knowledge. O PISA (2018) demonstrou que 55% dos estudantes brasileiros estão com um baixo rendimento em Ciências, evidenciando a necessidade de introduzir novas metodologias com vistas em ressignificar o ensino-aprendizagem da disciplina. Foi, então, proposta a construção de um livreto sobre o Controle Biológico de pragas pelos estudantes, temática que parte da realidade dos sujeitos. O objetivo foi construir conhecimento a partir da produção textual e ilustrativa sobre o tema, considerando o desenvolvimento dos indicadores de Alfabetização Científica: escrever em ciências, criar, articular ideias e problematizar. Foram realizados: levantamento bibliográfico; socialização; produção textual e ilustrativa em dupla; correção; produção textual e ilustrativa (segunda versão); organização e montagem do livreto para leitura complementar ao Livro Didático. Os estudantes demonstraram apropriação do conhecimento a partir das produções, corroborando que práticas de escrita e produção ilustrativa são favoráveis na disciplina de Ciências, sendo isto intensificado pela relação do conhecimento científico com conhecimento cotidiano.

Living cells have developed a set of complex signaling responses, which allow them to withstand different environmental challenges. Signaling pathways enable the cell to monitor external and internal states and to articulate the appropriate physiological responses. Cellular signal transmission requires the dynamic formation of spatiotemporal controlled molecular interactions. One of the most important signaling circuits in Saccharomyces cerevisiae is the one controlled by cAMP-Protein Kinase A (PKA). In budding yeast, extracellular glucose and a plethora of signals related with growth and stress conditions regulate the intracellular cAMP levels that modulate PKA activity which in turn regulates a broad range of cellular processes. The cAMP-PKA signaling output requires a controlled specificity of the PKA responses. In this review we discuss the molecular mechanisms that are involved in the establishment of the specificity in the cAMP-PKA signaling pathway in S.cerevisiae. Fil: Portela, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Rossi, Silvia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Segundo a Organização Mundial de Saúde, 170 milhões de pessoas de pessoas no mundo possuem algum tipo de deficiência visual. Destas, cerca de 40 milhões são cegas, enquanto que algo em torno de 130 milhões de pessoas possui limitações severas de visão. Ainda segundo este órgão, estes números irão dobrar até 2020, em decorrência destes dados, é de extrema importância para a sociedade a inclusão do deficiente visual. No Brasil, desde a década de 90 têm sido propostas e implementadas políticas de inclusão das pessoas com alguma deficiência, as quais abrangem direitos trabalhistas, acesso amplo ao lazer e à cultura, aprendizagem, etc. Entretanto, tendo em vista os inúmeros problemas existentes nos setores voltados à educação, percebe-se que os professores em geral enfrentam inúmeras dificuldades e desafios no seu dia-a-dia e não contam com uma capacitação adequada para atender aos anseios da sociedade pela inclusão das pessoas com deficiência, principalmente as que possuem limitações na percepção visual. Diante disso, ressalta-se a importância de conhecer melhor as características e as dificuldades escolares enfrentadas pela pessoa com deficiência visual, bem como o que tem sido feito em benefício delas ao redor do mundo e no Brasil. Assim, o presente estudo tem como objetivo principal apresentar um panorama geral sobre a deficiência visual no Brasil, enfocando metodologias e tecnologias que favorecem a inclusão da pessoa com deficiência visual, particularmente nas instituições de ensino. Serão realizadas buscas “online” por artigos periódicos científicos bem como por livros e outros materiais, com vistas a compor uma revisão bibliográfica crítica, demonstrando a viabilidade das metodologias e materiais de ensino existentes e sua aplicabilidade para a inclusão das pessoas com deficiência. SCHOOL INCLUSION MINORED IN VISUAL IMPAIRMENT ABSTRACT According to the World Health Organization, 170 million people around the world have any kind of visual impairment, 40 million are blind and 130 million have severe sight limitations. It is estimated that such situation tends to get worse, mostly when they enter in the educational system. The present paper aims to present an overview about visual impairment in Brazil, minoring methodologies and technologies that benefit visual impaired subjects, mainly at educational institutions. It was decided to perform “online” searches for articles in scientific journals as well as books and other material as a methodology to accomplish a narrative bibliographic review. According to the results obtained, it was verified that since the nineties, outlines and inclusion policies for visual impaired people have been implemented in Brazil; they cover labor rights, access to leisure and culture, education, etc. However, it can be noticed that teachers, in general, ignore several difficulties and challenges those individuals face in their daily routine added to the fact they do not receive proper trainee to assist the needs for the inclusion of impaired people, chiefly the visual impaired ones. Conversely, several education enabling mechanisms related to visual impairment were observed, such as equipment and technology as well as data from visual impaired people and the viability of the methodologies and teaching materials created to benefit those individuals. Keywords: Visual Impairment. Inclusion. Teaching. Teaching Resources. Inclusive Technology. Teacher’s Development.

Low complexity regions are involved in the assembly and disassembly of P-bodies (PBs). Saccharomyces cerevisiae contains three genes encoding the protein kinase A (PKA) catalytic subunit: TPK1, TPK2 and TPK3. Tpk2 and Tpk3 isoforms localize to PBs upon glucose starvation showing different mechanisms and kinetics of accumulation. In contrast to the other two isoforms, Tpk2 harbors a glutamine-rich prion-like domain (PrLD) at the N-terminus. Here we show that the appearance of Tpk2 foci in response to glucose starvation, heat stress or stationary phase was dependent on its PrLD. Moreover, the PrLD of Tpk2 was necessary for efficient PB and stress granule aggregation during stress conditions and in quiescent cells. Deletion of PrLD does not affect the in vitro and in vivo kinase activity of Tpk2 or its interaction with the regulatory subunit Bcy1. We present evidence that the PrLD of Tpk2 serves as a scaffold domain for PB assembly in a manner that is independent of Pat1 phosphorylation by PKA. In addition, a mutant strain where Tpk2 lacks PrLD showed a decrease of turnover of mRNA during glucose starvation. This work therefore provides new insight into the mechanism of stress-induced cytoplasmic mRNP assembly, and the role of isoform specific domains in the regulation of PKA catalytic subunit specificity and dynamic localization to cytoplasmic RNPs granules.

In response to environmental changes cells rapidly rearrange their gene expression pattern in order to adapt to the new conditions. Chromatin remodeling is critical for this process playing a major role in the induction of genes involved in stress responses. We demonstrated previously that TPK1, encoding one of the catalytic subunits of PKA from Saccharomyces cerevisiae, is upregulated under heat shock. Herein, we investigate the chromatin remodeling of the TPK1, TPK2 and TPK3 promoters under heat stress. The TPK1 promoter is the only one that presents three positioned nucleosomes. Upon heat stress or osmostress these nucleosomes are evicted in clear correlation with promoter activation and upregulation of TPK1 mRNA levels. We find that remodelers SWI/SNF, RSC, INO80 and ISW1 participate in chromatin remodeling of the TPK1 promoter under thermal stress conditions. RSC and INO80 are necessary for nucleosomes positioning and contribute to repression of the TPK1 promoter under normal conditions while SWI/SNF participates in the eviction of nucleosomes after heat stress. SWI/SNF complex is recruited to the TPK1 promoter upon heat shock in a Msn2/4-dependent manner. Finally, both Tpk1 and Tpk2 catalytic subunits are recruited to the TPK1 promoter with opposite association patterns. Tpk1 catalytic activity is necessary for nucleosome rearrangement on the TPK1 promoter while Tpk2 and Tpk3 inhibit the promoter activity and maintain a repressive chromatin conformation. This work enlightens the mechanism of regulation of TPK1 expression during heat-stress, contributing to the knowledge of specificity in fine-tuning the cAMP-PKA signaling circuit.

Yeast cells can adapt their growth in response to the nutritional environment. Glucose is the favourite carbon source of Saccharomyces cerevisiae, which prefers a fermentative metabolism despite the presence of oxygen. When glucose is consumed, the cell switches to the aerobic metabolism of ethanol, during the so-called diauxic shift. The difference between fermentative and aerobic growth is in part mediated by a regulatory mechanism called glucose repression. During glucose derepression a profound gene transcriptional reprogramming occurs and genes involved in the utilization of alternative carbon sources are expressed. Protein kinase A (PKA) controls different physiological responses following the increment of cAMP as a consequence of a particular stimulus. cAMP-PKA is one of the major pathways involved in the transduction of glucose signalling. In this work the regulation of the promoters of the PKA subunits during respiratory and fermentative metabolism are studied. It is demonstrated that all these promoters are upregulated in the presence of glycerol as carbon source through the Snf1/Cat8 pathway. However, in the presence of glucose as carbon source, the regulation of each PKA promoter subunits is different and only TPK1 is repressed by the complex Hxk2/Mig1 in the presence of active Snf1. Copyright © 2017 John Wiley & Sons, Ltd.

A Síndrome da Alienação Parental é geralmente definida como um distúrbio ou uma desordem que surge principalmente no contexto de disputas judiciais pela guarda dos filhos, sendo sua primeira manifestação a campanha injustificada de desmoralização contra o pai ou a mãe. Embora exista vasta literatura nacional no campo das Ciências Jurí­dicas, em relação ao campo da psicologia, o mesmo não se repete. Mais especificamente em relação a uma análise do fenômeno com viés do behaviorismo radical, a literatura é até o momento inexistente. O objetivo do presente artigo é fazer uma revisão bibliográfica narrativa acerca da Síndrome da Alienação Parental, por meio de documentos de dois dos autores mais significativos sobre o tema, a saber, Darnall e Gardner, estabelecendo a análise conceitual dos seus argumentos por meio da proposta teórica e filosófica de interpretação do Behaviorismo Radical sob três principais temas: o mentalismo, as emoções e sensações e o comportamento operante. Como principal resultado obtivera-se que ambos os autores citados, dentro dos documentos analisados, dispõem de dados insuficientes para que uma análise funcional detalhada e sólida dos comportamentos relacionados à Síndrome da Alienação Parental seja realizada, embora existam informações relevantes acerca das grandes classes de mecanismos comportamentais atuantes na síndrome e em suas descrições. Conclui-se que a análise funcional, por meio de relatos de dinâmicas parentais em que a alienação parental ocorre, assim como um estudo pormenorizado dos comportamentos verbais da síndrome podem ser ferramentas bastante relevantes na compreensão do funcionamento mais detalhado da alienação parental.

Cada vez mais os Sistemas de Gestão da Segurança e Saúde no Trabalho, fazem parte integrante do sistema global de gestão das organizações, dando resposta a um mercado global cada vez mais exigente e competitivo. Consequentemente, os Sistemas de SST assumem um papel estratégico no desenvolvimento das políticas, na prevenção, na produtividade, na reputação e na competitividade das empresas. O objetivo do presente trabalho é assim, através de um estudo de caso, conceber e implementar um modelo de Sistema Gestão Integrado QAS numa empresa de construção civil. Pretende que a implementação efetiva deste projeto contribua para que o Sistema de Gestão Integrado responda ao novo referencial normativo ISO 45001:2018, e que seja passível de ser certificado por entidade certificadora externa, de forma integrada até final do primeiro semestre do próximo ano. O estudo em causa procurou: identificar e caracterizar todas as áreas do Sistema Integrado QAS, que carecem de melhorias, face aos requisitos do novo referencial; sugerir um conjunto de soluções e propor uma estratégia de implementação de modelo de Sistema de Gestão Integrado QAS. Numa perspetiva de futuro foram tidos em consideração todos os requisitos da nova norma ISO 45001, bem como as das normas ISO 9001:2015 e ISO 14001:2015, com a finalidade da certificação integrada do sistema de gestão. No início do presente trabalho é abordada a temática dos sistemas de gestão da segurança e saúde no trabalho, as normas existentes e o novo referencial ISO 45001 que substituirá as atuais normas OHSAS 18001:2007 e a NP 4397:2008, pela qual a empresa é certificada de forma integrada. Seguidamente é efetuado um diagnóstico do sistema atual da empresa objeto de estudo, identificadas as necessidades de elaboração e/ou adaptação da documentação e procedimentos. Na parte final do trabalho e com base em Auditoria diagnóstico ao sistema implementado, é definida uma estratégia e linhas orientadoras, propondo as ações necessárias para a implementação dos novos requisitos que dão cumprimento ao estabelecido na ISO 45001. Foram elaborados alguns dos documentos chave do Sistema e identificados todos os outros que devem ser elaborados e/ou adaptados, com vista à preparação da organização para a certificação externa sob a forma integrada dos seus três sistemas. Nowadays increasingly Occupational Safety and Health Management Systems are an integral part of the global management system of the organizations, replying to much more demanding and competitive global market. As a result, OSH Systems play a strategic role in policy development, prevention, productivity, reputation and competitiveness of the companies. The objective of the present work is, through a case study, design and implement a Quality, Environment and Safety (QES) Integrated Management System model in a construction company. It intends that the effective implementation of this project will contribute for the Integrated Management System answer to the new normative reference ISO 45001: 2018 and allows it to be certified by an external certifying entity, in an integrated way until the end of the first six months of 2020. The study case focus on identify and characterize all areas of the QAS Integrated System, which need improvement, in view of the requirements of the new framework; suggest a set of solutions and propose a strategy for implementing QES Integrated Management System model. Moring forward, all requirements of the new ISO 45001 standard as well as ISO 9001: 2015 and ISO 14001: 2015 have been taken into account for the purpose of an integrated management system certification.. At the beginning of this project, it is approached the theme of occupational safety and health management systems, the existing standards and the new ISO 45001 standard will replace the current OHSAS 18001: 2007 and NP 4397: 2008, by which the company is certified in an integrated format to prepare de migration. Then a diagnosis of the current system of the company under study is made, identifying the needs of elaboration and/ or adaptation of the documentation and procedures. At the end of this project and based on Gap Analysis to the implemented system, a strategy and guidelines are defined, proposing the necessary actions for the implementation of the new requirements that comply with the established in ISO 45001. Some of the key documents of the System and others that need to be designed and / or adapted to prepare the organization for external certification in the integrated form of its three systems, were elaborated.

Glycolysis represents the fundamental metabolic pathway for glucose catabolism across biology, and glycolytic enzymes are amongst the most abundant proteins in cells. Their expression at such levels provides a particular challenge. Here we demonstrate that the glycolytic mRNAs are localized to granules in yeast and human cells. Detailed live cell and smFISH studies in yeast show that the mRNAs are actively translated in granules, and this translation appears critical for the localization. Furthermore, this arrangement is likely to facilitate the higher level organisation and control of the glycolytic pathway. Indeed, the degree of fermentation required by cells seems intrinsically connected to the extent of mRNA localization to granules. On this basis, we have termed the granules, Core Fermentation (CoFe) granules. Therefore, glycolytic mRNA granules likely represent a conserved means to maintain high-level co-ordinated enzyme synthesis for a critical metabolic pathway.

Pizzinga et al. show that mRNAs encoding a range of translation factors are localized to granules that get transported into the yeast daughter cell using the She2p/She3p machinery. This likely supports an intensification of protein synthetic activity to facilitate apical polarized growth., mRNA localization serves key functions in localized protein production, making it critical that the translation machinery itself is present at these locations. Here we show that translation factor mRNAs are localized to distinct granules within yeast cells. In contrast to many messenger RNP granules, such as processing bodies and stress granules, which contain translationally repressed mRNAs, these granules harbor translated mRNAs under active growth conditions. The granules require Pab1p for their integrity and are inherited by developing daughter cells in a She2p/She3p-dependent manner. These results point to a model where roughly half the mRNA for certain translation factors is specifically directed in granules or translation factories toward the tip of the developing daughter cell, where protein synthesis is most heavily required, which has particular implications for filamentous forms of growth. Such a feedforward mechanism would ensure adequate provision of the translation machinery where it is to be needed most over the coming growth cycle.

The therapeutic range in advanced and castration resistant prostate cancer is widening. Therapies must offer real clinical efficacy, and they also should be acceptable and desirable for patients, specially in advanced disease. We analyze the value of quality of life analysis in patients with advanced prostate cancer.We performed a bibliographic review (Pubmed) with the various health related quality of life scales available and different clinical trials on advanced prostate cancer.There are numerous therapeutic options but, due to variations in study design, a different evaluation of adverse events and different therapeutic regimens, comparisons are difficult. A common method to interpret results is not available, so most of the times that interpretation is left to statistical significance, which is not always well correlated with clinical significance.To propose the most adequate treatment in patient`s interest, we need results focused on patients that combine not only quantity or overall survival but also quality of life. Parameters such as QALY should be included in clinical trials as evaluation objectives in order to favor decision taking.

Yeast cells can adapt their growth in response to the nutritional environment. Glucose is the favourite carbon source of Saccharomyces cerevisiae, which prefers a fermentative metabolism despite the presence of oxygen. When glucose is consumed, the cell switches to the aerobic metabolism of ethanol, during the so-called diauxic shift. The difference between fermentative and aerobic growth is in part mediated by a regulatory mechanism called glucose repression. During glucose derepression a profound gene transcriptional reprogramming occurs and genes involved in the utilization of alternative carbon sources are expressed. Protein kinase A (PKA) controls different physiological responses following the increment of cAMP as a consequence of a particular stimulus. cAMP-PKA is one of the major pathways involved in the transduction of glucose signalling. In this work the regulation of the promoters of the PKA subunits during respiratory and fermentative metabolism are studied. It is demonstrated that all these promoters are upregulated in the presence of glycerol as carbon source through the Snf1/Cat8 pathway. However, in the presence of glucose as carbon source, the regulation of each PKA promoter subunits is different and only TPK1 is repressed by the complex Hxk2/Mig1 in the presence of active Snf1. Copyright © 2017 John WileySons, Ltd.

The cAMP-dependent protein kinase (PKA) signaling is a broad pathway that plays important roles in the transduction of environmental signals triggering precise physiological responses. However, how PKA achieves the cAMP-signal transduction specificity is still in study. The regulation of expression of subunits of PKA should contribute to the signal specificity. Saccharomyces cerevisiae PKA holoenzyme contains two catalytic subunits encoded by TPK1, TPK2 and TPK3 genes, and two regulatory subunits encoded by BCY1 gene. We studied the activity of these gene promoters using a fluorescent reporter synthetic genetic array screen, with the goal of systematically identifying novel regulators of expression of PKA subunits. Gene ontology analysis of the identified modulators showed enrichment not only in the category of transcriptional regulators, but also in less expected categories such as lipid and phosphate metabolism. Inositol, choline and phosphate were identified as novel upstream signals that regulate transcription of PKA subunit genes. The results support the role of transcription regulation of PKA subunits in cAMP specificity signaling. Interestingly, known targets of PKA phosphorylation are associated with the identified pathways opening the possibility of a reciprocal regulation. PKA would be coordinating different metabolic pathways and these processes would in turn regulate expression of the kinase subunits. Fil: Pautasso, María Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Reca, Sol Rita. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Chatfield Reed, Kate. University of Calgary; Canadá Fil: Chua, Gordon. University of Calgary; Canadá Fil: Galello, Fiorella Ariadna. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Portela, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Zaremberg, Vanina. University of Calgary; Canadá Fil: Rossi, Silvia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Our aim in this work was to further characterize the complexity and specificity of the three different isoforms (Tpk1, Tpk2 and Tpk3) of the catalytic and regulatory (Bcy1) subunits of PKA in Saccharomyces cerevisiae. We thus analyzed the subcellular localization of the PKA subunits in living cells by using strains carrying GFP (green fluorescent protein) fused to each PKA subunit. During exponential growth on glucose, both Bcy1 and Tpk2 localized in the nucleus, whereas Tpk1 and Tpk3 showed a mixed pattern of nucleo-cytoplasmic localization. During exponential growth on glycerol and during stationary phase, the PKA subunits showed mostly cytoplasmic localization, with the peculiarity that Tpk2 and Tpk3 but not Bcy1, were found associated to P-bodies and EGP bodies. Tpk3 was accumulated into P-bodies during glucose deprivation and hyper osmotic stress. Deletion of Tpk3 altered the kinetics of P-body formation. Analysis of protein expression showed that the relative expression pattern of each Tpk changes from low levels under fermentative metabolism to higher levels during stationary phase. The increase in Tpk levels produced an imbalance with Bcy1 levels. Our data suggest that the signaling specificity through PKA in yeast could be mediated by a particular subcellular localization of each isoform of Tpk.

A importância dos registos de enfermagem é actualmente reconhecida e indispensável para assegurar a continuidade dos cuidados, realçando a sua função de comunicação dos aspectos resultantes deste processo de cuidados. Sendo o ser humano o alvo da profissão de enfermagem, é imperativo construir uma linguagem em que o elemento humano esteja bem visível. Mas será que esta linguagem tradutora de sensibilidade e que identifica a pessoa como única está presente nos registos realizados? Esta questão surgiu como força impulsionadora para a realização deste estudo de carácter exploratório e descritivo, com uma abordagem qualitativa e teve como principal objectivo conhecer a expressão do agir pensado, da dimensão humana do cuidar, nos registos de evolução de enfermagem. Procedeu-se à análise das notas de evolução de dois clientes de cada um dos 18 serviços, num período de 24 horas. O tratamento dos dados foi realizado segundo a análise de conteúdo de Bardin. Deste estudo concluímos que, os enfermeiros reconhecem como grande finalidade dos registos a garantia da continuidade dos cuidados, realçando os aspectos técnicos e relacionados com as intervenções interdependentes. Os aspectos emocionais e os relativos à família não têm uma expressão significativa. As categorias, embora com menor número de unidades de registo, relacionadas com a atenção centrada na pessoa que está envolvida no processo de cuidar, a referência à sua autonomia e capacidade de decisão, faz sobressair na escrita a sua singularidade e a presença do enfermeiro nesta interacção de cuidar e ser cuidado.

Gene expression regulation by intracellular stimulus-activated protein kinases is essential for cell adaptation to environmental changes. There are three PKA catalytic subunits in Saccharomyces cerevisiae: Tpk1, Tpk2, and Tpk3 and one regulatory subunit: Bcy1. Previously, it has been demonstrated that Tpk1 and Tpk2 are associated with coding regions and promoters of target genes in a carbon source and oxidative stress dependent manner. Here we studied five genes, ALD6, SED1, HSP42, RPS29B, and RPL1B whose expression is regulated by saline stress. We found that PKA catalytic and regulatory subunits are associated with both coding regions and promoters of the analyzed genes in a stress dependent manner. Tpk1 and Tpk2 recruitment was completely abolished in catalytic inactive mutants. BCY1 deletion changed the binding kinetic to chromatin of each Tpk isoform and this strain displayed a deregulated gene expression in response to osmotic stress. In addition, yeast mutants with high PKA activity exhibit sustained association to target genes of chromatin-remodeling complexes such as Snf2-catalytic subunit of the SWI/SNF complex and Arp8-component of INO80 complex, leading to upregulation of gene expression during osmotic stress. Tpk1 accumulation in the nucleus was stimulated upon osmotic stress, while the nuclear localization of Tpk2 and Bcy1 showed no change. We found that each PKA subunit is transported into the nucleus by a different β-karyopherin pathway. Moreover, β-karyopherin mutant strains abolished the chromatin association of Tpk1 or Tpk2, suggesting that nuclear localization of PKA catalytic subunits is required for its association to target genes and properly gene expression. Fil: Baccarini, Leticia Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Martínez Montañés, Fernando. University of Fribourg; Suiza Fil: Rossi, Silvia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Proft, Markus. Instituto de Biomedicina; España. Consejo Superior de Investigaciones Científicas; España Fil: Portela, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Dez amostras de sêmen de dois reprodutores caprinos, da raça Parda-alpina, colhidas em vagina artificial, foram submetidas a quatro tratamentos para avaliação da eficiência do etilenoglicol e do glicerol, associados ou não ao EDTA, na criopreservação da célula espermática caprina. O diluente usado era à base de Tris-gema de ovo contendo 7% de glicerol (glicerol E glicerol+EDTA) ou 7% de etilenoglicol (etilenoglicol e etilenoglicol + EDTA), sendo que nos grupos glicerol+EDTA e etilenoglicol+EDTA foi associado ao diluente 0,1% de EDTA. As amostras foram mantidas por 60 minutos em geladeira a 40C, onde então era efetuada a congelação em nitrogênio (-1960C). A descongelação foi realizada em banho-maria a 370C por 30segundos. As médias obtidas para motilidade (%) a descongelação, para os grupos glicerol, glicerol+EDTA, etilenoglicol, etilenoglicol+EDTA, foram, respectivamente, 51%; 61%; 10% e 12%. Os grupos que utilizaram o glicerol como crioprotetor obtiveram melhores taxas de motilidade pós-descongelação, principalmente quando foi associado ao diluidor o EDTA (grupo glicerol+EDTA). Porém, esse resultado foi comprometido pelos maiores índices de alterações patológicas nos grupos que continham o glicerol. PALAVRAS-CHAVE: Caprinos, criopreservação, sêmen, glicerol, etilenoglicol, EDTA

Pyk1 (pyruvate kinase 1) from Saccharomyces cerevisiae was characterized as a substrate for PKA (protein kinase A) from bovine heart and yeast. By designing Pyk1 synthetic peptides containing potential PKA sequence targets (Ser22, Thr94 and Thr478) we determined that the peptide S22 was a substrate for PKA in vitro, with a Ksp* (specificity constant) 10-fold and 3-fold higher than Kemptide for bovine heart and yeast PKA respectively. In vitro phosphorylation of the Pyk1 S22A mutant protein was decreased by as much as 90% when compared with wild-type Pyk1 and the Pyk1 T94A mutant. The Ksp* values for Pyk1 and Pyk1 T94A were the same, indicating that both proteins are phosphorylated at the same site by PKA. Two-dimensional PAGE of Pyk1 and Pyk1 S22A indicates that in vivo the S22A mutation prevented the formation of one of the Pyk1 isoforms. We conclude that in yeast the major PKA phosphorylation site of Pyk1 is Ser22. Phosphorylation of Ser22 leads to a Pyk1 enzyme that is more active in the absence of FBP (fructose 1,6-bisphosphate). The specificity of yeast and mammalian PKA towards the S22 peptide and towards whole Pyk1 protein was measured and compared. The Ksp* for the S22 peptide is higher than that for Pyk1, indicating that the peptide modelled on Pyk1 is a much better substrate than Pyk1, regardless of which tissue was used as the source of PKA. However, the Km of Pyk1 protein is lower than that of the better substrate, the S22 peptide, indicating that ground-state substrate binding is not the major determinant of substrate specificity for PKA.

Protein kinase A (PKA) activity was measured in situ in permeabilised Saccharomyces cerevisiae cells in the absence and the presence of cAMP. Four strains genetically predicted to have differential PKA-dependent phenotypes were used: a wild-type strain and a strain containing a bcy1-14 mutation (with almost constitutively active PKA), and the same strains with overexpression of the wild-type or mutant BCY1 gene, respectively. Cells were grown on galactose or glucose. The measured phenotypic characteristics were: trehalose and glycogen levels and the activity of a reporter gene under control of the NTH1 promoter. The 'endogenous' PKA activity (measured in situ in the absence of cAMP) showed the best correlation with the PKA-dependent phenotypes determined in vivo. We propose that this parameter offers a good estimate for the degree of activation of PKA in vivo.

The presence and characteristics of androgen receptors (AR) have been des­cribed by our group in one human melanoma cell line. We now investigated their presence in two other human melanoma cell lines, IIB-MEL-LES and IIB-MEL-IAN, as well as in biopsies of human metastatic melanoma. Scatchard analysis revealed a single binding component for both cell lines, the apparent dissociation constant obtained being 15 nM, with a binding capacity of 280 fmol/mg total cell protein for IIB-MEL-LES cells and of 14 nM with a binding capacity of 206 fmol/mg total cell protein for IIB-MEL-IAN. When specifi­ci­ty was assessed, as seen before, not only androgen and anti­an­dro­gen, but also non-andro­ge­nic com­pounds were able to compete for [3H]-R1881 binding. When immu­nocyto­che­mis­try of IIB-MEL-LES and IIB-MEL-IAN cells was performed for AR, both cell lines were deeply stained in the nu­cleus whe­reas no staining was found for oestrogen and pro­ges­te­rone receptors. Every specimen of mela­no­ma metas­ta­ses tested for the presence of AR was deeply stained, and the intensity of the stai­ning was high in the majority. Several hormones and antihormones were tested for their ability to affect cell proli­fe­ration. On both cell lines, Testosterone, Dihydrotesterone, Oestradiol and Pro­ges­terone significantly stimulated cell proliferation, with reversion by Hydroxy-Flutamide, Casodex or Tamo­xi­fen. Fil: Morvillo, V.. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Luthy, Isabel Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Bravo, A. I.. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos ; Argentina Fil: Capurro, M. I.. Instituto Luis Federico Leloir; Fil: Portela, P.. Instituto Luis Federico Leloir; Fil: Calandra, Ricardo Saul. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina Fil: Mordoh, Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina

Active treatment in localized prostate cancer, in its various types, is assumed as a valid alternative. The effect of the possible overtreatment has raised that options such as active surveillance are offered as an alternative to active treatments, without evidence about its validity in many points. The objective of this study is to analyze the current controversies to define candidates to this alternative, follow up criteria, impact on quality of life and evidence bases to do it.We perform an analysis updating the Medline search with the terms localized prostate cancer and active surveillance, analyzing the articles and their evidence, as well as guidelines recommendations.Selection criteria for candidates to active surveillance are heterogeneous, without evidence of uniformity. Likewise, follow up and its criteria or progression are not well defined. The impact on progression, or delay in decision-making, have not been analyzed and we lack of studies of highest evidence including comparative studies for cancer specific or global survival results.Although AS seems to be a reasonable alternative in many patients with localized prostate cancer, we still need to define many features of inclusion and decision-making. Comparative studies are needed to better define selection and validity of active surveillance.

The influence of stress on total leukocyte count from bronchoalveolar lavage (BAL) was investigated in rats sensitized and challenged with ovalbumin (OVA). The animals were injected intraperitoneally with a suspension of OVA plus aluminum hydroxide in 0.9% NaCl (Day 0) and boosted at Day 7 with an identical OVA solution, administered subcutaneously. From the first to the 13th day after sensitization, rats were placed individually in a shuttle box where they received 50 escapable footshocks per day, always preceded by a sound signal (S); the responses that occurred during both S and shocks canceled the stressful stimulation. On Day 14, animals were submitted to a single session of 50 inescapable footshocks, preceded by the same S; immediately after, the animals were submitted to a 1% OVA-inhalation challenge. Results showed high levels of stress in the shocked animals as detected through both ultrasonic vocalizations (UVs) and social interaction test in an open field. Total leukocyte count in BAL from stressed animals (24 h post-OVA challenge) revealed a significant increase in the number of inflammatory cells in comparison to that measured in sensitized, nonstressed challenged rats. These data demonstrate that stress plays a relevant and important role on total bronchoalveolar cell count in OVA-sensitized rats.

The Lewis(x) (Le(x)) trisaccharide (CD15) linked to proteins and glycolipids is highly expressed on the surface of normal human polymorphonuclear neutrophils (PMN) and several human neoplasias, such as breast and gastrointestinal carcinomas and chronic myeloid leukemias. FC-2.15 is an IgM murine mAb that specifically recognizes Le(x) and has been previously shown to mediate the in vitro lysis of Le(x)(+) cells by human complement. In a phase I clinical trial of FC-2.15, a temporary neutropenia was the main toxicity, and antitumor responses were observed. In order to characterize FC-2.15 further and determine the physiological relevance of Le(x) binding, the reactivity of FC-2.15 on PMN was investigated under several conditions. Flow cytometry revealed a strong reactivity of FC-2.15 with almost 100% of PMN, and Scatchard analysis demonstrated an affinity constant of 5.14 x 10(9) M(-1) and 1.11 x 10(6) antigen sites/cell. In vitro, the binding of Le(x) epitopes by FC-2.15 induced PMN homotypic aggregation, only 28.4 +/- 4.1% remaining as single cells. When PMN and the Le(x)(+) MCF-7 breast cancer cells were co-incubated, FC-2.15 induced heterotypic aggregation. In 51Cr-release assays employing human complement, FC-2.15 lysed 93.4 +/- 7.9% of PMN and 87.8 +/- 10.7% of MCF-7 cells. However, when the effect of FC-2.15 was tested in ex vivo circulating blood, no lytic activity against PMN was detected, whereas MCF-7 cells were still lysed. Blood smears demonstrated that FC-2.15 induced PMN agglutination and heterotypic aggregates when MCF-7 cells were present. A pretreatment of PMN with colchicine impaired PMN agglutination both in vitro (single PMN = 81.15 +/- 4.35%) and in ex vivo circulating blood. In the latter condition, FC-2.15-lytic activity was restored, suggesting that PMN homotypic aggregation by FC-2.15, but not lysis, is dependent on microtubule integrity and that PMN agglutination hinders their lysis. Moreover, when 51Cr-release assays were performed following agglutination, FC-2.15 cytotoxicity was restricted to isolated PMN. It is suggested that crosslinking of Le(x) epitopes by FC-2.15 induces PMN to form homotypic aggregates. It is suggested that the neutropenia observed in FC-2.15-treated patients would be due to PMN agglutination and margination, rather than lysis. In addition, FC-2.15 appears to be able to lyse Le(x)(+) tumor cells in circulation.