Quantifying aggressive behavior in zebrafish

Aggression is a complex behavior that influences social relationships and can be seen as adaptive or maladaptive depending on the context and intensity of expression. A model organism suitable for genetic dissection of the underlying neural mechanisms of aggressive behavior is still needed. Zebrafish has already proven to be a powerful vertebrate model organism for the study of normal and pathological brain function. Despite the fact that zebrafish is a gregarious species that forms shoals, when allowed to interact in pairs, both males and females express aggressive behavior and establish dominance hierarchies. Here, we describe two protocols that can be used to quantify aggressive behavior in zebrafish, using two different paradigms: (1) staged fights between real opponents and (2) mirror-elicited fights. We also discuss the methodology for the behavior analysis, the expected results for both paradigms, and the advantages and disadvantages of each paradigm in face of the specific goals of the study.

Microfluidics-based single-cell functional proteomics microchip for portraying protein signal transduction networks within the framework of physicochemical principles, with applications in fundamental and translational cancer research

Single-cell functional proteomics assays can connect genomic information to biological function through quantitative and multiplex protein measurements. Tools for single-cell proteomics have developed rapidly over the past 5 years and are providing unique opportunities. This thesis describes an emerging microfluidics-based toolkit for single cell functional proteomics, focusing on the development of the single cell barcode chips (SCBCs) with applications in fundamental and translational cancer research.

The microchip designed to simultaneously quantify a panel of secreted, cytoplasmic and membrane proteins from single cells will be discussed at the beginning, which is the prototype for subsequent proteomic microchips with more sophisticated design in preclinical cancer research or clinical applications. The SCBCs are a highly versatile and information rich tool for single-cell functional proteomics. They are based upon isolating individual cells, or defined number of cells, within microchambers, each of which is equipped with a large antibody microarray (the barcode), with between a few hundred to ten thousand microchambers included within a single microchip. Functional proteomics assays at single-cell resolution yield unique pieces of information that significantly shape the way of thinking on cancer research. An in-depth discussion about analysis and interpretation of the unique information such as functional protein fluctuations and protein-protein correlative interactions will follow.

The SCBC is a powerful tool to resolve the functional heterogeneity of cancer cells. It has the capacity to extract a comprehensive picture of the signal transduction network from single tumor cells and thus provides insight into the effect of targeted therapies on protein signaling networks. We will demonstrate this point through applying the SCBCs to investigate three isogenic cell lines of glioblastoma multiforme (GBM).

The cancer cell population is highly heterogeneous with high-amplitude fluctuation at the single cell level, which in turn grants the robustness of the entire population. The concept that a stable population existing in the presence of random fluctuations is reminiscent of many physical systems that are successfully understood using statistical physics. Thus, tools derived from that field can probably be applied to using fluctuations to determine the nature of signaling networks. In the second part of the thesis, we will focus on such a case to use thermodynamics-motivated principles to understand cancer cell hypoxia, where single cell proteomics assays coupled with a quantitative version of Le Chatelier's principle derived from statistical mechanics yield detailed and surprising predictions, which were found to be correct in both cell line and primary tumor model.

The third part of the thesis demonstrates the application of this technology in the preclinical cancer research to study the GBM cancer cell resistance to molecular targeted therapy. Physical approaches to anticipate therapy resistance and to identify effective therapy combinations will be discussed in detail. Our approach is based upon elucidating the signaling coordination within the phosphoprotein signaling pathways that are hyperactivated in human GBMs, and interrogating how that coordination responds to the perturbation of targeted inhibitor. Strongly coupled protein-protein interactions constitute most signaling cascades. A physical analogy of such a system is the strongly coupled atom-atom interactions in a crystal lattice. Similar to decomposing the atomic interactions into a series of independent normal vibrational modes, a simplified picture of signaling network coordination can also be achieved by diagonalizing protein-protein correlation or covariance matrices to decompose the pairwise correlative interactions into a set of distinct linear combinations of signaling proteins (i.e. independent signaling modes). By doing so, two independent signaling modes – one associated with mTOR signaling and a second associated with ERK/Src signaling have been resolved, which in turn allow us to anticipate resistance, and to design combination therapies that are effective, as well as identify those therapies and therapy combinations that will be ineffective. We validated our predictions in mouse tumor models and all predictions were borne out.

In the last part, some preliminary results about the clinical translation of single-cell proteomics chips will be presented. The successful demonstration of our work on human-derived xenografts provides the rationale to extend our current work into the clinic. It will enable us to interrogate GBM tumor samples in a way that could potentially yield a straightforward, rapid interpretation so that we can give therapeutic guidance to the attending physicians within a clinical relevant time scale. The technical challenges of the clinical translation will be presented and our solutions to address the challenges will be discussed as well. A clinical case study will then follow, where some preliminary data collected from a pediatric GBM patient bearing an EGFR amplified tumor will be presented to demonstrate the general protocol and the workflow of the proposed clinical studies.

A novel methodology for simulating contact-line behavior in capillary-driven flows

Despite the wide swath of applications where multiphase fluid contact lines exist, there is still no consensus on an accurate and general simulation methodology. Most prior numerical work has imposed one of the many dynamic contact-angle theories at solid walls. Such approaches are inherently limited by the theory accuracy. In fact, when inertial effects are important, the contact angle may be history dependent and, thus, any single mathematical function is inappropriate. Given these limitations, the present work has two primary goals: 1) create a numerical framework that allows the contact angle to evolve naturally with appropriate contact-line physics and 2) develop equations and numerical methods such that contact-line simulations may be performed on coarse computational meshes.

Fluid flows affected by contact lines are dominated by capillary stresses and require accurate curvature calculations. The level set method was chosen to track the fluid interfaces because it is easy to calculate interface curvature accurately. Unfortunately, the level set reinitialization suffers from an ill-posed mathematical problem at contact lines: a ``blind spot'' exists. Standard techniques to handle this deficiency are shown to introduce parasitic velocity currents that artificially deform freely floating (non-prescribed) contact angles. As an alternative, a new relaxation equation reinitialization is proposed to remove these spurious velocity currents and its concept is further explored with level-set extension velocities.

To capture contact-line physics, two classical boundary conditions, the Navier-slip velocity boundary condition and a fixed contact angle, are implemented in direct numerical simulations (DNS). DNS are found to converge only if the slip length is well resolved by the computational mesh. Unfortunately, since the slip length is often very small compared to fluid structures, these simulations are not computationally feasible for large systems. To address the second goal, a new methodology is proposed which relies on the volumetric-filtered Navier-Stokes equations. Two unclosed terms, an average curvature and a viscous shear VS, are proposed to represent the missing microscale physics on a coarse mesh.

All of these components are then combined into a single framework and tested for a water droplet impacting a partially-wetting substrate. Very good agreement is found for the evolution of the contact diameter in time between the experimental measurements and the numerical simulation. Such comparison would not be possible with prior methods, since the Reynolds number Re and capillary number Ca are large. Furthermore, the experimentally approximated slip length ratio is well outside of the range currently achievable by DNS. This framework is a promising first step towards simulating complex physics in capillary-dominated flows at a reasonable computational expense.

Metastability and dynamics of stem cells: from direct observations to inference at the single cell level

Organismal development, homeostasis, and pathology are rooted in inherently probabilistic events. From gene expression to cellular differentiation, rates and likelihoods shape the form and function of biology. Processes ranging from growth to cancer homeostasis to reprogramming of stem cells all require transitions between distinct phenotypic states, and these occur at defined rates. Therefore, measuring the fidelity and dynamics with which such transitions occur is central to understanding natural biological phenomena and is critical for therapeutic interventions.

While these processes may produce robust population-level behaviors, decisions are made by individual cells. In certain circumstances, these minuscule computing units effectively roll dice to determine their fate. And while the 'omics' era has provided vast amounts of data on what these populations are doing en masse, the behaviors of the underlying units of these processes get washed out in averages.

Therefore, in order to understand the behavior of a sample of cells, it is critical to reveal how its underlying components, or mixture of cells in distinct states, each contribute to the overall phenotype. As such, we must first define what states exist in the population, determine what controls the stability of these states, and measure in high dimensionality the dynamics with which these cells transition between states.

To address a specific example of this general problem, we investigate the heterogeneity and dynamics of mouse embryonic stem cells (mESCs). While a number of reports have identified particular genes in ES cells that switch between 'high' and 'low' metastable expression states in culture, it remains unclear how levels of many of these regulators combine to form states in transcriptional space. Using a method called single molecule mRNA fluorescent in situ hybridization (smFISH), we quantitatively measure and fit distributions of core pluripotency regulators in single cells, identifying a wide range of variabilities between genes, but each explained by a simple model of bursty transcription. From this data, we also observed that strongly bimodal genes appear to be co-expressed, effectively limiting the occupancy of transcriptional space to two primary states across genes studied here. However, these states also appear punctuated by the conditional expression of the most highly variable genes, potentially defining smaller substates of pluripotency.

Having defined the transcriptional states, we next asked what might control their stability or persistence. Surprisingly, we found that DNA methylation, a mark normally associated with irreversible developmental progression, was itself differentially regulated between these two primary states. Furthermore, both acute or chronic inhibition of DNA methyltransferase activity led to reduced heterogeneity among the population, suggesting that metastability can be modulated by this strong epigenetic mark.

Finally, because understanding the dynamics of state transitions is fundamental to a variety of biological problems, we sought to develop a high-throughput method for the identification of cellular trajectories without the need for cell-line engineering. We achieved this by combining cell-lineage information gathered from time-lapse microscopy with endpoint smFISH for measurements of final expression states. Applying a simple mathematical framework to these lineage-tree associated expression states enables the inference of dynamic transitions. We apply our novel approach in order to infer temporal sequences of events, quantitative switching rates, and network topology among a set of ESC states.

Taken together, we identify distinct expression states in ES cells, gain fundamental insight into how a strong epigenetic modifier enforces the stability of these states, and develop and apply a new method for the identification of cellular trajectories using scalable in situ readouts of cellular state.

Propagandistas, vadios e outros manipuladores do direito na obra Quando o Diabo Reza de Mário de Carvalho

A leitura do romance Quando o diabo reza de Mário de Carvalho propõe uma procura da ética, da moral e do direito, que proporcione reflexão sobre os valores que constroem a pessoa e a projetam na sociedade e no Estado. Procura ver-se o direito na literatura, demonstrando-se que este texto literário tem a capacidade de apelar ao reconhecimento do direito, por nos apresentar histórias de procedimentos ilegais, criminosos e de tentativas de manipulação da leis para fins diversos dos valores éticos e morais que visam tutelar. O texto desenvolve várias ações desviantes do direito, criando personagens marginais inconformadas, não só com a sua vida, mas com as leis que não lhes permitem atingir os seus objetivos. Analisa os seus comportamentos, as suas justificações, evidenciando as contradições entre nós e os outros. O trabalho explora as relações entre direito e literatura, reconhecendo a retórica e processos de interpretação comuns, e a forma como o texto questiona princípios, procedimentos e institutos jurídicos, contribuindo para a construção da memória e do mundo. Aborda dum ponto de vista geral os estudos desenvolvidos pelo chamado Law and Literature Movement (Capítulo I), identifica e analisa as personagens, interpretando o seu posicionamento com a ética, a moral e o direito (Capítulo II), e reconhece os institutos do nosso ordenamento jurídico presentes na narrativa, e o seu papel na ação do romance (Capítulo III).

O esverdeamento da economia e os tributos verdes : um duro caminho rumo à sustentabilidade da gestão dos resíduos sólidos urbanos (RSU)

Tese (doutorado)—Universidade de Brasília, Centro de Desenvolvimento Sustentável, 2013.

Credit risk & forward price models

Doutoramento em Gestão

Os efeitos das perceções de instrumentalização da avaliação de desempenho nas respostas face à organização

Doutoramento em Gestão

História de aprendizagem e seus contextos : uma análise comportamental das aquisições corporativas

Tese (doutorado)—Universidade de Brasília, Faculdade de Economia, Administração e Contabilidade, Programa de Pós-Graduação em Administração, 2016.

Patterns of sedentary behavior : insights from observational and experimental studies on body composition and energy expenditure

It is recognized that sedentary behavior (SB) has deleterious effects on numerous health outcomes and it appears that physiological mechanisms underlying these harms are distinct from the ones explaining moderate-to-vigorous physical activity (MVPA) benefits. Sedentary behavior represents a large portion of human’s life and is increasing with technological development. A new current of opinion supports the idea that the manner SB is accumulated plays an important role. This dissertation presents six research studies conducted under the scope of SB. In the methodological area, the first study highlighted the magnitude of potential errors in estimating SB and its patterns from common alternative methods (accelerometer and heart rate monitor) compared to ActivPAL. This study presented the accelerometer as a valid method at a group level. Two studies (2 and 5) were performed in older adults (the most sedentary group in the population) to test the associations for SB patterns with abdominal obesity using accelerometry. The findings showed positive graded associations for prolonged sedentary bouts with abdominal obesity and showed that those who interrupted SB more frequently were less likely to present abdominal obesity. Therefore, public health recommendations regarding breaking up SB more often are expected to be relevant. The associations between sedentary patterns and abdominal obesity were independent of MVPA in older adults. However, the low MVPA in this group makes it unclear whether this independent relationship still exists if highly active persons are analysed. Study 3 inovates by examining the association of SB with body fatness in highly trained athletes and found SB to predict total fat mass and trunk fat mass, independently of age and weekly training time. Study 4 also brings novelty to this research field by quantifying the metabolic and energetic cost of the transition from sitting to standing and then sitting back down (a break), informing about the modest energetic costs (0.32 kcal·min−1). Finally, from a successful multicomponent pilot intervention to reduce and break up SB (study 6), an important behavioral resistance to make more sit/stand transitions despite successfully reducing sitting time (~ 1.85 hours·day-1) was found, which may be relevant to inform future behavioral modification programs. The present work provides observational and experimental evidence on the relation for SB patterns with body composition outcomes and energy regulation that may be relevant for public health interventions.

Relatório de estágio de intervenção comunitária

O presente relatório reporta-se ao estágio de intervenção comunitária, decorrido entre 16/9/2013 e 31/01/2014, na população idosa da Freguesia de Tramaga, Concelho de Ponte de Sor, Distrito de Portalegre, parte integrante de uma das regiões mais envelhecidas do país: Alto Alentejo. Constitui-se como instrumento para avaliação da manifesta Unidade Curricular, título de Mestre em Enfermagem e validar competências para atribuição do título de Enfermeiro Especialista em Enfermagem Comunitária e Saúde Pública pela Ordem dos Enfermeiros. A Intervenção Comunitária sustentou-se com base na metodologia do planeamento em saúde, metodologia de referência no âmbito da aquisição de competências da especialidade em Enfermagem Comunitária e Saúde Pública (Regulamento n.º 128/2011), das quais se distinguem capacidades para trabalhar em parceria com as comunidades, reconhecer as suas necessidades, planear cuidados adequados aos seus problemas mais prementes, enfatizar práticas conducentes a comportamentos e estilos de vida saudáveis, adotando a promoção da saúde e o empowerment como pedra basilar de intervenção. As intervenções dirigiram-se para os problemas priorizados, designadamente: quedas, comorbilidades, depressão, obesidade, polifarmácia e vulnerabilidades como: Viver só, isolamento, iliteracia e condição de habitação. Os objetivos gerais subjacentes tinham como desígnio sensibilizar população idosa e entidades políticas/locais/concelhias, sobre os problemas de saúde que influenciam negativamente a qualidade de vida destes idosos; contribuir para a sua minimização e melhoria da qualidade de vida, através de abordagens educativas, no âmbito de comportamentos e estilos de vida saudáveis, perspetivando o seu empowerment. Na sinergia entre Mestrandas, comunidade e organizações políticas/locais/concelhias houve possibilidade de discutir e analisar soluções para alguns problemas identificados. Foram desenvolvidas atividades de educação em saúde: ‘atelier da saúde’; Fórum comunitário; Sessão ‘Segurança do idoso’ (em parceria estratégica de ampliação de eficácia com GNR e Teleassistência); todas tiveram o cunho interativo da comunidade. Foram aditadas estratégias de transmissão e disponibilização de informação no domínio de comportamentos e adoção de estilos de vida saudáveis, difundida por panfletos e meios de comunicação social local, configurando complementaridade com ações educativas.Supõe-se que as ações desenvolvidas alteraram comportamentos e contribuíram para a melhoria da qualidade de vida dos idosos, uma vez que estimulam a participação, interação, integração e perspetivam o empowerment

Railgun

Com a presente evolução das railguns na Marinha dos Estados Unidos da América e possível instalação em seus navios num futuro muito próximo, outras marinhas se seguirão. Creio que será do interesse da Marinha Portuguesa acompanhar esta evolução tecnológica, considerando as vantagens que advém da adoção deste tipo de armamento. Neste documento são abordados os princípios básicos subjacentes ao funcionamento da railgun, com principal foco nas questões eletrodinâmicas. Pretende-se adquirir familiaridade com este novo tipo de armamento através do estudo crítico dos seus princípios de funcionamento. O princípio básico de funcionamento de uma railgun, à primeira vista, parece bastante simples, à luz da aplicação imediata da expressão da força de Lorentz sobre um condutor percorrido por corrente elétrica. No entanto, tudo se torna mais complicado no caso de uma variação rápida dos parâmetros envolvidos (regime transitório), que exige uma análise mais aprofundada do comportamento da corrente, campos elétrico e magnético, e todos os materiais envolvidos neste sistema. Este trabalho envolveu ainda a construção de duas railguns, uma primeira de dimensões mais pequenas para ganhar familiaridade com o sistema, e uma última de dimensões de laboratório na qual foram feitos vários disparos para testar diferentes tipos de material e dimensões de projétil. Em suma, é demonstrado neste documento uma análise, no domínio do tempo, da distribuição espacial do campo eletromagnético, corrente elétrica e consequente fluxo de energia, complementados por uma parte experimental.

The Effect of Stress Management Program Using Cognitive Behavior Approach on Mental Health of the Mothers of the Children With Attention Deficit Hyperactivity Disorder

Background: Attention deficit hyperactivity disorder is one of the most common psychiatric disorders in children. Objectives: The study aimed to evaluate the effectiveness of stress management program using cognitive behavior approach on mental health of the mothers of the children with attention deficit hyperactivity disorder. Patients and Methods: In this interventional study, 90 mothers of the children with attention deficit hyperactivity disorder were randomly allocated into three intervention, placebo, and control groups. The general health questionnaire was used to measure mental health. Besides, stress was assessed through the depression-anxiety-stress scale. The two instruments were completed at baseline, immediately after, and one month after the intervention by the mothers. Afterwards, within group comparisons were made using one-sample repeated measurement ANOVA. One-way ANOVA was used for inter group comparisons. Mothers in the placebo group only participated in meetings to talk and express feelings without receiving any interventions. Results: At the baseline, no significant difference was found among the three groups regarding the means of stress, anxiety, depression, and mental health. However, a significant difference was observed in the mean score of stress immediately after the intervention (P = 0.033). The results also showed a significant difference among the three groups regarding the mean score of mental health (P < 0.001). One month after the intervention, the mean difference of mental health score remained significant only in the intervention group (P < 0.001). Conclusions: The study findings confirmed the effectiveness of stress management program utilizing cognitive behavior approach in mental health of the mothers of the children with attention deficit hyperactivity disorder.

Knowledge of HIV transmission risk behavior, and perceptions of risk among Haitian women living in Miami, Florida

This study was conducted to gain a more comprehensive understanding of HIV risk among Haitian women. The variables measured were: knowledge of HIV transmission, sexual risk behaviors, and perceptions of risk among Haitian women. The sociocultural aspect of the Haitian women's lives with regard to their risky behaviors was also examined. A total of 101 Haitian women (aged 25-53) who attended two comprehensive health clinics were interviewed. A combined questionnaire derived from both the ARM-Q and the RBA was used. In general, the women had good knowledge of the sexual transmission of HIV I AIDS and indicated that they were susceptible to HIV infection. However, knowledge and perceptions of risk were not translated into sexual risk-reduction behaviors with their partners. Multiplicity of partners and low incidence of condom use were the two major sexual risk factors isolated in this study. Results indicate Haitian women were more likely to use condoms if they possessed greater HIV knowledge and their sexual partners held more positive attitudes toward using condoms. Also, Haitian women may have failed to protect themselves because behavior changes could have involved threats to their social and economic survival, relationships and culturally sanctioned roles. This suggests the need to include male partners in HIV prevention interventions with Haitian women. Future research should focus on preventing high-risk behavior by improving knowledge, altering the male partners' attitudes toward condoms, and enhancing communication and negotiation skills. Nursing implications and recommendations for culturally sensitive and relevant AIDS prevention efforts are discussed.

An Examination of Psychological Meaningfulness, Safety, and Availability as the Underlying Mechanisms linking Job Features and Personal Characteristics to Work Engagement

The present study tested a nomological net of work engagement that was derived from its extant research. Two of the main work engagement models that have been presented and empirically tested in the literature, the JD-R model and Kahn’s model, were integrated to test the effects that job features and personal characteristics can have on work engagement through the psychological conditions of meaningfulness, safety, and availability. In this study, safety refers to psychological perceptions of safety and not workplace safety behaviors. The job features that were tested in this model included person-job fit, autonomy, co-worker relations, supervisor support, procedural justice, and interactional justice, while the personal characteristics consisted of self-consciousness, self-efficacy, extraversion, and neuroticism. Thirty-four hypotheses and a conceptual model were tested in order to establish the viability of this nomological net of work engagement in which it was expected that meaningfulness would mediate the relationships between job features and work engagement, safety would mediate the relationships that job features and personal characteristics have with work engagement, and availability (physical, emotional, and cognitive resources) would mediate the relationships that personal characteristics have with work engagement. Furthermore, analyses were run in order to determine the factor structure of work engagement, assess whether or not it exhibits differential validity from organizational commitment and job satisfaction, and confirm that it is positively related to the outcome variable of organizational citizenship behavior (OCB). The final sample consisted of 500 workers from an online labor market who responded to a questionnaire composed of measures of all constructs included in this study. Findings show that work engagement is best represented as a three-factor construct, composed of vigor, dedication and absorption. Furthermore, support was found for the distinction of work engagement from the related constructs of organizational commitment and job satisfaction. With regard to the proposed model, meaningfulness proved to be the strongest predictor of work engagement. Results show that it partially mediates the relationships that all job features have with work engagement. Safety proved to be a partial mediator of the relationships that autonomy, co-worker relations, supervisor support, procedural justice, interactional justice, and self-efficacy have with work engagement, and fully mediate the relationship between neuroticism and work engagement. Findings also show that availability partially mediates the positive relationships that extraversion and self-efficacy have with work engagement, and fully mediates the negative relationship that neuroticism has with work engagement. Finally, a positive relationship was found between work engagement and OCB. Research and organizational implications are discussed.