Exchange protein directly activated by cyclic AMP (EPAC) activation reverses neutrophil dysfunction induced by β2-agonists, corticosteroids, and critical illness

Background Neutrophils play a role in the pathogenesis of asthma, chronic obstructive pulmonary disease, and pulmonary infection. Impaired neutrophil phagocytosis predicts hospital-acquired infection. Despite this, remarkably few neutrophil-specific treatments exist. 

Objectives We sought to identify novel pathways for the restoration of effective neutrophil phagocytosis and to activate such pathways effectively in neutrophils from patients with impaired neutrophil phagocytosis. 

Methods Blood neutrophils were isolated from healthy volunteers and patients with impaired neutrophil function. In healthy neutrophils phagocytic impairment was induced experimentally by using β₂-agonists. Inhibitors and activators of cyclic AMP (cAMP)-dependent pathways were used to assess the influence on neutrophil phagocytosis in vitro. 

Results β₂-Agonists and corticosteroids inhibited neutrophil phagocytosis. Impairment of neutrophil phagocytosis by β₂-agonists was associated with significantly reduced RhoA activity. Inhibition of protein kinase A (PKA) restored phagocytosis and RhoA activity, suggesting that cAMP signals through PKA to drive phagocytic impairment. However, cAMP can signal through effectors other than PKA, such as exchange protein directly activated by cyclic AMP (EPAC). An EPAC-activating analog of cAMP (8CPT-2Me-cAMP) reversed neutrophil dysfunction induced by β₂-agonists or corticosteroids but did not increase RhoA activity. 8CPT-2Me-cAMP reversed phagocytic impairment induced by Rho kinase inhibition but was ineffective in the presence of Rap-1 GTPase inhibitors. 8CPT-2Me-cAMP restored function to neutrophils from patients with known acquired impairment of neutrophil phagocytosis. 

Conclusions EPAC activation consistently reverses clinical and experimental impairment of neutrophil phagocytosis. EPAC signals through Rap-1 and bypasses RhoA. EPAC activation represents a novel potential means by which to reverse impaired neutrophil phagocytosis.

Effects of light pollution by car headlights on loggerhead hatchlings in Cape Verde

[EN] Many sea turtle nesting areas are experiencing a tremendous growth in tourism during the last decades that will likely continue in the near future. Many touristic activities involve light pollution by the increasing presence of vehicles close or even over the beaches. Vehicles can drive towards or along the beaches and even stay with the lights turned on illuminating during prolonged periods of time significant zones with sea turtle nesting activity. Thus, it is important to evaluate the impact of car light pollution on both nesting females and newborns in their search of the sea.

“It’s the People That Drive It”: Motivating and Supporting Collaborative Data Use Practices in Three Teacher Education Programs

Thesis (Ph.D.)--University of Washington, 2016-08

Searching for Masculine Capital: Experiences Leading to High Drive for Muscularity in Men

Studies on the drive for muscularity (DFM) have primarily been quantitative, focused on identifying correlates. Currently little is known about men’s experiences leading them to desire high levels of muscle and engage in behaviours to increase their masculine capital. Our purpose was to explore the stories of men with high DFM revealing the socio-cultural and personal factors leading to DFM and their search for masculine capital. In-depth life-history interviews and multiple in-the-field conversations were undertaken with twenty men (Mean age=28.45, SD=6.96, years) scoring ≥ 3 on the Drive for Muscularity Scale (Mean=4.30, SD=0.70). Men’s stories focused on a set of dysfunctional childhood and adolescent socio-cultural interactions, including forms of symbolic violence, between them and significant others. In these interactions men were exposed to dominant social narratives of masculinity, and through comparisons and reinforcement they identified discrepancies between themselves and these narratives. In late adolescence and early adulthood men came to believe that they lacked masculine capital. Men struggled to increase their masculine capital through engagement with other traditional masculine activities (e.g., sport) and driven by activating events, they compensated through DFM desires and behaviours. This study advances knowledge by revealing the socio-cultural and personal processes participants believed led to their high DFM. Findings disclose that men’s search for masculine capital may have led them to develop and maintain high levels of DFM.

Assessing variation in personal health service delivery and workplace smoking ban enforcement by local health departments

Thesis (Ph.D.)--University of Washington, 2016-08

The regulation of cancer cell invasive behaviour by Chloride Interacellular Channel 3 (CLIC3) and Transglutaminasae 2

A study into the role of secreted CLIC3 in tumour cell invasion. The initiation and progression of cancers is thought to be linked to their relationship with a population of activated fibroblasts, which are associated with tumours. I have used an organotypic approach, in which plugs of collagen I are preconditioned with fibroblastic cells, to characterise the mechanisms through which carcinoma-associated fibroblasts (CAFs) influence the invasive behaviour of tumour cells. I have found that immortalised cancer-associated fibroblasts (iCAFs) support increased invasiveness of cancer cells, and that this is associated with the ability of CAFs to increase the fibrillar collagen content of the extracellular matrix (ECM). To gain mechanistic insight into this phenomenon, an in-depth SILAC-based mass proteomic analysis was conducted, which allowed quantitative comparison of the proteomes of iCAFs and immortalised normal fibroblast (iNFs) controls. Chloride Intracellular Channel Protein 3 (CLIC3) was one of the most significantly upregulated components of the iCAF proteome. Knockdown of CLIC3 in iCAFs reduced the ability of these cells to remodel the ECM and to support tumour cell invasion through organotypic plugs. A series of experiments, including proteomic analysis of cell culture medium that had been preconditioned by iCAFs, indicated that CLIC3 itself was a component of the iCAF secretome that was responsible for the ability of iCAFs to drive tumour cell invasiveness. Moreover, addition of soluble recombinant CLIC3 (rCLIC3) was sufficient to drive the extension of invasive pseudopods in cancer cell lines, and to promote disruption of the basement membrane in a 3D in vitro model of the ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) transition. My investigation into the mechanism through which extracellular CLIC3 drives tumour cell invasiveness led me to focus on the relationship between CLIC3 and the ECM modifying enzyme, transglutaminase-2 (TG2). Through this, I have found that TG2 physically associates with CLIC3 and that TG2 is necessary for CLIC3 to drive tumour cell invasiveness. These data identifying CLIC3 as a key pro-invasive factor, which is secreted by CAFs, provides an unprecedented mechanism through which the stroma may drive cancer progression.

Factors that influence entrepreneurship activity by gender in the European Union

This research work aims to discuss the gender issue concerning entrepreneurship in European Union countries in a period of nine years, from 2007 to 2015, identifying the factors which drive individuals to be entrepreneurs. The study mainly concentrates on identifying and quantifying the personal, social, political and economic features which are motivating individuals, especially women, to be entrepreneurs, as well as the main difficulties they feel during the process of business creation. In order to explore the entrepreneurial activity across a set of developed countries the econometric methodology of panel data (in particular the fixed effects and random effects models) is applied to a data set of entrepreneurial statistical indicators calculated and made available by the Global Entrepreneurship Monitor. The results show that the knowledge of other start-up entrepreneurs, a desired career choice, the governmental support and the existence of public policies that promote entrepreneurship (specially within the framework of small and medium sized firms) and the transfer of R&D are factors influencing negatively on the rate of female entrepreneurship. None of the observed variables are barriers for male entrepreneurs. The perceived capabilities and opportunities, the entrepreneurial intention, the policies to lower taxes and bureaucracy and the social and cultural norms are identified drives for women for engaging in a process of running their own ventures. These findings offer a set of valid knowledge to understand which measures could be implemented or should be changed and improved at a political and managerial level for stimulating entrepreneurship, especially for women.

Factors that drive Female Entrepreneurship in Armenia

Entrepreneurs play a key role in any economy. Entrepreneurship includes creativity, innovation, risk taking, planning and management and is described as transferring ideas into action. Female entrepreneurship, in particular, is considered an important tool in enabling female empowerment and emancipation. In the light of recent world events, this has become a crucial area to study and understand, especially with respect to motivations, obstacles, constraints and consequences of female entrepreneurship. Having the previous framework in attention, this thesis focuses on female entrepreneurship in a developing country - Armenia – and proposes a conceptual framework of the phenomenon. A joint cooperation between the World Bank and the European Bank for Reconstruction and Development allowed to conduct an enterprise survey in the country and from that survey a microeconomic dataset was released and applied in this research study. A logistic regression econometric method is applied to the dataset to identify and measure the relationship between female entrepreneurship and several factors such as the location, size, legal status, market and obstacles faced by Armenian firms. The study concludes that women entrepreneurs in Armenia share many common features and obstacles with their male counterparts. Moreover, gender of the top managers, the firm’s location, size, main market type, the number of competitors and full-time employees, the adoption of new marketing methods, the access to land, the tax administration system and an inadequately educated workforce are found to be statistical significant factors in the explanation of female entrepreneurship in Armenia.

Effects of Drive Speed Modulation on Dynamics of Slender Rotating Structures

Slender rotating structures are used in many mechanical systems. These structures can suffer from undesired vibrations that can affect the components and safety of a system. Furthermore, since some these structures can operate in a harsh environment, installation and operation of sensors that are needed for closed-loop and collocated control schemes may not be feasible. Hence, the need for an open-loop non-collocated scheme for control of the dynamics of these structures. In this work, the effects of drive speed modulation on the dynamics of slender rotating structures are studied. Slender rotating structures are a type of mechanical rotating structures, whose length to diameter ratio is large. For these structures, the torsion mode natural frequencies can be low. In particular, for isotropic structures, the first few torsion mode frequencies can be of the same order as the first few bending mode frequencies. These situations can be conducive for energy transfer amongst bending and torsion modes. Scenarios with torsional vibrations experienced by rotating structures with continuous rotor-stator contact occur in many rotating mechanical systems. Drill strings used in the oil and gas industry are an example of rotating structures whose torsional vibrations can be deleterious to the components of the drilling system. As a novel approach to mitigate undesired vibrations, the effects of adding a sinusoidal excitation to the rotation speed of a drill string are studied. A portion of the drill string located within a borewell is considered and this rotating structure has been modeled as an extended Jeffcott rotor and a sinusoidal excitation has been added to the drive speed of the rotor. After constructing a three-degree-of-freedom model to capture lateral and torsional motions, the equations of motions are reduced to a single differential equation governing torsional vibrations during continuous stator contact. An approximate solution has been obtained by making use of the Method of Direct Partition of Motions with the governing torsional equation of motion. The results showed that for a rotor undergoing forward or backward whirling, the addition of sinusoidal excitation to the drive speed can cause an increase in the equivalent torsional stiffness, smooth the discontinuous friction force at contact, and reduce the regions of negative slope in the friction coefficient variation with respect to speed. Experiments with a scaled drill string apparatus have also been conducted and the experimental results show good agreement with the numerical results obtained from the developed models. These findings suggest that the extended Jeffcott rotordynamics model can be useful for studies of rotor dynamics in situations with continuous rotor-stator contact. Furthermore, the results obtained suggest that the drive speed modulation scheme can have value for attenuating drill-string vibrations.

Successional dynamics in inland dune shrub communities drive changes in functional diversity

Communities can be defined as assemblages of species coexisting under particular environments. The relationship between environment and species are regulated by both environmental requirements –which ultimately determine the species capacity to establish and survive in a particular environment– and the ecological interactions occurring during assembly processes –which also determine community composition by conditioning species coexistence. In this context, plant functional traits are attributes that represent ecological strategies and determine how plants respond to environmental factors and interact with other species. Therefore, the analysis of how traits vary through the dynamics of communities, such as along successions, can give insights about how environmental requirements and species interactions may determine the composition and functional structure of these communities. The xerophytic shrub communities inhabiting inland sand dunes in SW Portugal are characterized by successional processes that are mainly driven by local (edaphic gradients and human disturbance) and regional (climate) processes. Therefore, they constitute an appropriate system for studying species interactions and environmentcommunity co-variations based on functional terms. Using these communities as a model, we evaluate the hypothesis that successional community changes in species composition of xerophytic shrub communities can result in concurrent changes in functional diversity

Targeted gene modification in Fragaria vesca mediated by CRISPR/Cas9 system

Genome editing is becoming an important biotechnological tool for gene function analysis and crop improvement, being the CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR associated protein 9) system the most widely used. The natural CRISPR/Cas9 system has been reduced to two components: a single-guide RNA (sgRNA) for target recognition via RNA-DNA base pairing, which is commonly expressed using a promoter for small-RNAs (U6 promoter), and the Cas9 endonuclease for DNA cleavage (1). To validate the CRISPR/Cas9 system in strawberry plants, we designed two sgRNAs directed against the floral homeotic gene APETALA3 (sgRNA-AP3#1 and sgRNA-AP3#2). This gene was selected because ap3 mutations induce clear developmental phenotypes in which petals and stamens are missing or partially converted to sepals and carpels respectively (2). In this work, we used two different U6 promoters to drive the sgRNA-AP3s expression: AtU6-26 from Arabidopsis (4), and a U6 promoter from Fragaria vesca (FvU6) (this work). We also tested two different coding sequences of Cas9: a human- (hSpCas9) (3) and a plant-codon optimized (pSpCas9) (this work). Transient expression experiments using both CRISPR/Cas9 systems (AtU6-26:sgRNA-AP3#1_35S:hSpCas9_AtU6-26:sgRNA-AP3#2 and FvU6:sgRNA-AP3#1_35S:pSpCas9_FvU6:sgRNA-AP3#2) were performed infiltrating Agrobacterium tumefaciens into F. vesca fruits. PCR amplification and sequencing analyses across the target sites showed a deletion of 188-189 bp corresponding to the region comprised between the two cutting sites of Cas9, confirming that the CRISPR/Cas9 system is functional in F. vesca. Remarkably, the two systems showed different mutagenic efficiency that could be related to differences in expression of the U6 promoters as well as differences in the Cas9 transcripts stability and translation. Stable transformants for both F. vesca (2n) and Fragaria X anannassa (8n) are currently being established to test whether is possible to obtain heritable homozygous mutants derived from CRISPR/Cas9 strategies in strawberry. Thus, our work offers a promising tool for genome editing and gene functional analysis in strawberry. This tool might represent a more efficient alternative to the sometimes inefficient RNAi silencing methods commonly used in this species.

Export and import market-specific characteristics. How they drive the decision to trade and how much

Using a rich firm-level dataset on the Italian manufacturing industry, this paper provides a comprehensive analysis of the role that firms and market characteristics play in shaping firms’ trade activities. We enhance the previous analyses by considering firms’ engagement in international transactions, by focusing on either exports or imports. We show that the determinants of a firm’s export participation and value across countries also drive import behavior. Our research is consistent with the presence of country-specific sunk costs and with a qualitatively similar role of gravity forces and other country attributes on both sides of trading activities. Our evidence, however, militates in favor of a framework where variations in market characteristics have a larger impact on imports than exports.

Design Optimization of Modern Machine-drive Systems for Maximum Fault Tolerant and Optimal Operation

Modern electric machine drives, particularly three phase permanent magnet machine drive systems represent an indispensable part of high power density products. Such products include; hybrid electric vehicles, large propulsion systems, and automation products. Reliability and cost of these products are directly related to the reliability and cost of these systems. The compatibility of the electric machine and its drive system for optimal cost and operation has been a large challenge in industrial applications. The main objective of this dissertation is to find a design and control scheme for the best compromise between the reliability and optimality of the electric machine-drive system. The effort presented here is motivated by the need to find new techniques to connect the design and control of electric machines and drive systems. A highly accurate and computationally efficient modeling process was developed to monitor the magnetic, thermal, and electrical aspects of the electric machine in its operational environments. The modeling process was also utilized in the design process in form finite element based optimization process. It was also used in hardware in the loop finite element based optimization process. The modeling process was later employed in the design of a very accurate and highly efficient physics-based customized observers that are required for the fault diagnosis as well the sensorless rotor position estimation. Two test setups with different ratings and topologies were numerically and experimentally tested to verify the effectiveness of the proposed techniques. The modeling process was also employed in the real-time demagnetization control of the machine. Various real-time scenarios were successfully verified. It was shown that this process gives the potential to optimally redefine the assumptions in sizing the permanent magnets of the machine and DC bus voltage of the drive for the worst operating conditions. The mathematical development and stability criteria of the physics-based modeling of the machine, design optimization, and the physics-based fault diagnosis and the physics-based sensorless technique are described in detail. To investigate the performance of the developed design test-bed, software and hardware setups were constructed first. Several topologies of the permanent magnet machine were optimized inside the optimization test-bed. To investigate the performance of the developed sensorless control, a test-bed including a 0.25 (kW) surface mounted permanent magnet synchronous machine example was created. The verification of the proposed technique in a range from medium to very low speed, effectively show the intelligent design capability of the proposed system. Additionally, to investigate the performance of the developed fault diagnosis system, a test-bed including a 0.8 (kW) surface mounted permanent magnet synchronous machine example with trapezoidal back electromotive force was created. The results verify the use of the proposed technique under dynamic eccentricity, DC bus voltage variations, and harmonic loading condition make the system an ideal case for propulsion systems.

Autarcie radicale et autarcie dérivée chez Proclus : le problème de l’autarcie de l’âme par illumination

Louis-André Dorion, dans un article publié en 2006 « Platon, Proclus et l’autarcie du monde », affirme que le concept d’autarcie dérivée de Proclus est un non-sens. Nous montrerons dans cet article que, loin d’être aporétique, le concept d’autarcie dérivée est tout à fait cohérent avec le système philosophique de Proclus. Pour défendre notre thèse, nous nous intéresserons spécifiquement à l’autarcie de l’âme, à travers le phénomène d’illumination suivant le processus de purification de l’âme séparable chez Proclus, rendant compte du caractère autoconstitutif de l’âme. Ainsi, nous comprendrons en quoi l’ontologie proclusienne permet de saisir les hypostases (Un, intellect et âme), non pas dans un rapport de causalité unilatérale hiérarchique, mais plutôt selon une dialectique de procession centrifuge et de conversion centripète. Finalement, il nous sera possible d’exposer que l’idée de dérivation ne contredit pas l’essence de l’autarcie, mais en est plutôt sa propre condition de possibilité.

Autarcie radicale et autarcie dérivée chez Proclus : le problème de l’autarcie de l’âme par illumination

Louis-André Dorion, dans un article publié en 2006 « Platon, Proclus et l’autarcie du monde », affirme que le concept d’autarcie dérivée de Proclus est un non-sens. Nous montrerons dans cet article que, loin d’être aporétique, le concept d’autarcie dérivée est tout à fait cohérent avec le système philosophique de Proclus. Pour défendre notre thèse, nous nous intéresserons spécifiquement à l’autarcie de l’âme, à travers le phénomène d’illumination suivant le processus de purification de l’âme séparable chez Proclus, rendant compte du caractère autoconstitutif de l’âme. Ainsi, nous comprendrons en quoi l’ontologie proclusienne permet de saisir les hypostases (Un, intellect et âme), non pas dans un rapport de causalité unilatérale hiérarchique, mais plutôt selon une dialectique de procession centrifuge et de conversion centripète. Finalement, il nous sera possible d’exposer que l’idée de dérivation ne contredit pas l’essence de l’autarcie, mais en est plutôt sa propre condition de possibilité.