CalDAG-GEFI is at the nexus of calcium-dependent platelet activation.

The importance of the second messengers calcium (Ca(2+)) and diacylglycerol (DAG) in platelet signal transduction was established more than 30 years ago. Whereas protein kinase C (PKC) family members were discovered as the targets of DAG, little is known about the molecular identity of the main Ca(2+) sensor(s). We here identify Ca(2+) and DAG-regulated guanine nucleotide exchange factor I (CalDAG-GEFI) as a critical molecule in Ca(2+)-dependent platelet activation. CalDAG-GEFI, through activation of the small GTPase Rap1, directly triggers integrin activation and extracellular signal-regulated kinase-dependent thromboxane A(2) (TxA(2)) release. CalDAG-GEFI-dependent TxA(2) generation provides crucial feedback for PKC activation and granule release, particularly at threshold agonist concentrations. PKC/P2Y12 signaling in turn mediates a second wave of Rap1 activation, necessary for sustained platelet activation and thrombus stabilization. Our results lead to a revised model for platelet activation that establishes one molecule, CalDAG-GEFI, at the nexus of Ca(2+)-induced integrin activation, TxA(2) generation, and granule release. The preferential activation of CalDAG-GEFI over PKC downstream of phospholipase C activation, and the different kinetics of CalDAG-GEFI- and PKC/P2Y12-mediated Rap1 activation demonstrate an unexpected complexity to the platelet activation process, and they challenge the current model that DAG/PKC-dependent signaling events are crucial for the initiation of platelet adhesion.

A (des)necessidade do conceito de capital social no direito societário brasileiro: uma análise à luz do direito norte-americano e europeu

O trabalho analisa o tema do capital social no direito societário brasileiro. Seu objetivo é demonstrar, do ponto de vista jurídico, os malefícios e benefícios que o instituto promove. Apesar de ser tido como um conceito clássico e essencial para as sociedades com limitação de responsabilidade no Brasil, esse instituto vem sendo cada vez mais criticado no sentido de que não desempenha suas funções clássicas (organização, produção, e proteção de credores) de maneira efetiva nos dias atuais. Nesse contexto, direito societário moderno vem passando por uma evolução no sentido de questionar a efetividade de seus institutos. A análise aqui proposta do capital social segue esse raciocínio. Para auxiliar na interpretação do instituto no Brasil, serão utilizadas serão estudadas as lições e legislações dos ordenamentos europeu e norte-americano, onde o tema já foi amplamente debatido. O tratamento dado pelo Revised Model Business Corporation Act, legislação modelo norte americana, e da Segunda Diretiva do Capital da União Europeia aos instituto serão comparados com o tratamento da Lei das S.A. para o capital social. Por fim, são identificadas algumas particularidades do instituto do capital social em relação aos ordenamentos estrangeiros, que demonstram que uma eventual supressão do conceito de capital social no Brasil possuiria características próprias que não estão presentes na Europa e nos Estados Unidos. Nesse contexto, serão identificados os custos legislativos que uma eventual mudança do regime de capital social teria no sistema legislativo brasileiro.

La debida imparcialidad de la Administración Tributaria

Este trabajo de investigación, tiene por objeto el análisis de la posición de la Administración Tributaria en el marco conceptual de la relación jurídico-tributaria, y de las implicaciones que de cara al principio de imparcialidad, pueden tener los diversos posicionamientos conceptuales que de ésta se hagan, en procura de alcanzar una sana circunscripción de la naturaleza y el enfoque que se puede esperar y exigir de una Administración Tributaria moderna. Para lograrlo, se analizan los orígenes y alcances de la relación jurídico-tributaria, así como las múltiples y hasta divergentes acepciones doctrinales acerca de los actores directamente involucrados por el ordenamiento en tal relación, con especial atención a las divergentes acepciones del concepto de sujeto activo, y de la identificación de la Administración Tributaria en ese rol. Se aborda el estudio del panorama, las condiciones, el papel y los objetivos de la Administración Tributaria en la dinámica actual de la relación obligacional tributaria, de frente a la hipótesis de una concepción alternativa y más eficiente de la Administración Tributaria, como un garante del cumplimiento del Derecho Tributario, y ya no simplemente como un garante de la recaudación tributaria, lo que lleva casualmente a una revisión del modelo de “Relación Cooperativa” entre sujetos pasivos y Administración Tributaria, como un posible nuevo enfoque del paradigma de participación de ésta última en la relación, con evidencias funcionales y ejemplificativas, de institutos jurídicos tendentes a un nuevo paradigma, actualmente vigentes en España, Italia y Holanda.

Using small area estimation and geographic information systems technology to target health services for the uninsured

The three articles that comprise this dissertation describe how small area estimation and geographic information systems (GIS) technologies can be integrated to provide useful information about the number of uninsured and where they are located. Comprehensive data about the numbers and characteristics of the uninsured are typically only available from surveys. Utilization and administrative data are poor proxies from which to develop this information. Those who cannot access services are unlikely to be fully captured, either by health care provider utilization data or by state and local administrative data. In the absence of direct measures, a well-developed estimation of the local uninsured count or rate can prove valuable when assessing the unmet health service needs of this population. However, the fact that these are “estimates” increases the chances that results will be rejected or, at best, treated with suspicion. The visual impact and spatial analysis capabilities afforded by geographic information systems (GIS) technology can strengthen the likelihood of acceptance of area estimates by those most likely to benefit from the information, including health planners and policy makers. ^ The first article describes how uninsured estimates are currently being performed in the Houston metropolitan region. It details the synthetic model used to calculate numbers and percentages of uninsured, and how the resulting estimates are integrated into a GIS. The second article compares the estimation method of the first article with one currently used by the Texas State Data Center to estimate numbers of uninsured for all Texas counties. Estimates are developed for census tracts in Harris County, using both models with the same data sets. The results are statistically compared. The third article describes a new, revised synthetic method that is being tested to provide uninsured estimates at sub-county levels for eight counties in the Houston metropolitan area. It is being designed to replicate the same categorical results provided by a current U.S. Census Bureau estimation method. The estimates calculated by this revised model are compared to the most recent U.S. Census Bureau estimates, using the same areas and population categories. ^

Binding of the HIV-1 gp120 -V3 to cellular glycosphingolipids is necessary for CXCR4 recruitment and infection

Infection by human immunodeficiency virus type 1 (HIV-1) is a multi-step process, and detailed analyses of the various events critical for productive infection are necessary to clearly understanding the infection process and identifying novel targets for therapeutic interventions. Evidence from this study reveals binding of the viral envelope protein to host cell glycosphingolipids (GSLs) as a novel event necessary for the orderly progression of the host cell-entry and productive infection by HIV-1. Data obtained from co-immunoprecipitation analyses and confocal microscopy showed that the ability of viral envelope to interact with the co-receptor CXCR4 and productive infection of HIV-1 were inhibited in cells rendered GSL-deficient, while both these activities were restored after reconstitution of the cells with specific GSLs like GM3. Furthermore, evidence was obtained using peptide-inhibitors of HIV-1 infection to show that binding of a specific region within the V3-loop of the envelope protein gp120 to the host cell GSLs is the trigger necessary for the CD4-bound gp120 to recruit the CXCR4 co-receptor. Infection-inhibitory activity of the V3 peptides was compromised in GSL-deficient cells, but could be restored by reconstitution of GSLs. Based on these findings, a revised model for HIV-1 infection is proposed that accounts for the established interactions between the viral envelope and host cell receptors while enumerating the importance of the new findings that fill the gap in the current knowledge of the sequential events for the HIV-1 entry. According to this model, post-CD4 binding of the HIV-1 envelope surface protein gp120 to host cell GSLs, mediated by the gp120-V3 region, enables formation of the gp120-CD4-GSL-CXCR4 immune-complex and productive infection. The identification of cellular GSLs as an additional class of co-factors necessary for HIV-1 infection is important for enhancing the basic knowledge of the HIV-1 entry that can be exploited for developing novel antiviral therapeutic strategies. ^

Documentation of sediment cores

The presence of a complex bedform arrangement on the sea-floor of the continental shelf in the western Amundsen Sea Embayment, West Antarctica, indicates a multi-temporal record of flow related to the activity of one or more ice streams in the past. Mapping and division of the bedforms into distinct landform assemblages reveals their time-transgressive history, which implies that bedforms can neither be considered part of a single down-flow continuum nor a direct proxy for palaeo-ice velocity, as suggested previously. A main control on the bedform imprint is the geology of the shelf, which is divided broadly between rough bedrock on the inner shelf, and smooth, dipping sedimentary strata on the mid-to-outer shelf. Inner shelf bedform variability is well preserved, revealing information about local, complex basal-ice conditions, meltwater flow, and ice dynamics over time. These details, which are not apparent at the scale of regional morphological studies, indicate that past ice streams flowed across the entire shelf, at times, and often had onset zones that lay within the interior of the Antarctic Ice Sheet today. In contrast, highly elongated subglacial bedforms on sedimentary strata of the middle to outer shelf represent a timeslice snapshot of the last activity of ice-stream flow, and may be a truer representation of fast palaeo-ice flow in these locations. A revised model for ice streams on the shelf captures complicated multi-temporal bedform patterns associated with an Antarctic palaeo-ice stream for the first time, and confirms a strong substrate control on this ice stream system that drained the West Antarctic Ice Sheet during the Late Quaternary.

Dystrobrevin and dystrophin: An interaction through coiled-coil motifs

Dystrobrevin, a dystrophin-related and -associated protein, has been proposed to be important in the formation and maintenance of the neuromuscular junction. Dystrobrevin coprecipitates with both the acetylcholine receptor complex as well as the dystrophin glycoprotein complex. Although the nature of dystrobrevin’s association with the dystrophin glycoprotein complex remains unclear, it is known that dystrobrevin binds directly to the syntrophins, a heterologous group of dystrophin-associated proteins. Using the yeast two-hybrid system to identify protein–protein interactions, we present evidence for the heterodimerization of dystrobrevin directly with dystrophin. The C terminus of dystrobrevin binds specifically to the C terminus of dystrophin. We further refined this site of interaction to these proteins’ homologous coiled-coil motifs that flank their respective syntrophin-binding sites. We also show that the interaction between the dystrobrevin and dystrophin coiled-coil domains is specific and is not due to a nonspecific coiled-coil domain interaction. From the accumulated evidence of protein–protein interactions presented here and elsewhere, we propose a partially revised model of the organization of the dystrophin-associated glycoprotein complex.

Heme transfer to the heme chaperone CcmE during cytochrome c maturation requires the CcmC protein, which may function independently of the ABC-transporter CcmAB

Cytochrome c maturation in Escherichia coli requires the ccm operon, which encodes eight membrane proteins (CcmABCDEFGH). CcmE is a periplasmic heme chaperone that binds heme covalently and transfers it onto apocytochrome c in the presence of CcmF, CcmG, and CcmH. In this work we addressed the functions of the ccmABCD gene products with respect to holo-CcmE formation and the subsequent ligation of heme to apocytochrome c. In the absence of the ccmABCD genes, heme is not bound to CcmE. We report that CcmC is functionally uncoupled from the ABC transporter subunits CcmA and CcmB, because it is the only Ccm protein that is strictly required for heme transfer and attachment to CcmE. Site-directed mutagenesis of conserved histidines inactivates the CcmC protein, which is in agreement with the hypothesis that this protein interacts directly with heme. We also present evidence that questions the role of CcmAB as a heme exporter; yet, the transported substrate remains unknown. CcmD was found to be involved in stabilizing the heme chaperone CcmE in the membrane. We propose a heme-trafficking pathway as part of a substantially revised model for cytochrome c maturation in E. coli.

Identification of the vitamin K-dependent carboxylase active site: Cys-99 and Cys-450 are required for both epoxidation and carboxylation

The vitamin K-dependent carboxylase modifies and renders active vitamin K-dependent proteins involved in hemostasis, cell growth control, and calcium homeostasis. Using a novel mechanism, the carboxylase transduces the free energy of vitamin K hydroquinone (KH2) oxygenation to convert glutamate into a carbanion intermediate, which subsequently attacks CO2, generating the γ-carboxylated glutamate product. How the carboxylase effects this conversion is poorly understood because the active site has not been identified. Dowd and colleagues [Dowd, P., Hershline, R., Ham, S. W. & Naganathan, S. (1995) Science 269, 1684–1691] have proposed that a weak base (cysteine) produces a strong base (oxygenated KH2) capable of generating the carbanion. To define the active site and test this model, we identified the amino acids that participate in these reactions. N-ethyl maleimide inhibited epoxidation and carboxylation, and both activities were equally protected by KH2 preincubation. Amino acid analysis of 14C- N-ethyl maleimide-modified human carboxylase revealed 1.8–2.3 reactive residues and a specific activity of 7 × 108 cpm/hr per mg. Tryptic digestion and liquid chromatography electrospray mass spectrometry identified Cys-99 and Cys-450 as active site residues. Mutation to serine reduced both epoxidation and carboxylation, to 0.2% (Cys-99) or 1% (Cys-450), and increased the Kms for a glutamyl substrate 6- to 8-fold. Retention of some activity indicates a mechanism for enhancing cysteine/serine nucleophilicity, a property shared by many active site thiol enzymes. These studies, which represent a breakthrough in defining the carboxylase active site, suggest a revised model in which the glutamyl substrate indirectly coordinates at least one thiol, forming a catalytic complex that ionizes a thiol to initiate KH2 oxygenation.

Formulation of multiparticulate systems as lyophilised orally disintegrating tablets

The current study aimed to exploit the electrostatic associative interaction between carrageenan and gelatin to optimise a formulation of lyophilised orally disintegrating tablets (ODTs) suitable for multiparticulate delivery. A central composite face centred (CCF) design was applied to study the influence of formulation variables (gelatin, carrageenan and alanine concentrations) on the crucial responses of the formulation (disintegration time, hardness, viscosity and pH). The disintegration time and viscosity were controlled by the associative interaction between gelatin and carrageenan upon hydration which forms a strong complex that increases the viscosity of the stock solution and forms tablet with higher resistant to disintegration in aqueous medium. Therefore, the levels of carrageenan, gelatin and their interaction in the formulation were the significant factors. In terms of hardness, increasing gelatin and alanine concentration was the most effective way to improve tablet hardness. Accordingly, optimum concentrations of these excipients were needed to find the best balance that fulfilled all formulation requirements. The revised model showed high degree of predictability and optimisation reliability and therefore was successful in developing an ODT formulation with optimised properties that were able deliver enteric coated multiparticulates of omeprazole without compromising their functionality.

Maritime expressions:a corpus based exploration of maritime metaphors

This study uses a purpose-built corpus to explore the linguistic legacy of Britain’s maritime history found in the form of hundreds of specialised ‘Maritime Expressions’ (MEs), such as TAKEN ABACK, ANCHOR and ALOOF, that permeate modern English. Selecting just those expressions commencing with ’A’, it analyses 61 MEs in detail and describes the processes by which these technical expressions, from a highly specialised occupational discourse community, have made their way into modern English. The Maritime Text Corpus (MTC) comprises 8.8 million words, encompassing a range of text types and registers, selected to provide a cross-section of ‘maritime’ writing. It is analysed using WordSmith analytical software (Scott, 2010), with the 100 million-word British National Corpus (BNC) as a reference corpus. Using the MTC, a list of keywords of specific salience within the maritime discourse has been compiled and, using frequency data, concordances and collocations, these MEs are described in detail and their use and form in the MTC and the BNC is compared. The study examines the transformation from ME to figurative use in the general discourse, in terms of form and metaphoricity. MEs are classified according to their metaphorical strength and their transference from maritime usage into new registers and domains such as those of business, politics, sports and reportage etc. A revised model of metaphoricity is developed and a new category of figurative expression, the ‘resonator’, is proposed. Additionally, developing the work of Lakov and Johnson, Kovesces and others on Conceptual Metaphor Theory (CMT), a number of Maritime Conceptual Metaphors are identified and their cultural significance is discussed.

Extending the Theory of Planned Behavior in the exercise domain: A study of community college students in an urban multicultural setting

The purpose of this study was to assess the intention to exercise among ethnically and racially diverse community college students using the Theory of Planned Behavior (TPB). In addition to identifying the variables associated with motivation or intention of college students to engage in physical activity, this study tested the model of the Theory of Planned Behavior, asking: Does the TPB model explain intention to exercise among a racially/ethnically diverse group of college students? ^ The relevant variables were the TPB constructs (behavioral beliefs, normative beliefs, and control beliefs), which combined to form a measure of intention to exercise. Structural Equation Modeling was used to test the predictive power of the TPB constructs for predicting intention to exercise. Following procedures described by Ajzen (2002), the researcher developed a questionnaire encompassing the external variables of student demographics (age, gender, work status, student status, socio-economic status, access to exercise facilities, and past behavior), major constructs of the TPB, and two questions from the Godin Leisure Time Questionnaire (GLTQ; Godin & Shephard, 1985). Participants were students (N = 255) who enrolled in an on-campus wellness course at an urban community college. ^ The demographic profile of the sample revealed a racially/ethnically diverse study population. The original model that was used to reflect the TPB as developed by Ajzen was not supported by the data analyzed using SEM; however, a revised model that the researcher thought was theoretically a more accurate reflection of the causal relations between the TPB constructs was supported. The GLTQ questions were problematic for some students; those data could not be used in the modeling efforts. The GLTQ measure, however, revealed a significant correlation with intention to exercise (r = .27, p = .001). Post-hoc comparisons revealed significant differences in normative beliefs and attitude toward exercising behavior between Black students and Hispanic students. Compared to Black students, Hispanic students were more likely to (a) perceive “friends” as approving of them being physically active and (b) rate being physically active for 30 minutes per day as “beneficial”. No statistically significant difference was found among groups on overall intention to exercise. ^

Extending the Theory of Planned Behavior in the Exercise Domain: A Study of Community College Students in an Urban Multicultural Setting

The purpose of this study was to assess the intention to exercise among ethnically and racially diverse community college students using the Theory of Planned Behavior (TPB). In addition to identifying the variables associated with motivation or intention of college students to engage in physical activity, this study tested the model of the Theory of Planned Behavior, asking: Does the TPB model explain intention to exercise among a racially/ethnically diverse group of college students? The relevant variables were the TPB constructs (behavioral beliefs, normative beliefs, and control beliefs), which combined to form a measure of intention to exercise. Structural Equation Modeling was used to test the predictive power of the TPB constructs for predicting intention to exercise. Following procedures described by Ajzen (2002), the researcher developed a questionnaire encompassing the external variables of student demographics (age, gender, work status, student status, socio-economic status, access to exercise facilities, and past behavior), major constructs of the TPB, and two questions from the Godin Leisure Time Questionnaire (GLTQ; Godin & Shephard, 1985). Participants were students (N = 255) who enrolled in an on-campus wellness course at an urban community college. The demographic profile of the sample revealed a racially/ethnically diverse study population. The original model that was used to reflect the TPB as developed by Ajzen was not supported by the data analyzed using SEM; however, a revised model that the researcher thought was theoretically a more accurate reflection of the causal relations between the TPB constructs was supported. The GLTQ questions were problematic for some students; those data could not be used in the modeling efforts. The GLTQ measure, however, revealed a significant correlation with intention to exercise (r = .27, p = .001). Post-hoc comparisons revealed significant differences in normative beliefs and attitude toward exercising behavior between Black students and Hispanic students. Compared to Black students, Hispanic students were more likely to (a) perceive “friends” as approving of them being physically active and (b) rate being physically active for 30 minutes per day as “beneficial”. No statistically significant difference was found among groups on overall intention to exercise.

Lifetime and distribution of ozone and related pollutants in the eastern United States

Most major cities in the eastern United States have air quality deemed unhealthy by the EPA under a set of regulations known as the National Ambient Air Quality Standards (NAAQS). The worst air quality in Maryland is measured in Edgewood, MD, a small community located along the Chesapeake Bay and generally downwind of Baltimore during hot, summertime days. Direct measurements and numerical simulations were used to investigate how meteorology and chemistry conspire to create adverse levels of photochemical smog especially at this coastal location. Ozone (O3) and oxidized reactive nitrogen (NOy), a family of ozone precursors, were measured over the Chesapeake Bay during a ten day experiment in July 2011 to better understand the formation of ozone over the Bay and its impact on coastal communities such as Edgewood. Ozone over the Bay during the afternoon was 10% to 20% higher than the closest upwind ground sites. A combination of complex boundary layer dynamics, deposition rates, and unaccounted marine emissions play an integral role in the regional maximum of ozone over the Bay. The CAMx regional air quality model was assessed and enhanced through comparison with data from NASA’s 2011 DISCOVER-AQ field campaign. Comparisons show a model overestimate of NOy by +86.2% and a model underestimate of formaldehyde (HCHO) by –28.3%. I present a revised model framework that better captures these observations and the response of ozone to reductions of precursor emissions. Incremental controls on electricity generating stations will produce greater benefits for surface ozone while additional controls on mobile sources may yield less benefit because cars emit less pollution than expected. Model results also indicate that as ozone concentrations improve with decreasing anthropogenic emissions, the photochemical lifetime of tropospheric ozone increases. The lifetime of ozone lengthens because the two primary gas-phase sinks for odd oxygen (Ox ≈ NO2 + O3) – attack by hydroperoxyl radicals (HO2) on ozone and formation of nitrate – weaken with decreasing pollutant emissions. This unintended consequence of air quality regulation causes pollutants to persist longer in the atmosphere, and indicates that pollutant transport between states and countries will likely play a greater role in the future.

The Issue with Issues Management: An Engagement Approach to Integrate Gender and Emotion into Issues Management

Using sexual assault on college campuses as a context for interrogating issues management, this study offers a normative model for inclusive issues management through an engagement approach that can better account for the gendered and emotional dimensions of issues. Because public relations literature and research have offered little theoretical or practical guidance for how issues managers can most effectively deal with issues such as sexual assault, this study represents a promising step forward. Results for this study were obtained through 32 in-depth interviews with university issues managers, six focus groups with student populations, and approximately 92 hours of participant observation. By focusing on inclusion, this revised model works to have utility for an array of issues that have previously fallen outside of the dominant masculine and rationale spheres that have worked to silence marginalized publics’ experiences. Through adapting previous issues management models to focus on inclusion at the heart of a strategic process, and engagement as the strategy for achieving this, this study offers a framework for ensuring more voices are heard—which enables organizations to more effectively communicate with their publics. Additionally, findings from this research may also help practitioners at different types of organizations develop better, and proactive, communication strategies for handling emotional and gendered issues as to avoid negative media attention and work to change organizational culture.