Fatty acids and hypolipidemic drugs regulate peroxisome proliferator-activated receptors α- and γ-mediated gene expression via liver fatty acid binding protein: A signaling path to the nucleus

Peroxisome proliferator-activated receptor α (PPARα) is a key regulator of lipid homeostasis in hepatocytes and target for fatty acids and hypolipidemic drugs. How these signaling molecules reach the nuclear receptor is not known; however, similarities in ligand specificity suggest the liver fatty acid binding protein (L-FABP) as a possible candidate. In localization studies using laser-scanning microscopy, we show that L-FABP and PPARα colocalize in the nucleus of mouse primary hepatocytes. Furthermore, we demonstrate by pull-down assay and immunocoprecipitation that L-FABP interacts directly with PPARα. In a cell biological approach with the aid of a mammalian two-hybrid system, we provide evidence that L-FABP interacts with PPARα and PPARγ but not with PPARβ and retinoid X receptor-α by protein–protein contacts. In addition, we demonstrate that the observed interaction of both proteins is independent of ligand binding. Final and quantitative proof for L-FABP mediation was obtained in transactivation assays upon incubation of transiently and stably transfected HepG2 cells with saturated, monounsaturated, and polyunsaturated fatty acids as well as with hypolipidemic drugs. With all ligands applied, we observed strict correlation of PPARα and PPARγ transactivation with intracellular concentrations of L-FABP. This correlation constitutes a nucleus-directed signaling by fatty acids and hypolipidemic drugs where L-FABP acts as a cytosolic gateway for these PPARα and PPARγ agonists. Thus, L-FABP and the respective PPARs could serve as targets for nutrients and drugs to affect expression of PPAR-sensitive genes.

Least activation path for protein folding: investigation of staphylococcal nuclease folding by stopped-flow circular dichroism.

Is the pathway of protein folding determined by the relative stability of folding intermediates, or by the relative height of the activation barriers leading to these intermediates? This is a fundamental question for resolving the Levinthal paradox, which stated that protein folding by a random search mechanism would require a time too long to be plausible. To answer this question, we have studied the guanidinium chloride (GdmCl)-induced folding/unfolding of staphylococcal nuclease [(SNase, formerly EC 3.1.4.7; now called microbial nuclease or endonuclease, EC 3.1.31.1] by stopped-flow circular dichroism (CD) and differential scanning microcalorimetry (DSC). The data show that while the equilibrium transition is a quasi-two-state process, kinetics in the 2-ms to 500-s time range are triphasic. Data support the sequential mechanism for SNase folding: U3 <--> U2 <--> U1 <--> N0, where U1, U2, and U3 are substates of the unfolded protein and N0 is the native state. Analysis of the relative population of the U1, U2, and U3 species in 2.0 M GdmCl gives delta-G values for the U3 --> U2 reaction of +0.1 kcal/mol and for the U2 --> U1 reaction of -0.49 kcal/mol. The delta-G value for the U1 --> N0 reaction is calculated to be -4.5 kcal/mol from DSC data. The activation energy, enthalpy, and entropy for each kinetic step are also determined. These results allow us to make the following four conclusions. (i) Although the U1, U2, and U3 states are nearly isoenergetic, no random walk occurs among them during the folding. The pathway of folding is unique and sequential. In other words, the relative stability of the folding intermediates does not dictate the folding pathway. Instead, the folding is a descent toward the global free-energy minimum of the native state via the least activation path in the vast energy landscape. Barrier avoidance leads the way, and barrier height limits the rate. Thus, the Levinthal paradox is not applicable to the protein-folding problem. (ii) The main folding reaction (U1 --> N0), in which the peptide chain acquires most of its free energy (via van der Waals' contacts, hydrogen bonding, and electrostatic interactions), is a highly concerted process. These energy-acquiring events take place in a single kinetic phase. (iii) U1 appears to be a compact unfolded species; the rate of conversion of U2 to U1 depends on the viscosity of solution. (iv) All four relaxation times reported here depend on GdmCl concentrations: it is likely that none involve the cis/trans isomerization of prolines. Finally, a mechanism is presented in which formation of sheet-like chain conformations and a hydrophobic condensation event precede the main-chain folding reaction.

Codominance and toxins: a path to drugs of nearly unlimited selectivity.

The effectiveness of drugs is often limited by their insufficient selectivity. I propose designs of therapeutic agents that address this problem. The key feature of these reagents, termed comtoxins (codominance-mediated toxins), is their ability to utilize codominance, a property characteristic of many signals in proteins, including degradation signals (degrons) and nuclear localization signals. A comtoxin designed to kill cells that express intracellular proteins P1 and P2 but to spare cells that lack P1 and/or P2 is a multidomain fusion containing a cytotoxic domain and two degrons placed within or near two domains P1* and P2* that bind, respectively, to P1 and P2. In a cell containing both P1 and P2, these proteins would bind to the P1* and P2* domains of the comtoxin and sterically mask the nearby (appropriately positioned) degrons, resulting in a long-lived and therefore toxic drug. By contrast, in a cell lacking P1 and/or P2, at least one of the comtoxin's degrons would be active (unobstructed), yielding a short-lived and therefore nontoxic drug. A comtoxin containing both a degron and a nuclear localization signal can be designed to kill exclusively cells that contain P1 but lack P2. Analogous strategies yield comtoxins sensitive to the presence (or absence) of more than two proteins in a cell. Also considered is a class of comtoxins in which a toxic domain is split by a flexible insert containing binding sites for the target proteins. The potentially unlimited, combinatorial selectivity of comtoxins may help solve the problem of side effects that bedevils present-day therapies, for even nonselective delivery of a comtoxin would not affect cells whose protein "signatures" differ from the targeted one.

Populismos y democracias defectuosas en El Ecuador (1996-2012): continuidades y diferencia

From 1995 to 2015, Ecuador experienced one of its longest periods of deep political, social and economic crisis. During this interval, three democratically elected governments (Bucaram, 1997; Mahuad, 2000 and Gutiérrez, 2005) were overthrown and a critical juncture arose in 2006 as a result. Since 2007, and as a consequence of these chaotic circumstances, new populist strongmen ascended and, amid the biggest bonanza of oil revenues in Ecuadorian history, established a defective democracy. The gradual escalation of authoritarian tendencies during the three consecutive terms in which Rafael Correa has acted as President, have resulted in the severe weakening of the country’s democratic institutions, since Correa’s has strived to perpetuate himself in power through continual re-election into office, instead of building an institutional quality-democracy. This study aims to clarify the historical foundations of the recurrence of caudillistas, populist and authoritarian governments in Ecuador, revealing the basis of the specific path dependence of Ecuadorian politics. We also explore the Jungian theory, specifically the “pseudo-hero myth”, as the political narrative which Correa’s regime successfully employed to establish its hegemony. Additionally, we perform a psychological-political case analysis by examining the social psychology components underlying the Ecuadorian path dependence towards authoritarian and populist caudillos: Specifically, our case study is framed within historical institutionalism, which focuses on methodological individualism to attend various political science and psychological-political theories...

On Becoming a Working Visual Artist: Opportunities and Challenges Along the Path

Thousands of students graduate from colleges and art schools every year with the goal of becoming working visual artists. The majority of them, however, find that earning a living as a working artist is a tough and competitive career path. This Capstone Project, through an extensive literature review and interviews, examines the factors and characteristics that influence whether an individual will make the leap to becoming a working visual artist. Research results indicate that primary factors in achieving the status of working visual artists include specific personality traits, experiential and career-oriented arts education, and development of strong business skills.

Efficient tool path computation using multi-core GPUs

Tool path generation is one of the most complex problems in Computer Aided Manufacturing. Although some efficient strategies have been developed, most of them are only useful for standard machining. However, the algorithms used for tool path computation demand a higher computation performance, which makes the implementation on many existing systems very slow or even impractical. Hardware acceleration is an incremental solution that can be cleanly added to these systems while keeping everything else intact. It is completely transparent to the user. The cost is much lower and the development time is much shorter than replacing the computers by faster ones. This paper presents an optimisation that uses a specific graphic hardware approach using the power of multi-core Graphic Processing Units (GPUs) in order to improve the tool path computation. This improvement is applied on a highly accurate and robust tool path generation algorithm. The paper presents, as a case of study, a fully implemented algorithm used for turning lathe machining of shoe lasts. A comparative study will show the gain achieved in terms of total computing time. The execution time is almost two orders of magnitude faster than modern PCs.

Teaching-learning process of historical heritage: a Journey as a beginning, path and end

The implantation of new university degrees within the European Higher Education Area implies the need of innovative methodologies in teaching and learning to improve the skills and competencies of students and to answer the growing needs that society continuously demands to heritage management experts. The present work shows an application of the teaching methodology proposed during the international workshop entitled “I International Planning Preservation Workshop. Learning from Al Andalus”, which included the participation of the University of Alicante and Granada, Università Politecnico di Milano and Hunter College City University of New York; where we tried to dissolve traditional boundaries derived of interuniversity cooperation programs. The main objective of the workshop was to discuss and debate the role of urban Historical Centers within the Global Heritage by the integrated work through multidisciplinary teams and the creation of a permanent international working group between these universities to both teach and research. The methodology of this workshop was very participatory and considered the idea of a new learning process generated by "a journey experience." A trip from global to local (from the big city to the small village) but also a trip from the local (historical) part of a big city to the global dimension of contemporary historical villages identified by the students through a system of exhibition panels in affinity groups, specific projects proposed by lecturers and teachers or the generation of publications in various areas (texts, photographs, videos, etc.). So, the participation of the students in this multidisciplinary meeting has enhanced their capacity for self-criticism in several disciplines and has promoted their ability to perform learning and research strategies in an autonomous way. As a result, it has been established a permanent international work structure for the development of projects of the Historical City. This relationship has generated the publication of several books whose contents have reflected the conclusions developed in the workshop and several teaching proposals shared between those institutions. All these aspects have generated a new way of understanding the teaching process through a journey, in order to study the representative role of university in the historical heritage and to make students (from planning, heritage management, architecture, geography, sociology, history or engineering areas) be compromised on searching strategies for sustainable development in the Contemporary City.

Tracing the Origins of Athlete Development Models in Sport: a Citation Path Analysis

Reviews of the sport psychology literature have identified a number of models of athlete development in sport (Alfermann & Stambulova, 2007; Durand-Bush &Salmela, 2001). However, minimal research has investigated the origins of knowledge from which each model was developed. The purpose of this study was to systematically examine the influential texts responsible for providing the basis of athlete development models in sport. A citation path analysis of the sport psychology literature was used to generate a knowledge development path of seven athlete development models in sport. The analysis identified influential texts and authors in the conceptualization of athlete development. The popula-tion of 229 texts (articles, books, book chapters) was selected in two phases. Phase1 texts were articles citing seven articles depicting models of athlete development(n  75). Phase 2 included texts cited three or more times by Phase 1 articles (n  154). The analysis revealed how the scholarship of Benjamin Bloom (1985) has been integrated into the field of sport psychology, and how two articles appearing in 1993 and 2003 helped shape present conceptualizations of athlete development