Selective and Specific Activation of Rab5 during Endocytosis of Receptor Tyrosine Kinases

The Rab family of proteins are low molecular weight GTPases that have the ability to switch between GTP- (active) and GDP- (inactive) bound form, and in that sense act as molecular switches. Through distinct localization on various vesicles and organelles and by cycling through GTP/GDP bound forms, Rabs are able to recruit and activate numerous effector proteins, both spatially and temporally, and hence behave as key regulators of trafficking in both endocytic and biosynhtetic pathways. The Rab5 protein has been shown to regulate transport from plasma membrane to the early endosome as well as activate signaling pathways from the early endosome. This dissertation focused on understanding Rab5 activation via endocytosis of receptor tyrosine kinases (RTKs). First, tyrosine kinase activity of RTKs was linked to endosome fusion by demonstrating that tyrosine kinase inhibitors block endosome fusion and activation of Rab5, and a constitutively active form of Rab5 is able to rescue endosome fusion. However, depending on how much ligand is available at the cell surface, the receptor-ligand complexes can be internalized via a number of distinct pathways. Similarly, Rab5 was activated in a ligand-dependent concentration dependent manner via clathrin- and caveolin-mediated pathways, as well as a pathway independent of both. However, overexpression Rabex-5, a nucleotide exchange factor for Rab5, is able to rescue activation even when all of the pathways of EGF-receptor internalization were blocked. Next, the three naturally occurring splice variants of Rabex-5 selectively activated Rab5. Lastly, Rabex-5 inhibits differentiation of 3T3-L1 and PC12 cells through 1) degradation of signaling endosome via Rab5-dependent fusion with the early endosome, 2) and inhibition of signaling cascade via ubiquitination of Ras through the ZnF domain at the N-terminus of Rabex-5. In conclusion, these data shed light on complexity of the endosomal trafficking system where tyrosine kinase activity of the receptor is able to affect endosome fusion; how different endocytic pathways affect activation of one of the key regulators of early endocytic events; and how selective activation of Rab5 via Rabex-5 can control adipogenesis and neurogenesis.

Pseudomonas Aeruginosa AmpR Transcriptional Regulatory Network

In Enterobacteriaceae, the transcriptional regulator AmpR, a member of the LysR family, regulates the expression of a chromosomal β-lactamase AmpC. The regulatory repertoire of AmpR is broader in Pseudomonas aeruginosa, an opportunistic pathogen responsible for numerous acute and chronic infections including cystic fibrosis. Previous studies showed that in addition to regulating ampC, P. aeruginosa AmpR regulates the sigma factor AlgT/U and production of some quorum sensing (QS)-regulated virulence factors. In order to better understand the ampR regulon, the transcriptional profiles generated using DNA microarrays and RNA-Seq of the prototypic P. aeruginosa PAO1 strain with its isogenic ampR deletion mutant, PAO∆ampR were analyzed. Transcriptome analysis demonstrates that the AmpR regulon is much more extensive than previously thought influencing the differential expression of over 500 genes. In addition to regulating resistance to β-lactam antibiotics via AmpC, AmpR also regulates non-β-lactam antibiotic resistance by modulating the MexEF-OprN efflux pump. Virulence mechanisms including biofilm formation, QS-regulated acute virulence, and diverse physiological processes such as oxidative stress response, heat-shock response and iron uptake are AmpR-regulated. Real-time PCR and phenotypic assays confirmed the transcriptome data. Further, Caenorhabditis elegans model demonstrates that a functional AmpR is required for full pathogenicity of P. aeruginosa. AmpR, a member of the core genome, also regulates genes in the regions of genome plasticity that are acquired by horizontal gene transfer. The extensive AmpR regulon included other transcriptional regulators and sigma factors, accounting for the extensive AmpR regulon. Gene expression studies demonstrate AmpR-dependent expression of the QS master regulator LasR that controls expression of many virulence factors. Using a chromosomally tagged AmpR, ChIP-Seq studies show direct AmpR binding to the lasR promoter. The data demonstrates that AmpR functions as a global regulator in P. aeruginosa and is a positive regulator of acute virulence while negatively regulating chronic infection phenotypes. In summary, my dissertation sheds light on the complex regulatory circuit in P. aeruginosa to provide a better understanding of the bacterial response to antibiotics and how the organism coordinately regulates a myriad of virulence factors.

HRM and innovation:looking across levels

Studies are starting to explore the role of HRM in fostering organizational innovation but empirical evidence remains contradictory and theory fragmented. This is partly because extant literature by and large adopts a unitary level of analysis, rather than reflecting on the multi-level demands that innovation presents. Building on an emergent literature focused on HRM’s role in shaping innovation, we shed light on the question of whether, and how, HRM might influence employees’ innovative behaviours in the direction of strategically important goals. Drawing upon institutional theory, our contributions are three-fold: to bring out the effect of two discrete HRM configurations- one underpinned by a control and the other by an entrepreneurial ethos, on attitudes and behaviours at the individual level; to reflect the way in which employee innovative behaviours arising from these HRM configurations coalesce to shape higher-level phenomena, such as organizational-level innovation; and to bring out two distinct patterns of bottom-up emergence, one driven primarily by composition and the other by both composition and compilation.

Lecturas de la prensa popular: Un estudio de recepción del periódico Q'hubo en la localidad de Kennedy en Bogotá

En este trabajo de grado se aborda la relación entre las audiencias y los medios a partir de un estudio de recepción del diario Q’hubo en los sectores populares de Bogotá, específicamente en la localidad de Kennedy de esta ciudad. El estudio reconoce que las formas determinadas de interactuar con los medios están atadas a los contextos, las instituciones y los entornos culturales y sociales desde los que lector construye su realidad y se inserta en la sociedad actual. Por lo mismo, se nutre de estos elementos para entender los procesos de recepción a través de las formas cotidianas de consumo del diario Q’hubo frente al contexto mediático actual. Se sustenta que el Q’hubo se comunica con sus lectores combinando aspectos clásicos de la prensa popular y adaptando aspectos de diseño y lenguaje aportados por los nuevos medios. Lo que a su vez se relaciona con apropiaciones, usos y modos de leer de la audiencia, que reivindica interacciones tradicionales y desarrolla interacciones emergentes gracias a la convergencia de las tecnologías con la cultura letrada, oral y visual.

¿Quién es terrorista? Aproximación a las nociones de terrorismo en la política de seguridad democrática durante el gobierno de Álvaro Uribe Vélez

El objetivo del presente trabajo de grado es estudiar el discurso de la lucha contra el terrorismo en Colombia y dar luces al debate que giró alrededor de quien era señalado como terrorista durante el gobierno de Álvaro Uribe Vélez. Indaga sobre la instrumentalización que se ha dado al discurso de la lucha contra el terrorismo por actores políticos, quienes, en contraposición a los actores jurídicos, buscan mantener la ambigüedad del mencionado debate. Partiendo de los planteamientos de Donatella Della Porta y Charles Tilly, quienes ponen en tela de juicio la validez del terrorismo como concepto dentro de las ciencias sociales y, utilizando casos para ejemplificar cómo los actores políticos se benefician de la ambigüedad del discurso configurado alrededor del terrorismo, el presente trabajo muestra cómo se implementó este discurso en Colombia enfocándose en los actores que podían modificar la realidad a partir de su posición en el mencionado debate.

Caracterización paraclínica de trabajadores con patologías respiratorias de un centro de referencia neumológico, Bucaramanga, 2014-2016

Introducción: La enfermedad respiratoria ocupacional es causada por la exposición a diferentes agentes en el trabajo. Las pruebas objetivas realizadas en sospecha de enfermedad respiratoria de origen laboral, son importantes herramientas que permiten realizar un adecuado diagnóstico, una detección precoz de la enfermedad respiratoria ocupacional, disminuye el progreso rápido de la patología, la morbilidad de los trabajadores y el impacto negativo sobre su futuro laboral. Objetivo: Caracterizar las pruebas paraclínicas de las patologías respiratorias de trabajadores, en un centro de referencia neumológico de Bucaramanga año 2014-2016. Materiales y métodos: Se realizó un estudio descriptivo retrospectivo con datos secundarios de 96 trabajadores que laboran en diferentes actividades económicas. Se incluyeron variables sociodemográficas, laborales, ayudas imagenológicas y pruebas de función pulmonar, realizando 3 grupos de acuerdo a su patología que fueron: Asma, síndrome de disfunción reactiva de la vía aérea y neumoconiosis. En el análisis estadístico se emplearon medidas de tendencia central y dispersión. Resultados: De los 96 trabajadores 84.4% son hombres, las actividades económicas más frecuentes fueron la industria del petróleo y gas en un 27.1% y trabajadores en materiales de construcción en un 19.8%. En la caracterización paraclínica por grupo de patología, para asma predominó la obstrucción en la espirometría (46.9%) y los volúmenes pulmonares con atrapamiento aéreo (95.5%), en RADS (síndrome de disfunción de vías aéreas reactivas) los volúmenes pulmonares con atrapamiento aéreo (77%) y en las neumoconiosis para Rx de tórax (90.3%) y Tac de tórax (100%) reportaron alteraciones parenquimatosas, espirometría con obstrucción (54.8%) y volúmenes pulmonares con atrapamiento aéreo (62.5 %).Discusión y Conclusiones: Las ocupaciones de mayor riesgo para desarrollo de neumopatías de origen ocupacional fuero, , son la minería y construcción y para asma la agricultura y manufacturas. Para asma se evidenció que no hay significancia diagnóstica para estudios imagenológicos pero sí para las pruebas de función pulmonar. Para neumoconiosis el estudio imagenológico es el de mayor importancia ya que en las radiografías se presentan cambios incluso mucho antes de la afectación de la función pulmonar. Para RADS se concluyó que la realización de un test de provocación con metacolina sería el Gold estándar para el diagnóstico. Las pruebas de función respiratoria son de vital importancia para determinar la enfermedad ocupacional en trabajadores expuestos para vigilancia y detección precoz, es conveniente la realización de protocolos para la evaluación y diagnóstico de la enfermedad respiratoria de origen ocupacional. Palabras claves: Neumoconiosis, asma ocupacional, función pulmonar, radiografía de tórax, ocupación, Colombia.

Evoluzione delle celle solari al silicio

Negli ultimi anni si è assistito nel panorama mondiale ad un notevole ed importante cambiamento nell’asset della produzione dell’energia elettrica. L’aumentare del costo nell'estrazione dei combustibili fossili ed il conseguente impatto ambientale che questi hanno nell'atmosfera ha influito notevolmente sullo sviluppo e sulla diffusione della tecnologia fotovoltaica. L’evoluzione di questa tecnologia ha permesso di raggiungere la “grid parity”, ossia l’uguaglianza tra il costo di 1 kWh prodotto dal fotovoltaico con 1 kWh prodotto da fonti fossili, per molte aree del pianeta. Tale tecnologia non è recente, già negli anni ‘60 si svilupparono i primi studi a riguardo, ma negli ultimi anni ha subito notevoli migliorie che l’hanno portata ad essere competitiva nel settore dell’energia elettrica. Il presente lavoro di tesi riporta una rassegna delle celle solari a base di silicio, a partire dal silicio monocristallino, al silicio poli e multi cristallino, fino ad arrivare a celle a film sottile o celle costituite da materiali multi-fase contenenti strutture quantiche e nanocristalli di Si. Negli ultimi anni tutti gli studi si stanno concentrando su un nuovo tipo di fotovoltaico, quello di terza generazione, il cui scopo è quello di ottenere dispositivi che, ad un prezzo molto contenuto dovuto ad un utilizzo di materiali economici come i Si-NCs (nano cristalli di silicio), possano garantire elevate efficienze in modo tale che la tecnologia fotovoltaica sia in grado di affermarsi definitivamente nel settore dell’energia.

Discriminating stromatolite formation modes using rare earth element geochemistry: Trapping and binding versus in situ precipitation of stromatolites from the Neoproterozoic Bitter Springs Formation, Northern Territory, Australia

Stromatolites consist primarily of trapped and bound ambient sediment and/or authigenic mineral precipitates, but discrimination of the two constituents is difficult where stromatolites have a fine texture. We used laser ablation-inductively coupled plasma-mass spectrometry to measure trace element (rare earth element – REE, Y and Th) concentrations in both stromatolites (domical and branched) and closely associated particulate carbonate sediment in interspaces (spaces between columns or branches) from bioherms within the Neoproterozoic Bitter Springs Formation, central Australia. Our high resolution sampling allows discrimination of shale-normalised REE patterns between carbonate in stromatolites and immediately adjacent, fine-grained ambient particulate carbonate sediment from interspaces. Whereas all samples show similar negative La and Ce anomalies, positive Gd anomalies and chondritic Y/Ho ratios, the stromatolites and non-stromatolite sediment are distinguishable on the basis of consistently elevated light REEs (LREEs) in the stromatolitic laminae and relatively depleted LREEs in the particulate sediment samples. Additionally, concentrations of the lithophile element Th are higher in ambient sediment samples than in stromatolites, consistent with accumulation of some fine siliciclastic detrital material in the ambient sediment but a near absence in the stromatolites. These findings are consistent with the stromatolites consisting dominantly of in situ carbonate precipitates rather than trapped and bound ambient sediment. Hence, high resolution trace element (REE + Y, Th) geochemistry can discriminate fine-grained carbonates in these stromatolites from coeval non-stromatolitic carbonate sediment and demonstrates that the sampled stromatolites formed primarily from in situ precipitation, presumably within microbial mats/biofilms, rather than by trapping and binding of ambient sediment. Identification of the source of fine carbonate in stromatolites is significant, because if it is not too heavily contaminated by trapped ambient sediment, it may contain geochemical biosignatures and/or direct evidence of the local water chemistry in which the precipitates formed.

Desorption electrospray ionisation mass spectrometry reveals in situ modification of a hindered amine light stabiliser resulting from direct N–OR bond cleavage

Detection and characterisation of structural modifications of a hindered amine light stabiliser (HALS) directly from a polyester-based coil coating have been achieved by desorption electrospray ionisation mass spectrometry (DESI-MS) for the first time. In situ detection is made possible by exposing the coating to an acetone vapour atmosphere prior to analysis. This is a gentle and non-destructive treatment that allows diffusion of analyte to the surface without promoting lateral migration. Using this approach a major structural modification of the HALS TINUVIN®123 (bis(1-octyloxy-2,2,6,6-tetramethyl-4-piperidyl) sebacate) was discovered where one N-ether piperidine moiety (N-OC8H17) is converted to a secondary piperidine (N–H). With the use of 2-dimensional DESI-MS imaging the modification was observed to arise during high curing temperatures (ca. 260 °C) and under simulated physiological conditions (80 °C, full solar spectrum). It is proposed that the secondary piperidine derivative is a result of a highly reactive aminyl radical intermediate produced by N–O homolytic bond cleavage. The nature of the bond cleavage is also suggested by ESR spin-trapping experiments employing α-phenyl-N-tert-butyl nitrone (PBN) in toluene at 80 °C. The presence of a secondary piperidine derivative in situ and the implication of N–OR competing with NO–R bond cleavage suggest an alternative pathway for generation of the nitroxyl radical—an essential requirement in anti-oxidant activity that has not previously been described for the N-ether sub-class of HALS.

Solvation dynamics, energy distribution and trapping of a light solute ion

In the theoretical treatments of the dynamics of solvation of a newly created ion in a dipolar solvent, the self-motion of the solute is usually ignored. Recently, it has been shown that for a light ion the translational motion of the ion can significantly enhance its own rate of solvation. Therefore, solvation itself may not be the rate determining step in the equilibration. Instead, the rate determining step is the search of the low energy configuration which serves to localize the light ion. In this article a microscopic calculation of the probability distribution of the interaction energy of the nascent charge with the dipolar solvent molecules is presented in order to address this problem of solute trapping. It is found that to a good approximation, this distribution is Gaussian and the second moment of this distribution is exactly equal to the half of its own solvation energy. It is shown that this is in excellent agreement with the simulation results that are available for the model Brownian dipolar lattice and for liquid acetonitrile. If the distortion of the solvent by the ion is negligible then the same relation gives the energy distribution for the solvated ion, with the average centered at the final equilibrium solvation energy. These results are expected to be useful in understanding various chemical processes in dipolar liquids. Another interesting outcome of the present study is a simple dynamic argument that supports Onsager's ''inverse snow-ball'' conjecture of solvation of a light ion. A simple derivation of the semi-phenomenological relation between the solvation time correlation function and the single particle orientation, reported recently by Maroncelli et al. (J. Phys. Chem. 97 (1993) 13), is also presented.

Instability in threshold voltage and subthreshold behavior in Hf-In-Zn-O thin film transistors induced by bias-and light-stress

Electrical bias and light stressing followed by natural recovery of amorphous hafnium-indium-zinc-oxide (HIZO) thin film transistors with a silicon oxide/nitride dielectric stack reveals defect density changes, charge trapping and persistent photoconductivity (PPC). In the absence of light, the polarity of bias stress controls the magnitude and direction of the threshold voltage shift (Δ VT), while under light stress, VT consistently shifts negatively. In all cases, there was no significant change in field-effect mobility. Light stress gives rise to a PPC with wavelength-dependent recovery on time scale of days. We observe that the PPC becomes more pronounced at shorter wavelengths. © 2010 American Institute of Physics.

Electron trapping materials for use in a picosecond infrared streak camera

We describe our research on the employment of an infrared upconversion screen made of electron trapping material (ETM) in combination with the high sensitivity of the S-20 photocathode responsive to visible radiation to produce a streak camera arrangement capable of viewing and recording infrared incident pulses. The ETM-based upconversion screen converts 800-1600 nm infrared radiation to visible light which is viewed or recorded by the S-20 photocathode. The peak values of the upconversion efficiency are located at 1165 nm for CaS:Eu, Sm and 1060 nm for CaS:Ce, Sm. The present experiment showed time resolution was 12.3 ps for a CaS:Eu, Sm screen and 8.4 ps for a CaS:Ce, Sm screen. The minimum detectability is 4.8 x 10(-9) J/mm(2) (minimum detectability of the coupled visible streak camera is 8.3x10(-10) J/mm(2)). Other parameters, such as spatial resolution and dynamic range, have also been measured and analyzed. The results show ETM can be used in the measurement of infrared ultrafast phenomena up to picosecond time domain. In consideration of the limited number of trapped electrons in ETM, the infrared-sensitive streak camera consisting of an ETM-based upconversion screen is suitable to operate in the single shot mode. (C) 1999 American Institute of Physics. [S0034-6748(99)00112-4].

Change of the dominant luminescent mechanism with increasing current density in molecularly doped organic light-emitting devices

We have fabricated and measured a series of electroluminescent devices with the structure of ITO/TPD/Eu(TTA)(3)phen (x):CBP/BCP/ ALQ/LiF/Al, where x is the weight percentage of Eu(TTA)3phen (from 0% to 6%). At very low current density, carrier trapping is the dominant luminescent mechanism and the 4% doped device shows the highest electroluminescence (EL) efficiency among all these devices. With increasing current density, Forster energy transfer participates in EL process. At the current density of 10.0 and 80.0mA/ cm(2), 2% and 3% doped devices show the highest EL efficiency, respectively. From analysis of the EL spectra and the EL efficiency-current density characteristics, we found that the EL efficiency is manipulated by Forster energy transfer efficiency at high current density. So we suggest that the dominant luminescent mechanism changes gradually from carrier trapping to Forster energy transfer with increasing current density. Moreover, the conversion of dominant EL mechanism was suspected to be partly responsible for the EL efficiency roll-off because of the lower EL quantum efficiency of Forster energy transfer compared with carrier trapping.

Improved color purity and efficiency by a coguest emitter system in doped red light-emitting devices

We demonstrate red organic light-emitting diodes (OLEDs) with improved color purity and electroluminescence (EL) efficiency by codoping a green fluorescent sensitizer 10-(2-benzothiazolyl)-2,3,6,7-tetrahydro-1, 1, 7,7-tetramethyl-1H, 5H, 11H-(1)-benzopyropyrano(6,7-8-ij)quinolizin-11-one (C545T) as the second dopant and a red fluorescent dye 4-(dicyanomethylene)-2-t-butyl-6(1,1,7,7tetramethyljulolidyl-9-enyl)-4H-pyran (DCJTB) as the lumophore into tris(8-hydroquinoline) aluminum (Alq(3)) host. It was found that the C545 T dopant did not by itself emit but assisted the carrier trapping from the host Alq(3) to the red emitting dopant. The red OLEDs realized by this approach not only kept the purity of the emission color, but also significantly improved the EL efficiency. The current efficiency and power efficiency, respectively, reached 12 cd/A at a current density of 0.3 mA/cm(2) and 10lm/W at a current density of 0.02 mA/cm(2), which are enhanced by 1.4 and 2.6 times compared with devices where the emissive layer is composed of the DCJTB doped Alq(3), and a stable red emission (chromaticity coordinates: x = 0.64, y = 0.36) was obtained in a wide range of voltage. Our results indicate that the coguest system is a promising method for obtaining high-efficiency red OLEDs.