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Heterologous expression of enzymes can generate a background-free environment that facilitates investigation of enzyme properties, for instance to focus on particular isoforms in case of gene families, or on individual splicing variants. If a proper host can be found, in vivo assays are often simpler than overexpression and purification, followed by in vitro measurements, would be. We expressed plant ubiquitin ligase PRT6 in the budding yeast Saccharomyces cerevisiae for studies on activity and substrate preferences. Expression of this large enzyme profits from the eukaryotic folding catalysis provided by budding yeast, and from the presence of endogenous ubiquitin activating enzyme. While yeast encodes a ubiquitin ligase, Ubr1, that is functionally related to PRT6, a strain with deletion of the UBR1 gene offers a background-free host. Two different substrates were analyzed. One was a model substate, and the other one a natural substrate fused to a reporter. Two different methods were compared for assessment of protein stability. A method based on internal standardization via tandem fluorescent timer measurement turned out to be complementary to standardization based on cell culture density.
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Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Ubiquitinación , Ubiquitina/metabolismoRESUMEN
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
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Dual-energy CT has opened up a new level of quantitative X-ray imaging for many diagnostic applications. The energy dependence of the X-ray attenuation is the key to quantitative material decomposition of the volume under investigation. This material decomposition allows the calculation of virtual native images in contrast enhanced angiography, virtual monoenergetic images for beam-hardening artifact reduction and quantitative material maps, among others. These visualizations have been proven beneficial for various diagnostic questions. Here, we demonstrate a new method of 'virtual dual-energy CT' employing grating-based phase-contrast for quantitative material decomposition. Analogue to the measurement at two different energies, the applied phase-contrast measurement approach yields dual information in form of a phase-shift and an attenuation image. Based on these two image channels, all known dual-energy applications can be demonstrated with our technique. While still in a preclinical state, the method features the important advantages of direct access to the electron density via the phase image, simultaneous availability of the conventional attenuation image at the full energy spectrum and therefore inherently registered image channels. The transfer of this signal extraction approach to phase-contrast data multiplies the diagnostic information gained within a single CT acquisition. The method is demonstrated with a phantom consisting of exemplary solid and fluid materials as well as a chicken heart with an iodine filled tube simulating a vessel. For this first demonstration all measurements have been conducted at a compact laser-undulator synchrotron X-ray source with a tunable X-ray energy and a narrow spectral bandwidth, to validate the quantitativeness of the processing approach.
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Random number generation is a task that engages working memory and executive processes within the domain of number representation. In the present study we address the role of language in number processing by switching languages during random number generation (numbers 1-9), using German (L1) and English (L2), and alternating L1/L2. Results indicate large correspondence between performance in L1 and L2. In contrast to nonswitching performance, randomization with alternating languages showed a significant increase of omitted responses, whereas the random sequences were less stereotyped, showing significantly less repetition avoidance and cycling behavior. During an intentional switch between languages, errors in language sequence appeared in 23% of responses on the average, independently of the quality of randomization but associated with a clear persistence of L2. These results indicate that random number generation is more closely linked to auditory-phonological representation of numerals than to visual arabic notation.
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Atención/fisiología , Lóbulo Frontal/fisiología , Lenguaje , Memoria/fisiología , Multilingüismo , Desempeño Psicomotor/fisiología , Conducta Verbal/fisiología , Adulto , Análisis de Varianza , Femenino , Humanos , Masculino , Matemática , Pruebas Neuropsicológicas , Solución de Problemas/fisiologíaRESUMEN
The effects of different levels of language proficiency on random number generation were examined in this study. 16 healthy right-handed students (7 women, 9 men; aged 22 to 25 years, M=23.8, SD=.83) attempted to generate a random sequence of the digits 1 to 9 at pacing frequencies of 1, 1.5, and 2 Hz. Randomization was done in German (native language L1), English (first foreign language L2), and French (second foreign language L3). There was a pattern of redundancy and seriation tendencies, increasing with speed of generation for all languages (L1-L3). While using L2 and L3, responses slowed and the number of errors committed increased. Further, there was a peculiar pattern of dissociation in nonrandom performance with an increase of habitual counting in ones and a strong reduction of counting in twos. All effects were most pronounced when subjects used L3 and 2-Hz pacing rates. Slowing and nonrandomness was not correlated with self-assessment parameters regarding language proficiency. We suggest that in a task involving number activation in a nonnative language, lack of proficiency will interfere with random number generation, leading to interruptions and rule breaking, at least when reaching the limits of attentional capacity at higher pacing rates.