Dimitris Pikionis: Roads of the Times

Dimitris Pikionis: Roads of the Times

The relationship between young people's transit use and their perceptions of equity concepts in transit service provision

This study investigates the effect of price and travel mode fairness and spatial equity in transit provision on the perceived transit service quality, willingness to pay, and habitual frequency of use. Based on the theory of planned behavior, we developed a web-based questionnaire for revealed preferences data collection. The survey was administered among young people in Copenhagen and Lisbon to explore the transit perceptions and use under different economic and transit provision conditions. The survey yielded 499 questionnaires, analyzed by means of structural equation models. Results show that higher perceived fairness relates positively to higher perceived quality of transit service and higher perceived ease of paying for transit use. Higher perceived spatial equity in service provision is associated with higher perceived service quality. Higher perceived service quality relates to higher perceived ease of payment, which links to higher frequency of transit use.

Caracterización y análisis del coste de mejora de la calidad de servicio en transporte urbano en autobús, basado en la metodología ABC (activity based costing)

El estudio de la calidad de servicio en transporte público ha tenido distintos enfoques, tanto en la investigación como en sus aplicaciones prácticas, tales como el desarrollo del método SERVQUAL, los proyectos QUATTRO y EQUIP de la Comisión Europea, el Transit Capacity and Quality of Service Manual y finalmente el estándar EN 13816, siendo la consideración principal la calidad desde el punto de vista del usuario, es decir, la calidad percibida, cuyo objetivo en la práctica trata de obtener un Índice de Satisfacción del Cliente como medida de la calidad global. Esta aproximación deja de lado aspectos tanto sobre calidad suministrada como sobre los suministradores del servicio, las administraciones y los operadores, particularmente en cuanto a las implicaciones económicas. El objetivo principal de la presente tesis es la estimación del impacto económico real de la mejora de atributos de calidad para un operador de autobús urbano. En el capítulo 2, se lleva a cabo un análisis del estado del arte sobre el estudio de la calidad, tanto de los métodos de estudio, como de su aplicación al campo de transporte público, y, específicamente, del estudio de costes. En los capítulos 3 y 4 se propone la aplicación de la metodología ABC (Activity Based Costing) para el caso de un contrato de servicio de transporte público en autobús en la Comunidad de Madrid, concretamente la conexión de Tres Cantos. Para ello se utilizan datos de encuestas de calidad esperada y percibida por los usuarios, junto con información económica y de actividad, así como entrevistas a personal de la empresa, conformando todo ello una metodología compacta de investigación y análisis. Los resultados de las encuestas y los análisis posteriores identificaron la frecuencia de servicio, la puntualidad de los autobuses y la seguridad de la conducción como las tres variables mas importantes desde el punto de vista del usuario, y ha sido en referencia a estas tres variables sobre las que se ha desarrollado la aplicación del método ABC contenida en el capítulo 5. Los resultados descritos en el capítulo 6 muestran la consistencia y la aplicabilidad de esta metodología, superando las simplificaciones de los procedimientos tradicionales contables. En cuanto a los resultados obtenidos, se ha comprobado que una aproximación tradicional basada en costes por kilometro puede llegar a sobrevalorar los costes asociados a mejoras de calidad hasta tres veces los costes calculados según el método ABC. Finalmente en el capítulo 7 se recogen las principales conclusiones y se apuntan algunas líneas futuras de investigación basadas en mejoras sobre la aplicación y transferibilidad de los resultados. The study of service quality and its implication for transport contracts has several approaches in research and practical applications, such as the development of the SERVQUAL method, the UE QUATTRO and EQUIP research projects, the Transit Capacity and Quality of Service Manual and finally the EN 13816 standard. In all cases the main stream is the consideration of quality from the user’s point of view, thus obtaining a Customer Satisfaction Index as a measurement of the overall service quality. This approach ignores quality consequences for transport providers, authorities and operators, particularly economic implications. The main target of this thesis is to estimate the real economic impact of improving quality attributes for a bus operator. Chapter 2 includes a thorough analysis of the state of the art of general methodologies on service quality, their applications for public transport, and particularly for quality costing. In chapters 3 and 4, the Activity Based Costing (ABC) methodology is proposed for a suburban bus contract in Madrid, Tres Cantos. Perceived and expected quality surveys are used together with economic and performance information, as well as interviews with key staff from the operator. The whole set of surveys, interviews and ABC methodology puts together a comprehensive approach for analysis and research. Surveys and subsequent analysis shows service headway, punctuality and safety as the three most important service attributes from the user’s point of view. The proposed methodology has been applied to these three variables, as described in chapter 5. Results obtained in chapter 6 show the consistency and practicality of this methodology, overcoming simplifications from traditional accounting procedures. According to the latter, a calculation of costs based on mileage can overvalue quality improvements up to three times the results following ABC methodology. Finally, in chapter 7, the main conclusions are highlighted and some future research is suggested in terms of application improvements as well as results transferability.

Regulation of chloroplast protein import through a protochlorophyllide-responsive transit peptide

NADPH:protochlorophyllide (Pchlide) oxidoreductase (POR) is the key enzyme of chlorophyll biosynthesis in angiosperms. In barley, two POR enzymes, termed PORA and PORB, exist. Both are nucleus-encoded plastid proteins that must be imported posttranslationally from the cytosol. Whereas the import of the precursor of PORA, pPORA, previously has been shown to depend on Pchlide, the import of pPORB occurred constitutively. To study this striking difference, chimeric precursor proteins were constructed in which the transit sequences of the pPORA and pPORB were exchanged and fused to either their cognate polypeptides or to a cytosolic dihydrofolate reductase (DHFR) reporter protein of mouse. As shown here, the transit peptide of the pPORA (transA) conferred the Pchlide requirement of import onto both the mature PORB and the DHFR. By contrast, the transit peptide of the pPORB directed the reporter protein into both chloroplasts that contained or lacked translocation-active Pchlide. In vitro binding studies further demonstrated that the transit peptide of the pPORA, but not of the pPORB, is able to bind Pchlide. We conclude that the import of the authentic pPORA and that of the transA-PORB and transA-DHFR fusion proteins is regulated by a direct transit peptide-Pchlide interaction, which is likely to occur in the plastid envelope, a major site of porphyrin biosynthesis.

Dominant transformation by mutated human ras genes in vitro requires more than 100 times higher expression than is observed in cancers

The gene-mutation-cancer hypothesis holds that mutated cellular protooncogenes, such as point-mutated proto-ras, “play a dominant part in cancer,” because they are sufficient to transform transfected mouse cell lines in vitro [Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, K. & Watson, J. D. (1994) Molecular Biology of the Cell (Garland, New York)]. However, in cells transformed in vitro mutated human ras genes are expressed more than 100-fold than in the cancers from which they are isolated. In view of the discrepancy between the very low levels of ras transcription in cancers and the very high levels in cells transformed in vitro, we have investigated the minimal level of human ras expression for transformation in vitro. Using point-mutated human ras genes recombined with different promoters from either human metallothionein-IIA or human fibronectin or from retroviruses we found dominant in vitro transformation of the mouse C3H cell line only with ras genes linked to viral promoters. These ras genes were expressed more than 120-fold higher than are native ras genes of C3H cells. The copy number of transfected ras genes ranged from 2–6 in our system. In addition, nondominant transformation was observed in a small percentage (2–7%) of C3H cells transfected with ras genes that are expressed less than 20 times higher than native C3H ras genes. Because over 90% of cells expressing ras at this moderately enhanced level were untransformed, transformation must follow either a nondominant ras mechanism or a non-ras mechanism. We conclude that the mutated, but normally expressed, ras genes found in human and animal cancers are not likely to “play a dominant part in cancer.” The conclusion that mutated ras genes are not sufficient or dominant for cancer is directly supported by recent discoveries of mutated ras in normal animals, and in benign human tissue, “which has little potential to progress” [Jen, J., Powell, S. M., Papadopoulos, N., Smith, K. J., Hamilton, S. R., Vogelstein, B. & Kinzler, K. W. (1994) Cancer Res. 54, 5523–5526]. Even the view that mutated ras is necessary for cancer is hard to reconcile with (i) otherwise indistinguishable cancers with and without ras mutations, (ii) metastases of the same human cancers with and without ras mutations, (iii) retroviral ras genes that are oncogenic without point mutations, and (iv) human tumor cells having spontaneously lost ras mutation but not tumorigencity.

Determining divergence times with a protein clock: Update and reevaluation

A recent study of the divergence times of the major groups of organisms as gauged by amino acid sequence comparison has been expanded and the data have been reanalyzed with a distance measure that corrects for both constraints on amino acid interchange and variation in substitution rate at different sites. Beyond that, the availability of complete genome sequences for several eubacteria and an archaebacterium has had a great impact on the interpretation of certain aspects of the data. Thus, the majority of the archaebacterial sequences are not consistent with currently accepted views of the Tree of Life which cluster the archaebacteria with eukaryotes. Instead, they are either outliers or mixed in with eubacterial orthologs. The simplest resolution of the problem is to postulate that many of these sequences were carried into eukaryotes by early eubacterial endosymbionts about 2 billion years ago, only very shortly after or even coincident with the divergence of eukaryotes and archaebacteria. The strong resemblances of these same enzymes among the major eubacterial groups suggest that the cyanobacteria and Gram-positive and Gram-negative eubacteria also diverged at about this same time, whereas the much greater differences between archaebacterial and eubacterial sequences indicate these two groups may have diverged between 3 and 4 billion years ago.

In vivo NMR and MRI using injection delivery of laser-polarized xenon

Because xenon NMR is highly sensitive to the local environment, laser-polarized xenon could be a unique probe of living tissues. Realization of clinical and medical science applications beyond lung airspace imaging requires methods of efficient delivery of laser-polarized xenon to tissues, because of the short spin-lattice relaxation times and relatively low concentrations of xenon attainable in the body. Preliminary results from the application of a polarized xenon injection technique for in vivo 129Xe NMR/MRI are extrapolated along with a simple model of xenon transit to show that the peak local concentration of polarized xenon delivered to tissues by injection may exceed that delivered by respiration by severalfold.

Characterization of the Golgi Complex Cleared of Proteins in Transit and Examination of Calcium Uptake Activities

To characterize endogenous molecules and activities of the Golgi complex, proteins in transit were >99% cleared from rat hepatocytes by using cycloheximide (CHX) treatment. The loss of proteins in transit resulted in condensation of the Golgi cisternae and stacks. Isolation of a stacked Golgi fraction is equally efficient with or without proteins in transit [control (CTL SGF1) and cycloheximide (CHX SGF1)]. Electron microscopy and morphometric analysis showed that >90% of the elements could be positively identified as Golgi stacks or cisternae. Biochemical analysis showed that the cis-, medial-, trans-, and TGN Golgi markers were enriched over the postnuclear supernatant 200- to 400-fold with and 400- to 700-fold without proteins in transit. To provide information on a mechanism for import of calcium required at the later stages of the secretory pathway, calcium uptake into CTL SGF1 and CHX SGF1 was examined. All calcium uptake into CTL SGF1 was dependent on a thapsigargin-resistant pump not resident to the Golgi complex and a thapsigargin-sensitive pump resident to the Golgi. Experiments using CHX SGF1 showed that the thapsigargin-resistant activity was a plasma membrane calcium ATPase isoform in transit to the plasma membrane and the thapsigargin-sensitive pump was a sarcoplasmic/endoplasmic reticulum calcium ATPase isoform. In vivo both of these calcium ATPases function to maintain millimolar levels of calcium within the Golgi lumen.

Intramolecular vibrational dephasing obeys a power law at intermediate times

Experimental intramolecular vibrational dephasing transients for several large organic molecules are reanalyzed. Fits to the experimental data, as well as full numerical quantum calculations with a factorized potential surface for all active degrees of freedom of fluorene indicate that power law decays, not exponentials, occur at intermediate times. The results support a proposal that power law decays describe vibrational dephasing dynamics in large molecules at intermediate times because of the local nature of energy flow.

Using rare mutations to estimate population divergence times: A maximum likelihood approach

In this paper we propose a method to estimate by maximum likelihood the divergence time between two populations, specifically designed for the analysis of nonrecurrent rare mutations. Given the rapidly growing amount of data, rare disease mutations affecting humans seem the most suitable candidates for this method. The estimator RD, and its conditional version RDc, were derived, assuming that the population dynamics of rare alleles can be described by using a birth–death process approximation and that each mutation arose before the split of a common ancestral population into the two diverging populations. The RD estimator seems more suitable for large sample sizes and few alleles, whose age can be approximated, whereas the RDc estimator appears preferable when this is not the case. When applied to three cystic fibrosis mutations, the estimator RD could not exclude a very recent time of divergence among three Mediterranean populations. On the other hand, the divergence time between these populations and the Danish population was estimated to be, on the average, 4,500 or 15,000 years, assuming or not a selective advantage for cystic fibrosis carriers, respectively. Confidence intervals are large, however, and can probably be reduced only by analyzing more alleles or loci.