Immunolocalization of ion transport proteins in human autosomal dominant polycystic kidney epithelial cells.

The kidneys of patients with autosomal dominant polycystic kidney disease become massively enlarged due to the progressive expansion of myriad fluid-filled cysts. The epithelial cells that line the cyst walls are responsible for secreting the cyst fluid, but the mechanism through which this secretion occurs is not well established. Recent studies suggest that renal cyst epithelial cells actively secrete Cl across their apical membranes, which in turn drives the transepithelial movement of Na and water. The characteristics of this secretory flux suggest that it is dependent upon the participation of an apical cystic fibrosis transmembrane conductance regulator (CFTR)-like Cl channel and basolateral Na,K-ATPase. To test this hypothesis, we have immunolocalized the CFTR and Na,K-ATPase proteins in intact cysts and in cyst epithelial cells cultured in vitro on permeable filter supports. In both settings, cyst epithelial cells were found to possess Na,K-ATPase exclusively at their basolateral surfaces; apical labeling was not detected. The CFTR protein was present at the apical surfaces of cyst epithelial cells that had been stimulated to secrete through incubation in forskolin. CFTR was detected in intracellular structures in cultured cyst epithelial cells that had not received the forskolin treatment. These results demonstrate that the renal epithelial cells that line cysts in autosomal dominant polycystic kidney disease express transport systems with the appropriate polarity to mediate active Cl and fluid secretion.

Glycoprotein 330/megalin: probable role in receptor-mediated transport of apolipoprotein J alone and in a complex with Alzheimer disease amyloid beta at the blood-brain and blood-cerebrospinal fluid barriers.

A soluble form of Alzheimer disease amyloid beta-protein (sA beta) is transported in the blood and cerebrospinal fluid mainly complexed with apolipoprotein J (apoJ). Using a well-characterized in situ perfused guinea pig brain model, we recently obtained preliminary evidence that apoJ facilitates transport of sA beta (1-40)-apoJ complexes across the blood-brain barrier and the blood-cerebrospinal fluid barrier, but the mechanisms remain poorly understood. In the present study, we examined the transport process in greater detail and investigated the possible role of glycoprotein 330 (gp330)/megalin, a receptor for multiple ligands, including apoJ. High-affinity transport systems with a Km of 0.2 and 0.5 nM were demonstrated for apoJ at the blood-brain barrier and the choroid epithelium in vivo, suggesting a specific receptor-mediated mechanism. The sA beta (1-40)-apoJ complex shared the same transport mechanism and exhibited 2.4- to 10.2-fold higher affinity than apoJ itself. Binding to microvessels, transport into brain parenchyma, and choroidal uptake of both apoJ and sA beta (1-40)-apoJ complexes were markedly inhibited (74-99%) in the presence of a monoclonal antibody to gp330/megalin and were virtually abolished by perfusion with the receptor-associated protein, which blocks binding of all known ligands to gp330. Western blot analysis of cerebral microvessels with the monoclonal antibody to gp330 revealed a protein with a mass identical to that in extracts of kidney membranes enriched with gp330/megalin, but in much lower concentration. The findings suggest that gp330/megalin mediates cellular uptake and transport of apoJ and sA beta (1-40)-apoJ complex at the cerebral vascular endothelium and choroid epithelium.

The EU regime on biofuels in transport: Still in search of sustainability. Egmont Paper No. 68, July 2014

Like other regions of the world, the EU is developing biofuels in the transport sector to reduce oil consumption and mitigate climate change. To promote them, it has adopted favourable legislation since the 2000s. In 2009 it even decided to oblige each Member State to ensure that by 2020 the share of energy coming from renewable sources reached at least 10% of their final consumption of energy in the transport sector. Biofuels are considered the main instrument to reach that percentage since the development of other alternatives (such as hydrogen and electricity) will take much longer than expected. Meanwhile, these various legislative initiatives have driven the production and consumption of biofuels in the EU. Biofuels accounted for 4.7% of EU transport fuel consumption in 2011. They have also led to trade and investment in biofuels on a global scale. This large-scale expansion of biofuels has, however, revealed numerous negative impacts. These stem from the fact that first-generation biofuels (i.e., those produced from food crops), of which the most important types are biodiesel and bioethanol, are used almost exclusively to meet the EU’s renewable 10% target in transport. Their negative impacts are: socioeconomic (food price rises), legal (land-grabbing), environmental (for instance, water stress and water pollution; soil erosion; reduction of biodiversity), climatic (direct and indirect land-use effects resulting in more greenhouse gas emissions) and public finance issues (subsidies and tax relief). The extent of such negative impacts depends on how biofuel feedstocks are produced and processed, the scale of production, and in particular, how they influence direct land use change (DLUC) and indirect land use change (ILUC) and the international trade. These negative impacts have thus provoked mounting debates in recent years, with a particular focus on ILUC. They have forced the EU to re-examine how it deals with biofuels and submit amendments to update its legislation. So far, the EU legislation foresees that only sustainable biofuels (produced in the EU or imported) can be used to meet the 10% target and receive public support; and to that end, mandatory sustainability criteria have been defined. Yet they have a huge flaw. Their measurement of greenhouse gas savings from biofuels does not take into account greenhouse gas emissions resulting from ILUC, which represent a major problem. The Energy Council of June 2014 agreed to set a limit on the extent to which firstgeneration biofuels can count towards the 10% target. But this limit appears to be less stringent than the ones made previously by the European Commission and the European Parliament. It also agreed to introduce incentives for the use of advanced (second- and third-generation) biofuels which would be allowed to count double towards the 10% target. But this again appears extremely modest by comparison with what was previously proposed. Finally, the approach chosen to take into account the greenhouse gas emissions due to ILUC appears more than cautious. The Energy Council agreed that the European Commission will carry out a reporting of ILUC emissions by using provisional estimated factors. A review clause will permit the later adjustment of these ILUC factors. With such legislative orientations made by the Energy Council, one cannot consider yet that there is a major shift in the EU biofuels policy. Bolder changes would have probably meant risking the collapse of the high-emission conventional biodiesel industry which currently makes up the majority of Europe’s biofuel production. The interests of EU farmers would have also been affected. There is nevertheless a tension between these legislative orientations and the new Commission’s proposals beyond 2020. In any case, many uncertainties remain on this issue. As long as solutions have not been found to minimize the important collateral damages provoked by the first generation biofuels, more scientific studies and caution are needed. Meanwhile, it would be wise to improve alternative paths towards a sustainable transport sector, i.e., stringent emission and energy standards for all vehicles, better public transport systems, automobiles that run on renewable energy other than biofuels, or other alternatives beyond the present imagination.

Travel behaviour change in old age : the role of critical incidents in public transport

Older people’s travel behaviour is affected by negative or positive critical incidents in the public transport environment. With the objective of identifying such inci- dents during whole trips and examining how travel beha- viour had changed, we have conducted in-depth interviews with 30 participants aged 65–91 years in the County of Stockholm, Sweden. Out of 469 incidents identified, 77 were reported to have resulted in travel behaviour change, 67 of them in a negative way. Most critical incidents were encountered in the physical environment on-board vehicles and at stations/stops as well as in pricing/ticketing. The findings show that more personal assistance, better driving behaviour, and swift maintenance of elevators and escala- tors are key facilitators that would improve predictability in travelling and enhance vulnerable older travellers’ feeling of security. The results demonstrate the benefit of involving different groups of end users in future planning and design, such that transport systems would meet the various needs of its end users.

Arsenic transport by zebrafish aquaglyceroporins

Background Arsenic is one of the most ubiquitous toxins and endangers the health of tens of millions of humans worldwide. It is a mainly a water-borne contaminant. Inorganic trivalent arsenic (AsIII) is one of the major species that exists environmentally. The transport of AsIII has been studied in microbes, plants and mammals. Members of the aquaglyceroporin family have been shown to actively conduct AsIII and its organic metabolite, monomethylarsenite (MAsIII). However, the transport of AsIII and MAsIII in in any fish species has not been characterized. Results In this study, five members of the aquaglyceroporin family from zebrafish (Danio rerio) were cloned, and their ability to transport water, glycerol, and trivalent arsenicals (AsIII and MAsIII) and antimonite (SbIII) was investigated. Genes for at least seven aquaglyceroporins have been annotated in the zebrafish genome project. Here, five genes which are close homologues to human AQP3, AQP9 and AQP10 were cloned from a zebrafish cDNA preparation. These genes were namedaqp3, aqp3l, aqp9a, aqp9b and aqp10 according to their similarities to the corresponding human AQPs. Expression of aqp9a, aqp9b, aqp3, aqp3l and aqp10 in multiple zebrafish organs were examined by RT-PCR. Our results demonstrated that these aquaglyceroporins exhibited different tissue expression. They are all detected in more than one tissue. The ability of these five aquaglyceroporins to transport water, glycerol and the metalloids arsenic and antimony was examined following expression in oocytes from Xenopus leavis. Each of these channels showed substantial glycerol transport at equivalent rates. These aquaglyceroporins also facilitate uptake of inorganic AsIII, MAsIII and SbIII. Arsenic accumulation in fish larvae and in different tissues from adult zebrafish was studied following short-term arsenic exposure. The results showed that liver is the major organ of arsenic accumulation; other tissues such as gill, eye, heart, intestine muscle and skin also exhibited significant ability to accumulate arsenic. The zebrafish larvae also accumulate considerable amounts of arsenic. Conclusion This is the first molecular identification of fish arsenite transport systems and we propose that the extensive expression of the fish aquaglyceroporins and their ability to transport metalloids suggests that aquaglyceroporins are the major pathways for arsenic accumulation in a variety of zebrafish tissues. Uptake is one important step of arsenic metabolism. Our results will contribute to a new understanding of aquatic arsenic metabolism and will support the use of zebrafish as a new model system to study arsenic associated human diseases.

Perceived Accessibility of Public Transport as a Potential Indicator of Social Inclusion

Perceived accessibility has been acknowledged as an important aspect of transport policy since the 70s. Nevertheless, very few empirical studies have been conducted in this field. When aiming to improve social inclusion, by making sus-tainable transport modes accessible to all, it is important to understand the factors driving perceived accessibility. Un-like conventional accessibility measures, perceived accessibility focuses on the perceived possibilities and ease of en-gaging in preferred activities using different transport modes. We define perceived accessibility in terms of how easy it is to live a satisfactory life with the help of the transport system, which is not necessarily the same thing as the objec-tive standard of the system. According to previous research, perceived accessibility varies with the subjectively-rated quality of the mode of transport. Thus, improvements in quality (e.g. trip planning, comfort, or safety) increase the per-ceived accessibility and make life easier to live using the chosen mode of transport. This study (n=750) focuses on the perceived accessibility of public transport, captured using the Perceived Accessibility Scale PAC (Lättman, Olsson, & Fri-man, 2015). More specifically, this study aims to determine how level of quality affects the perceived accessibility in public transport. A Conditional Process Model shows that, in addition to quality, feeling safe and frequency of travel are important predictors of perceived accessibility. Furthermore, elderly and those in their thirties report a lower level of perceived accessibility to their day-to-day activities using public transport. The basic premise of this study is that sub-jective experiences may be as important as objective indicators when planning and designing for socially inclusive transport systems.

Failure detection for transport processes on networks

Diffusion on networks is a convenient framework to describe transport systems of different nature (from biological transport systems to urban mobility). The mathematical models are based on master equations that describe the diffusion processes by means of the weighted Laplacian matrix that connects the nodes. The link weight represent the coupling strength between the nodes. In this thesis we cope with the problem of localizing a single-edge failure that occurs in the network. An edge failure is meant to be as a sudden decrease of its transport capacities. An incomplete observation of the dynamical state of the network is available. An optimal clustering procedure based on the correlation properties among the node states is proposed. The network dimensionality is then reduced introducing representative nodes for each cluster, whose dynamical state is observed. We check the efficiency of the failure localization for our clustering method in comparison with more traditional techniques, using different graph configurations.

Study of an advanced transport system exploiting magnetic levitation: the Beckhoff XPlanar

The transport system is one of the most important components to be chosen in the design of an automatic machine. There is a wide variety of different choices that can be made in picking this element, each one having its own strengths and its own drawbacks. If it is desired to obtain some elaborate behaviour from the transport system, it is a good idea to think about some flexible and advanced solutions. Among these transport systems, the newest is the Beckhoff XPlanar. This transport system exploits magnetic levitation to move some passive magnetic movers on a completely customizable plane, in an entirely contact-free way. This provides a fast, clean, and noiseless motion, which is extremely desirable in a modern automatic machine. The purpose of this Thesis is to analyse the potentialities and the problems of this new device, starting from the basics. After having presented in detail the topic, an analysis on the hardware components needed to build this system is performed. Then, it is conducted a study on the concepts needed to know how to build a controller having the purpose of dealing with this system. After that, the various types of motion are studied and executed and, later on, some experiments on the real kit are carried out. These studies start from the diagnostic and involve other analyses that are used to test the limits of this transport system. In performing these analyses, it is noticed how the kit presents some problems in reaching the limits of the dynamics. Finally, two different types of station cycle are implemented, which are useful to get a rough idea on the potentialities of this new advanced transport system.

Endophytic population of Pantoea agglomerans in citrus plants and development of a cloning vector for endophytes

Harmless bacteria inhabiting inner plant tissues are termed endophytes. Population fluctuations in the endophytic bacterium Pantoea agglomerans associated with two species of field cultured citrus plants were monitored over a two-year period. The results demonstrated that populations of P. agglomerans fluctuated in Citrus reticulata but not C. sinensis. A cryptic plasmid pPA3.0 (2.9 kb) was identified in 35 out of 44 endophytic isolates of P. agglomerans and was subsequently sequenced. The origins of replication were identified and nine out of 18 open reading frames (ORFs) revealed homology with described proteins. Notably, two ORFs were related to cellular transport systems and plasmid maintenance. Plasmid pPA3.0 was cloned and the gfp gene inserted to generate the pPAGFP vector. The vector was introduced into P. agglomerans isolates and revealed stability was dependent on the isolate genotype, ninety-percent stability values were reached after 60 hours of bacterial cultivation in most evaluated isolates. In order to definitively establish P. agglomerans as an endophyte, the non-transformed bacterium was reintroduced into in vitro cultivated seedlings and the density of inner tissue colonization in inoculated plants was estimated by bacterium re-isolation, while the tissue niches preferred by the bacterium were investigated by scanning electronic microscopy (SEM). Cells from P. agglomerans (strain ARB18) at similar densities were re-isolated from roots, stems and leaves and colonization of parenchyma and xylem tissues were observed. Data suggested that P. agglomerans is a ubiquitous citrus endophyte harboring cryptic plasmids. These characteristics suggest the potential to use the bacterium as a vehicle to introduce new genes in host plants via endophytic bacterial transformation.

Development and evaluation of neural network freeway incident detection models using field data

This paper discusses a multi-layer feedforward (MLF) neural network incident detection model that was developed and evaluated using field data. In contrast to published neural network incident detection models which relied on simulated or limited field data for model development and testing, the model described in this paper was trained and tested on a real-world data set of 100 incidents. The model uses speed, flow and occupancy data measured at dual stations, averaged across all lanes and only from time interval t. The off-line performance of the model is reported under both incident and non-incident conditions. The incident detection performance of the model is reported based on a validation-test data set of 40 incidents that were independent of the 60 incidents used for training. The false alarm rates of the model are evaluated based on non-incident data that were collected from a freeway section which was video-taped for a period of 33 days. A comparative evaluation between the neural network model and the incident detection model in operation on Melbourne's freeways is also presented. The results of the comparative performance evaluation clearly demonstrate the substantial improvement in incident detection performance obtained by the neural network model. The paper also presents additional results that demonstrate how improvements in model performance can be achieved using variable decision thresholds. Finally, the model's fault-tolerance under conditions of corrupt or missing data is investigated and the impact of loop detector failure/malfunction on the performance of the trained model is evaluated and discussed. The results presented in this paper provide a comprehensive evaluation of the developed model and confirm that neural network models can provide fast and reliable incident detection on freeways. (C) 1997 Elsevier Science Ltd. All rights reserved.

Vascular biology of magnesium and its transporters in hypertension

Magnesium may influence blood pressure by modulating vascular tone and structure through its effects on myriad biochemical reactions that control vascular contraction/dilation, growth/apoptosis, differentiation and inflammation. Magnesium acts as a calcium channel antagonist, it stimulates production of vasodilator prostacyclins and nitric oxide and it alters vascular responses to vasoconstrictor agents. Mammalian cells regulate Mg(2+) concentration through special transport systems that have only recently been characterized. Magnesium efflux occurs via Na(2+)-dependent and Na(2+)-independent pathways. Mg(2+) influx is controlled by recently cloned transporters including Mrs2p, SLC41A1, SLC41A2, ACDP2, MagT1, TRPM6 and TRPM7. Alterations in some of these systems may contribute to hypomagnesemia and intracellular Mg(2+) deficiency in hypertension and other cardiovascular pathologies. In particular, increased Mg(2+) efflux through dysregulation of the vascular Na(+)/Mg(2+) exchanger and decreased Mg(2+) influx due to defective vascular and renal TRPM6/7 expression/activity may be important in altered vasomotor tone and consequently in blood pressure regulation. The present review discusses the role of Mg(2+) in vascular biology and implications in hypertension and focuses on the putative transport systems that control magnesium homeostasis in the vascular system. Much research is still needed to clarify the exact mechanisms of cardiovascular Mg(2+) regulation and the implications of aberrant cellular Mg(2+) transport and altered cation status in the pathogenesis of hypertension and other cardiovascular diseases.

Amino acids and their transporters in the retina

This review provides an overview of the distributions, properties and roles of amino acid transport systems in normal and pathological retinal tissues and discusses the roles of specific identified transporters in the mammalian retina. The retina is used in this context as a vehicle for describing neuronal and glial properties. which are in semi, but not all cases comparable to those found elsewhere an the brain. Where significant departures are noted, these are discussed in the context of functional specialisations of the retina and its relationship to adjacent supporting tissues such as the retinal pigment epithelium. Specific examples are given where immunocytochemical labelling for amino acid transporters may yield inaccurate results, possibly because of activity-dependent conformation changes of epitopes in these proteins which render the epitopes more or less accessible to antibodies. (C) 2001 Elsevier Science Ltd. All rights reserved.

Modelagem e simulação de sistema logístico de distribuição de carne de frango

O sistema logístico para distribuição de produtos acabados caracteriza-se pela integração dos serviços de comunicação, transporte e financeiros com a finalidade de atender às demandas do consumidor final. Estima-se que no estado do Espírito Santo, o consumo de carne de frango seja de 44,4 quilos per capita por ano. Para atender a esta demanda, o estado conta com matadouros-frigoríficos distribuídos pelo seu território, bem como, com a participação de outras empresas localizadas no país. Em sistemas de transportes, são característicos Problemas de Roteamento de Veículos (VRP), que precisam ser estudados, caracterizados e otimizados, normalmente, através de rotinas computacionais, que permitem avaliar maior quantidade de variáveis. O presente trabalho teve por objetivo caracterizar um VRP de um matadouro-frigorífico da região do Sul do Espírito Santo e desenvolver um aplicativo computacional que seja suporte para os gestores de logística, servindo para avaliar e propor rotas, e analisar parâmetros logísticos do processo de distribuição de produtos. No desenvolvimento do aplicativo computacional foi necessário caracterizar o sistema logístico da empresa, coletar e analisar os dados das operações logísticas, desenvolver as rotinas computacionais que representassem o sistema em estudo, verificar a confiabilidade dos resultados fornecidos pelo aplicativo, validá-lo e então, poder realizar as experimentações. O aplicativo desenvolvido permitiu reproduzir dados do sistema estudado e avaliar rotas segundo parâmetros logísticos. Pode-se concluir que o aplicativo computacional desenvolvido é útil aos gestores de logística, permitindo a avaliação das rotas praticadas e de novas configurações de rotas.

Mobi-system: assistente pessoal de viagem

Este trabalho sugere uma solução de integração de dados em tempo real no contexto dos transportes públicos. Com o aumento das alternativas oferecidas aos utilizadores dos transportes públicos é importante que estes conheçam todas as alternativas com base em informação em tempo real para que realizem a escolha que melhor se enquadre às suas necessidades. Por outro lado, os operadores de transportes públicos deverão ser capazes de disponibilizar toda a informação pretendida com o mínimo de esforço ou de alterações ao sistema que têm implementado. Neste trabalho serão utilizadas ferramentas que permitem fornecer uma visão homogénea das várias fontes de dados heterogéneas, sendo essa homogeneidade o ponto de integração de todas as fontes de dados com as aplicações cliente.