Detergent-free purification of ABC (ATP-binding-cassette) transporters

ABC (ATP-binding-cassette) transporters carry out many vital functions and are involved in numerous diseases, but study of the structure and function of these proteins is often hampered by their large size and membrane location. Membrane protein purification usually utilizes detergents to solubilize the protein from the membrane, effectively removing it from its native lipid environment. Subsequently, lipids have to be added back and detergent removed to reconstitute the protein into a lipid bilayer. In the present study, we present the application of a new methodology for the extraction and purification of ABC transporters without the use of detergent, instead, using a copolymer, SMA (polystyrene-co-maleic acid). SMA inserts into a bilayer and assembles into discrete particles, essentially solubilizing the membrane into small discs of bilayer encircled by a polymer, termed SMALPs (SMA lipid particles). We show that this polymer can extract several eukaryotic ABC transporters, P-glycoprotein (ABCB1), MRP1 (multidrug-resistance protein 1; ABCC1), MRP4 (ABCC4), ABCG2 and CFTR (cystic fibrosis transmembrane conductance regulator; ABCC7), from a range of different expression systems. The SMALP-encapsulated ABC transporters can be purified by affinity chromatography, and are able to bind ligands comparably with those in native membranes or detergent micelles. A greater degree of purity and enhanced stability is seen compared with detergent solubilization. The present study demonstrates that eukaryotic ABC transporters can be extracted and purified without ever being removed from their lipid bilayer environment, opening up awide range of possibilities for the future study of their structure and function. © The Authors Journal compilation © 2014 Biochemical Society.

Managing enterprise resource planning and multi-organisational enterprise governance:a new contingency framework for the enterprisation of operations

This research has been undertaken to determine how successful multi-organisational enterprise strategy is reliant on the correct type of Enterprise Resource Planning (ERP) information systems being used. However there appears to be a dearth of research as regards strategic alignment between ERP systems development and multi-organisational enterprise governance as guidelines and frameworks to assist practitioners in making decision for multi-organisational collaboration supported by different types of ERP systems are still missing from theoretical and empirical perspectives. This calls for this research which investigates ERP systems development and emerging practices in the management of multi-organisational enterprises (i.e. parts of companies working with parts of other companies to deliver complex product-service systems) and identify how different ERP systems fit into different multi-organisational enterprise structures, in order to achieve sustainable competitive success. An empirical inductive study was conducted using the Grounded Theory-based methodological approach based on successful manufacturing and service companies in the UK and China. This involved an initial pre-study literature review, data collection via 48 semi-structured interviews with 8 companies delivering complex products and services across organisational boundaries whilst adopting ERP systems to support their collaborative business strategies – 4 cases cover printing, semiconductor manufacturing, and parcel distribution industries in the UK and 4 cases cover crane manufacturing, concrete production, and banking industries in China in order to form a set of 29 tentative propositions that have been validated via a questionnaire receiving 116 responses from 16 companies. The research has resulted in the consolidation of the validated propositions into a novel concept referred to as the ‘Dynamic Enterprise Reference Grid for ERP’ (DERG-ERP) which draws from multiple theoretical perspectives. The core of the DERG-ERP concept is a contingency management framework which indicates that different multi-organisational enterprise paradigms and the supporting ERP information systems are not the result of different strategies, but are best considered part of a strategic continuum with the same overall business purpose of multi-organisational cooperation. At different times and circumstances in a partnership lifecycle firms may prefer particular multi-organisational enterprise structures and the use of different types of ERP systems to satisfy business requirements. Thus the DERG-ERP concept helps decision makers in selecting, managing and co-developing the most appropriate multi-organistional enterprise strategy and its corresponding ERP systems by drawing on core competence, expected competitiveness, and information systems strategic capabilities as the main contingency factors. Specifically, this research suggests that traditional ERP(I) systems are associated with Vertically Integrated Enterprise (VIE); whilst ERPIIsystems can be correlated to Extended Enterprise (EE) requirements and ERPIII systems can best support the operations of Virtual Enterprise (VE). The contribution of this thesis is threefold. Firstly, this work contributes to a gap in the extant literature about the best fit between ERP system types and multi-organisational enterprise structure types; and proposes a new contingency framework – the DERG-ERP, which can be used to explain how and why enterprise managers need to change and adapt their ERP information systems in response to changing business and operational requirements. Secondly, with respect to a priori theoretical models, the new DERG-ERP has furthered multi-organisational enterprise management thinking by incorporating information system strategy, rather than purely focusing on strategy, structural, and operational aspects of enterprise design and management. Simultaneously, the DERG-ERP makes theoretical contributions to the current IS Strategy Formulation Model which does not explicitly address multi-organisational enterprise governance. Thirdly, this research clarifies and emphasises the new concept and ideas of future ERP systems (referred to as ERPIII) that are inadequately covered in the extant literature. The novel DERG-ERP concept and its elements have also been applied to 8 empirical cases to serve as a practical guide for ERP vendors, information systems management, and operations managers hoping to grow and sustain their competitive advantage with respect to effective enterprise strategy, enterprise structures, and ERP systems use; referred to in this thesis as the “enterprisation of operations”.

CamSim:a distributed smart camera network simulator

Smart cameras allow pre-processing of video data on the camera instead of sending it to a remote server for further analysis. Having a network of smart cameras allows various vision tasks to be processed in a distributed fashion. While cameras may have different tasks, we concentrate on distributed tracking in smart camera networks. This application introduces various highly interesting problems. Firstly, how can conflicting goals be satisfied such as cameras in the network try to track objects while also trying to keep communication overhead low? Secondly, how can cameras in the network self adapt in response to the behavior of objects and changes in scenarios, to ensure continued efficient performance? Thirdly, how can cameras organise themselves to improve the overall network's performance and efficiency? This paper presents a simulation environment, called CamSim, allowing distributed self-adaptation and self-organisation algorithms to be tested, without setting up a physical smart camera network. The simulation tool is written in Java and hence allows high portability between different operating systems. Relaxing various problems of computer vision and network communication enables a focus on implementing and testing new self-adaptation and self-organisation algorithms for cameras to use.

Procedure of Formalization of the Indices of Banks’ Stable Functioning in Comparative Estimates of Their Development

The advisability of analyzing the banks liquidity and profitability as the key factor when building the comparative estimate of their functioning is considered. The procedure of formal description of the bank stable functioning indices is substantiated. Fuzzy interpretation of the bank management efficiency estimation is offered. The possibility to formalize the bank functioning estimates on the basis of the corresponding fuzzy set levels hierarchy is analyzed. The comparative estimate of different bank systems functioning is given.

The interconnectedness of ageing:does the convoy principal apply?

This is an excerpt from the content: The convoy principal states that any system is only as functional as its ‘slowest’ unit. As organisms are made up of interconnected networks of physiological systems, it is possible that this principle applies to the biology of ageing. Often biogerontology will focus either on organismal ageing (mechanisms associated with increased longevity of a lower model organism for example), ageing of an individual organ system (such as the cardiovascular/musculoskeletal/immune) or ageing at the cellular level (from telomere length to cellular senescence, with many different cell types being studied) without considering the interconnectedness between the three and importantly, between the separate units of the convoy; the different organ systems. Conceptually, research that aims to identify ‘anti-ageing’ therapies is often deemed to be reaching for a panacea that will arrest or slow down the ageing process as a whole, whereas a more realistic aim is to first identify how we can improve the perfor ...

Investigating ocular allergy:can we determine a better measurement?

Objective: Ocular allergy is a broad group of allergic conditions involving inflammation of the conjunctiva and the most common forms are seasonal allergic conjunctivitis (SAC; 90% of cases) and perennial allergic conjunctivitis (PAC; 5% of cases). The main symptom is ocular itching caused by mast cell degranulation leading to the release of histamine and other mediators such as tryptase. Tryptase is a neutral protease that is selectively concentrated in the secretory granules of human mast cells and has been shown to be a sensitive and specific marker of type I hypersensitivity reaction. The objective was to ascertain the best assay method for determining the tryptase levels in tear samples and whether this can be used to determine the efficacy of non-pharmacological treatments compared to no treatment or their combined effect with anti-allergic medication for SAC and PAC. Method: Thirty patients with a history of SAC were recruited into a randomised blind study during winter months when all the patients were asymptomatic. Suitability was determined by skin prick and conjunctival provocation tests. Patients were randomly assigned to either a non-pharmacological or a pharmacological Intervention group and received each test condition assigned to their group in a randomly assigned order. Symptoms were provoked by exposure to pollen in an environmental test chamber where the temperature, humidity and grass pollen levels were set to a high pollen count day. Tear samples were taken set intervals during the visit and then processed by enzyme linked immunosorbent assay (ELISA) for the detection of tryptase levels. Preliminary results: Results are still being analysed but the preliminary optimisation experiments tested four different ELISA systems; two indirect assays and two capture ‹sandwich› assays. The results suggest that in both sandwich assay systems non-specific binding occurred which could not be easily overcome. The indirect assay systems both showed specific reactions, and the sensitivity achieved was greater with the monoclonal than the polyclonal antibody. Using these findings the indirect assay system was optimised to provide a standardised system for measuring tryptase. Initial trials using human tear samples displayed tryptase levels between 23.1 and 175.1 ng/ml; levels which fall within the anticipated range for patients with SAC. Further statistical work is needed to determine whether tryptase levels vary between the treatments 75.

Europol and the EU’s fight against organized crime: exploring the potential of experimentalist governance

While issues relating to the development, legitimacy and accountability of the European Police Office, Europol, have been intensively discussed in political and academic circles, the actual impact of Europol on policy-making in the European Union has yet to receive scholarly attention. By investigating the evolution and the role of Europol's organized crime reports, this article elaborates on whether Europol has been able to exert an influence beyond its narrowly defined mandate. Theoretically informed by the assumptions of experimentalist governance, the article argues that the different legal systems and policing traditions of EU member states have made it difficult for the EU to agree on a common understanding on how to fight against organized crime. This lack of consensus, which has translated into a set of vague and broadly formulated framework goals and guidelines, has enabled Europol to position its Organized Crime Threat Assessments as the point of reference in the respective EU policy-making area. Europol's interest in improving its institutional standing thereby converged with the interest of different member states to use Europol as a socialization platform to broadcast their ideas and to ‘Europeanize’ their national counter-organized crime policy.

Detection and quantification of protein oxidation in sarcopenic models:a mass spectrometry study

Oxidised biomolecules in aged tissue could potentially be used as biomarkers for age-related diseases; however, it is still unclear whether they causatively contribute to ageing or are consequences of the ageing process. To assess the potential of using protein oxidation as markers of ageing, mass spectrometry (MS) was employed for the identification and quantification of oxidative modifications in obese (ob/ob) mice. Lean muscle mass and strength is reduced in obesity, representing a sarcopenic model in which the levels of oxidation can be evaluated for different muscular systems including calcium homeostasis, metabolism and contractility. Several oxidised residues were identified by tandem MS (MS/MS) in both muscle homogenate and isolated sarcoplasmic reticulum (SR), an organelle that regulates intracellular calcium levels in muscle. These modifications include oxidation of methionine, cysteine, tyrosine, and tryptophan in several proteins such as sarcoplasmic reticulum calcium ATPase (SERCA), glycogen phosphorylase, and myosin. Once modifications had been identified, multiple reaction monitoring MS (MRM) was used to quantify the percentage modification of oxidised residues within the samples. Preliminary data suggests proteins in ob/ob mice are more oxidised than the controls. For example SERCA, which constitutes 60-70% of the SR, had approximately a 2-fold increase in cysteine trioxidation of Cys561 in the obese model when compared to the control. Other obese muscle proteins have also shown a similar increase in oxidation for various residues. Further analysis with complex protein mixtures will determine the potential diagnostic use of MRM experiments for analysing protein oxidation in small biological samples such as muscle needle biopsies.

Az állam szerepe a turizmusban (The role of government in tourism)

Az állam szerepe napjainkban átalakulóban van, ami az egyes gazdasági területeken más és más szabályozórendszer alkalmazását igényli a szakterület sajátosságainak és az iparág igényeinek, nemzetközi környezetének megfelelően. A turizmus azon gazdasági területek közé tartozik, amelynek szerepe az elmúlt időszakban dinamikusan növekedett, többek között jelentős bruttó hazai termék-előállító, munkaigényes jellegéből, valamint jelentős beruházást gerjesztő hatásából adódóan. A turizmus mint üzleti tevékenység nem nélkülözheti az állam szerepét. Számos olyan externália társul hozzá, ami államilag koordinált esetben a fejlődést magasabb pályára állítja, és társadalmilag hatékonyabb szinten valósul meg. Napjainkban az állam szerepének újraértelmezése, feladatainak újbóli átgondolása és a megfelelő súlypontok megtalálása elengedhetetlenül szükséges annak érdekében, hogy a turizmus pozitív hatásai hosszú távon érvényesüljenek. _______________ Nowadays the role of the government is changing, which needs the usage of different regulatory systems regarding the requirements of the industry and the international environment. Tourism belongs to the economics fields which importance are growing very rapidly because of its effects such as GDP contribution effect, workforce effect and investment exciting effect. Tourism as a business activity needs the significant role of the government. Tourism is in association with a lot of externalise, therefore can secure the coordination of the state the development and the efficiency at a higher level. Nowadays the rethinking of the task of the government is needed on the field of tourism in the interest of the enforcement of its positive effects.

A high efficient biometrics approach for unconstrained iris segmentation and recognition

This dissertation develops an innovative approach towards less-constrained iris biometrics. Two major contributions are made in this research endeavor: (1) Designed an award-winning segmentation algorithm in the less-constrained environment where image acquisition is made of subjects on the move and taken under visible lighting conditions, and (2) Developed a pioneering iris biometrics method coupling segmentation and recognition of the iris based on video of moving persons under different acquisitions scenarios. The first part of the dissertation introduces a robust and fast segmentation approach using still images contained in the UBIRIS (version 2) noisy iris database. The results show accuracy estimated at 98% when using 500 randomly selected images from the UBIRIS.v2 partial database, and estimated at 97% in a Noisy Iris Challenge Evaluation (NICE.I) in an international competition that involved 97 participants worldwide involving 35 countries, ranking this research group in sixth position. This accuracy is achieved with a processing speed nearing real time. The second part of this dissertation presents an innovative segmentation and recognition approach using video-based iris images. Following the segmentation stage which delineates the iris region through a novel segmentation strategy, some pioneering experiments on the recognition stage of the less-constrained video iris biometrics have been accomplished. In the video-based and less-constrained iris recognition, the test or subject iris videos/images and the enrolled iris images are acquired with different acquisition systems. In the matching step, the verification/identification result was accomplished by comparing the similarity distance of encoded signature from test images with each of the signature dataset from the enrolled iris images. With the improvements gained, the results proved to be highly accurate under the unconstrained environment which is more challenging. This has led to a false acceptance rate (FAR) of 0% and a false rejection rate (FRR) of 17.64% for 85 tested users with 305 test images from the video, which shows great promise and high practical implications for iris biometrics research and system design.

Isso é o nome das coisas: a palavra-canção em Arnaldo Antunes

This thesis aims to analyze a corpus containing some hybrid poems and some images related to these poems because the omnipresence of some of them in different semiotic systems and even different books of Arnaldo Antunes. The poems in question will be drawn from two of his books: Things and Name; We will also consider some excerpts from songs, which are present in the same author career discs; as well as observe the embodiment of the poem video coming out of the paper support and enters on the TV screen through the VHS / DVD project Name. Our work focuses on this corpus, mainly observing a recurrent feature already observed at Masters level that is the hallmark of firstness, theoretical category developed by Charles Sanders Peirce. In addition to observe the semiotic aspect, we will also be a discussion of the relationship of verbal texts with visual and its nuances with changing media. The semiotic theory is basically anchored in Peirce vision studied by Lucia Santaella on the headquarters of language and thought (noise, verbal and visual). And with regard to the study of the songs, we use the theory of Luiz Tatit, which discusses the verbal intonation and musical indices as the party responsible for global understanding of Song.

O antagonismo do receptor de serotonina do tipo 7 da substância cinzenta periaquedutal dorsal reduz a ansiedade experimental basal, mas não aquela gerada pela retirada do etanol em ratos

Ethanol-dependent individuals who reduce or discontinue its use may present Alcohol Withdrawal Syndrome, which is characterized by unpleasant signs and symptoms, such as anxiety, that may trigger relapses. Ethanol, a psychotropic drug, is able to promote behavioral and neurophysiological changes, acting on different neurotransmitter systems, including the serotonergic, which has also been directly associated with aversive states, including anxiety. This study aimed to investigate the participation of type 7 serotonin receptor (5-HT7) of the dorsal periaqueductal gray (DPAG) on basal experimental anxiety and that caused by ethanol withdrawal. For this, 75-100 days old Wistar rats were subjected to two experiments. On the first one, animals underwent stereotactic surgery for implantation of guide cannulas used for administration of the drug directly into the DPAG. After seven days, the animals received doses of 2.5; 5 and 10 nmols of type 7 receptor antagonist SB269970 (SB) or vehicle intra-DPAG and, ten minutes after, they were exposed to elevated plus maze (EPM). In the following day, the animals were submitted to the same treatment and tested in the open field (OF). In the second experiment, animals received increasing concentrations (2%, 4%, 6%) of ethanol as the only source of liquid diet or water (control group), both with free access to chow. Seventy two hours and ninety six hours after the ethanol withdrawal, animals received SB (2.5 and 5.0 nmols) intraDPAG ten minutes before the test in the LCE and OF, respectively. In experiment 1, the dose of antagonist 10 nmols was able of reversing the anxiety generated by EPM. In the experiment 2, ineffective SB doses on the LCE (2.5 and 5.0 nmol) were not able to reverse the anxiety caused by the ethanol withdrawal in the EPM, although the dose of 2.5 nmols of SB has reversed its hipolocomotor effect in this test. This result suggests that the 5-HT7 receptor is involved in the modulation of the basal experimental anxiety in rats, but not in the anxiety caused by ethanol withdrawal in the DPAG.

Estratégias para obtenção de adequada iluminação natural em escolas: uma análise de sistemas de aberturas para Natal/RN

The purpose of this research is to analyze different daylighting systems in schools in the city of Natal/RN. Although with the abundantly daylight available locally, there are a scarce and diffuse architectural recommendations relating sky conditions, dimensions of daylight systems, shading, fraction of sky visibility, required illuminance, glare, period of occupation and depth of the lit area. This research explores different selected apertures systems to explore the potential of natural light for each system. The method has divided into three phases: The first phase is the modeling which involves the construction of three-dimensional model of a classroom in Sketchup software 2014, which is featured in follow recommendations presented in the literature to obtain a good quality of environmental comfort in school settings. The second phase is the dynamic performance computer simulation of the light through the Daysim software. The input data are the climate file of 2009 the city of Natal / RN, the classroom volumetry in 3ds format with the assignment of optical properties of each surface, the sensor mapping file and the user load file . The results produced in the simulation are organized in a spreadsheet prepared by Carvalho (2014) to determine the occurrence of useful daylight illuminance (UDI) in the range of 300 to 3000lux and build graphics illuminance curves and contours of UDI to identify the uniformity of distribution light, the need of the minimum level of illuminance and the occurrence of glare.

A neuromodulação do córtex pré-frontal dorsolateral na percepção de tempo em contexto neutro ou emocionalmente ativo

The time perception is critical for environmental adaptation in humans and other species. The temporal processing, has evolved through different neural systems, each responsible for processing different time scales. Among the most studied scales is that spans the arrangement of seconds to minutes. Evidence suggests that the dorsolateral prefrontal (DLPFC) cortex has relationship with the time perception scale of seconds. However, it is unclear whether the deficit of time perception in patients with brain injuries or even "reversible lesions" caused by transcranial magnetic stimulation (TMS) in this region, whether by disruption of other cognitive processes (such as attention and working memory) or the time perception itself. Studies also link the region of DLPFC in emotional regulation and specifically the judgment and emotional anticipation. Given this, our objective was to study the role of the dorsolateral prefrontal cortex in the time perception intervals of active and emotionally neutral stimuli, from the effects of cortical modulation by transcranial direct current stimulation (tDCS), through the cortical excitation (anodic current), inhibition (cathode current) and control (sham) using the ranges of 4 and 8 seconds. Our results showed that there is an underestimation when the picture was presented by 8 seconds, with the anodic current in the right DLPFC, there is an underestimation and with cathodic current in the left DLPFC, there is an overestimation of the time reproduction with neutral ones. The cathodic current over the left DLPFC leads to an inverse effect of neutral ones, an underestimation of time with negative pictures. Positive or negative pictures improved estimates for 8 second and positive pictures inhibited the effect of tDCS in DLPFC in estimating time to 4 seconds. With this work, we conclude that the DLPFC plays a key role in the o time perception and largely corresponds to the stages of memory and decision on the internal clock model. The left hemisphere participates in the perception of time in both active and emotionally neutral contexts, and we can conclude that the ETCC and an effective method to study the cortical functions in the time perception in terms of cause and effect.

AWI Bathymetric Chart of the Gakkel Ridge (AWI BCGR) (Scale 1:150,000)

The 1 : 1,500,000 AWI Bathymetric Chart of the Gakkel Ridge (AWI BCGR) has been developed from multibeam data measured during the Arctic Mid-Ocean Ridge Expedition in 2001 (AMORE 2001, ARK-XVII/2). This expedition was conducted to investigate the Gakkel Ridge in the Arctic Ocean and was carried out by the icebreaking research vessels RV Polarstern and USCGC Healy. Polarstern is equipped with the multibeam sonar system Hydrosweep DS-2, whereas Healy carries Seabeam 2112. During the expedition an area of 8890 km length and 18 - 46 km width, situated between 82°N/8°W and 87°N/75°E, was surveyed simultaneously by both vessels. Water depths ranged from 566 to 5673 meters. Dense sea ice cover derogated the sonar measurements and decreased data quality. Data errors were corrected in an extensive post-processing. The data of two different sonar systems had to be consolidated in order to derive a high resolution bathymetry of the Gakkel Ridge. Final result was a digital terrain model (DTM) with a grid spacing of 100 meters, which was utilized for generating the map series AWI Bathymetric Chart of the Gakkel Ridge, consisting of ten map sheets.