An endophilin-dynamin complex promotes budding of clathrin-coated vesicles during synaptic vesicle recycling

Clathrin-mediated vesicle recycling in synapses is maintained by a unique set of endocytic proteins and interactions. We show that endophilin localizes in the vesicle pool at rest and in spirals at the necks of clathrin-coated pits (CCPs) during activity in lamprey synapses. Endophilin and dynamin colocalize at the base of the clathrin coat. Protein spirals composed of these proteins on lipid tubes in vitro have a pitch similar to the one observed at necks of CCPs in living synapses, and lipid tubules are thinner than those formed by dynamin alone. Tubulation efficiency and the amount of dynamin recruited to lipid tubes are dramatically increased in the presence of endophilin. Blocking the interactions of the endophilin SH3 domain in situ reduces dynamin accumulation at the neck and prevents the formation of elongated necks observed in the presence of GTPγS. Therefore, endophilin recruits dynamin to a restricted part of the CCP neck, forming a complex, which promotes budding of new synaptic vesicles.

Amphiphysin is a component of clathrin coats formed during synaptic vesicle recycling at the lamprey giant synapse

Amphiphysin is a protein enriched at mammalian synapses thought to function as a clathrin accessory factor in synaptic vesicle endocytosis. Here we examine the involvement of amphiphysin in synaptic vesicle recycling at the giant synapse in the lamprey. We show that amphiphysin resides in the synaptic vesicle cluster at rest and relocates to sites of endocytosis during synaptic activity. It accumulates at coated pits where its SH3 domain, but not its central clathrin/AP-2-binding (CLAP) region, is accessible for antibody binding. Microinjection of antibodies specifically directed against the CLAP region inhibited recycling of synaptic vesicles and caused accumulation of clathrin-coated intermediates with distorted morphology, including flat patches of coated presynaptic membrane. Our data provide evidence for an activity-dependent redistribution of amphiphysin in intact nerve terminals and show that amphiphysin is a component of presynaptic clathrin-coated intermediates formed during synaptic vesicle recycling.

Colocalization of synapsin and actin during synaptic vesicle recycling

It has been hypothesized that in the mature nerve terminal, interactions between synapsin and actin regulate the clustering of synaptic vesicles and the availability of vesicles for release during synaptic activity. Here, we have used immunogold electron microscopy to examine the subcellular localization of actin and synapsin in the giant synapse in lamprey at different states of synaptic activity. In agreement with earlier observations, in synapses at rest, synapsin immunoreactivity was preferentially localized to a portion of the vesicle cluster distal to the active zone. During synaptic activity, however, synapsin was detected in the pool of vesicles proximal to the active zone. In addition, actin and synapsin were found colocalized in a dynamic filamentous cytomatrix at the sites of synaptic vesicle recycling, endocytic zones. Synapsin immunolabeling was not associated with clathrin-coated intermediates but was found on vesicles that appeared to be recycling back to the cluster. Disruption of synapsin function by microinjection of antisynapsin antibodies resulted in a prominent reduction of the cytomatrix at endocytic zones of active synapses. Our data suggest that in addition to its known function in clustering of vesicles in the reserve pool, synapsin migrates from the synaptic vesicle cluster and participates in the organization of the actin-rich cytomatrix in the endocytic zone during synaptic activity.

Cement As a Waste Form for Nuclear Fission Products: The Case of 90Sr and Its Daughters

One of the main challenges faced by the nuclear industry is the long-term confinement of nuclear waste. Because it is inexpensive and easy to manufacture, cement is the material of choice to store large volumes of radioactive materials, in particular the low-level medium-lived fission products. It is therefore of utmost importance to assess the chemical and structural stability of cement containing radioactive species. Here, we use ab initio calculations based on density functional theory (DFT) to study the effects of ⁹⁰Sr insertion and decay in C-S-H (calcium-silicate-hydrate) in order to test the ability of cement to trap and hold this radioactive fission product and to investigate the consequences of its β-decay on the cement paste structure. We show that ⁹⁰Sr is stable when it substitutes the Ca²⁺ ions in C-S-H, and so is its daughter nucleus ⁹⁰Y after β-decay. Interestingly, ⁹⁰Zr, daughter of ⁹⁰Y and final product in the decay sequence, is found to be unstable compared to the bulk phase of the element at zero K but stable when compared to the solvated ion in water. Therefore, cement appears as a suitable waste form for ⁹⁰Sr storage.

The Risk Implications of Globalisation: An Exploratory Analysis of 105 Major Industrial Incidents (1971–2010)

This paper revisits work on the socio-political amplification of risk, which predicts that those living in developing countries are exposed to greater risk than residents of developed nations. This prediction contrasts with the neoliberal expectation that market driven improvements in working conditions within industrialising/developing nations will lead to global convergence of hazard exposure levels. It also contradicts the assumption of risk society theorists that there will be an ubiquitous increase in risk exposure across the globe, which will primarily affect technically more advanced countries. Reviewing qualitative evidence on the impact of structural adjustment reforms in industrialising countries, the export of waste and hazardous waste recycling to these countries and new patterns of domestic industrialisation, the paper suggests that workers in industrialising countries continue to face far greater levels of hazard exposure than those of developed countries. This view is confirmed when a data set including 105 major multi-fatality industrial disasters from 1971 to 2000 is examined. The paper concludes that there is empirical support for the predictions of socio-political amplification of risk theory, which finds clear expression in the data in a consistent pattern of significantly greater fatality rates per industrial incident in industrialising/developing countries.

Modelling of Evaporator in Waste Heat Recovery System using Finite Volume Method and Fuzzy Technique

The evaporator is an important component in the Organic Rankine Cycle (ORC)-based Waste Heat Recovery (WHR) system since the effective heat transfer of this device reflects on the efficiency of the system. When the WHR system operates under supercritical conditions, the heat transfer mechanism in the evaporator is unpredictable due to the change of thermo-physical properties of the fluid with temperature. Although the conventional finite volume model can successfully capture those changes in the evaporator of the WHR process, the computation time for this method is high. To reduce the computation time, this paper develops a new fuzzy based evaporator model and compares its performance with the finite volume method. The results show that the fuzzy technique can be applied to predict the output of the supercritical evaporator in the waste heat recovery system and can significantly reduce the required computation time. The proposed model, therefore, has the potential to be used in real time control applications.

Control design for multicast-aware class-based networks

The expectations of citizens from the Information Technologies (ITs) are increasing as the ITs have become integral part of our society, serving all kinds of activities whether professional, leisure, safety-critical applications or business. Hence, the limitations of the traditional network designs to provide innovative and enhanced services and applications motivated a consensus to integrate all services over packet switching infrastructures, using the Internet Protocol, so as to leverage flexible control and economical benefits in the Next Generation Networks (NGNs). However, the Internet is not capable of treating services differently while each service has its own requirements (e.g., Quality of Service - QoS). Therefore, the need for more evolved forms of communications has driven to radical changes of architectural and layering designs which demand appropriate solutions for service admission and network resources control. This Thesis addresses QoS and network control issues, aiming to improve overall control performance in current and future networks which classify services into classes. The Thesis is divided into three parts. In the first part, we propose two resource over-reservation algorithms, a Class-based bandwidth Over-Reservation (COR) and an Enhanced COR (ECOR). The over-reservation means reserving more bandwidth than a Class of Service (CoS) needs, so the QoS reservation signalling rate is reduced. COR and ECOR allow for dynamically defining over-reservation parameters for CoSs based on network interfaces resource conditions; they aim to reduce QoS signalling and related overhead without incurring CoS starvation or waste of bandwidth. ECOR differs from COR by allowing for optimizing control overhead minimization. Further, we propose a centralized control mechanism called Advanced Centralization Architecture (ACA), that uses a single state-full Control Decision Point (CDP) which maintains a good view of its underlying network topology and the related links resource statistics on real-time basis to control the overall network. It is very important to mention that, in this Thesis, we use multicast trees as the basis for session transport, not only for group communication purposes, but mainly to pin packets of a session mapped to a tree to follow the desired tree. Our simulation results prove a drastic reduction of QoS control signalling and the related overhead without QoS violation or waste of resources. Besides, we provide a generic-purpose analytical model to assess the impact of various parameters (e.g., link capacity, session dynamics, etc.) that generally challenge resource overprovisioning control. In the second part of this Thesis, we propose a decentralization control mechanism called Advanced Class-based resource OverpRovisioning (ACOR), that aims to achieve better scalability than the ACA approach. ACOR enables multiple CDPs, distributed at network edge, to cooperate and exchange appropriate control data (e.g., trees and bandwidth usage information) such that each CDP is able to maintain a good knowledge of the network topology and the related links resource statistics on real-time basis. From scalability perspective, ACOR cooperation is selective, meaning that control information is exchanged dynamically among only the CDPs which are concerned (correlated). Moreover, the synchronization is carried out through our proposed concept of Virtual Over-Provisioned Resource (VOPR), which is a share of over-reservations of each interface to each tree that uses the interface. Thus, each CDP can process several session requests over a tree without requiring synchronization between the correlated CDPs as long as the VOPR of the tree is not exhausted. Analytical and simulation results demonstrate that aggregate over-reservation control in decentralized scenarios keep low signalling without QoS violations or waste of resources. We also introduced a control signalling protocol called ACOR Protocol (ACOR-P) to support the centralization and decentralization designs in this Thesis. Further, we propose an Extended ACOR (E-ACOR) which aggregates the VOPR of all trees that originate at the same CDP, and more session requests can be processed without synchronization when compared with ACOR. In addition, E-ACOR introduces a mechanism to efficiently track network congestion information to prevent unnecessary synchronization during congestion time when VOPRs would exhaust upon every session request. The performance evaluation through analytical and simulation results proves the superiority of E-ACOR in minimizing overall control signalling overhead while keeping all advantages of ACOR, that is, without incurring QoS violations or waste of resources. The last part of this Thesis includes the Survivable ACOR (SACOR) proposal to support stable operations of the QoS and network control mechanisms in case of failures and recoveries (e.g., of links and nodes). The performance results show flexible survivability characterized by fast convergence time and differentiation of traffic re-routing under efficient resource utilization i.e. without wasting bandwidth. In summary, the QoS and architectural control mechanisms proposed in this Thesis provide efficient and scalable support for network control key sub-systems (e.g., QoS and resource control, traffic engineering, multicasting, etc.), and thus allow for optimizing network overall control performance.

Propriedades intrínsecas e de processamento de termoplásticos elastoméricos incorporando borracha de pneu reciclada

O trabalho apresentado tem por objetivo contribuir para a valorização da borracha proveniente de pneus em fim de vida, assente em princípios de sustentabilidade ambiental. A abordagem adotada para a concretização deste objetivo consiste na incorporação de borracha de pneus em formulações de base termoplástica e elastomérica (TPE), adequadas ao processo de moldação por injeção. São desenvolvidos estudos sobre a morfologia, propriedades mecânicas, térmicas e reológicas das ligas poliméricas à base de granulado de borracha de pneu (GTR). A falta de adesão entre o GTR e a matriz polimérica leva à degradação das propriedades mecânicas dos materiais obtidos. A estratégia explorada passa pela utilização de um elastómero para promover o encapsulamento do GTR e, desta forma, procurar obter ligas com propriedades mecânicas características de um TPE. São analisadas ligas ternárias (TPEGTR) compostas por polipropileno (PP) de elevada fluidez, GTR e elastómero virgem. O efeito da presença de diferentes elastómeros nas ligas é analisado neste trabalho: um elastómero de etilenopropileno- dieno (EPDM), e um novo elastómero de etileno-propileno (EPR) obtido por catálise metalocénica. O estudo da morfologia das ligas obtidas mostra haver interação entre os materiais, sendo possível inferir a viabilidade da estratégia adotada para promover a adesão do GTR. A incorporação de elastómero promove o aumento da resistência ao impacto e da extensão na rotura nas ligas, o que é atribuído, fundamentalmente, ao encapsulamento do GTR e ao aumento da tenacidade da matriz termoplástica. Com o objetivo de avaliar a influência da estrutura cristalina das ligas TPEGTR no seu comportamento mecânico, procede-se à análise do processo de cristalização sob condições isotérmicas e não isotérmicas. Neste estudo, é avaliado o efeito da presença dos materiais que constituem a fase elastomérica na cinética de cristalização. Para cada uma das ligas desenvolvidas, recorre-se ao modelo de Avrami para avaliar o efeito da temperatura no mecanismo de nucleação, na morfologia das estruturas cristalinas e na taxa de cristalização. Recorre-se à reometria capilar para estudar, sob condições estacionárias, o comportamento reológico das ligas TPEGTR. O modelo de Cross-WLF é utilizado para avaliar o comportamento reológico de todos os materiais, obtendo-se resultados similares àqueles obtidos experimentalmente. O comportamento reológico dos polímeros PP, EPR e EPDM é do tipo reofluidificante, tendo o EPR um comportamento reológico similar ao do PP e o EPDM um comportamento reo-fluidificante mais pronunciado. Em todas as ligas analisadas o comportamento reológico revela-se do tipo reo-fluidificante, sendo que a presença de GTR promove o aumento da viscosidade. Os parâmetros obtidos do modelo de Cross-WLF são utilizados para realizar a simulação da etapa de injeção recorrendo a um software comercial. Os resultados obtidos são validados experimentalmente pelo processo de moldação por injeção, evidenciando uma boa adequabilidade da aplicação deste modelo a estas ligas. O trabalho desenvolvido sobre ligas TPEGTR, constitui um contributo para a valorização da borracha proveniente de pneus em fim de vida, assente em princípios de sustentabilidade ambiental.

Sustainability study of the application of geosynthetic clay liners in hostile and aggressive environments

This paper discusses the sustainable performance of geosynthetic clay liners (GCLs) which are popularly specified as “leachate retaining” or as “water proofing” membranes in the geo-environmental construction industry. Geosynthetic clay liners (GCLs) are composite matting comprising of bentonite clay with two covering geosynthetics. These are innovative labour saving construction material, developed over the last three decades. The paper outlines the variety of Geosynthetic Clay Liners (GCLs) can be classified essentially into two distinctly different forms viz; (a) air dry (< 8% m/c) with granular or powdered bentonite or (b) bentonite cake factory prehydrated to a moisture content (~40% m/c) beyond its shrinkage limit and vacuum extruded as a clay cake to enhance its sustainable performance. The dominant mineral in bentonite clay is the three-layered (2:1) clay mineral montmorillonite. High quality bentonites need to be used in the GCL manufacture. Sodium montmorillonite has the desired characteristic of high swelling capacity, high cation exchange capacity and the consequently very low hydraulic conductivity, providing the basis for the hydraulic sealing medium in GCLs. These encapsulate the active montmorillonite clay minerals which depend on the water and chemical balance between the sealing element and the surrounding geo environment. Quantitative mineralogical analyses and an assessment of the adsorbed cation regime, diffusion coefficients and clay leachate compatibility must necessarily be an integral part of the site appraisal to ensure acceptable long term sustainability and performance. Factors influencing the desired performance of bentonite in the GCLs placed in difficult construction and hostile chemical environments are discussed in this paper. Accordingly, the performance specifications for GCLs are identified and the appropriateness of enhancing the cation exchange capacity with polymer treatment and the need for factory prehydration of the untreated sodium bentonite is emphasised. The advantage of factory prehydrating the polymer treated bentonite to fluid content beyond its shrinkage limit and subsequently factory processing it to develop laminated clay is to develop a GCL that has enviable sealing characteristics with a greater resistance to geochemical attack and cracking. Since clay liners are buried in the ground as base liners, capping layer or as structural water proofing membrane, they can easily avoid strict quality and performance monitoring being “out of sight, out of mind!”. It is very necessary that barrier design for leachate containment must necessarily be in accordance with legislative requirement Assessment of long term hydraulic conductivities and clay-leachate compatibility assessment is deemed necessary. The derogatory factors affecting the sustainable performance of the bentonite in GCLs placed in difficult construction and hostile chemical environments are discussed. Sustainability concepts incorporated in waste management practice must aim to achieve 100% recycling and fully implement the handling of solid waste in developing countries with relatively lower labour costs.