Involvement of the Ubiquitin/Proteasome System in Sorting of the Interleukin 2 Receptor β Chain to Late Endocytic Compartments

Down-regulation of cell surface growth factor receptors plays a key role in the tight control of cellular responses. Recent reports suggest that the ubiquitin system, in addition to participating in degradation by the proteasome of cytosolic and nuclear proteins, might also be involved in the down-regulation of various membrane receptors. We have previously characterized a signal in the cytosolic part of the interleukin 2 receptor β chain (IL2Rβ) responsible for its targeting to late endosomes/lysosomes. In this report, the role of the ubiquitin/proteasome system on the intracellular fate of IL2Rβ was investigated. Inactivation of the cellular ubiquitination machinery in ts20 cells, which express a thermolabile ubiquitin-activating enzyme E1, leads to a significant decrease in the degradation rate of IL2Rβ, with little effect on its internalization. In addition, we show that a fraction of IL2Rβ can be monoubiquitinated. Furthermore, mutation of the lysine residues of the cytosolic region of a chimeric receptor carrying the IL2Rβ targeting signal resulted in a decreased degradation rate. When cells expressing IL2Rβ were treated either by proteasome or lysosome inhibitors, a significant decrease in receptor degradation was observed. Our data show that ubiquitination is required for the sorting of IL2Rβ toward degradation. They also indicate that impairment of proteasome function might more generally affect intracellular routing.

Nitric oxide partitioning into mitochondrial membranes and the control of respiration at cytochrome c oxidase

An emerging and important site of action for nitric oxide (NO) within cells is the mitochondrial inner membrane, where NO binds to and inhibits members of the electron transport chain, complex III and cytochrome c oxidase. Although it is known that inhibition of cytochrome c oxidase by NO is competitive with O2, the mechanisms that underlie this phenomenon remain unclear, and the impact of both NO and O2 partitioning into biological membranes has not been considered. These properties are particularly interesting because physiological O2 tensions can vary widely, with NO having a greater inhibitory effect at low O2 tensions (<20 μM). In this study, we present evidence for a consumption of NO in mitochondrial membranes in the absence of substrate, in a nonsaturable process that is O2 dependent. This consumption modulates inhibition of cytochrome c oxidase by NO and is enhanced by the addition of exogenous membranes. From these data, it is evident that the partition of NO into mitochondrial membranes has a major impact on the ability of NO to control mitochondrial respiration. The implications of this conclusion are discussed in the context of mitochondrial lipid:protein ratios and the importance of NO as a regulator of respiration in pathophysiology.

Differential Control of Xanthophylls and Light-Induced Stress Proteins, as Opposed to Light-Harvesting Chlorophyll a/b Proteins, during Photosynthetic Acclimation of Barley Leaves to Light Irradiance

Barley (Hordeum vulgare L.) plants were grown at different photon flux densities ranging from 100 to 1800 μmol m−2 s−1 in air and/or in atmospheres with reduced levels of O2 and CO2. Low O2 and CO2 partial pressures allowed plants to grow under high photosystem II (PSII) excitation pressure, estimated in vivo by chlorophyll fluorescence measurements, at moderate photon flux densities. The xanthophyll-cycle pigments, the early light-inducible proteins, and their mRNA accumulated with increasing PSII excitation pressure irrespective of the way high excitation pressure was obtained (high-light irradiance or decreased CO2 and O2 availability). These findings indicate that the reduction state of electron transport chain components could be involved in light sensing for the regulation of nuclear-encoded chloroplast gene expression. In contrast, no correlation was found between the reduction state of PSII and various indicators of the PSII light-harvesting system, such as the chlorophyll a-to-b ratio, the abundance of the major pigment-protein complex of PSII (LHCII), the mRNA level of LHCII, the light-saturation curve of O2 evolution, and the induced chlorophyll-fluorescence rise. We conclude that the chlorophyll antenna size of PSII is not governed by the redox state of PSII in higher plants and, consequently, regulation of early light-inducible protein synthesis is different from that of LHCII.

Phenotypic knockout of the high-affinity human interleukin 2 receptor by intracellular single-chain antibodies against the alpha subunit of the receptor.

The experimental manipulation of peptide growth hormones and their cellular receptors is central to understanding the pathways governing cellular signaling and growth control. Previous work has shown that intracellular antibodies targeted to the endoplasmic reticulum (ER) can be used to capture specific proteins as they enter the ER, preventing their transport to the cell surface. Here we have used this technology to inhibit the cell surface expression of the alpha subunit of the high-affinity interleukin 2 receptor (IL-2R alpha). A single-chain variable-region fragment of the anti-Tac monoclonal antibody was constructed with a signal peptide and a C-terminal ER retention signal. Intracellular expression of the single-chain antibody was found to completely abrogate cell surface expression of IL-2R alpha in stimulated Jurkat T cells. IL-2R alpha was detectable within the Jurkat cells as an immature 40-kDa form that was sensitive to endoglycosidase H, consistent with its retention in a pre- or early Golgi compartment. A single-chain antibody lacking the ER retention signal was also able to inhibit cell surface expression of IL-2R alpha although the mechanism appeared to involve rapid degradation of the receptor chain within the ER. These intracellular antibodies will provide a valuable tool for examining the role of IL-2R alpha in T-cell activation, IL-2 signal transduction, and the deregulated growth of leukemic cells which overexpress IL-2R alpha.

Calibration of the length of a chain of single gold atoms

Using a scanning tunnelling microscope or mechanically controllable break junction it has been shown that it is possible to control the formation of a wire made of single gold atoms. In these experiments an interatomic distance between atoms in the chain of ∼3.6 Å was reported which is not consistent with recent theoretical calculations. Here, using precise calibration procedures for both techniques, we measure the length of the atomic chains. Based on the distance between the peaks observed in the chain length histogram we find the mean value of the interatomic distance before chain rupture to be 2.5±0.2 Å. This value agrees with the theoretical calculations for the bond length. The discrepancy with the previous experimental measurements was due to the presence of He gas, that was used to promote the thermal contact, and which affects the value of the work function that is commonly used to calibrate distances in scanning tunnelling microscopy and mechanically controllable break junctions at low temperatures.

Handling of Uncertainty in Life Cycle Inventory by Correlated Multivariate Lognormal Distributions: Application to the Design of Supply Chain Networks

Poster presented in the 24th European Symposium on Computer Aided Process Engineering (ESCAPE 24), Budapest, Hungary, June 15-18, 2014.

Handling of Uncertainty in Life Cycle Inventory by Correlated Multivariate Lognormal Distributions: Application to the Design of Supply Chain Networks

In this work, we analyze the effect of incorporating life cycle inventory (LCI) uncertainty on the multi-objective optimization of chemical supply chains (SC) considering simultaneously their economic and environmental performance. To this end, we present a stochastic multi-scenario mixed-integer linear programming (MILP) coupled with a two-step transformation scenario generation algorithm with the unique feature of providing scenarios where the LCI random variables are correlated and each one of them has the desired lognormal marginal distribution. The environmental performance is quantified following life cycle assessment (LCA) principles, which are represented in the model formulation through standard algebraic equations. The capabilities of our approach are illustrated through a case study of a petrochemical supply chain. We show that the stochastic solution improves the economic performance of the SC in comparison with the deterministic one at any level of the environmental impact, and moreover the correlation among environmental burdens provides more realistic scenarios for the decision making process.

Modelling a biomass supply chain through discrete-event simulation

The organizational structure of the companies in the biomass energy sector, regarding the supply chain management services, can be greatly improved through the use of software decision support tools. These tools should be able to provide real-time alternative scenarios when deviations from the initial production plans are observed. To make this possible it is necessary to have representative production chain process models where several scenarios and solutions can be evaluated accurately. Due to its nature, this type of process is more adequately represented by means of event-based models. In particular, this work presents the modelling of a typical biomass production chain using the computing platform SIMEVENTS. Throughout the article details about the conceptual model, as well as simulation results, are provided

Forest-based supply chain modelling using the SimPy simulation framework

Proper management of supply chains is fundamental in the overall system performance of forestbased activities. Usually, efficient management techniques rely on a decision support software, which needs to be able to generate fast and effective outputs from the set of possibilities. In order to do this, it is necessary to provide accurate models representative of the dynamic interactions of systems. Due to forest-based supply chains’ nature, event-based models are more suited to describe their behaviours. This work proposes the modelling and simulation of a forestbased supply chain, in particular the biomass supply chain, through the SimPy framework. This Python based tool allows the modelling of discrete-event systems using operations such as events, processes and resources. The developed model was used to access the impact of changes in the daily working plan in three situations. First, as a control case, the deterministic behaviour was simulated. As a second approach, a machine delay was introduced and its implications in the plan accomplishment were analysed. Finally, to better address real operating conditions, stochastic behaviours of processing and driving times were simulated. The obtained results validate the SimPy simulation environment as a framework for modelling supply chains in general and for the biomass problem in particular.

(Table T1) Long-chain alkenones, UK'37 and derived sea surface temperatures of ODP Sites 184-1147 and 184-1148

A reconnaissance study of alkenone stratigraphy for the past 35 m.y. in the northern South China Sea (SCS) using sediments from Sites 1147 and 1148 of Ocean Drilling Program (ODP) Leg 184 has been completed. Alkenones were not detected in sediment samples older than ~31 Ma. However, C37:2 appeared in the sedimentary record between ~8 and 31 Ma and both C37:2 and C37:3 were present between 0 and 8 Ma. These changes in alkenone occurrences may signal a response to global-scale Neogene cooling as well as to monsoon intensification and sea level changes over time as a result of Himalayan uplift and the opening of the SCS. Alternatively, they may be related to an evolutionary record of the development of temperature control on alkenone production in coccolithophores. The Uk'37 index for 0-8 Ma produces sea-surface temperatures (SST) of 19°-26°C, which are in the range of previously determined glacial-interglacial values for the northern SCS. Before the late Pleistocene (~1.2 Ma), the SST range is between 23° and 26°C with less variation. This change in variability may signify the early stage of intensified winter monsoons where cold wind and waters from the north may not yet have had a significant effect on SST or it may be the evolutionary link between the early development of unsaturated alkenones in coccolithophores and modern temperature control of alkenone production. We believe a long-term alkenone record is useful for further understanding of global-scale neogene cooling, the development of the East Asian monsoon system, and the evolutionary development of temperature control on alkenone unsaturation. Our data indicate that a high-resolution Uk'37 record for at least the last ~8 Ma is feasible for the northern SCS.

Design strategies for controlling the molecular weight and rate using reversible addition-fragmentation chain transfer mediated living radical polymerization

Living radical polymerization has allowed complex polymer architectures to be synthesized in bulk, solution, and water. The most versatile of these techniques is reversible addition-fragmentation chain transfer (RAFT), which allows a wide range of functional and nonfunctional polymers to be made with predictable molecular weight distributions (MWDs), ranging from very narrow to quite broad. The great complexity of the RAFT mechanism and how the kinetic parameters affect the rate of polymerization and MWD are not obvious. Therefore, the aim of this article is to provide useful insights into the important kinetic parameters that control the rate of polymerization and the evolution of the MWD with conversion. We discuss how a change in the chain-transfer constant can affect the evolution of the MWD. It is shown how we can, in principle, use only one RAFT agent to obtain a poly-mer with any MWD. Retardation and inhibition are discussed in terms of (1) the leaving R group reactivity and (2) the intermediate radical termination model versus the slow fragmentation model. (c) 2005 Wiley Periodicals, Inc.

Developing an enterprise simulator to support electronic supply chain for B2B electronic process

The application of any e-Solution promises significant returns. In particular, using internet technologies both within enterprises and across the supply (value) chain provides real opportunity, not only for operational improvement but also for innovative strategic positioning. However, significant questions obscure potential investment; how any value will actually be created and, importantly, how this value will be shared across the value chain is not clear. This paper will describe a programme of research that is developing an enterprise simulator that will provide a more fundamental understanding of the impact of e-Solutions across operational supply chains, in terms of both standard operational and financial measures of performance. An efficient supply chain reduces total costs of operations by sharing accurate real-time information and coordinating inter-organizational business processes. This form of electronic link between organizations is known as business-to-business (B2B) e-Business. The financial measures go beyond simple cost calculations to real bottom-line performance by modelling the financial transactions that business processes generate. The paper will show how this enterprise simulator allows for a complete supply chain to be modelled in this way across four key applications: control system design, virtual enterprises, pan-supply-chain performance metrics and supporting e-Supply-chain design methodology.

Implementing collaborative practices in the healthcare supply chain:insights into hospital-vendor operations

The paper discusses the characteristics of healthcare supply chains, and puts particular emphasis on the implementation of VMI/CMI in this sector specific context. By the means of case study research the paper provides empirical data on the benefits of the above collaborative practices for both the hospital and vendors. The paper contributes to the stream of research on VMI/CMI in the healthcare sector, where limited research attempts have been conducted so far. In contrast to other surveys this case study shows that specific and measurable cost reductions exist, in addition to other improvements such as better control over the inventories, and also in reduction of administrative work. Results obtained may be also relevant to other hospitals and vendors and as they can form a basis for comparisons. Copyright © 2013 Inderscience Enterprises Ltd.

Green supply chain performance measurement using the analytic hierarchy process:a comparative analysis of manufacturing organisations

The main purpose of this research is to develop and deploy an analytical framework for measuring the environmental performance of manufacturing supply chains. This work's theoretical bases combine and reconcile three major areas: supply chain management, environmental management and performance measurement. Researchers have suggested many empirical criteria for green supply chain (GSC) performance measurement and proposed both qualitative and quantitative frameworks. However, these are mainly operational in nature and specific to the focal company. This research develops an innovative GSC performance measurement framework by integrating supply chain processes (supplier relationship management, internal supply chain management and customer relationship management) with organisational decision levels (both strategic and operational). Environmental planning, environmental auditing, management commitment, environmental performance, economic performance and operational performance are the key level constructs. The proposed framework is then applied to three selected manufacturing organisations in the UK. Their GSC performance is measured and benchmarked by using the analytic hierarchy process (AHP), a multiple-attribute decision-making technique. The AHP-based framework offers an effective way to measure and benchmark organisations’ GSC performance. This study has both theoretical and practical implications. Theoretically it contributes holistic constructs for designing a GSC and managing it for sustainability; and practically it helps industry practitioners to measure and improve the environmental performance of their supply chain. © 2013 Copyright Taylor and Francis Group, LLC. CORRIGENDUM DOI 10.1080/09537287.2012.751186 In the article ‘Green supply chain performance measurement using the analytic hierarchy process: a comparative analysis of manufacturing organisations’ by Prasanta Kumar Dey and Walid Cheffi, Production Planning & Control, 10.1080/09537287.2012.666859, a third author is added which was not included in the paper as it originally appeared. The third author is Breno Nunes.