Novel indicators to better monitor the collection and recovery of (critical) raw materials in WEEE: Focus on screens.

Currently, in the European Union (EU), e-waste chain performance is assessed by technical indicators that aim to ensure system compliance with collection and recovery targets set by the WEEE Directive. This study proposes indicators to improve WEEE flow monitoring beyond the current overall weight-based approach, including complementary flows and treatment performance. A case study focused on the screen category in France is presented. In 2017, the collection rate of cathode-ray tube screens (CRT) was 68%, while for flat panel display (FPD) generated only 14% was collected. CRT screens have less precious and critical materials than FDP. Thus, elements like cobalt and gold highly concentrated in FPD, have a collection rate two to four times lower than elements such as copper (37%) which represents a high proportion in CRTs. Recycling is the main treatment in France. Nevertheless, the recycling rate per element varies significantly due to the low collection, and also the lack of technology and/or secondary raw materials market. The elements with higher recycling rates are base metals such as copper (28%), followed by precious metals like silver (23%), and gold (13%). Except for palladium, the recycling rate of the critical raw materials targeted in the study ranged from 6% (cobalt) to 0% (e.g. neodymium and indium). The results stress the need for indicators to support the development of WEEE chain from waste management to secondary (critical) raw materials suppliers.

Influence of scope definition in recycling rate calculation for European e-waste extended producer responsibility.

Efficiency indicators have been frequently used to assess end-of-life chain performance, mostly. In terms of the percentage of mass sent to re-use, recycling, and/or energy recovery facilities. While legislation gives a standard definition for recycling and recovery rates, stakeholders sometimes redefine them to better fit their own scopes and objectives. Therefore, to accurately interpret the results of an efficiency indicator, during a decision-making process, it's necessary to fully understand the scope definition used to calculate it. This work discusses the influence of scope definition when establishing performance rates. It does this by introducing further alternative scope definitions and comparing them to those defined by legislation and stakeholders. As a case study, the proposed complementary scopes are applied to the recycling chain of flat panel displays in France.

A regional approach for the calculation of characteristic toxicity factors using the USEtox model.

The lack of the spatial coverage as one of the main limitations of the Life-cycle impact assessment (LCIA) models leads to disagreement between their results. The USEtox model is only model that provides 8 continental and 17 subcontinental zones but does not consider the wind and water transfers affected areas around the source of pollution. Current investigation proposes the way to reduce this limitation by using the results of chemical analysis (instrumental neutron activation analysis "INAA") of pork meat as a regional indicator of anthropogenic influence. The concentration coefficient of Cr by replacing the Bioaccumulation factor (BAF) is extrapolated into the calculation of Exposure factor (XF) to modify Characterization factor (CF). Impacted and clean areas of Tomsk district (Russia) placed around Northern industrial hub (Seversk city) are studied. Neither area is located directly in the industrial hub, but the impacted area is under an anthropogenic influence due to air and water transfer of pollution. Results of our investigation present the difference between results of own investigation and default values of USEtox. Probably the model can minimize the impact because of lack of experiment data in the database. The database can be extended more with other analytical results for wide range of metals and geographical locations.

Wiring Together Synthetic Bacterial Consortia to Create a Biological Integrated Circuit.

The promise of adapting biology to information processing will not be realized until engineered gene circuits, operating in different cell populations, can be wired together to express a predictable function. Here, elementary biological integrated circuits (BICs), consisting of two sets of transmitter and receiver gene circuit modules with embedded memory placed in separate cell populations, were meticulously assembled using live cell lithography and wired together by the mass transport of quorum-sensing (QS) signal molecules to form two isolated communication links (comlinks). The comlink dynamics were tested by broadcasting "clock" pulses of inducers into the networks and measuring the responses of functionally linked fluorescent reporters, and then modeled through simulations that realistically captured the protein production and molecular transport. These results show that the comlinks were isolated and each mimicked aspects of the synchronous, sequential networks used in digital computing. The observations about the flow conditions, derived from numerical simulations, and the biofilm architectures that foster or silence cell-to-cell communications have implications for everything from decontamination of drinking water to bacterial virulence.

Preassembled Single-Stranded RNA-Argonaute Complexes: A Novel Method to Silence Genes in Cryptosporidium.

Cryptosporidiosis is a common cause of diarrhea morbidity and mortality worldwide. Research progress on this infection has been slowed by lack of methods to genetically manipulate Cryptosporidium parasites. Small interfering RNA (siRNA) is widely used to study gene function, but Cryptosporidium species lack the enzymes necessary to process siRNA. By preassembling complexes with the human enzyme Argonaute 2 (hAgo2) and Cryptosporidium single-stranded RNA (ssRNA), we induced specific slicing in Cryptosporidium RNA targets. We demonstrated the reduction in expression of target genes at the mRNA and protein levels by transfecting live parasites with ssRNA-hAgo2 complexes. Furthermore we used this method to confirm the role of selected molecules during host cell invasion. This novel method provides a novel means of silencing Cryptosporidium genes to study their role in host-parasite interactions and as potential targets for chemotherapy.

Biological noise abatement: coordinating the responses of autonomous bacteria in a synthetic biofilm to a fluctuating environment using a stochastic bistable switch.

Noise is inherent to single cell behavior. Its origins can be traced to the stochasticity associated with a few copies of genes and low concentrations of protein and ligands. We have studied the mechanisms by which the response of noisy elements can be entrained for biological signal processing. To elicit predictable biological function, we have engineered a gene environment that incorporates a gene regulatory network with the stringently controlled microenvironment found in a synthetic biofilm. The regulatory network leverages the positive feedback found in quorum-sensing regulatory components of the lux operon, which is used to coordinate cellular responses to environmental fluctuations. Accumulation of the Lux receptor in cells, resulting from autoregulation, confers a rapid response and enhanced sensitivity to the quorum-sensing molecule that is retained after cell division as epigenetic memory. The memory of the system channels stochastic noise into a coordinated response among quorum-sensing signal receivers in a synthetic biofilm in which the noise diminishes with repeated exposure to noisy transmitters on the input of a signaling cascade integrated into the same biofilm. Thus, gene expression in the receivers, which are autonomous and do not communicate with each other, is synchronized to fluctuations in the environment.

Epigenetic memory emerging from integrated transcription bursts.

Some autonomous bacteria coordinate their actions using quorum-sensing (QS) signals to affect gene expression. However, noise in the gene environment can compromise the cellular response. By exercising precise control over a cell's genes and its microenvironment, we have studied the key positive autoregulation element by which the lux QS system integrates noisy signals into an epigenetic memory. We observed transcriptional bursting of the lux receptor in cells stimulated by near-threshold levels of QS ligand. The bursts are integrated over time into an epigenetic memory that confers enhanced sensitivity to the ligand. An emergent property of the system is manifested in pattern formation among phenotypes within a chemical gradient.

Synthetic networks: oscillators and toggle switches for Escherichia coli.

Bacterial synthetic gene networks are constructed by manipulating the regulation of genes inside a cell, with the purpose of eliciting novel regulatory behaviors. The methods for manipulating genes and gene regulation in E. coli are well established, making it the preferred host for basic studies of synthetic networks. We focus our work on constructing two kinds of synthetic gene networks: toggle switches (bistable systems) and oscillators. Toggle switches are capable of exhibiting two stable steady states of gene expression (OFF and ON) without stable intermediate states; the steady state reached by the system depends on the previous history of the system. Biological oscillators exhibit regular cycles in gene expression around an unstable steady state. Studying these two kinds of synthetic networks helps advance our understanding of natural bistable systems and oscillators, such as the circadian oscillators controlling gene expression in many types of cells, and the genetic systems controlling the cell cycle and differentiation in metazoans.

A comparison of random vs. chemotaxis-driven contacts of T cells with dendritic cells during repertoire scanning.

Generating adaptive immunity after infection or immunization requires physical interactions within a lymph node (LN) T-zone between antigen-bearing dendritic cells (DCs) that arrive from peripheral tissues and rare cognate T cells entering via high endothelial venules (HEVs). This interaction results in activation of cognate T cells, expansion of that T cell lineage and their exit from the LN T-zone via efferent lymphatics (ELs). How antigen-specific T cells locate DCs within this complex environment is controversial, and both random T cell migration and chemotaxis have been proposed. We developed an agent-based computational model of a LN that captures many features of T cell and DC dynamics observed by two-photon microscopy. Our simulations matched in vivo two-photon microscopy data regarding T cell speed, short-term directional persistence of motion and cell motility. We also obtained in vivo data regarding density of T cells and DCs within a LN and matched our model environment to measurements of the distance from HEVs to ELs. We used our model to compare chemotaxis with random motion and showed that chemotaxis increased total number of T cell DC contacts, but decreased unique contacts, producing fewer activated T cells. Our results suggest that, within a LN T-zone, a random search strategy is optimal for a rare cognate T cell to find its DC match and maximize production of activated T cells.

Using two-component systems and other bacterial regulatory factors for the fabrication of synthetic genetic devices.

Synthetic biology is an emerging field in which the procedures and methods of engineering are extended living organisms, with the long-term goal of producing novel cell types that aid human society. For example, engineered cell types may sense a particular environment and express gene products that serve as an indicator of that environment or affect a change in that environment. While we are still some way from producing cells with significant practical applications, the immediate goals of synthetic biology are to develop a quantitative understanding of genetic circuitry and its interactions with the environment and to develop modular genetic circuitry derived from standard, interoperable parts that can be introduced into cells and result in some desired input/output function. Using an engineering approach, the input/output function of each modular element is characterized independently, providing a toolkit of elements that can be linked in different ways to provide various circuit topologies. The principle of modularity, yet largely unproven for biological systems, suggests that modules will function appropriately based on their design characteristics when combined into larger synthetic genetic devices. This modularity concept is similar to that used to develop large computer programs, where independent software modules can be independently developed and later combined into the final program. This chapter begins by pointing out the potential usefulness of two-component signal transduction systems for synthetic biology applications and describes our use of the Escherichia coli NRI/NRII (NtrC/NtrB) two-component system for the construction of a synthetic genetic oscillator and toggle switch for E. coli. Procedures for conducting measurements of oscillatory behavior and toggle switch behavior of these synthetic genetic devices are described. It then presents a brief overview of device fabrication strategy and tactics and presents a useful vector system for the construction of synthetic genetic modules and positioning these modules onto the bacterial chromosome in defined locations.

Toward a multiscale model of antigen presentation in immunity.

A functioning immune system and the process of antigen presentation in particular encompass events that occur at multiple length and time scales. Despite a wealth of information in the biological literature regarding each of these scales, no single representation synthesizing this information into a model of the overall immune response as it depends on antigen presentation is available. In this article, we outline an approach for integrating information over relevant biological and temporal scales to generate such a representation for major histocompatibility complex class II-mediated antigen presentation. In addition, we begin to address how such models can be used to answer questions about mechanisms of infection and new strategies for treatment and vaccines.

Experimental validation of a critical domain size in reaction-diffusion systems with Escherichia coli populations.

In a one-variable, finite size reaction-diffusion system, the existence of a minimal domain size required for the existence of a non-zero steady state is predicted, provided that the reaction-diffusion variable has a fixed value of zero at the boundaries of the domain (Dirichlet boundary conditions). This type of reaction diffusion model can be applied in population biology, in which the finite domain of the system represents a refuge where individuals can live normally immersed in a desert, or region where the conditions are so unfavourable that individuals cannot live in it. Building on a suggestion by Kenkre and Kuperman, and using non-chemotactic E. coli populations and a quasi-one-dimensional experimental design, we were able to find a minimal size (approximately 0.8 cm) for a refuge immersed in a region irradiated with intense UV light. The observed minimal size is in reasonable agreement with theory.