Electrodialytic processes in solid matrices. New insights into batteries recycling. A review.

Electrodialytic Remediation has been widely applied to the recovery of different contaminants from numerous solid matrices solving emerging issues of environmental concern. Results and conclusions reported in studies about real contaminated matrices are summarizes in this work. The influence of the pH value on the treatment effectiveness has been widely proved highlighting the phenomenon “water splitting” in the membrane surface. This dissociation of water molecules is related to the “limiting current” which is desirable to be exceed at the Anion Exchange Membrane in order to produce the entering of protons toward solid matrix. Other important parameters for the optimization of the technique, such as the current density and the liquid to solid ratio, are also discussed through the revision of studies using real solid matrices. This work also focusses on the pioneer proposal of electrokinetic technologies for the recycling of lithium ion batteries considering the relevance of waste properties in the design and optimization of the technique. From a thorough literature revision, it could be concluded that further experimental results are needed to allow an optimal application of the technique to the rising problem of residues from batteries. The main aim of this work is to take the first steps in the recovery of valuable metals from spent batteries, such as Li and Co, incorporating principles of green chemistry.The authors acknowledge the financial support from the “Plan Propio de Investigación de la Universidad de Málaga with Project numbers: PPIT.UMA.B1.2017/20 and PPIT.UMA.B5.2018/17 and the European project THROUGH H2020-MSCA-RISE- 2017-778045. The first author also acknowledge the postdoctoral contract obtained from University of Malaga

A Genetically Encoded AND Gate for Cell-Targeted Metabolic Labeling of Proteins

We describe a genetic AND gate for cell-targeted metabolic labeling and proteomic analysis in complex cellular systems. The centerpiece of the AND gate is a bisected methionyl-tRNA synthetase (MetRS) that charges the Met surrogate azidonorleucine (Anl) to tRNAMet. Cellular protein labeling occurs only upon activation of two different promoters that drive expression of the N- and C-terminal fragments of the bisected MetRS. Anl-labeled proteins can be tagged with fluorescent dyes or affinity reagents via either copper-catalyzed or strain-promoted azide–alkyne cycloaddition. Protein labeling is apparent within 5 min after addition of Anl to bacterial cells in which the AND gate has been activated. This method allows spatial and temporal control of proteomic labeling and identification of proteins made in specific cellular subpopulations. The approach is demonstrated by selective labeling of proteins in bacterial cells immobilized in the center of a laminar-flow microfluidic channel, where they are exposed to overlapping, opposed gradients of inducers of the N- and C-terminal MetRS fragments. The observed labeling profile is predicted accurately from the strengths of the individual input signals

A nonplanar slow rupture episode during the 2000 Miyakejima dike intrusion

Magmatic intrusions release extensional strain in the Earth's crust upon availability of magma. Intrusions are typically accompanied by earthquake swarms and by surface faulting that is often larger than what is expected from the magnitude of the induced earthquakes. The 2000 Miyakejima dike intrusion triggered the largest volcanic earthquake swarm monitored so far, with five Ml>6 earthquakes. We analyze the seismicity and deformation induced by the Miyakejima dike with the aim of constraining the timescale and mechanisms of slow strain release during the episode. In six earthquake bursts lasting few hours and migrating at 3c1 km h 121 we find candidates for slow earthquakes. Each burst nucleated at the tips of previous bursts, suggesting stress interaction. The variability of fault plane solutions indicates that the bursts occurred on a complex system of fractures, consistent with weakly consolidated surface layers strained by spatially inhomogneous stresses that change in time, such as those induced by a dike. Based on dislocation models, we find that deformation is best explained by aseismic slip (in addition to the seismic burst), with a moment 1.3 to 2.3 times larger than the earthquakes' seismic moment, and opening of 0.20 \ub1 0.07 m on the dike. The aseismic slip occurred over a few hours, with moment, duration, and migration velocity consistent with that of previously observed slow slip events. We argue that the seismic bursts are likely driven by slow slip, sharing most properties with tectonic slow slip events and swarms, but occurring on a set of nonaligned faults

A 'resource allocator' for transcription based on a highly fragmented T7 RNA polymerase

Synthetic genetic systems share resources with the host, including machinery for transcription and translation. Phage RNA polymerases (RNAPs) decouple transcription from the host and generate high expression. However, they can exhibit toxicity and lack accessory proteins (σ factors and activators) that enable switching between different promoters and modulation of activity. Here, we show that T7 RNAP (883 amino acids) can be divided into four fragments that have to be co‐expressed to function. The DNA‐binding loop is encoded in a C‐terminal 285‐aa ‘σ fragment’, and fragments with different specificity can direct the remaining 601‐aa ‘core fragment’ to different promoters. Using these parts, we have built a resource allocator that sets the core fragment concentration, which is then shared by multiple σ fragments. Adjusting the concentration of the core fragment sets the maximum transcriptional capacity available to a synthetic system. Further, positive and negative regulation is implemented using a 67‐aa N‐terminal ‘α fragment’ and a null (inactivated) σ fragment, respectively. The α fragment can be fused to recombinant proteins to make promoters responsive to their levels. These parts provide a toolbox to allocate transcriptional resources via different schemes, which we demonstrate by building a system which adjusts promoter activity to compensate for the difference in copy number of two plasmids.United States. Office of Naval Research (N00014‐13‐1‐0074)National Institutes of Health (U.S.) (5R01GM095765)National Science Foundation (U.S.) (Synthetic Biology Engineering Research Center (SA5284‐11210))United States. Dept. of Defense (National Defense Science and Engineering Graduate Fellowship (NDSEG) Program))Hertz Foundation (Fellowship

Tandem repeat coupled with endonuclease cleavage (TREC): a seamless modification tool for genome engineering in yeast

The complete synthetic Mycoplasma genitalium genome (∼583 kb) has been assembled and cloned as a circular plasmid in the yeast Saccharomyces cerevisiae. Attempts to engineer the cloned genome by standard genetic methods involving the URA3/5-fluoroorotic acid (5-FOA) counter-selection have shown a high background of 5-FOA resistant clones derived from spontaneous deletions of the bacterial genome maintained in yeast. Here, we report a method that can seamlessly modify the bacterial genome in yeast with high efficiency. This method requires two sequential homologous recombination events. First, the target region is replaced with a mutagenesis cassette that consists of a knock-out CORE (an18-bp I-SceI recognition site, the SCEI gene under the control of the GAL1 promoter, and the URA3 marker) and a DNA fragment homologous to the sequence upstream of the target site. The replacement generates tandem repeat sequences flanking the CORE. Second, galactose induces the expression of I-SceI, which generates a double-strand break (DSB) at the recognition site. This DSB promotes intra-molecular homologous recombination between the repeat sequences, and leads to an excision of the CORE. As a result, a seamless modification is generated. This method can be adapted for a variety of genomic modifications and may provide an important tool to modify and design natural or synthetic genomes propagated in yeast

A transposase strategy for creating libraries of circularly permuted proteins

A simple approach for creating libraries of circularly permuted proteins is described that is called PERMutation Using Transposase Engineering (PERMUTE). In PERMUTE, the transposase MuA is used to randomly insert a minitransposon that can function as a protein expression vector into a plasmid that contains the open reading frame (ORF) being permuted. A library of vectors that express different permuted variants of the ORF-encoded protein is created by: (i) using bacteria to select for target vectors that acquire an integrated minitransposon; (ii) excising the ensemble of ORFs that contain an integrated minitransposon from the selected vectors; and (iii) circularizing the ensemble of ORFs containing integrated minitransposons using intramolecular ligation. Construction of a Thermotoga neapolitana adenylate kinase (AK) library using PERMUTE revealed that this approach produces vectors that express circularly permuted proteins with distinct sequence diversity from existing methods. In addition, selection of this library for variants that complement the growth of Escherichia coli with a temperature-sensitive AK identified functional proteins with novel architectures, suggesting that PERMUTE will be useful for the directed evolution of proteins with new functions

Modular control of multiple pathways using engineered orthogonal T7 polymerases

Synthetic genetic sensors and circuits enable programmable control over the timing and conditions of gene expression. They are being increasingly incorporated into the control of complex, multigene pathways and cellular functions. Here, we propose a design strategy to genetically separate the sensing/circuitry functions from the pathway to be controlled. This separation is achieved by having the output of the circuit drive the expression of a polymerase, which then activates the pathway from polymerase-specific promoters. The sensors, circuits and polymerase are encoded together on a ‘controller’ plasmid. Variants of T7 RNA polymerase that reduce toxicity were constructed and used as scaffolds for the construction of four orthogonal polymerases identified via part mining that bind to unique promoter sequences. This set is highly orthogonal and induces cognate promoters by 8- to 75-fold more than off-target promoters. These orthogonal polymerases enable four independent channels linking the outputs of circuits to the control of different cellular functions. As a demonstration, we constructed a controller plasmid that integrates two inducible systems, implements an AND logic operation and toggles between metabolic pathways that change Escherichia coli green (deoxychromoviridans) and red (lycopene). The advantages of this organization are that (i) the regulation of the pathway can be changed simply by introducing a different controller plasmid, (ii) transcription is orthogonal to host machinery and (iii) the pathway genes are not transcribed in the absence of a controller and are thus more easily carried without invoking evolutionary pressure.United States. Office of Naval Research (Award number N00014-10-1-0245)National Science Foundation (U.S.). (CCF-0943385)National Institutes of Health (U.S.) (AI067699)National Science Foundation (U.S.). Graduate Research FellowshipAmerican Society for Engineering Education. National Defense Science and Engineering Graduate FellowshipHertz Foundation. Graduate Fellowshi

B cell priming for extrafollicular antibody responses requires Bcl-6 expression by T cells

Bcl-6 expression in CD4+ T cells is required to generate extrafollicular antibody responses