Unusual Stability Research Articles

Optical conductivity and superconductivity in highly overdoped La2−xCaxCuO4 thin films

We have used atomic layer-by-layer oxide molecular beam epitaxy to grow epitaxial thin films of [Formula: see text] with x up to 0.5, greatly exceeding the solubility limit of Ca in bulk systems ([Formula: see text]). A comparison of the optical conductivity measured by spectroscopic ellipsometry to prior predictions from dynamical mean-field theory demonstrates that the hole concentration p is approximately equal to x. We find superconductivity with [Formula: see text] of 15 to 20 K up to the highest doping levels and attribute the unusual stability of superconductivity in [Formula: see text] to the nearly identical radii of La and Ca ions, which minimizes the impact of structural disorder. We conclude that careful disorder management can greatly extend the "superconducting dome" in the phase diagram of the cuprates.

Improving the alkaline stability of pepsin through rational protein design using renin, an alkaline-stable aspartic protease, as a structural and functional reference

The present study sought to identify the structural determinants of aspartic protease structural stability and activity at elevated pH. Various hypotheses have been published regarding the features responsible for the unusual alkaline structural stability of renin, however, few structure-function studies have verified these claims. Using pepsin as a model system, and renin as a template for functional and structural alkaline stability, a rational re-design of pepsin was undertaken to identify residues contributing to the alkaline instability of pepsin-like aspartic proteases in regards to both structure and function. We constructed 13 mutants based on this strategy. Among them, mutants D159 L and D60A led to an increase in activity at elevated pH levels (p ≤ 0.05) and E4V and H53F were shown to retain native-like structure at elevated pH (p ≤ 0.05). Previously suggested carboxyl groups Asp11, Asp118, and Glu13 were individually shown not to be responsible for the structural instability or lack of activity at neutral pH in pepsin. The importance of the β-barrel to structural stability was highlighted as the majority of the stabilizing residues identified, and 39% of the weakly conserved residues in the N-terminal lobe, were located in β-sheet strands of the barrel. The results of the present study indicate that alkaline stabilization of pepsin will require reduction of electrostatic repulsions and an improved understanding of the role of the hydrogen bonding network of the characteristic β-barrel.

Marriage of Peroxides and Nitrogen Heterocycles: Selective Three-Component Assembly, Peroxide-Preserving Rearrangement, and Stereoelectronic Source of Unusual Stability of Bridged Azaozonides.

Stable bridged azaozonides can be selectively assembled via a catalyst-free three-component condensation of 1,5-diketones, hydrogen peroxide, and an NH-group source such as aqueous ammonia or ammonium salts. This procedure is scalable and can produce gram quantities of bicyclic stereochemically rich heterocycles. The new azaozonides are thermally stable and can be stored at room temperature for several months without decomposition and for at least 1 year at -10 °C. The chemical stability of azaozonides was explored for their subsequent selective transformations including the first example of an aminoperoxide rearrangement that preserves the peroxide group. The amino group in aminoperoxides has remarkably low nucleophilicity and does not participate in the usual amine alkylation and acylation reactions. These observations and the 15 pKa units decrease in basicity in comparison with a typical dialkyl amine are attributed to the strong hyperconjugative nN→σ*C-O interaction with the two antiperiplanar C-O bonds. Due to the weakness of the complementary nO→σ*C-N donation from the peroxide oxygens (a consequence of "inverse α-effect"), this interaction depletes electron density from the NH moiety, protects it from oxidation, and makes it similar in properties to an amide.

Synthesis of Novel Trimethoprim Complexes and Their Analysis by Ultraviolet Derivative Spectroscopy

In this research, the actions of antibiotic trimethoprim alone and with metals combained. Then assayed through formation of base transition metal compounds as ligands have the chance to achieve an unusual arrangement and stability complexes of coordination. We show advancement in using transtional metal compounds medications for treamentt manyl human illnesses such as carcinomas, lymphomas, control of infections, anti-inflammatory disorders, diabetes, with neurological conditions. This combination with transition metal observed the interaction can be separated by derivative spectroscopic method and measure the characterization of compound by IR and UV spectroscopy.

Size-controlled nanocrystals reveal spatial dependence and severity of nanoparticle coalescence and Ostwald ripening in sintering phenomena.

A major aim in the synthesis of nanomaterials is the development of stable materials for high-temperature applications. Although the thermal coarsening of small and active nanocrystals into less active aggregates is universal in material deactivation, the atomic mechanisms governing nanocrystal growth remain elusive. By utilizing colloidally synthesized Pd/SiO2 powder nanocomposites with controlled nanocrystal sizes and spatial arrangements, we unravel the competing contributions of particle coalescence and atomic ripening processes in nanocrystal growth. Through the study of size-controlled nanocrystals, we can uniquely identify the presence of either nanocrystal dimers or smaller nanoclusters, which indicate the relative contributions of these two processes. By controlling and tracking the nanocrystal density, we demonstrate the spatial dependence of nanocrystal coalescence and the spatial independence of Ostwald (atomic) ripening. Overall, we prove that the most significant loss of the nanocrystal surface area is due to high-temperature atomic ripening. This observation is in quantitative agreement with changes in the nanocrystal density produced by simulations of atomic exchange. Using well-defined colloidal materials, we extend our analysis to explain the unusual high-temperature stability of Au/SiO2 materials up to 800 °C.

ER/K-link-Leveraging a native protein linker to probe dynamic cellular interactions.

ER/K α-helices are a subset of single alpha helical domains, which exhibit unusual stability as isolated protein secondary structures. They adopt an elongated structural conformation, while regulating the frequency of interactions between proteins or polypeptides fused to their ends. Here we review recent advances on the structure, stability and function of ER/K α-helices as linkers (ER/K linkers) in native proteins. We describe methodological considerations in the molecular cloning, protein expression and measurement of interaction strengths, using sensors incorporating ER/K linkers. We highlight biological insights obtained over the last decade by leveraging distinct biophysical features of ER/K-linked sensors. We conclude with the outlook for the use of ER/K linkers in the selective modulation of dynamic cellular interactions.

An unconventional route to an ambipolar azaheterocycle and its in situ generated radical anion.

Herein we report the synthesis, characterization and application of an azaheterocycle 4 obtained via an unprecedented C-N coupling. The neutral azaheterocycle undergoes one-electron reduction to form an air-stable radical anion in situ, which provides added benefit towards operational stability of the device during n-type charge transport. The unusual stability of this radical anion is due to the fact that the fused cyclopentane ring upon reduction forms aromatic cyclopentadienyl anion, and the negative charge delocalizes over the nearly planar azaheterocycle core. The present azaheterocycle can be considered as a mimic of a fullerene fragment, which shows balanced ambipolar charge transport in space charge limited current (SCLC) devices with moderate hole (μh) and electron (μe) mobilities (μh = 2.96 × 10-3 cm2 V-1 s-1 and μe = 1.11 × 10-4 cm2 V-1 s-1). Theoretical studies such as nucleus independent chemical shifts (NICS) calculations, anisotropy of the induced current density (ACID) plots, spin density mapping and anisotropic mobility calculations were performed to corroborate the experimental findings.

Non-sterical stabilization of one-electron-oxidized NiSalen complex by thiophene core

A combined theoretical/experimental study of the new thiophene-based NiSalen complex with unconjugated bridging fragment. This complex demonstrates unusual stability of the oxidized form, which is not typical for this class of compounds.

Evaluation of the in Vivo Aphrodisiac Activity of a Cyclotide Extract from Hybanthus enneaspermus.

Hybanthus enneaspermus is an Indian folk medicinal herb that has been widely used as a libido enhancer. This plant belongs to the Violaceae plant family, which ubiquitously contains disulfide-rich cyclic peptides named cyclotides. Cyclotides are an expanding plant-derived peptide family with numerous interesting bioactivities, and their unusual stability against proteolysis has attracted much attention in drug design applications. Recently, H. enneaspermus has been reported to be a rich source of cyclotides, and hence, it was of interest to investigate whether cyclotides contribute to its aphrodisiac activity. In this study, we evaluated the in vivo aphrodisiac activity of the herbal powder, extract, and the most abundant cyclotide, hyen D, extracted from H. enneaspermus on rats in a single dose regimen. After dosing, the sexual behaviors of male rats were observed, recorded, analyzed, and compared with those of the vehicle group. The results show that the extract and hyen D significantly decreased the intromission latency of sexually naïve male rats and the extract improved a range of other measured sexual parameters. The results suggest that the extract could enhance libido as well as facilitate erectile function in male rats and that the cyclotide hyen D could contribute to the libido-enhancing activity of this ethnomedicinal herb.

Scalable Fabrication of Scratch-Proof Transparent Al/F-SnO2 Hybrid Electrodes with Unusual Thermal and Environmental Stability.

Fabrication protocols of transparent conducting electrodes (TCEs), including those which produce TCEs of high values of figure of merit, often fail to address issues of scalability, stability, and cost. When it comes to working with high-temperature stable electrodes, one is left with only one and that too, an expensive choice, namely, fluorine-doped SnO2 (FTO). It is rather difficult to replace FTO with a low-cost TCE due to stability issues. In the present work, we have shown that an Al nanomesh fabricated employing the crack template method exhibits extreme thermal stability in air even at 500 °C, compared with that of FTO. In order to fill in the non-conducting island regions present in between the mesh wires, a moderately conducting material SnO2 layer was found adequate. The innovative step employed in the present work relates to the SnO2 deposition without damaging the underneath Al, which is a challenge in itself, as the commonly used precursor, SnCl2 solution, is quite corrosive toward Al. Optimization of spray coating of the precursor while the Al mesh on a glass substrate held at an appropriate temperature was the key to form a stable hybrid electrode. The resulting Al/SnO2 electrode exhibited an excellent transparency of ∼83% at 550 nm and a low sheet resistance of 5.5 Ω/□. SnO2 coating additionally made the TCE scratch-proof and mechanically stable, as the adhesion tape test showed only 8% change in sheet resistance after 1000 cycles. Further, to give FTO-like surface finish, the SnO2 surface was fluorinated by treating with a Selectfluor solution. As a result, the Al/F-SnO2 hybrid film exhibited one order higher surface conductivity with negligible sensitivity toward humidity and volatile organics, while becoming robust toward neutral electrochemical environments. Finally, a custom-designed projection lithography technique was used to pixelate the Al/SnO2 hybrid film for optoelectronic device applications.

Axially and Helically Chiral Cationic Radical Bicarbazoles: SOMO-HOMO Level Inversion and Chirality Impact on the Stability of Mono- and Diradical Cations.

We report persistent chiral organic mono- and diradical cations based on bicarbazole molecular design with an unprecedented stability dependence on the type of chirality, namely, axial versus helical. An unusual chemical stability was observed for sterically unprotected axial bicarbazole radical in comparison with monocarbazole and helical bicarbazole ones. Such results were experimentally and theoretically investigated, revealing an inversion in energy of the singly occupied molecular orbital (SOMO) and the highest (doubly) occupied molecular orbital (HOMO) in both axial and helical bicarbazole monoradicals along with a subtle difference of electronic coupling between the two carbazole units, which is modulated by their relative dihedral angle and related to the type of chirality. Such findings allowed us to explore in depth the SOMO-HOMO inversion (SHI) in chiral radical molecular systems and provide new insights regarding its impact on the stability of organic radicals. Finally, these specific electronic properties allowed us to prepare a persistent, intrinsically chiral, diradical which notably displayed near-infrared electronic circular dichroism responses up to 1100 nm and almost degenerate singlet-triplet ground states with weak antiferromagnetic interactions evaluated by magnetometry experiments.

Methane Bi-reforming for direct ethanol production over smart Cu/Mn- ferrite catalysts

The main goal of this research is direct synthesis of ethanol from methane bi-reforming reaction (H2O/CO2 methane reforming, SCMR) in a single step catalytic process. Smart nano-structured spinel ferrite catalysts with small ratio of Cu or Mn in composite with Fe cluster were prepared using ultrasonic assistance technique. Characterization data indicate the stability of the prepared ferrite structure during calcination step, so it can use for high temperature reaction. From XPS and Raman spectra, cupper ferrite shows deviation from ideal spinel structure, which increases the migration of bulk oxygen into surface to share in catalytic activity. The catalytic activity test was performed at moderate reaction condition (700 °C) using CO2 as soft oxidant to avoid methane combustion reaction and steam to release the alcoholic products. The two catalysts show unusual high catalytic activity and stability due to strong synergetic effect between metal and Fe. Cu–Fe gives high selectivity (∼50%) toward liquid product (ethanol), while Mn–Fe is more selective (∼70%) toward gas product (syngas). Furthermore, the chemical looping step is used to obtain ultra-stable SCMR reaction. The steam activation steps were performed for re-oxidation of reduced catalyst in addition to removal of carbon deposited.

Exploring the Complexation of Counterion in Novel Family of Polyelectrolytes with Unexpected Solubility Behaviour

In the present paper we study the effect of complexation in linear negatively charged polyelectrolytes with different alkali ions. With combination of IR-spectroscopy, X-ray diffraction and nanocalorimetry, we attempted to explain unusual solubility, crystallinity and thermal stability of these polymers. The increase of thermal stability and insolubility in water in series of semi-crystalline polysalts as K+ ≤ H+ <Na+ was explained by effectiveness of formation of chelating complex. Insoluble in water sodium salt shows the highest thermal stability of crystal phase up to . In contrast, well soluble in water amorphous lithium salt does not self-organize in chelating complex and is presented in ionic form.

REACTIVITY OF NUCLEOPHILES AND α-EFFECT IN SUBSTITUTION PROCESSES AT ELECTRON - DEFICIENCY CENTERS (Part 2)

The review analyzes issues related to the reactivity of nucleophiles and the manifestation of the α-effect in substitution processes at electron-deficient centers. The fundamental aspects of this phenomenon, as well as the possibilities and prospects of using α-nucleophiles in systems for the highly efficient degradation of substrates - ecotoxicants of various natures, are discussed. In the first part of the review such aspects were observed: inorganic α-nucleophiles as the most effective class of reagents for the decomposition of organic phosphorus compounds, hydroxylamine, its N-alkyl derivatives, oximes, and hydroxamic acids, reactivity of the НОО– anion in the processes of acyl group transfer, reactivity of oximate ions, inorganic α-nucleophiles as the basis of formulations for the degradation of neurotoxins, vesicants, and organophosphorus pesticides, design of inhibited acetylcholinesterase reactivators based on hydroxylamine derivatives, ways of structural modification of α-nucleophiles and systems based on them. The data on the reactivity of typical inorganic α-nucleophiles in the cleavage of acyl-containing substrates, including phosphorus acid esters, which provide abnormally high reaction rates in comparison with other supernucleophiles, are analyzed. Various types of such α-nucleophiles, features of their structure and reactivity are considered. It was shown that an important feature of hydroxylamine, oximes, and hydroxamic acids is the presence of a fragment with adjacent O and N (–N – O – H) atoms containing one or more lone electron pairs, which determines their belonging to the class of α-nucleophiles. It has been shown that a many of factors can be responsible for the manifestation of the α-effect and its magnitude, the main of which is the destabilization of the ground state of the nucleophile due to repulsion of lone electron pairs, stabilization of the transition state, the unusual thermodynamic stability of reaction products, solvation effects of the solvent, type of hybridization of the electrophilic center, etc.

Soft Dynamic Confinement of Membrane Proteins by Dehydrated Trehalose Matrices: High-Field EPR and Fast-Laser Studies

In memory of the 85th birthday of Yakov S. Lebedev (Moscow), who died in 1996, we start this Review on soft-glass matrix effects in donor–acceptor complexes with an appreciation of his pioneering work on high-field EPR spectroscopy on tribochemically generated donor–acceptor complexes. The mechanochemical activation of polycrystalline mixtures of porphyrins (and other donors) and quinone acceptors was found to produce large concentrations of triplet donor molecules and donor–acceptor radical pairs with unusual stability. The Review is continued with reporting on W-band high-field EPR and fast-laser studies on disaccharide matrix effects on structure and dynamics of donor–acceptor protein complexes related to photosynthesis, including the non-oxygenic bacterial reaction center (RC) and the oxygenic RCs Photosystem I (PS I) and Photosystem II (PS II, preliminary results). Some organisms can survive complete dehydration and high temperatures by adopting an anhydrobiotic state in which the intracellular medium contains large amounts of disaccharides, in particular trehalose and sucrose. Trehalose is most effective in protecting biostructures, both in vivo and in vitro. To clarify the molecular mechanisms of disaccharide bioprotection, structure and dynamics of sucrose and trehalose matrices at different controlled hydration levels were probed by perdeuterated nitroxide spin labels and native cofactor intermediates in their charge-separated states. Trehalose forms a homogeneous amorphous phase in which the hosted molecules are uniformly distributed. Notably, their rotational mobility at room temperature is dramatically impaired by the trehalose H-bonding network confinement to an extent that in normal protein–matrix systems is only observed at low temperatures around 150 K. From the experimental results, formation of an extended H-bonding network of trehalose with protein molecules is inferred, involving both bulk and local water molecules. The H-bond network extends homogeneously over the whole matrix integrating and immobilizing the hosted protein. Taken together, these observations suggest that in photosystems, such as bacterial RCs and PS I complexes, of different size and complexity regarding subunit composition and oligomeric organization, the molecular configuration of the cofactors involved in the primary processes of charge separation is not significantly distorted by incorporation into trehalose glass, even under extensive dehydration. By means of pulsed W-band high-field multiresonance EPR spectroscopies, such as ELDOR-detected NMR and ENDOR, in conjunction with using isotope labeled water (D2O and H217O), the biologically important issue of sensing and quantification of local water in proteins is addressed. The bacterial RC embedded into the trehalose glass matrix is used as model system. The two native radical cofactor ions of the primary electron donor and acceptor as well as an artificial nitroxide spin label site-specifically attached to the protein surface are studied in the experiments. The three paramagnetic reporter groups probe distinctly different local environments. They sense water molecules via their magnetic hyperfine and quadrupole interactions with either deuterons or 17O nuclei. It is shown that by using oxygen-17 labeled water, quantitative conclusions can be drawn differentiating between local and bulk water. It is concluded that dry trehalose operates as anhydrobiotic protein stabilizer by means of selective changes in the first solvation shell of the protein upon trehalose–matrix dehydration with subsequent changes in the hydrogen-bonding network. Such changes usually have an impact on the global function of a biological system. Finally, preliminary results of optical and W-band EPR experiments on the extremolytes ectoine and its derivative hydroxyectoine are reported; these compounds appear to share several stress-protecting properties with trehalose in terms of stabilizing protein matrices. For instance, they display remarkable stabilizing capabilities towards sensitive proteins and enzymes with respect to freeze-thawing, heat-treatment, and freeze-drying procedures. Moreover, hydroxyectoine is a good glass-forming compound and exhibits a remarkable bioprotective effect against desiccation and heat denaturation of functional protein complexes.

REACTIVITY OF NUCLEOPHILES AND α-EFFECT IN SUBSTITUTION PROCESSES AT ELECTRON - DEFICIENCY CENTERS

The review analyzes issues related to the reactivity of nucleophiles and the manifestation of the α-effect in substitution processes at electron-deficient centers. The fundamental aspects of this phenomenon, as well as the possibilities and prospects of using α-nucleophiles in systems for the highly efficient degradation of substrates - ecotoxicants of various natures, are discussed. In the first part of the review such aspects were observed: inorganic α-nucleophiles as the most effective class of reagents for the decomposition of organic phosphorus compounds, hydroxylamine, its N-alkyl derivatives, oximes, and hydroxamic acids, reactivity of the НОО– anion in the processes of acyl group transfer, reactivity of oximate ions, inorganic α-nucleophiles as the basis of formulations for the degradation of neurotoxins, vesicants, and organophosphorus pesticides, design of inhibited acetylcholinesterase reactivators based on hydroxylamine derivatives, ways of structural modification of α-nucleophiles and systems based on them. The data on the reactivity of typical inorganic α-nucleophiles in the cleavage of acyl-containing substrates, including phosphorus acid esters, which provide abnormally high reaction rates in comparison with other supernucleophiles, are analyzed. Various types of such α-nucleophiles, features of their structure and reactivity are considered. It was shown that an important feature of hydroxylamine, oximes, and hydroxamic acids is the presence of a fragment with adjacent O and N (–N – O – H) atoms containing one or more lone electron pairs, which determines their belonging to the class of α-nucleophiles. It has been shown that a many of factors can be responsible for the manifestation of the α-effect and its magnitude, the main of which is the destabilization of the ground state of the nucleophile due to repulsion of lone electron pairs, stabilization of the transition state, the unusual thermodynamic stability of reaction products, solvation effects of the solvent, type of hybridization of the electrophilic center, etc.

Silver Adsorption on Calcium Niobate(001) Nanosheets: Calorimetric Energies That Explain Sinter-Resistant Support.

Metal nanoparticles deposited on oxide supports are essential to many technologies, including catalysts, fuel cells, and electronics. Therefore, understanding the chemical bonding strength between metal nanoparticles and oxide surfaces is of great interest. The adsorption energetics, adhesion energy, and adsorbate structure of Ag on dehydrated HCa2Nb3O10(001) nanosheets at 300 K have been studied using metal adsorption calorimetry and surface spectroscopies. These dehydrated ("dh") calcium niobate nanosheets (dh-HCa2Nb3O10(001)) have the stoichiometry Ca4Nb6O19. They impart unusual stability to metal nanoparticles when used as catalyst supports and are easy-to-prepare by Langmuir-Blodgett (LB) techniques, highly ordered, and essentially single-crystal surfaces of mixed oxides with a huge ratio of terrace to edge sites. Below the monolayer coverage, Ag grows on dh-HCa2Nb3O10(001) as 2D islands of thickness ∼2 layers. The differential heat of Ag adsorption is initially ∼303 kJ/mol, increasing slowly to ∼338 kJ/mol by 0.8 ML. At higher coverages, Ag atoms mainly add on top of these 2D islands, growing 3D nanoparticles of increasing thickness, as the heat decreases asymptotically toward silver's heat of sublimation (285 kJ/mol). The adhesion energy of Ag(s) to this Ca niobate surface is estimated to be 4.33 J/m2, larger than that on any oxide surface previously measured. This explains the sinter resistance reported for metal nanoparticles on this support. Electron transfer from Ag into the calcium niobate is also measured. These results demonstrate an easy way to do single-crystal-type surface science studies-and especially thermochemical measurements-on the complex surfaces of mixed oxides: using LB-deposited perovskite nanosheets and ultrahigh-vacuum annealing in O2.

Escape of TLR5 Recognition by Leptospira spp.: A Rationale for Atypical Endoflagella.

Leptospira (L.) interrogans are invasive bacteria responsible for leptospirosis, a worldwide zoonosis. They possess two periplasmic endoflagellae that allow their motility. L. interrogans are stealth pathogens that escape the innate immune recognition of the NOD-like receptors NOD1/2, and the human Toll-like receptor (TLR)4, which senses peptidoglycan and lipopolysaccharide (LPS), respectively. TLR5 is another receptor of bacterial cell wall components, recognizing flagellin subunits. To study the contribution of TLR5 in the host defense against leptospires, we infected WT and TLR5 deficient mice with pathogenic L. interrogans and tracked the infection by in vivo live imaging of bioluminescent bacteria or by qPCR. We did not identify any protective or inflammatory role of murine TLR5 for controlling pathogenic Leptospira. Likewise, subsequent in vitro experiments showed that infections with different live strains of L. interrogans and L. biflexa did not trigger TLR5 signaling. However, unexpectedly, heat-killed bacteria stimulated human and bovine TLR5, but did not, or barely induced stimulation via murine TLR5. Abolition of TLR5 recognition required extensive boiling time of the bacteria or proteinase K treatment, showing an unusual high stability of the leptospiral flagellins. Interestingly, after using antimicrobial peptides to destabilize live leptospires, we detected TLR5 activity, suggesting that TLR5 could participate in the fight against leptospires in humans or cattle. Using different Leptospira strains with mutations in the flagellin proteins, we further showed that neither FlaA nor Fcp participated in the recognition by TLR5, suggesting a role for the FlaB. FlaB have structural homology to Salmonella FliC, and possess conserved residues important for TLR5 activation, as shown by in silico analyses. Accordingly, we found that leptospires regulate the expression of FlaB mRNA according to the growth phase in vitro, and that infection with L. interrogans in hamsters and in mice downregulated the expression of the FlaB, but not the FlaA subunits. Altogether, in contrast to different bacteria that modify their flagellin sequences to escape TLR5 recognition, our study suggests that the peculiar central localization and stability of the FlaB monomers in the periplasmic endoflagellae, associated with the downregulation of FlaB subunits in hosts, constitute an efficient strategy of leptospires to escape the TLR5 recognition and the induced immune response.

Molecular genetics of the virulence plasmids of pathogenic Escherichia coli O104:H4

In 2011 a large outbreak of enterohemorrhagic gastroenteritis and haemolytic uremic syndrome (HUS) throughout Europe resulted in almost 4,000 infections, 845 cases of HUS and 54 fatalities. This was due to a dangerous exchange of mobile genetic elements (MGE) resulting in a hybrid strain of E. coli O104:H4. This strain carried an unusual combination of EAEC- and STEC-associated virulence factors on a plasmid and phage respectively. In vitro the virulence plasmid has exhibited unusual stability under a wide range of environmental stresses, contrasting with rapid plasmid loss in the human gut. This project will characterise plasmid encoded maintenance systems responsible for its unique stability. Current investigations focus on toxin-antitoxin (TA) systems involved in postsegregationalkilling which contribute to plasmid maintenance therefore resulting in increased virulence. By inducing expression of putative TA genes cloned onto lab-made plasmid vectors, we have analysed their function in the cell. Once characterised we will further investigate the effects of various environmental conditions able to disrupt these TA systems, ultimately resulting in plasmid loss. This atypical strain displayed heightened pathogenicity and providedun foreseen treatment challenges. We aim to further our understanding of MGE carriage in O104:H4 as a model to predict and combat future outbreaks of hybrid pathovars.

Effect of 2,2,6,6-Tetramethylpiperidinyl-Oxide (TEMPO) as an Electrolyte Additive and its SEI Formation on MCMB-Electrode

In this study, we developed the benefit of TEMPO additive for the suppression of the solvent/additive decomposition by trapping the radicals formed. With unusual stability as a neutral radical the 2 % TEMPO as an additive into STD + 5% FEC electrolyte resulted in the modification of the interphase. Through in-situ DRIFTS results we can find out and investigate the mechanism for the decomposition of TEMPO. The studies described herein also demonstrate that MCMB-electrodes in STD + 5% FEC solution containing TEMPO additive can be utilized together with high voltage cathode.