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Vibrio species play a crucial role in maintaining the carbon and nitrogen balance between the oceans and the land through their ability to employ chitin as a sole source of energy. This study describes the structural basis for the action of the GH20 β‐N‐acetylglucosaminidase (VhGlcNAcase) in chitin metabolism by Vibrio campbellii (formerly V. harveyi) strain ATCC BAA‐1116. Crystal structures of wild‐type VhGlcNAcase in the absence and presence of the sugar ligand, and of the unliganded D437A mutant, were determined. VhGlcNAcase contains three distinct domains: an N‐terminal carbohydrate‐binding domain linked to a small α+β domain and a C‐terminal (β/α)8 catalytic domain. The active site of VhGlcNAcase has a narrow, shallow pocket that is suitable for accommodating a small chitooligosaccharide. VhGlcNAcase is a monomeric enzyme of 74 kDa, but its crystal structures show two molecules of enzyme per asymmetric unit, in which Gln16 at the dimeric interface of the first molecule partially blocks the entrance to the active site of the neighboring molecule. The GlcNAc unit observed in subsite −1 makes exclusive hydrogen bonds to the conserved residues Arg274, Tyr530, Asp532 and Glu584, while Trp487, Trp546, Trp582 and Trp505 form a hydrophobic wall around the −1 GlcNAc. The catalytic mutants D437A/N and E438A/Q exhibited a drastic loss of GlcNAcase activity, confirming the catalytic role of the acidic pair (Asp437–Glu438)., Crystal structures of a GH20 β‐N‐acetylglucosaminidase from V. campbellii reveal substrate specificity in chitin utilization.

Gravity thickening by deep cone thickener is popularly applied in cemented paste backfill, which is essential for tailings management and green mining. To avoid rake bogging, high concentration and high flowability are required for thickened tailings slurry. These properties depend critically on the yield stress of the slurry. The yield stress, in turn, can be influenced by the flocculant dosage and the acidity (pH) that form the flocculation condition. Experiments have been performed to eventually quantify the dependence of yield stress on these two variables. The yield stress test of flocculated and thickened copper mineral tailings slurry, and the analysis of flocculant adsorption, were conducted at different pH values and flocculant dosages. It turns out that the amount of flocculant adsorbed on the tailings particles surface has a maximum with respect to pH but increases with the increasing flocculant dosage. In turn, the amount of flocculant adsorbed affects the yield stress, a macroscopic property of the slurry, through the total interactions (microscopic) between polymer-coated tailings particles. Bridging interaction is essential for total interactions. It is obtained that the yield stress varies analogously to the absolute of the minimum value of the total interactions at different pH values and flocculant dosages. Therefore, interparticle interactions should be adjusted by control pH and flocculant dosage to achieve high-concentration and high-flowability underflow to avoid rake bogging in a deep cone thickener.

A phosphine-catalyzed oligomerization of arynes using selenocyanates was developed. The use of JohnPhos as a bulky phosphine is the key to accessing α,ω-bisfunctionalized oligo(ortho-arylenes) with RSe as the substituent at one terminus and CN as the substituent at the other. The in situ formation of R3PSeR' cations, serving as sterically encumbered electrophiles, hinders the immediate reaction that affords the 1,2-bisfunctionalization product and instead opens a competitive pathway leading to oligomerization. Various optimized conditions for the predominant formation of dimers, but also for higher oligomers such as trimers and tetramers, were developed. Depending on the electronic properties of the electrophilic reaction partner, even compounds up to octamers were isolated. Optimization experiments revealed that a properly tuned phosphine as catalyst is of crucial importance. Mechanistic studies demonstrated that the cascade starts with the attack of cyanide; aryne insertion into n-mers leading to (n+1)-mers was ruled out.

Recently, a rare type of relapse was reported upon treating a B cell acute lymphoblastic leukemia (B-ALL) patient with anti-CD19 chimeric antigen receptor (CAR)-T cells caused by unintentional transduction of residual malignant B cells (CAR-B cells). We show that anti-CD19 and anti-CD20 CARs are presented on the surface of lentiviral vectors (LVs), inducing specific binding to the respective antigen. Binding of anti-CD19 CAR-encoding LVs containing supernatant was reduced by CD19-specific blocking antibodies in a dose-dependent manner, and binding was absent for unspecific LV containing supernatant. This suggests that LVs bind via displayed CAR molecules to CAR antigen-expressing cells. The relevance for CAR-T cell manufacturing was evaluated when PBMCs and B-ALL malignant B cells were mixed and transduced with anti-CD19 or anti-CD20 CAR-displaying LVs in clinically relevant doses to mimic transduction conditions of unpurified patient leukapheresis samples. Malignant B cells were transduced at higher levels with LVs displaying anti-CD19 CARs compared to LVs displaying non-binding control constructs. Stability of gene transfer was confirmed by applying a potent LV inhibitor and long-term cultures for 10 days. Our findings provide a potential explanation for the emergence of CAR-B cells pointing to safer manufacturing procedures with reduced risk of this rare type of relapse in the future., Graphical abstract, Schaser and colleagues found that lentiviral vectors encoding B cell-specific CARs display the respective CAR on the particle surface. The LV-displayed CARs retained their antigen-binding potential and specifically promoted transduction of malignant B cells. These findings provide a potential explanation for the emergence of CAR-expressing malignant cells in B-ALL patients.

Emerging applications in spectroscopy-related bioanalytics demand for integrated devices with small geometric footprints and fast response times. While hollow core waveguides principally provide such conditions, currently used approaches include limitations such as long diffusion times, limited light-matter interaction, substantial implementation efforts, and difficult waveguide interfacing. Here, we introduce the concept of the optofluidic light cage that allows for fast and reliable integrated spectroscopy using a novel on-chip hollow core waveguide platform. The structure, implemented by 3D nanoprinting, consists of millimeter-long high-aspect-ratio strands surrounding a hollow core and includes the unique feature of open space between the strands, allowing analytes to sidewise enter the core region. Reliable, robust, and long-term stable light transmission via antiresonance guidance was observed while the light cages were immersed in an aqueous environment. The performance of the light cage related to absorption spectroscopy, refractive index sensitivity, and dye diffusion was experimentally determined, matching simulations and thus demonstrating the relevance of this approach with respect to chemistry and bioanalytics. The presented work features the optofluidic light cage as a novel on-chip sensing platform with unique properties, opening new avenues for highly integrated sensing devices with real-time responses. Application of this concept is not only limited to absorption spectroscopy but also includes Raman, photoluminescence, or fluorescence spectroscopy. Furthermore, more sophisticated applications are also conceivable in, e.g., nanoparticle tracking analysis or ultrafast nonlinear frequency conversion.

Due to the high-reflective characteristic of copper materials, welding with infrared lasers can be challenging and tends to a low process stability. New high-power laser sources and system technologies got introduced in the last years, promising better process stability and increased weld penetration for copper welding. Currently there are different approaches applied in industry: Wobble welding with a fast scanner; a variable, superimposed intensity distribution with a 2-in-1 fiber; or the use of laser sources with a visible wavelength (e.g. green). The current state of the art of these three technologies and their direct comparison with regard to the weldability of copper is discussed in this paper. The weld seams are compared in terms of seam quality, maximum welding depth and process speed. In addition, the possibility of a combination of considered system technologies is discussed.

In laser beam welding of aluminum alloy there is a high susceptibility of pore formation at low feed rates. Porosity can be improved by application of several laser spots to form the shape of the vapor capillary. In this regard, flexible, concentrically arranged laser spots with variable power distribution have recently been developed. Nevertheless, the influence of power distribution on weld quality and geometry has not been investigated. Therefore, this paper presents investigations regarding the influence of power distribution on the weld seam properties like penetration depth, seam width and porosity. Welding trials are carried out with three different aluminum alloys EN AW-1050, EN AW-6082 and EN AW-5083. Compared to a mono-spot the porosity can be reduced from 12 % to 8 % with power distribution of 75/25 of inner core and outer ring.

A large body of literature is available on wound healing in humans. Nonetheless, a standardized ex vivo wound model without disruption of the dermal compartment has not been put forward with compelling justification. Here, we present a novel wound model based on application of negative pressure and its effects for epidermal regeneration and immune cell behaviour. Importantly, the basement membrane remained intact after blister roof removal and keratinocytes were absent in the wounded area. Upon six days of culture, the wound was covered with one to three-cell thick K14+Ki67+ keratinocyte layers, indicating that proliferation and migration were involved in wound closure. After eight to twelve days, a multi-layered epidermis was formed expressing epidermal differentiation markers (K10, filaggrin, DSG-1, CDSN). Investigations about immune cell-specific manners revealed more T cells in the blister roof epidermis compared to normal epidermis. We identified several cell populations in blister roof epidermis and suction blister fluid that are absent in normal epidermis which correlated with their decrease in the dermis, indicating a dermal efflux upon negative pressure. Together, our model recapitulates the main features of epithelial wound regeneration, and can be applied for testing wound healing therapies and investigating underlying mechanisms.

Internal alkynes substituted by aliphatic or aromatic moieties or by heteroatoms were converted into sulphur-substituted acrylonitrile derivatives. Key is the use of Pd catalysis, which allows the addition of aromatic and aliphatic thiocyanates in an intra- and intermolecular manner. Substrates with several alkyne units underwent further carbopalladation steps after the initial thiopalladation step, thus generating in a cascade-like fashion an oligoene unit with sulphur at one terminus and the cyano group at the other., The intra- and intermolecular Pd-catalysed cyanosulfenylation of internal alkynes enables the formation of tetrasubstituted thioacrylonitriles and is extended to oligoyne systems leading to oligoenes and a cyanide transfer over four or six atoms.

Trimeric photosystem I from the cyanobacterium Thermosynechococcus elongatus (TePSI) is an intrinsic membrane protein, which converts solar energy into electrical energy by oxidizing the soluble redox mediator cytochrome c6 (Cyt c6) and reducing ferredoxin. Here, we use cryo-electron microscopy and small angle neutron scattering (SANS) to characterize the transient binding of Cyt c6 to TePSI. The structure of TePSI cross-linked to Cyt c6 was solved at a resolution of 2.9 A and shows additional cofactors as well as side chain density for 84% of the peptide chain of subunit PsaK, revealing a hydrophobic, membrane intrinsic loop that enables binding of associated proteins. Due to the poor binding specificity, Cyt c6 could not be localized with certainty in our cryo-EM analysis. SANS measurements confirm that Cyt c6 does not bind to TePSI at protein concentrations comparable to those for cross-linking. However, SANS data indicate a complex formation between TePSI and the non-native mitochondrial cytochrome from horse heart (Cyt cHH). Our study pinpoints the difficulty of identifying very small binding partners (less than 5% of the overall size) in EM structures when binding affinities are poor. We relate our results to well resolved co-structures with known binding affinities and recommend confirmatory methods for complexes with KM values higher than 20 μM.

Zinc oxide tetrapods (ZnO TP) have reached a high level of interest in the last years due to their broad fields of applications. Here, the combination of the adhesion promoting properties of both ZnO TP and poly(acrylic acid) (PAA) is demonstrated. A very rapid spray coating of water-based dispersions has been used to deposit homogenous thin nanocomposite films on poly(propylene) (PP) foils considered as model system. The characterization of ZnO TPs has been carried out by field-emission scanning electron microscopy (FE-SEM), high-resolution X-ray photoelectron spectroscopy (XPS) and transmission electron microscopy (TEM). The increased shear strength values exhibited by PP/PP laminates with interfacial ZnO TP/PAA films could be attributed to mechanical interlocking of ZnO TP and the macromolecular adhesion promotion by PAA. FE-SEM imaging after shear tests indicated a failure in the core of the ZnO TP. The fast application method via plasma activation of the PP foils and spray coating of the composite films with low concentrations of 0.085 mM PAA and 8 mg/ml ZnO TP makes this technique rather interesting for industrial applications.

In fusion devices, the retention of the fusion fuel deuterium (D) and tritium (T) in plasma-facing components (PFCs) is a major concern. Measurement of their hydrogen isotope content gives insight into the retention physics. In FREDIS, two methods of thermal desorption are used for retention measurements: In Thermal Desorption Spectrometry (TDS) the samples are heated by 6 infrared lamps up to 1433 K with linear temperature ramps of up to 1.67 K/s. The desorbed gases are detected up to 100 amu/e with a double-QMS (Quadrupole Mass Spectrometer) that can distinguish between helium and D2 and uses an innovative differential pumping system. In a connected vacuum chamber, a ∅3 mm spot can be heated on the sample surface by a high energy Nd:YAG laser pulse (E0

Plants are under relentless challenge by pathogenic bacteria, fungi, and oomycetes, for whom they provide a resource of living space and nutrients. Upon detection of pathogens, plants carry out multiple layers of defense response, orchestrated by a tightly organized network of hormones. In this review, we provide an overview of the phytohormones involved in immunity and the ways pathogens manipulate their biosynthesis and signaling pathways. We highlight recent developments, including the discovery of a defense signaling molecule, new insights into hormone biosynthesis, and the increasing importance of signaling hubs at which hormone pathways intersect.

As constituents of cellular membranes, lecithins feature high biocompatibility and great emulsifying properties due to their amphiphilicity. Additionally, there are expectations that these naturally occurring emulsifying agents can replace other skin damaging emulsifiers like sodium dodecyl sulfate or sodium laureth sulfate. However, cytotoxicity data of lecithin-based formulations on primary human skin cells are scarce. Thus, we developed nanoemulsions with different kinds of surfactants (amphoteric, anionic and non-ionic), studied the skin permeation of a model drug from this formulations employing Franz-type diffusion cells and monitored their cytotoxicity potential on primary human keratinocytes and fibroblasts using a cell proliferation assay. The skin diffusion studies demonstrated that the amphoteric lecithin-based emulsifiers were superior to non-ionic surfactants in terms of skin permeation, but inferior to anionic emulsifiers. Further, we found that the nanoemulsions containing the amphoteric lecithins as emulsifying agents lead to significantly higher viability rates of both epidermal keratinocytes and dermal fibroblasts than the investigated anionic and non-ionic surfactants. This renders them a promising alternative to conventional emulsifiers used in daily products.

COPD patients have an increased susceptibility to bacterial airway infections that can induce exacerbations. In response to infections, circulating monocytes become recruited to the infected tissue and secrete cytokines. We hypothesized that this cytokine response is reduced in COPD. Cultured peripheral blood monocytes of never smokers (NS) and smokers without (S) and with COPD (3 study populations, n = 36-37) were stimulated with extracts of Haemophilus influenzae, Staphylococcus aureus, or Streptococcus pneumoniae or with four different pathogen-associated molecular patterns (PAMPs). Four cytokines and 9 PAMP-related signaling molecules were measured and compared between the groups. Granulocyte-macrophage-colony-stimulating-factor responses to all stimulants were reduced in S and COPD compared to NS. Tumor-necrosis-factor-α responses to all bacterial extracts, peptidoglycan, and lipopolysaccharide were reduced in S and/or COPD. Interleukin-10 responses to S. aureus and lipoteichoic acid were increased in COPD. Correlations to pack-years and lung function were found. The peptidoglycan-receptor NOD2 and the mRNA of the lipopolysaccharide-receptor TLR4 were reduced in S and COPD. Cytokine responses of monocytes to bacteria are suppressed by smoking and in COPD possibly due to NOD2 and TLR4 reduction and/or interleukin-10 increase. This might help to explain the increased susceptibility to bacterial infections. These systemic molecular pathologies might be targets for therapeutic strategies to prevent infection-induced exacerbations. KEY MESSAGES: COPD subjects have an increased susceptibility to bacterial infections. This implies defects in the immune response to bacteria and is critical for disease progression. The cytokine response of monocytes to bacteria is reduced in COPD. This might be due to a reduced NOD2 and TLR4 and an increased IL-10 expression. This can explain the increased susceptibility to infections and help to identify drug targets.

We present a superconducting bolometer fabricated by a rolled-up technology that allows one to combine the two-dimensionality (2D) of the superconducting layer with a helical spiral curvature. The bolometer is formed as a free-standing Nb nanohelix acting as an ultrathin transition-edge sensor (TES) and having a negligible thermal contact to the substrate. We demonstrate the functionality of the thin-film TES by examining its microwave-detection performance in comparison with a commercial cryogenic bolometer from QMC Instruments. The nanohelix has been revealed to feature a noise equivalent power (NEP) of about 2 × 10

With hydrogen becoming more and more important as storage and carrier for renewable energy, there is an increasing need for flexible and efficient storage technologies. However, existing technologies, such as liquefaction or compression, often require a significant share of the hydrogens lower heating value. High-temperature metal hydrides (HT-MHs), such as magnesium hydride, are a promising alternative. Due to high operation temperatures, their application is challenging. A novel adiabatic hydrogen storage reactor based on the combination of a HT-MH with a thermochemical energy storage system (TCSS), such as Mg(OH)2/MgO + H2O, can be a solution. In this work, the previously published numerical simulations for hydrogen absorption are extended to the desorption process. A two-dimensional model for the hydrogen release was set up. The performance of the storage reactor is strongly dependent on the thermodynamic equilibrium of the reactions involved and less dependent on the reaction kinetics. Dehydrogenation is possible within 132 min, which is in the vicinity of the hydrogenation time. To enhance the dehydrogenation process, the water vapor pressure can be adjusted aiming for higher temperatures during the MgO hydration. Hydrogen can either be provided at constant pressure or constant mass flow rate.

This study discusses the synthesis and preparation of ionically cross-linked acid-base blend membranes based on phosphonated poly(pentafluorostyrene) and poly(benzimidazole) for vanadium redox flow battery application (VRFB). Poly(pentafluorostyrene) was obtained by emulsion polymerization and reacted with tris(trimethylsilyphosphite) to obtain highly phosphonated poly(pentafluorostyrene) (PWN). For obtaining chemically and mechanically stable membrane PWN was blended with poly(benzimidazole) (F6PBI) in different weight ratios. The blend membranes showed high resistance to heat (above 400 °C) and the harsh conditions of the VRFB (highly acid and oxidizing conditions). Among the blends, the membrane consisting of PWN/F6PBI (9/1) weight ratio was identified as the most suitable separator due to its high ion-exchange capacity and conductivity. Additionally, this membrane showed the highest Coulomb efficiency (99%) proving its excellent separation ability for VRFB electrolytes. A more detailed study of this blend membrane revealed low self-discharge rate of about 1.6 mV h−1 being almost an order of magnitude lower than those of Nafion®212 (10.7 mV h−1) and lasting above 120 h to the voltage drop. Additionally, the membrane showed no lost of capacity until 600 charge-discharge cycles and only 30% after the 1600 cycles. Thus, in this study, we were able to show for the first time the potentials of a phosphonic acid based electrolyte separator for the application in VRFB.

The formation of dislocation substructures in up to 10 µm deep subsurface regions of two aluminium alloys, Al-5Mg and Al-11Zn, was investigated under conditions of high homologous temperature reciprocal sliding wear (HT-RSW). Under creep conditions, Al-5Mg shows a solid solution type of inverse primary creep. In contrast, Al-11Zn creeps obstacle controlled and exhibits normal primary creep. These two materials were subjected to reciprocal sliding wear at 200 and 300 °C for 100 and 1000 cycles. Flat polished disks were exposed to the 1 mm reciprocal movements of a spherical aluminium oxide counterbody under normal forces of 5 and 10 N at an oscillation frequency of 1 Hz. Using focused ion beam (FIB) micromachining thin electron transparent foils were prepared from the surface regions of the as received and worn material states. Transmission electron microscopy (TEM) was used to study the evolution of nano and micro grain sizes in the surface regions. Despite the different creep behavior, the two materials behave similar under conditions of reciprocal sliding wear. The results obtained in the present work show that subgrain sizes decrease with increasing numbers of wear cycles and increasing normal forces. Subgrain sizes also increase with increasing temperature. At 300 °C, dynamic recrystallization was observed in both Al-alloys. The results of the present work are discussed in the light of previous results reported in the literature. Areas in need of further work are highlighted.

In previous studies, lecithin-based nanoemulsions (NEs) have been shown to be skin friendly drug carrier systems. Due to their nontoxic properties, NEs might also be suitable as wound healing agents. Hence, different O/W NEs based on lecithin Lipoid® S 75 and plant oils or medium chain triglycerides were produced and characterised. Two lipophilic natural wound healing agents, a betulin-enriched extract from birch bark (BET) and a purified spruce balm (PSB), were successfully incorporated and their effects on primary human skin cells were studied in vitro. MTT, BrdU and scratch assays uncovered the positive influence of the drug-loaded NEs on cell viability, proliferation and potential wound closure. Compared to control formulations, the NEs loaded with either BET or PSB led to higher cell viability rates of fibroblasts and keratinocytes. Higher proliferative activity of keratinocytes and fibroblasts was observed after the treatment, which is a prerequisite for wound closure. Indeed, in scratch assays NEs with PSB and notably BET showed significantly ameliorated wound closure rates than the negative control (unloaded NEs) and the positive control (NEs with dexpanthenol). Our findings suggest that BET and PSB are outstanding wound healing drugs and their incorporation into lecithin-based NEs may represent a valid strategy for wound care.

SUMMARYThe melanocortin-4 receptor (MC4R), a hypothalamic master regulator of energy homeostasis and appetite, is a G-protein coupled receptor and a prime target for the treatment of obesity. Here, we present cryo-electron microscopy structures of MC4R− Gs-protein complexes with two recently FDA-approved drugs, the peptide agonists NDP-α-MSH and setmelanotide, with 2.9 Å and 2.6 Å resolution. Together with signaling data, the complex structures reveal the agonist-induced origin of transmembrane helix (TM) 6 regulated receptor activation. In both structures, different ligand binding modes of NDP-α-MSH, a high-affinity variant of the endogenous agonist, and setmelanotide, an anti-obesity drug with biased signaling, underline the key role of TM3 for ligand-specific interactions and of calcium ion as a ligand-adaptable cofactor. The agonist-TM3 interplay subsequently impacts the receptor− Gs-protein interfaces, mainly at intracellular loop 2. These structures reveal mechanistic details of MC4R activation or inhibition and provide important insights into receptor selectivity that will facilitate the development of tailored anti-obesity drugs.

Hydralloy C5 (Ti0.95Zr0.05Mn1.46V0.45Fe0.09) is a very fast reacting metal hydride and is therefore investigated for two high thermal power applications in vehicles at room temperature and down to −20 °C: preheating and air conditioning. The experimental findings in a ~1 kg of hydride material scale are evaluated with regard to their implications for potential technical systems. In order to preheat components, Hydralloy C5 can be used to supply hydrogen to LaNi4.85Al0.15, which then releases thermal energy at a high thermal power even from temperatures as low as −20 °C. A main result is that, for the present reactor design, this coupled reaction shows a significant dependency of power on the ambient temperature. In fact, the reaction rate of Hydralloy C5 represents the main limitation on thermal power at low temperature. Nonetheless, the performance of at least 0.5 kW/kgMH allows the preheating of 2 L of lubricant by 20 K within 120 s by 2 kg of metal hydride in 0.8 L of volume. For air conditioning, the endothermal hydrogen desorption performance of Hydralloy C5 was investigated at 10–20 °C. The experiments performed for the considered open system with external hydrogen supply and release show thermal mean powers of up to 2 kW/kgMH. Overall, it is shown that the desorption reaction between 10 and 20 °C is thermally limited in the considered reactor design. Based on the power achieved, less than 4.5 kg of Hydralloy C5 would be sufficient to cover the air conditioning demand in conventional vehicles.

Summary: In this paper, we show that the essential Hsp90 co-chaperone Sgt1 is a member of a general protein quality control network that links folding and degradation through its participation in the degradation of misfolded proteins both in the cytosol and the endoplasmic reticulum (ER). Sgt1-dependent protein degradation acts in a parallel pathway to the ubiquitin ligase (E3) and ubiquitin chain elongase (E4), Hul5, and overproduction of Hul5 partly suppresses defects in cells with reduced Sgt1 activity. Upon proteostatic stress, Sgt1 accumulates transiently, in an Hsp90- and proteasome-dependent manner, with quality control sites (Q-bodies) of both yeast and human cells that co-localize with Vps13, a protein that creates organelle contact sites. Misfolding disease proteins, such as synphilin-1 involved in Parkinson’s disease, are also sequestered to these compartments and require Sgt1 for their clearance.

How to stop RNA polymerase Timely and tunable cessation of RNA synthesis is vital for cellular homeostasis. RNA helicases such as the archetypal termination factor r actively dismantle transcription complexes, but the transitory nature of termination makes the process hard to study structurally. Said et al. assembled ρ-bound transcription complexes and studied them using cryo–electron microscopy with an approach that captured a series of functional states en route to termination. They found an extensive and dynamic network of r interactions with RNA polymerase, nucleic acids, and accessory Nus factors. ρ mediates stepwise rearrangements of these contacts, transforming an actively transcribing complex into a moribund pretermination intermediate. Science , this issue p. 44

Octenidine dihydrochloride (OCT) is a widely used antiseptic molecule, promoting skin wound healing accompanied with improved scar quality after surgical procedures. However, the mechanisms by which OCT is contributing to tissue regeneration are not yet completely clear. In this study, we have used a superficial wound model by tape stripping of ex vivo human skin. Protein profiles of wounded skin biopsies treated with OCT-containing hydrogel and the released secretome were analyzed using liquid chromatography-mass spectrometry (LC–MS) and enzyme-linked immunosorbent assay (ELISA), respectively. Proteomics analysis of OCT-treated skin wounds revealed significant lower levels of key players in tissue remodeling as well as reepithelization after wounding such as pro-inflammatory cytokines (IL-8, IL-6) and matrix-metalloproteinases (MMP1, MMP2, MMP3, MMP9) when compared to controls. In addition, enzymatic activity of several released MMPs into culture supernatants was significantly lower in OCT-treated samples. Our data give insights on the mode of action based on which OCT positively influences wound healing and identified anti-inflammatory and protease-inhibitory activities of OCT.

Conformational isomerism plays a crucial role in defining the physical and chemical properties and biological activity of molecules ranging from simple organic compounds to complex biopolymers. However, it is often a significant challenge to differentiate and separate these isomers experimentally as they can easily interconvert due to their low rotational energy barrier. Here, we use the momentum correlation of fragment ions produced after inner-shell photoionization to distinguish conformational isomers of 1,2-dibromoethane (C2H4Br2). We demonstrate that the three-body breakup channel, C2H4+ + Br+ + Br+, contains signatures of both sequential and concerted breakup, which are decoupled to distinguish the geometries of two conformational isomers and to quantify their relative abundance. The sensitivity of our method to quantify these yields is established by measuring the relative abundance change with sample temperature, which agrees well with calculations. Our study paves the way for using Coulomb explosion imaging to track subtle molecular structural changes.

WFDC proteins such as peptidase inhibitor 3 and SLPI inhibit proteases in the epidermis and other tissues. In this study, we tested the hypothesis that further WFDC protein family members might contribute to epidermal homeostasis. We found that in addition to peptidase inhibitor 3 and SLPI, WFDC5 and WFDC12 were expressed in human epidermis. In contrast to WFDC5, the expression of WFDC12 was induced during the late differentiation of keratinocytes and was restricted to the outermost layer of live cells. Single-cell RNA sequencing demonstrated that WFDC12-positive keratinocytes were characterized by the upregulation of LCE mRNA expression and downregulated the expression of keratins and claudins. Immunogold-electron microscopy revealed the colocalization of WFDC12 with corneodesmosomes in the lower stratum corneum. WFDC12 was elevated in the affected skin of patients with psoriasis, atopic dermatitis, and Darier disease. By contrast, WFDC12 expression was strongly upregulated not only in the affected but even more so in clinically normal-appearing skin of patients with Netherton syndrome. Finally, functional analysis showed distinct inhibitory activity of WFDC12 on neutrophil elastase and epidermal kallikrein‒related peptidase. Altogether, our study identified WFDC12 as a marker of the last stage of epidermal keratinocyte differentiation and suggests that WFDC12 contributes to the control of protease activity in the stratum corneum.

The intra- and intermolecular synthesis of selenium-substituted acyclic and heterocyclic acrylonitrile derivatives is presented. The 1,2-difunctionalization of several internal alkynes substituted not only by aliphatic and aromatic residues but also by heteroelements is realized by the Pd-catalyzed activation of aromatic and aliphatic selenocyanates. A high functional group tolerance allows straightforward access to a broad scope of tetrasubstituted olefins. X-ray studies of some products reveal noncovalent chalcogen-chalcogen interactions between oxygen and selenium.

70 kDa heat shock proteins (Hsp70) are essential chaperones of the protein quality control network; vital for cellular fitness and longevity. The four cytosolic Hsp70’s in yeast, Ssa1-4, are thought to be functionally redundant but the absence of Ssa1 and Ssa2 causes a severe reduction in cellular reproduction and accelerates replicative aging. In our efforts to identify which Hsp70 activities are most important for longevity assurance, we systematically investigated the capacity of Ssa4 to carry out the different activities performed by Ssa1/2 by overproducing Ssa4 in cells lacking these Hsp70 chaperones. We found that Ssa4, when overproduced in cells lacking Ssa1/2, rescued growth, mitigated aggregate formation, restored spatial deposition of aggregates into protein inclusions, and promoted protein degradation. In contrast, Ssa4 overproduction in the Hsp70 deficient cells failed to restore the recruitment of the disaggregase Hsp104 to misfolded/aggregated proteins, to fully restore clearance of protein aggregates, and to bring back the formation of the nucleolus-associated aggregation compartment. Exchanging the nucleotide-binding domain of Ssa4 with that of Ssa1 suppressed this ‘defect’ of Ssa4. Interestingly, Ssa4 overproduction extended the short lifespan of ssa1Δ ssa2Δ mutant cells to a lifespan comparable to, or even longer than, wild type cells, demonstrating that Hsp104-dependent aggregate clearance is not a prerequisite for longevity assurance in yeast., Author summary All organisms have proteins that network together to stabilize and protect the cell throughout its lifetime. One of these types of proteins are the Hsp70s (heat shock protein 70). Hsp70 proteins take part in folding other proteins to their functional form, untangling proteins from aggregates, organize aggregates inside the cell and ensure that damaged proteins are destroyed. In this study, we investigated three closely related Hsp70 proteins in yeast; Ssa1, 2 and 4, in an effort to describe the functional difference of Ssa4 compared to Ssa1 and 2 and to answer the question: What types of cellular stress protection are necessary to reach a normal lifespan? We show that Ssa4 can perform many of the same tasks as Ssa1 and 2, but Ssa4 doesn’t interact in the same manner as Ssa1 and 2 with other types of proteins. This leads to a delay in removing protein aggregates created after heat stress. Ssa4 also cannot ensure that misfolded proteins aggregate correctly inside the nucleus of the cell. However, this turns out not to be necessary for yeast cells to achieve a full lifespan, which shows us that as long as cells can prevent aggregates from forming in the first place, they can reach a full lifespan.

We have investigated ferroelectric charged domains in polycrystalline hexagonal yttrium manganite thin films (Y1Mn1O3, Y0.95Mn1.05O3, Y1Mn0.99Ti0.01O3, and Y0.94Mn1.05Ti0.01O3) by scanning electron microscopy (SEM) in secondary electron emission mode with a small acceleration voltage. Using SEM at an acceleration voltage of 1.0 kV otherwise homogenous surface charging effects are reduced, polarization charges can be observed and polarization directions (±Pz) of the ferroelectric domains in the polycrystalline thin films can be identified. Thin films of different chemical composition have been deposited by pulsed laser deposition on Pt/SiO2/Si structures under otherwise same growth conditions. Using SEM it has been shown that different charged domain density networks are existing in polycrystalline yttrium manganite thin films.

Metal-containing formate dehydrogenases (FDH) catalyse the reversible oxidation of formate to carbon dioxide at their molybdenum or tungsten active site. They display a diverse subunit and cofactor composition, but structural information on these enzymes is limited. Here we report the cryo-electron microscopic structures of the soluble Rhodobacter capsulatus FDH (RcFDH) as isolated and in the presence of reduced nicotinamide adenine dinucleotide (NADH). RcFDH assembles into a 360 kDa dimer of heterotetramers revealing a putative interconnection of electron pathway chains. In the presence of NADH, the RcFDH structure shows charging of cofactors, indicative of an increased electron load., Rhodobacter capsulatus NAD+ dependent formate dehydrogenase (RcFDH) is a molybdoenzyme that catalyses the reversible oxidation of formate to carbon dioxide, and is of interest for biotechnological applications. Here the authors present the cryo-EM structures of RcFDH as isolated from R. capsulatus and in the reduced state with bound NADH, and discuss the enzyme mechanism.

SUMMARYProtein synthesis must be finely tuned in the nervous system, as it represents an essential feature of neurodevelopmental gene expression, and dominant pathology in neurological disease. However, the architecture of ribosomal complexes in the developing mammalian brain has not been analyzed at high resolution. This study investigates the architecture of ribosomesex vivofrom the embryonic and perinatal mouse neocortex, revealing Ebp1 as a 60S peptide tunnel exit binding factor at near-atomic resolution by multiparticle cryo-electron microscopy. The impact of Ebp1 on the neuronal proteome was analyzed by pSILAC and BONCAT coupled mass spectrometry, implicating Ebp1 in neurite outgrowth proteostasis, within vivoembryonic Ebp1 knockdown resulting in dysregulation of neurite outgrowth. Our findings reveal Ebp1 as a central component of neocortical protein synthesis, and the 60S peptide tunnel exit as a focal point of gene expression control in the molecular specification of neuronal morphology.Graphical abstract

The Type III Secretion Systems (T3SS) needle complex is a conserved syringe-shaped protein translocation nanomachine with a mass of about 3.5 MDa essential for the survival and virulence of many Gram-negative bacterial pathogens. This system is composed of a membrane-embedded basal body and an extracellular needle that deliver effector proteins into host cells. High-resolution structures of the T3SS from different organisms and infection stages are needed to understand the underlying molecular mechanisms of effector translocation. Here, we present the cryo-electron microscopy structure of the isolated Shigella T3SS needle complex. The inner membrane (IM) region of the basal body adopts 24-fold rotational symmetry and forms a channel system that connects the bacterial periplasm with the export apparatus cage. The secretin oligomer adopts a heterogeneous architecture with 16- and 15-fold cyclic symmetry in the periplasmic N-terminal connector and C-terminal outer membrane ring, respectively. Two out of three IM subunits bind the secretin connector via a β-sheet augmentation. The cryo-EM map also reveals the helical architecture of the export apparatus core, the inner rod, the needle and their intervening interfaces., Author summary Diarrheal diseases evoke about 2.2. million dead people annually and are the second leading cause of postneonatal child mortality worldwide. Shigella causing dysentery utilizes the type 3-secretion system (T3SS) to inject virulence factors into the gut cells. The T3SS needle complex is a syringe-shaped nanomachine consisting of two membrane-embedded ring systems that sheath a central export apparatus and a hollow needle-like structure through which the virulence factors are transported. We present here the structure of the Shigella T3SS needle complex obtained by high-end electron microscopy. The outer membrane (OM) ring system adopts a mixed 15- and 16-fold cyclic symmetry and the near-atomic structure shows the connection of the inner membrane (IM) and OM rings. Conserved channels in the IM ring connect the bacterial periplasm with the central export apparatus. Similar to the Salmonella flagellar system, the export apparatus and its connected needle-like structure assemble in a helical manner. This study advances our understanding of the role of essential structural elements in the T3SS assembly and function.

Single crystal Ni-based superalloys have long been an essential material for gas turbines in aero engines and power plants due to their outstanding high temperature creep, fatigue and oxidation resistance. A turning point was the addition of only 3 wt.% Re in the second generation of single crystal Ni-based superalloys which almost doubled the creep lifetime. Despite the significance of this improvement, the mechanisms underlying the so-called “Re effect” have remained controversial. Here, we provide direct evidence of Re enrichment to crystalline defects formed during creep deformation, using combined transmission electron microscopy, atom probe tomography and phase field modelling. We reveal that Re enriches to partial dislocations and imposes a drag effect on dislocation movement, thus reducing the creep strain rate and thereby improving creep properties. These insights can guide design of better superalloys, a quest which is key to reducing CO2 emissions in air-traffic., Adding minute amounts of rhenium to Ni-based single crystal superalloys extends their high temperature performance in engines, but the reasons behind that are still unclear. Here, the authors combine high resolution imaging and modelling to show that rhenium enriches and slows down partial dislocations to improve creep performance.

Block copolymer (BCP) self-assembly is a promising tool for next generation lithography as microphase separated polymer domains in thin films can act as templates for surface nanopatterning with sub-20 nm features. The replicated patterns can, however, only be as precise as their templates. Thus, the investigation of the morphology of polymer domains is of great importance. Commonly used analytical techniques (neutron scattering, scanning force microscopy) either lack spatial information or nanoscale resolution. Using advanced analytical (scanning) transmission electron microscopy ((S)TEM), we provide real space information on polymer domain morphology and interfaces between polystyrene (PS) and polymethylmethacrylate (PMMA) in cylinder- and lamellae-forming BCPs at highest resolution. This allows us to correlate the internal structure of polymer domains with line edge roughnesses, interface widths and domain sizes. STEM is employed for high-resolution imaging, electron energy loss spectroscopy and energy filtered TEM (EFTEM) spectroscopic imaging for material identification and EFTEM thickness mapping for visualisation of material densities at defects. The volume fraction of non-phase separated polymer species can be analysed by EFTEM. These methods give new insights into the morphology of polymer domains the exact knowledge of which will allow to improve pattern quality for nanolithography.

This work demonstrates the suitability of nitrogen removal during anaerobic acidification in batch configuration for a more efficient pre-treatment of chicken manure prior to anaerobic digestion. High loading rates corresponding to a total nitrogen input between 6.3 and 9.5 g L−1 allowed successful suppression of methanogenic archaea. To eliminate nitrogen, NH3-stripping and MAP (magnesium ammonium phosphate hexahydrate) precipitation were compared. In spite of decreased cell quantities detected using qPCR, removal of nitrogen caused an increase in volatile fatty acid (VFA) formation from 13 to 19%. The highest nitrogen removal during acidification (up to 29%) was achieved with three consecutives MAP precipitation steps, however, conductivity values were affected too, reaching 53.3 and 53.1 mS cm−1 after the three consecutive MAP precipitations. Additionally, MAP-precipitation reduced the concentration of important trace elements and 16S-rRNA amplicon sequencing revealed an altered taxonomic pattern, in which especially the bacterial families Marinilabiliaceae, Bacteroidales UCG-001, M2PB4-65 termite group and Idiomarinaceae were impaired. However, in spite of these inhibitory effects, nitrogen removal proved able to prevent unwanted methanogenesis and to enhance the yield of VFAs, and this strategy thus holds great potential for the optimized production of biogas in a two-phase system., We are thankful for the funding received from the German Ministry of Economic Affairs and Energy (grant no. 16KN041338, 16KN041342, 16KN041339 and 16KN041341).

High pressure storage of hydrogen is the established storage technology for automotive systems. However, around 15% of the lower heating value of hydrogen is spent to compress hydrogen up to the pressure of 700 bar. Since this energy is available on board but so far wasted, an open air-conditioning system based on metal hydrides is promising to reutilize this compression work. Here we present the experimental demonstration of a first of its kind system. The setup consists of two alternately operating plate reactors, each filled with around 1.5 kg of Hydralloy C2 ( T i 0.98 Z r 0.02 V 0.41 F e 0.09 C r 0.05 M n 1.46 ), coupled to a polymer electrolyte membrane fuel cell. The demonstration at an electrical power of 5 kW shows that the fuel cell operation is not affected by the alternately H2 desorbing reactors (half-cycle duration of 150 s). The system’s average cooling power was 662 W for an ambient temperature of 30 °C and a cooling temperature of 20 °C, reaching of specific cooling power of 227 W k g M H - 1 . Related to the maximum obtainable cooling power of 18.3% of the electrical fuel cell power, the cooling efficiency corresponds to 75%. As an innovative hydrogen pressure transducer the presented system can be transferred to all applications where an unused hydrogen pressure difference is available.

The Si nanowire growth can be well explained by the classical vapor–liquid–solid (VLS) process taking into account the respective Au–Si phase diagram. For oxide-based compound materials, no phase d...

Currently, additive manufacturing (AM) experiences significant attention in nearly all industrial sectors. AM is already well established in fields such as medicine or spare part production. Nevertheless, processing of high-performance nickel-based superalloys and especially single crystalline alloys such as CMSX-4® is challenging due to the difficulty of intense crack formation. Selective electron beam melting (SEBM) takes place at high process temperatures (~ 1000 °C) and under vacuum conditions. Current work has demonstrated processing of CMSX-4® without crack formation. In addition, by using appropriate AM scan strategies, even single crystals (SX SEBM CMSX-4®) develop directly from the powder bed. In this contribution, we investigate the mechanical properties of SX SEBM CMSX-4® prepared by SEBM in the as-built condition and after heat treatment. The focus is on hardness, strength, low cycle fatigue, and creep properties. These properties are compared with conventional cast and heat-treated material.

We present a detailed analysis of glyphosate-based herbicide (GBH) use patterns with a dataset of herbicide use on arable farms in five districts of Northern Germany. Two aspects are focused: (1) The relationship between GBH and selective herbicides used on a specific field in the same cropping period and (2) the interplay of tillage, GBH and selective herbicide use in weed management. The most interesting result of our analysis is that the intensity of selective herbicide use for in-crop weed management was not related to GBH treatment prior to the crop (except maize). GBH treatments increased the total herbicide intensity on a field. Hence, using GBH pre-crop does not necessarily save herbicides in the following crop. GBH were additional treatments, not replacing selective herbicides. Our data was collected from different districts in Northern Germany extending from West to East. In the Eastern districts, a larger proportion of fields were treated with GBH. When a field was treated with GBH, it was applied with a similar intensity, regardless of the district. Hence, differences in GBH use patterns were caused only by the decision to treat pre-crop, while the intensity of these GBH treatments is hardly varied. In the Eastern districts, the higher proportion of fields treated with GBH is associated with larger proportion of fields with non-plough tillage. Non-plough fields were also treated more intensively with selective herbicides. Nevertheless, the use of both GBH and selective herbicides on the same field was more frequent and intensive in the Western districts of Northern Germany. We attribute the differences in pre-crop GBH use and in-crop selective herbicide use to regional production differences. Small to medium proportions of fields with both ploughing for primary tillage and pre-crop GBH use in all districts and most prominent in winter barley, winter wheat and maize indicate a luxurious, easy reducible use of GBH. We propose to promote farms with non- and low-use of GBH as an example of good practice.

RTN1 is an endoplasmic reticulum-associated protein that was initially identified in neuronal tissues. Here we show that the main isoform RTN1A is a marker for dendritic cells. In the skin, HLA-DR+CD1ahighCD207+CD11cweak Langerhans cells were the only cells in the epidermis, and HLA-DR+CD11c+ dendritic cells were the main cells in the dermis, expressing this protein. RTN1A+ dendritic cells were also found in gingiva, trachea, tonsil, thymus, and peripheral blood. During differentiation of MUTZ-3 cells into Langerhans cells, expression of RTN1A mRNA and protein preceded established Langerhans cell markers CD1a and CD207, and RTN1A protein partially co-localized with the endoplasmic reticulum marker protein disulfide isomerase. In line with this observation, we found that RTN1A was expressed by around 80% of Langerhans cell precursors in human embryonic skin. Our findings show that RTN1A is a marker for cells of the dendritic lineage, including Langerhans cells and dermal dendritic cells. This unexpected finding will serve as a starting point for the elucidation of the, until now, elusive functional roles of RTN1A in both the immune and the nervous system.