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Linarite, PbCuSO4(OH)2, crystallizes as secondary mineral in form of clusters of crystals in Mine du Pradet at Cap Garonne in France. A tiny single crystal was isolated and its structure was refined from synchrotron diffraction data (λ = 0.049592 nm): P121/m1, a = 467.75(2), b = 563.48(4), c = 967.64(5) pm, β = 102.647(5)°, wR = 0.0544, 819 F 2 values and 56 variables. The proton positions were also refined. The Cu2+ cations are located within Cu(OH)2 ribbons and exhibit pronounced Jahn-Teller distortion (d(Cu–O): 2 × 191.9, 2 × 197.3 and 2 × 252.8 pm). The Cu(OH)2 ribbons are condensed with the sulfate tetrahedra and the lead cations. The latter are coordinated to eight oxygen atoms in a slightly anisotropic manner. Calculations of the electron localization function (ELF) of PbO (litharge), PbSO4 (anglesite) and PbCuSO4(OH)2 (linarite) show pronounced lone-pair character for PbO but rather isotropic ELF values around the lead cations in PbSO4 and PbCuSO4(OH)2.

Based on high-resolution tandem mass spectrometry (HR-MS2) and global natural products social molecular networking (GNPS), we found that plant-derived daidzein and genistein derivatives are polyhalogenated by termite-associated Actinomadura species RB99. MS-guided purification from extracts of bacteria grown under optimized conditions led to the isolation of eight polychlorinated isoflavones, including six unreported derivatives, and seven novel polybrominated derivatives, two of which showed antimicrobial activity.

Microcystins, cyclic nonribosomal heptapeptides, are the most well-known cyanobacterial toxins. They are exceptionally well studied, but open questions remain concerning their physiological role for the producing microorganism or their suitability as lead compounds for anticancer drug development. One means to study specialized metabolites in more detail is the introduction of functional groups that make a compound amenable for bioorthogonal, so-called click reactions. Although it was reported that microcystins cannot be derivatized by precursor-directed biosynthesis, we successfully used this approach to prepare clickable microcystins. Supplementing different azide- or terminal alkyne containing amino acid analogues into the cultivation medium of microcystin-producing cyanobacteria strains, we found that these strains differ strongly in their substrate acceptance. Exploiting this flexibility, we generated more than 40 different clickable microcystins. We conjugated one of these derivatives with a fluorogenic dye and showed that neither incorporation of the unnatural amino acid analogue nor attachment of the fluorescent label significantly affects the cytotoxicity against cell lines expressing the human organic anion transporting polypeptides 1B1 or 1B3. Using time-lapse microscopy, we observed that the fluorescent microcystin is rapidly taken up into eukaryotic cells expressing these transporters.

High‐affinity fluorescent derivatives of cyclic adenosine and guanosine monophosphate are powerful tools for investigating their natural targets. Cyclic nucleotide‐regulated ion channels belong to these targets and are vital for many signal transduction processes, such as vision and olfaction. The relation of ligand binding to activation gating is still challenging, and there is a need for fluorescent probes that enable the process to be broken down to the single‐molecule level. This inspired us to prepare fluorophore‐labeled cyclic nucleotides, which are composed of a bright dye and a nucleotide derivative with a thiophenol motif at position 8 that has already been shown to enable superior binding affinity. These bioconjugates were prepared by a novel cross‐linking strategy that involves substitution of the nucleobase with a modified thiophenolate in good yield. Both fluorescent nucleotides are potent activators of different cyclic nucleotide‐regulated ion channels with respect to the natural ligand and previously reported substances. Molecular docking of the probes excluding the fluorophore reveals that the high potency can be attributed to additional hydrophobic and cation‐π interactions between the ligand and the protein. Moreover, the introduced substances have the potential to investigate related target proteins, such as cAMP‐ and cGMP‐dependent protein kinases, exchange proteins directly activated by cAMP or phosphodiesterases., Selecting channels: Fluorescent cyclic nucleotide derivatives synthesized with a thiophenol motif‐containing heterobifunctional cross‐linking reagent efficiently activate CNG as well as HCN channels in the nanomolar range and are potential candidates to study related target proteins.

Growth from spores activated a biosynthetic gene cluster in Actinomadura sp. RB29, resulting in the identification of two novel groups of halogenated polyketide natural products, named maduralactomycins and actinospirols. The unique tetracyclic and spirocyclic structures were assigned based on a combination of NMR analysis, chemoinformatic calculations, X-ray crystallography, and 13C labeling studies. On the basis of HRMS2 data, genome mining, and gene expression studies, we propose an underlying noncanonical angucycline biosynthesis and extensive post-polyketide synthase (PKS) oxidative modifications.

RNAi-mediated knockdown of DICER1 and DROSHA, enzymes critically involved in miRNA biogenesis, has been postulated to affect the homeostasis and the angiogenic capacity of human endothelial cells. To re-evaluate this issue, we reduced the expression of DICER1 or DROSHA by RNAi-mediated knockdown and subsequently investigated the effect of these interventions on the angiogenic capacity of human umbilical vein endothelial cells (HUVEC) in vitro (proliferation, migration, tube formation, endothelial cell spheroid sprouting) and in a HUVEC xenograft assay in immune incompetent NSGTM mice in vivo. In contrast to previous reports, neither knockdown of DICER1 nor knockdown of DROSHA profoundly affected migration or tube formation of HUVEC or the angiogenic capacity of HUVEC in vivo. Furthermore, knockdown of DICER1 and the combined knockdown of DICER1 and DROSHA tended to increase VEGF-induced BrdU incorporation and induced angiogenic sprouting from HUVEC spheroids. Consistent with these observations, global proteomic analyses showed that knockdown of DICER1 or DROSHA only moderately altered HUVEC protein expression profiles but additively reduced, for example, expression of the angiogenesis inhibitor thrombospondin-1. In conclusion, global reduction of miRNA biogenesis by knockdown of DICER1 or DROSHA does not inhibit the angiogenic capacity of HUVEC. Further studies are therefore needed to elucidate the influence of these enzymes in the context of human endothelial cell-related angiogenesis.

Ionotropic purinergic receptors (P2X receptors) are non-specific cation channels that are activated by the binding of ATP at their extracellular side. P2X receptors contribute to multiple functions, including the generation of pain, inflammation, or synaptic transmission. The channels are trimers and structural information on several of their isoforms is available. In contrast, the cooperation of the subunits in the activation process is poorly understood. We synthesized a novel fluorescent ATP derivative, 2-[DY-547P1]-AET-ATP (fATP) to unravel the complex activation process in P2X2 and mutated P2X2 H319K channels with enhanced apparent affinity by characterizing the relation between ligand binding and activation gating. fATP is a full agonist with respect to ATP that reports the degree of binding by bright fluorescence. For quantifying the binding, a fast automated algorithm was employed on human embryonic kidney cell culture images. The concentrations of half maximum occupancy and activation as well as the respective Hill coefficients were determined. All Hill coefficients exceeded unity, even at an occupancy

The gold-rich intermetallic compounds REAu4Cd2 (RE = La–Nd, Sm) were synthesized from the elements in sealed tantalum ampoules. Their characterization by X-ray powder and single crystal data confirmed the tetragonal YbAl4Mo2 type, space group I4/mmm. The basic building units are Au4 squares (278 pm Au–Au in CeAu4Cd2) and infinite linear cadmium chains (275 pm Cd–Cd in CeAu4Cd2). We exemplarily studied the solid solution CeAu4+ x Cd2− x for x = 0–1 up to CeAu5Cd. Electron diffraction patterns on a CeAu5Cd sample confirm the single crystal data. They give no hint for complete gold-cadmium ordering. Temperature-dependent magnetic susceptibility measurements of CeAu4Cd2, CeAu5Cd, PrAu4Cd2 and NdAu4Cd2 show stable trivalent rare earth ions and give no hint for magnetic ordering above 3 K.

Significance G protein-coupled receptors (GPCRs) form a family of signaling molecules in the membrane of cells that plays a key role in transduction of cellular responses. Little is known about how rapidly GPCRs can be activated. While the “light receptor” rhodopsin in the eye activates within 1 ms, other GPCRs are thought to activate much slower. We use two entirely different techniques with advanced time resolution to activate a dimeric metabotropic glutamate GPCR: UV light-triggered uncaging of ligand in intact cells and piezo-driven ligand application in outside-out patches. We demonstrate initial conformational rearrangements within ≈1 ms that are followed by much slower (≈20 ms) activation in the transmembrane domain. Thus, the initial activation of a nonvisual GPCR proceeds with millisecond speed., G protein-coupled receptors (GPCRs) are key biological switches that transmit both internal and external stimuli into the cell interior. Among the GPCRs, the “light receptor” rhodopsin has been shown to activate with a rearrangement of the transmembrane (TM) helix bundle within ∼1 ms, while all other receptors are thought to become activated within ∼50 ms to seconds at saturating concentrations. Here, we investigate synchronous stimulation of a dimeric GPCR, the metabotropic glutamate receptor type 1 (mGluR1), by two entirely different methods: (i) UV light-triggered uncaging of glutamate in intact cells or (ii) piezo-driven solution exchange in outside-out patches. Submillisecond FRET recordings between labels at intracellular receptor sites were used to record conformational changes in the mGluR1. At millimolar ligand concentrations, the initial rearrangement between the mGluR1 subunits occurs at a speed of τ1 ∼ 1–2 ms and requires the occupancy of both binding sites in the mGluR1 dimer. These rapid changes were followed by significantly slower conformational changes in the TM domain (τ2 ∼ 20 ms). Receptor deactivation occurred with time constants of ∼40 and ∼900 ms for the inter- and intrasubunit conformational changes, respectively. Together, these data show that, at high glutamate concentrations, the initial intersubunit activation of mGluR1 proceeds with millisecond speed, that there is loose coupling between this initial step and activation of the TM domain, and that activation and deactivation follow a cyclic pathway, including—in addition to the inactive and active states—at least two metastable intermediate states.

Natural lead dioxide samples (β modification) from Cap Garonne, Mine du Pradet, France and Mount Trevasco, Italy were studied by single crystal X-ray and electron diffraction, clearly manifesting the rutile-type structure. A needle from the Mount Trevasco sample was investigated on a single crystal diffractometer: P42/mnm, a = 495.81(10), c = 338.66(7) pm, wR = 0.0513, 117 F 2 values (all data) and nine variables (293 K data). An additional data set at T = 90 K gave no hint for structural distortions.

Significance Membrane receptors control innumerous essential functions of living cells and are proven drug targets for pharmacological interventions. Many receptors are composed of a number of subunits and contain multiple binding sites for ligands, resulting in intricate mechanisms of subunit activation. To understand these mechanisms, the subunit action is usually modeled by allosteric schemes assuming a fixed subunit cooperativity. Here, we use subunit concatenation, binding-site mutations, and global mathematical analyses to study the interaction of the individual subunits in heterooligomeric olfactory cyclic nucleotide-gated ion channels, which are composed of three types of subunits. Our analyses provide a detailed thermodynamic profile for these interactions upon channel activation, and they can be applied to any other oligomeric ion channel and receptor., Cyclic nucleotide-gated (CNG) ion channels of olfactory neurons are tetrameric membrane receptors that are composed of two A2 subunits, one A4 subunit, and one B1b subunit. Each subunit carries a cyclic nucleotide-binding domain in the carboxyl terminus, and the channels are activated by the binding of cyclic nucleotides. The mechanism of cooperative channel activation is still elusive. Using a complete set of engineered concatenated olfactory CNG channels, with all combinations of disabled binding sites and fit analyses with systems of allosteric models, the thermodynamics of microscopic cooperativity for ligand binding was subunit- and state-specifically quantified. We show, for the closed channel, that preoccupation of each of the single subunits increases the affinity of each other subunit with a Gibbs free energy (ΔΔG) of ∼−3.5 to ∼−5.5 kJ ⋅ mol−1, depending on the subunit type, with the only exception that a preoccupied opposite A2 subunit has no effect on the other A2 subunit. Preoccupation of two neighbor subunits of a given subunit causes the maximum affinity increase with ΔΔG of ∼−9.6 to ∼−9.9 kJ ⋅ mol−1. Surprisingly, triple preoccupation leads to fewer negative ΔΔG values for a given subunit as compared to double preoccupation. Channel opening increases the affinity of all subunits. The equilibrium constants of closed–open isomerizations systematically increase with progressive liganding. This work demonstrates, on the example of the heterotetrameric olfactory CNG channel, a strategy to derive detailed insights into the specific mutual control of the individual subunits in a multisubunit membrane receptor.

P2X7 receptors are trimeric ion channels activated by extracellular ATP. Upon activation, they trigger cytolysis and apoptosis but also control cell proliferation. To shed more light on channel gating and the underlying function of the individual subunits, receptors of concatenated subunits were built containing a defined number of functional binding sites. The currents evoked by ATP were obtained in the outside-out configuration of the patch-clamp technique, and steady-state activation, as well as time courses, were analyzed. Our results show that each occupied binding site contributes to channel activation. While the occupation of a single binding site can already activate the channels, three bound ligands maximally stabilize the open state. Hence, P2X7 receptors can be described by a stepwise activation process.

Emerging evidence places small proteins (≤ 50 amino acids) more centrally in physiological processes. Yet, the identification of functional small proteins and the systematic genome annotation of their cognate small open reading frames (smORFs) remains challenging both experimentally and computationally. Ribosome profiling or Ribo-Seq (that is a deep sequencing of ribosome-protected fragments) enables detecting of actively translated open-reading frames (ORFs) and empirical annotation of coding sequences (CDSs) using the in-register translation pattern that is characteristic for genuinely translating ribosomes. Multiple identifiers of ORFs that use 3-nt periodicity in Ribo-Seq data sets have been successful in eukaryotic smORF annotation. Yet, they have difficulties evaluating prokaryotic genomes due to the unique architecture of prokaryotic genomes (e.g. polycistronic messages, overlapping ORFs, leaderless translation, non-canonical initiation etc.). Here, we present our new algorithm, smORFer, which performs with high accuracy in prokaryotic organisms in detecting smORFs. The unique feature of smORFer is that it uses integrated approach and considers structural features of the genetic sequence along with in-register translation and uses Fourier transform to convert these parameters into a measurable score to faithfully select smORFs. The algorithm is executed in a modular way and dependent on the data available for a particular organism allows using different modules for smORF search.

Urate homeostasis in humans is a complex and highly heritable process that involves i.e., metabolic urate biosynthesis, renal urate reabsorption, as well as renal and extrarenal urate excretion. Importantly, disturbances in urate excretion are a common cause of hyperuricemia and gout. The majority of urate is eliminated by glomerular filtration in the kidney followed by an, as yet, not fully elucidated interplay of multiple transporters involved in the reabsorption or excretion of urate in the succeeding segments of the nephron. In this context, genome-wide association studies and subsequent functional analyses have identified the ATP-binding cassette (ABC) transporter ABCG2 as an important urate transporter and have highlighted the role of single nucleotide polymorphisms (SNPs) in the pathogenesis of reduced cellular urate efflux, hyperuricemia, and early-onset gout. Recent publications also suggest that ABCG2 is particularly involved in intestinal urate elimination and thus may represent an interesting new target for pharmacotherapeutic intervention in hyperuricemia and gout. In this review, we specifically address the involvement of ABCG2 in renal and extrarenal urate elimination. In addition, we will shed light on newly identified polymorphisms in ABCG2 associated with early-onset gout.

Cerebral polymer coating embolism from intravascular devices represents a potentially serious complication to endovascular therapy (EVT). We report two cases of neuroendovascular treatment where filamentous polymer fragments were noted possibly due to damage of the surface coating during manipulation and backloading of microguidewires. As the exact origin of the debris was initially not known, microguidewires and fragments were examined with light microscopy, stereomicroscopy, scanning electron microscopy and attenuated-total-reflection Fourier transform infrared spectroscopy. Fragments consisted of polytetrafluoroethylene and silicone oil stemming from the proximal shaft of a standard microguidewire. To our knowledge, this is the first report of polytetrafluoroethylene coating fragments created during EVT. Future studies should assess the mechanism of polymer coating delamination and its potential consequences during EVT including inadvertent fragment migration into the cerebral circulation.

Abstract Adaptations of animal cells to growth in suspension culture concern in particular viral vaccine production, where very specific aspects of virus-host cell interaction need to be taken into account to achieve high cell specific yields and overall process productivity. So far, the complexity of alterations on the metabolism, enzyme, and proteome level required for adaptation is only poorly understood. In this study, for the first time, we combined several complex analytical approaches with the aim to track cellular changes on different levels and to unravel interconnections and correlations. Therefore, a Madin-Darby canine kidney (MDCK) suspension cell line, adapted earlier to growth in suspension, was cultivated in a 1-L bioreactor. Cell concentrations and cell volumes, extracellular metabolite concentrations, and intracellular enzyme activities were determined. The experimental data set was used as the input for a segregated growth model that was already applied to describe the growth dynamics of the parental adherent cell line. In addition, the cellular proteome was analyzed by liquid chromatography coupled to tandem mass spectrometry using a label-free protein quantification method to unravel altered cellular processes for the suspension and the adherent cell line. Four regulatory mechanisms were identified as a response of the adaptation of adherent MDCK cells to growth in suspension. These regulatory mechanisms were linked to the proteins caveolin, cadherin-1, and pirin. Combining cell, metabolite, enzyme, and protein measurements with mathematical modeling generated a more holistic view on cellular processes involved in the adaptation of an adherent cell line to suspension growth. Key points • Less and more efficient glucose utilization for suspension cell growth • Concerted alteration of metabolic enzyme activity and protein expression • Protein candidates to interfere glycolytic activity in MDCK cells

Morphotype switches frequently occur in Actinobacteria and are often associated with disparate natural product production. Here, we report on differences in the secondary metabolomes of two morphotypes of a Streptomyces species, including the discovery of a novel antimicrobial glycosylated macrolide, which we named termidomycin A. While exhibiting an unusual 46-member polyene backbone, termidomycin A (1) shares structural features with the clinically important antifungal agents amphotericin B and nystatin A1. Genomic analyses revealed a biosynthetic gene cluster encoding for a putative giant type I polyketide synthase (PKS), whose domain structure allowed us to propose the relative configuration of the 46-member macrolide. The architecture of the biosynthetic gene cluster was different in both morphotypes, thus leading to diversification of the product spectrum. Given the high frequency of genomic rearrangements in Streptomycetes, the metabolic analysis of distinct morphotypes as exemplified in this study is a promising approach for the discovery of bioactive natural products and pathways of diversification.

Hyperpolarization-activated and cyclic nucleotide (HCN) modulated channels are tetrameric cation channels. In each of the four subunits, the intracellular cyclic nucleotide-binding domain (CNBD) is coupled to the transmembrane domain via a helical structure, the C-linker. High-resolution channel structures suggest that the C-linker enables functionally relevant interactions with the opposite subunit, which might be critical for coupling the conformational changes in the CNBD to the channel pore. We combined mutagenesis, patch-clamp technique, confocal patch-clamp fluorometry, and molecular dynamics (MD) simulations to show that residue K464 of the C-linker is relevant for stabilizing the closed state of the mHCN2 channel by forming interactions with the opposite subunit. MD simulations revealed that in the K464E channel, a rotation of the intracellular domain relative to the channel pore is induced, which is similar to the cAMP-induced rotation, weakening the autoinhibitory effect of the unoccupied CL-CNBD region. We suggest that this CL-CNBD rotation is considerably involved in activation-induced affinity increase but only indirectly involved in gate modulation. The adopted poses shown herein are in excellent agreement with previous structural results.

Abstract The novel influenza A virus (IAV) defective interfering particle “OP7” inhibits IAV replication in a co-infection and was previously suggested as a promising antiviral agent. Here, we report a batch-mode cell culture-based production process for OP7. In the present study, a seed virus containing standard virus (STV) and OP7 was used. The yield of OP7 strongly depended on the production multiplicity of infection. To inactivate infectious STV in the OP7 material, which may cause harm in a potential application, UV irradiation was used. The efficacy of OP7 in this material was preserved, as shown by an in vitro interference assay. Next, steric exclusion chromatography was used to purify and to concentrate (~ 13-fold) the UV-treated material. Finally, administration of produced OP7 material in mice did not show any toxic effects. Furthermore, all mice infected with a lethal dose of IAV survived the infection upon OP7 co-treatment. Thus, the feasibility of a production workflow for OP7 and its potential for antiviral treatment was demonstrated. Key points • OP7 efficacy strongly depended on the multiplicity of infection used for production • Purification by steric exclusion chromatography increased OP7 efficacy • OP7-treated mice were protected against a lethal infection with IAV

Opening of hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels is controlled by membrane hyperpolarization and binding of cyclic nucleotides to the tetrameric cyclic nucleotide-binding domain (CNBD), attached to the C-linker disk (CL). Confocal patch-clamp fluorometry revealed a pronounced cooperativity of ligand binding among protomers. However, by which pathways allosteric signal transmission occurs remained elusive. Here, we investigate how changes in the structural dynamics of the CL- CNBD of mouse HCN2 upon cAMP binding relate to inter- and intrasubunit signal transmission. Applying a rigidity theory-based approach, we identify two intersubunit and one intrasubunit pathways that differ in allosteric coupling strength between cAMP binding sites or towards the CL. These predictions agree with results from electrophysiological and patch-clamp fluorometry experiments. Our results map out distinct routes within the CL-CNBD that modulate different cAMP binding responses in HCN2 channels. They signify that functionally relevant submodules may exist within and across structurally discernable subunits in HCN channels.

The soil microbiome is inherently complex with high biological diversity, and spatial heterogeneity typically occurring on the submillimetre scale. To study the microbial ecology of soils, and other microbiomes, biomolecules, that is, nucleic acids and proteins, must be efficiently and reliably co-recovered from the same biological samples. Commercial kits are currently available for the co-extraction of DNA, RNA and proteins but none has been developed for soil samples. We present a new protocol drawing on existing phenol-chloroform-based methods for nucleic acids co-extraction but incorporating targeted precipitation of proteins from the phenol phase. The protocol is cost-effective and robust, and easily implemented using reagents commonly available in laboratories. The method is estimated to be eight times cheaper than using disparate commercial kits for the isolation of DNA and/or RNA, and proteins, from soil. The method is effective, providing good quality biomolecules from a diverse range of soil types, with clay contents varying from 9.5% to 35.1%, which we successfully used for downstream, high-throughput gene sequencing and metaproteomics. Additionally, we demonstrate that the protocol can also be easily implemented for biomolecule co-extraction from other complex microbiome samples, including cattle slurry and microbial communities recovered from anaerobic bioreactors, as well as from Gram-positive and Gram-negative pure cultures.

The optimized superconducting stellarator device Wendelstein 7-X (with major radius , minor radius , and plasma volume) restarted operation after the assembly of a graphite heat shield and 10 inertially cooled island divertor modules. This paper reports on the results from the first high-performance plasma operation. Glow discharge conditioning and ECRH conditioning discharges in helium turned out to be important for density and edge radiation control. Plasma densities of with central electron temperatures were routinely achieved with hydrogen gas fueling, frequently terminated by a radiative collapse. In a first stage, plasma densities up to were reached with hydrogen pellet injection and helium gas fueling. Here, the ions are indirectly heated, and at a central density of a temperature of with was transiently accomplished, which corresponds to with a peak diamagnetic energy of and volume-averaged normalized plasma pressure . The routine access to high plasma densities was opened with boronization of the first wall. After boronization, the oxygen impurity content was reduced by a factor of 10, the carbon impurity content by a factor of 5. The reduced (edge) plasma radiation level gives routinely access to higher densities without radiation collapse, e.g. well above line integrated density and central temperatures at moderate ECRH power. Both X2 and O2 mode ECRH schemes were successfully applied. Core turbulence was measured with a phase contrast imaging diagnostic and suppression of turbulence during pellet injection was observed.

The giant panda is known worldwide for having successfully moved to a diet almost exclusively based on bamboo. Provided that no lignocellulose-degrading enzyme was detected in panda’s genome, bamboo digestion is believed to depend on its gut microbiome. However, pandas retain the digestive system of a carnivore, with retention times of maximum 12 h. Cultivation of their unique gut microbiome under controlled laboratory conditions may be a valid tool to understand giant pandas’ dietary habits, and provide valuable insights about what component of lignocellulose may be metabolized. Here, we collected gut microbiomes from fresh fecal samples of a giant panda (either entirely green or yellow stools) and supplied them with green leaves or yellow pith (i.e., the peeled stem). Microbial community composition was substrate dependent, and resulted in markedly different fermentation profiles, with yellow pith fermented to lactate and green leaves to lactate, acetate and ethanol, the latter to strikingly high concentrations (∼3%, v:v, within 3.5 h). Microbial metaproteins pointed to hemicellulose rather than cellulose degradation. The alpha-amylase from the giant panda (E.C. 3.2.1.1) was the predominant identified metaprotein, particularly in reactors inoculated with pellets derived from fecal samples (up to 60%). Gut microbiomes assemblage was most prominently impacted by the change in substrate (either leaf or pith). Removal of soluble organics from inocula to force lignocellulose degradation significantly enriched Bacteroides (in green leaf) and Escherichia/Shigella (in yellow pith). Overall, different substrates (either leaf or pith) markedly shaped gut microbiome assemblies and fermentation profiles. The biochemical profile of fermentation products may be an underestimated factor contributing to explain the peculiar dietary behavior of giant pandas, and should be implemented in large scale studies together with short-term lab-scale cultivation of gut microbiomes.

Xylan phenyl carbonate (XPC) derivatives were prepared and characterized comprehensively. By conversion of xylan with phenyl chloroformate either in dipolar aprotic solvents with LiCl or in an ionic liquid, XPC with degrees of substitution (DS) of up to 2.0, i.e., fully functionalized derivatives, could be obtained. The synthesis was studied with respect to the influence of different reaction parameters. It was found that the reaction medium as well as the type of starting xylan strongly affected the efficiency of the derivatization. The derivatives obtained were characterized by FT-IR- and NMR spectroscopy. Surprisingly, it was found that C-3 is the most reactive position in this particular reaction while substitution in position C-2 only occurred if the neighboring position C-3 already carried a phenyl carbonate group. XPC were found to form spherical nanoparticles (NP) of well-defined shape with diameters around 158 nm. These materials possess unique potential as activated NP for advanced applications.

Taxonomic and functional characterization of microbial communities from diverse environments such as the human gut or biogas plants by multi-omics methods plays an ever more important role. Researchers assign all identified genes, transcripts, or proteins to biological pathways to better understand the function of single species and microbial communities. However, due to the versality of microbial metabolism and a still-increasing number of newly biological pathways, linkage to standard pathway maps such as the KEGG central carbon metabolism is often problematic. We successfully implemented and validated a new user-friendly, stand-alone web application, the MPA_Pathway_Tool. It consists of two parts, called ‘Pathway-Creator’ and ‘Pathway-Calculator’. The ‘Pathway-Creator’ enables an easy set-up of user-defined pathways with specific taxonomic constraints. The ‘Pathway-Calculator’ automatically maps microbial community data from multiple measurements on selected pathways and visualizes the results. The MPA_Pathway_Tool is implemented in Java and ReactJS.

Metaproteomics, the mass spectrometry-based analysis of proteins from multispecies samples faces severe challenges concerning data analysis and results interpretation. To overcome these shortcomings, we here introduce the MetaProteomeAnalyzer (MPA) Portable software. In contrast to the original server-based MPA application, this newly developed tool no longer requires computational expertise for installation and is now independent of any relational database system. In addition, MPA Portable now supports state-of-the-art database search engines and a convenient command line interface for high-performance data processing tasks. While search engine results can easily be combined to increase the protein identification yield, an additional two-step workflow is implemented to provide sufficient analysis resolution for further postprocessing steps, such as protein grouping as well as taxonomic and functional annotation. Our new application has been developed with a focus on intuitive usability, adherence to data standards, and adaptation to Web-based workflow platforms. The open source software package can be found at https://github.com/compomics/meta-proteome-analyzer .

A polycrystalline sample of the TiNiSi type low-temperature (LT) modification of ScRuSi was synthesized by arc-melting. Longer annealing in a sealed silica tube (6 weeks at 1270 K) followed by quenching led to the high-temperature (HT) phase. HT-ScRuSi adopts the ZrNiAl structure type: P 6 ¯ 2 m , a = 688.27(9), c = 336.72(5) pm, w R 2 = 0.0861, 260 F 2 values, 14 variables. The striking structural building units are regular, tricapped trigonal prisms Si1@Ru 3 Sc 6 and Si2@Ru 6 Sc 3 . Both polymorphs have been characterized by 29 Si and 45 Sc MAS-NMR spectroscopy. The local scandium environments in the two polymorphs are easily distinguished by their electric field gradient tensor values, in agreement with theoretically calculated values.

Polycrystalline samples of Sr 0.95 Eu 0.05 Ba 8 [BN 2 ] 6 and Eu 0.937 Ba 8 [BN 2 ] 6 were synthesized via conventional solid-state reaction from the binary precursor compounds Sr 3 N 2 , EuN, Ba 3 N 2 and BN at 1000 °C. The structure of Eu 0.937 Ba 8 [BN 2 ] 6 was refined from single crystal X-ray diffractometer data: Eu 0.937 Ba 8 [BN 2 ] 6 type, Fd 3 ¯ m, a = 1594.54(9) pm, w R 2 = 0.0654, 380 F 2 values and 23 variables. The 8 a europium site shows an occupancy of only 93.7(9) % suggesting partial oxidation of europium to fulfil an electron-precise description, i. e. Eu II 0.81 Eu III 0.13 Ba 8 [BN 2 ] 6 . The mixed europium valence was confirmed by magnetic susceptibility measurements (reduced magnetic moment of 7.28 μ B per europium atom) and 151 Eu Mossbauer spectroscopy (Eu II : Eu III = 82: 18). The nitridoborate anions are coordinated by slightly distorted, mono-capped (europium) square prisms formed by the barium atoms. All metal cations are hexa-coordinated by nitrogen atoms. The EPR spectra of Eu x Sr 1-x Ba 8 [BN 2 ] 6 samples (0.001 ≤ x ≤ 0.01) suggest close to cubic local symmetry of the Eu 2+ dopant ions and reveal some highly unusual features: Magnetic hyperfine splitting is only observed with one of the Eu nuclear isotopes, and the coupling constant of 243.6 MHz is extremely large compared to values (90–100 MHz) typically observed in the literature for Eu 2+ doped crystalline materials.

We report the thin film deposition of copper iodide with zincblende type structure (γ-CuI) via metal-organic chemical vapor deposition (MOCVD). Single crystals and thin films of Cu and γ-CuI could be formed on various substrates from cyclopentadienylcopper triethylphosphine (CpCuPEt 3 ) and ethyliodide (C 2 H 5 I) as precursors for copper and iodine, respectively. Additionally, the chemical reaction behavior of thermally evaporated and MOCVD deposited elemental copper films with C 2 H 5 I is examined. SiO 2 glass and various single crystalline oxides and semiconductors were used as substrates. For all cases X-ray diffraction measurements revealed polycrystalline γ-CuI with zincblende type structure; other CuI phases could not be detected. Photoluminescence measurements show a broadened peak at 420 nm due to donor-acceptor pair recombination. Intense peaks at shorter wavelengths are attributed to excitonic emissions.

The new equiatomic silicides NbOsSi and TaOsSi as well as ZrOsSi, T IrSi ( T = Zr, Hf, Nb, Ta) and T PtSi ( T = Nb, Ta) were prepared from the elements by arc-melting. These silicides crystallize with the orthorhombic TiNiSi type structure, space group Pnma . Irregularly shaped crystals of ZrOsSi, NbOsSi, TaOsSi, ZrIrSi and HfIrSi were separated from the annealed samples and investigated by single-crystal X-ray diffraction ( a = 640.46(7), b = 404.07(5), c = 743.66(8) pm, wR 2 = 0.0285, 390 F 2 values, 20 variables for ZrOsSi; a = 629.78(6), b = 388.72(4), c = 727.48(7) pm, wR 2 = 0.0350, 397 F 2 values, 20 variables for NbOsSi, a = 626.80(6), b = 389.36(4), c = 726.22(7) pm, wR 2 = 0.0501, 385 F 2 values, 20 variables for TaOsSi, a = 653.48(8), b = 395.35(4), c = 739.19(8) pm, wR 2 = 0.0427, 413 F 2 values, 20 variables for ZrIrSi and a = 646.34(12), b = 393.57(7), c = 736.8(14) pm, wR 2 = 0.0582, 371 F 2 values, 20 variables for HfIrSi). The striking structural motifs in the new osmium compounds are three-dimensional [OsSi] networks (Os–Si: 240–251 pm) in which the osmium atoms have strongly distorted tetrahedral silicon coordination. High-pressure/high-temperature experiments (9.5 GPa/1520 K) on TaOsSi gave no hint for a structural phase transition. Temperature dependent measurements of the magnetic susceptibility and the electrical conductivity of NbOsSi and TaOsSi showed superconductivity below T C = 3.5 and 5.5 K, respectively. 29 Si solid state MAS NMR investigations of the prepared silicides approved the structural models and showed a correlation between the observed 29 Si resonance shifts and the electronegativity of the involved refractory metal.

The intermetallic compound CaAu2Al2, and the members of the solid solutions SrAu2–xAl2+x (0 ≤ x ≤ 0.33) and Ba3Au5+xAl6–x (x = 0, 0.14, 0.49) were synthesized from the elements in sealed tantalum ampoules. The Ca compound crystallizes with the orthorhombic ThRu2P2 type structure, whereas the targeted SrAu2Al2 was found to form a solid solution according to SrAu2–xAl2+x. For the Ba system no “BaAu2Al2” was found, however, Ba3Au5+xAl6–x was discovered to crystallize in the monoclinic space group C2/c with its own structure type. The samples were investigated by powder X-ray diffraction and their crystal structures were refined on the basis of single-crystal X-ray diffraction data. All compounds were characterized furthermore by susceptibility measurements. The crystallographic aluminum sites of CaAu2Al2 and Ba3Au5Al6 can be differentiated by 27Al solid state NMR spectra on the basis of their different electric field gradients, in agreement with theoretical calculations. The electron transfer from the alkaline earth metals and the aluminum atoms onto the gold atoms was investigated by X-ray photoelectron spectroscopy (XPS) classifying these intermetallics as aurides, in full agreement with the calculated Bader charges.

The ordered Laves phases RE 2Rh3Ga (RE=Y, La–Nd, Sm, Gd–Er) were synthesized by arc-melting of the elements and subsequent annealing. The samples were characterized by powder X-ray diffraction (XRD). They crystallize with the rhombohedral Mg2Ni3Si type structure, space group R3̅m. Three structures were refined from single crystal X-ray diffractometer data: a=557.1(1), c=1183.1(2), wR2=0.0591, 159 F 2 values, 10 variables for Y2Rh3Ga, a=562.5(2), c=1194.4(2) pm, wR2=0.0519, 206 F 2 values, 11 variables for Ce2Rh3Ga and a=556.7(2), c=1184.1(3) pm, wR2=0.0396, 176 F 2 values, 11 variables for Tb2Rh3Ga. The Rh3Ga tetrahedra are condensed via common corners and the large cavities left by the network are filled by the rare earth atoms. The RE 2Rh3Ga Laves phases crystallize with a translationengleiche subgroup of the cubic RERh2 Laves phases with MgCu2 type. Magnetic susceptibility measurements reveal Pauli paramagnetism for Y2Rh3Ga and La2Rh3Ga. Ce2Rh3Ga shows intermediate cerium valence while all other RE 2Rh3Ga phases are Curie–Weiss paramagnets which order magnetically at low temperatures. The 89Y and 71Ga solid state nuclear magnetic resonance (NMR) spectra of the diamagnetic representative Y2Rh3Ga show well-defined single resonances in agreement with an ordered bulk phase. In comparison to the binary Laves phase YRh2 a strongly increased 89Y resonance frequency is observed owing to a higher s-electron spin density at the 89Y nuclei as proven by density of states (DOS) calculations.

Acetate is a characteristic by-product of Escherichia coli K-12 growing in batch cultures with glucose, both under aerobic as well as anaerobic conditions. While the reason underlying aerobic acetate production is still under discussion, during anaerobic growth acetate production is important for ATP generation by substrate level phosphorylation. Under both conditions, acetate is produced by a pathway consisting of the enzyme phosphate acetyltransferase (Pta) producing acetyl-phosphate from acetyl-coenzyme A, and of the enzyme acetate kinase (AckA) producing acetate from acetyl-phosphate, a reaction that is coupled to the production of ATP. Mutants in the AckA-Pta pathway differ from each other in the potential to produce and accumulate acetyl-phosphate. In the publication at hand, we investigated different mutants in the acetate pathway, both under aerobic as well as anaerobic conditions. While under aerobic conditions only small changes in growth rate were observed, all acetate mutants showed severe reduction in growth rate and changes in the by-product pattern during anaerobic growth. The AckA– mutant showed the most severe growth defect. The glucose uptake rate and the ATP concentration were strongly reduced in this strain. This mutant exhibited also changes in gene expression. In this strain, the atoDAEB operon was significantly upregulated under anaerobic conditions hinting to the production of acetoacetate. During anaerobic growth, protein acetylation increased significantly in the ackA mutant. Acetylation of several enzymes of glycolysis and central metabolism, of aspartate carbamoyl transferase, methionine synthase, catalase and of proteins involved in translation was increased. Supplementation of methionine and uracil eliminated the additional growth defect of the ackA mutant. The data show that anaerobic, fermentative growth of mutants in the AckA-Pta pathway is reduced but still possible. Growth reduction can be explained by the lack of an important ATP generating pathway of mixed acid fermentation. An ackA deletion mutant is more severely impaired than pta or ackA-pta deletion mutants. This is most probably due to the production of acetyl-P in the ackA mutant, leading to increased protein acetylation.

Although metaproteomics, the study of the collective proteome of microbial communities, has become increasingly powerful and popular over the past few years, the field has lagged behind on the availability of user-friendly, end-to-end pipelines for data analysis. We therefore describe the connection from two commonly used metaproteomics data processing tools in the field, MetaProteomeAnalyzer and PeptideShaker, to Unipept for downstream analysis. Through these connections, direct end-to-end pipelines are built from database searching to taxonomic and functional annotation.

ABCG2 belongs to the ABC transporter superfamily and functions as a poly-specific efflux pump. As it can transport a broad spectrum of substrates out of cells, ABCG2 is thought to alter the pharmacokinetics of drugs applied to treat certain diseases. Especially, its potential to induce resistance to chemotherapy is currently the object of intense research. To foster understanding of mechanisms relevant for substrate recognition and selection of ABCG2 substrates and to finally develop selective therapeutic modulators (e.g. inhibitors) of ABCG2 transport activity, it is important to further explore the precise 3D structure of the transporter. While efforts to elucidate the three-dimensional structure of ABCG2 using X-ray crystal structure analysis have not been successful so far, high-resolution cryo-electron microscopy-based investigations have revealed exciting new insights into the structure and function of the transporter. In this review, we will focus on these seminal publications to summarize the current understanding of tertiary and quaternary structure, homodimerization or oligomerization, and functions of the ABCG2 transporter protein.

Petroleum hydrocarbons reach the deep-sea following natural and anthropogenic factors. The process by which they enter deep-sea microbial food webs and impact the biogeochemical cycling of carbon and other elements is unclear. Hydrostatic pressure (HP) is a distinctive parameter of the deep sea, although rarely investigated. Whether HP alone affects the assembly and activity of oil-degrading communities remains to be resolved. Here we have demonstrated that hydrocarbon degradation in deep-sea microbial communities is lower at native HP (10 MPa, about 1000 m below sea surface level) than at ambient pressure. In long-term enrichments, increased HP selectively inhibited obligate hydrocarbon-degraders and downregulated the expression of beta-oxidation-related proteins (i.e., the main hydrocarbon-degradation pathway) resulting in low cell growth and CO2 production. Short-term experiments with HP-adapted synthetic communities confirmed this data, revealing a HP-dependent accumulation of citrate and dihydroxyacetone. Citrate accumulation suggests rates of aerobic oxidation of fatty acids in the TCA cycle were reduced. Dihydroxyacetone is connected to citrate through glycerol metabolism and glycolysis, both upregulated with increased HP. High degradation rates by obligate hydrocarbon-degraders may thus be unfavourable at increased HP, explaining their selective suppression. Through lab-scale cultivation, the present study is the first to highlight a link between impaired cell metabolism and microbial community assembly in hydrocarbon degradation at high HP. Overall, this data indicate that hydrocarbons fate differs substantially in surface waters as compared to deep-sea environments, with in situ low temperature and limited nutrients availability expected to further prolong hydrocarbons persistence at deep sea.

Type 2 diabetes is a complex, heterogeneous, and polygenic disease. Currently, available drugs for treating type 2 diabetes predominantly include sulfonylureas, α-glucosidase inhibitors, and biguanides. However, long-term treatment with these therapeutic drugs is often accompanied by undesirable side effects, which have driven interest in the development of more effective and safer antidiabetic agents. To address the urgent need for new chemical solutions, we focused on the analysis of structurally novel and/or biologically new metabolites produced by insect-associated microbes as they have recently been recognized as a rich source of natural products. Comparative LC/MS-based analysis of Actinomadura sp. RB99, isolated from a fungus-growing termite, led to the identification of the type II polyketide synthase-derived fridamycin A. The structure of fridamycin A was confirmed by 1H NMR data and LC/MS analysis. The natural microbial product, fridamycin A, was examined for its antidiabetic properties in 3T3-L1 adipocytes, which demonstrated that fridamycin A induced glucose uptake in 3T3-L1 cells by activating the AMP-activated protein kinase (AMPK) signaling pathway but did not affect adipocyte differentiation, suggesting that the glucose uptake took place through activation of the AMPK signaling pathway without inducing adipogenesis. Our results suggest that fridamycin A has potential to induce fewer side effects such as weight gain compared to rosiglitazone, a commonly used antidiabetic drug, and that fridamycin A could be a novel potential therapeutic candidate for the management of type 2 diabetes.

Leaching of polymetallic sulfides is often challenging due to the complexity of these systems. The main issues are related to the formation of surface by-products or to the presence of different metals that require a greater consumption of chemicals compared to that needed to extract the target metal during leaching. This latter case is followed in the present work using X-ray photoelectron spectroscopy (XPS) during the leaching of a silver-containing polymetallic (Mn-Fe-Pb) sulfide under three different cyanidation procedures: (1) conventional leaching, (2) ultrasound-assisted leaching, and (3) leaching of a sample pretreated in alkaline media. In all cases, leaching results in the build-up of Pb compounds (oxides/hydroxides) on the surface of the minerals, suggesting a mechanism in which Pb is first leached by hydroxide and cyanide and later is re-adsorbed as an external layer on the minerals. On the other hand, the effect of each process on Mn was quite distinct: 1) Conventional leaching leads to the oxidation of the sulfide (pyrite) surface and a slight increase of surface Mn species; 2) Ultrasound-assisted leaching decreases surface Mn by dissolving oxidized species, which exposes fresh sulfide surfaces and increases Ag extraction; and 3) An alkaline pretreatment at high temperature exposed fresh sulfide surfaces and decreased the surface concentration of Mn. The fact that ultrasound-assisted leaching and leaching after an alkaline pretreatment allow for a higher Ag extraction (60% and 75% of total Ag, respectively, as opposed to the 25% extracted via conventional leaching), indicates that leaching is more efficient if the surface is not oxidized and if Mn is removed from the samples, either before or during leaching.

The ternary antimonide YIrSb was synthesized from the binary precursor YIr and elemental antimony by a diffusion controlled solid-state reaction. Single crystals were obtained by a flux technique with elemental bismuth as an inert solvent. The YIrSb structure (TiNiSi type, space group Pnma) was refined from single-crystal X-ray diffractometer data: a = 711.06(9), b = 447.74(5), c = 784.20(8) pm, wR2 = 0.0455, 535 F2 values, 20 variables. 89Y solid state MAS NMR and 121Sb Mossbauer spectra show single resonance lines in agreement with single-crystal X-ray data. YIrSb is a Pauli paramagnet.

Twenty new rare-earth metal rich intermetallic aluminium compounds, RE10TAl3 (RE = Y, Ho, Tm, Lu; T = Fe, Co, Ni, Ru, Rh, Pd, Os, Ir, Pt), were synthesized by arc melting the elements. The compounds crystallize, in analogy to e.g. the respective Cd representatives, with a ternary ordered structure as anti-type to the hexagonal Co2Al5 type, with the space group P63/mmc. The three crystallographically independent rare-earth metal sites occupy the aluminium positions of the aristotype, while the transition metal and aluminium atoms are ordered on the two cobalt sites. Like other rare-earth rich compounds the RE10TX3 members also exhibit transition-metal-centred T@RE6 trigonal prisms as striking structural building units. The prepared compounds have been investigated by susceptibility measurements and 27Al solid-state MAS-NMR measurements conducted on the Pauli-paramagnetic Y and Lu compounds. Some compounds show a certain amount of disorder as seen from the single crystal structure analysis and from signal broadening in the NMR investigations. By separating Knight shifts from second-order quadrupolar shifts via field dependent measurements, monotonic trends can be discerned regarding the effect of the T atom valence electron concentration and period number, as well as the effect of the closed 4f shell contributed in the Lu compounds. The results confirm that a comparison of Knight shifts within a series of isotypic compounds can reveal important electronic structure information in intermetallic systems.

With today's growing focus on on-demand energy production, the dynamic operation of biogas plants is becoming more and more important. Detecting unacceptable load changes and so preventing process failures is vital. In the present contribution, we propose an operating scheme that includes the detection of disturbances and intervention. This is then evaluated systematically with the help of a mathematical model for step changes in the organic loading rate. We summarize the qualitative results for each diagnosis variable in an operation diagram, enabling an intuitive, comprehensive comparison. Of the diagnosis variables analyzed, we find that the most promising for identifying incipient process failures in biogas plants are the volatile fatty acids to total alkalinity ratio (VFA/TA), followed by the ratio of methane to carbon dioxide γ in the biogas. In addition, the combination of the online measured variables pH value and gas composition can reveal disturbances early on in a wide range of different conditions.

The synthesis of the new chiral amidine (S,S)-N,N′-bis(1-phenylethyl)pivalamidine ((S)-HPETA) and its corresponding lithium salt (S)-LiPETA are reported, and their solid-state structures were investigated by single-crystal X-ray diffraction. Depending on the stoichiometric ratio and the ion radius of the rare-earth metal, the reaction of (S)-LiPETA with anhydrous lanthanide trihalides (Ln = Sc, Y, La, Nd, Sm, Lu) afforded mono-, bis-, and tris(amidinate) complexes. The mono(amidinate) compound [{(S)-PETA}LaI4Li2(thf)4], the bis(amidinate) complexes [({(S)-PETA}2Ln-μ-Cl)n] (Ln = Sc, Y, Nd, Sm, Lu), and the tris(amidinate) compound [{(S)-PETA}3Y] were isolated and structurally characterized by single-crystal X-ray diffraction. For the bis(amidinate) compounds, either monomeric or chloro-bridged dimeric structures were observed in the solid state. Furthermore, chiral bis(amidinate)-amido and -alkyl complexes [{(S)-PETA}2Ln{E(SiMe3)2}] (E = N, Ln = Y; E = CH, Ln = Sc, Y, Lu) were synthesized by salt metathesi...

The silicides ScIr4Si2 and RERh4Si2 (RE=Sc, Y, Tb-Lu) and silicide stannides RERh4Si2–x Sn x (RE=Y, Nd, Sm, Gd-Lu) were synthesized from the elements by arc-melting and subsequent annealing. The new compounds crystallize with the orthorhombic YRh4Ge2 type structure, space group Pnma. They were characterized by X-ray powder patterns and several structures were refined from single crystal X-ray diffractometer data. The main structural motifs of this series of silicides are tricapped trigonal prisms formed by the transition metal and rare earth atoms. One of the two crystallographically independent silicon sites allows for formation of solid solutions with tin, exemplarily studied for ErRh4Si2–x Sn x . Electronic structure calculations reveal strong covalent Rh–Si bonding as the main stability factor. Multinuclear (29Si, 45Sc, and 89Y) magic-angle spinning (MAS) NMR spectra of the structure representatives with diamagnetic rare-earth elements (Sc, Y, Lu) are found to be consistent with the crystallographic data and specifically confirm the selective substitution of Sn in the Si2 sites in the quaternary compounds YRh4SiSn and LuRh4SiSn.

The new equiatomic scandium transition metal aluminides ScTAl for T = Cr, Ru, Ag, Re, Pt, and Au were obtained by arc-melting of the elements followed by subsequent annealing for crystal growth. The samples were studied by powder and single crystal X-ray diffraction. The structures of three compounds were refined from single crystal X-ray diffractometer data: ScCrAl, MgZn2 type, P63/mmc, a = 525.77(3), c = 858.68(5) pm, R 1 = 0.0188, wR 2 = 0.0485, 204 F 2 values, 13 variables, ScPtAl, TiNiSi type, Pnma, a = 642.83(4), b = 428.96(2), c = 754.54(5) pm, R 1 = 0.0326, wR 2 = 0.0458, 448 F 2 values, 20 variables and ScAuAl, HfRhSn type, P6̅2c, a = 722.88(4), c = 724.15(4) pm, R 1 = 0.0316, wR 2 = 0.0653, 512 F 2 values, 18 variables. Phase pure samples of all compounds were furthermore investigated by magnetic susceptibility measurements, and Pauli-paramagnetism but no superconductivity was observed down to 2.1 K for all of them. The local structural features and disordering phenomena have been characterized by 27Al and 45Sc magic angle spinning (MAS) and static NMR spectroscopic investigations.

Phase pure polycrystalline PbP7 was prepared from the elements via a lead flux. Crystalline pieces with edge-lengths up to 1 mm were obtained. The assignment of the previously published P-31 solid state NMR spectrum to the seven distinct crystallographic sites was accomplished by radio-frequency driven dipolar recoupling (RFDR) experiments. As commonly found in other solid polyphosphides there is no obvious correlation between the P-31 chemical shift and structural parameters. PbP7 decomposes incongruently under release of phosphorus forming liquid lead as remainder. The thermal decomposition starts at T > 550 K with a vapor pressure almost similar to that of red phosphorus. Electronic structure calculations reveal PbP7 as a semiconductor according to the Zintl description and clearly shows the stereo active Pb-6s(2) lone pairs in the electron localization function ELF. (C) 2015 ELSEVIER. All rights reserved.

To overcome the disadvantages seen with current long-acting injectable formulations for risperidone (i.e., nonlinear release with initial lag time), an alternative delivery system was developed. The use of biodegradable PEG-PLGA polymers enabled us to formulate microparticles with a linear release over one month with no observable lag time. Diblock copolymers with PEG contents of 5 and 10% were transformed into risperidone-loaded highly porous microparticles with median particle sizes of 20–30 μm and narrow size distributions as confirmed by laser diffraction, electron and confocal microscopy. In parallel to characterizing the in vitro drug release behavior of the formulations, we studied their degradation/erosion profile by a couple of complementary techniques in detail. Compared to drug-free microparticles, the presence of risperidone led to a significantly faster polymer degradation accompanied by a more rapid mass loss and release of lactic acid. Proton nuclear magnetic resonance spectroscopy revealed a constant PEG content for the PEG-PLGA containing 5% of PEG over the course of incubation, while a decrease was observed for the diblock copolymer having an initial PEG content of 10%. In conclusion, PEG-PLGA polymers are also useful materials for the prolonged drug delivery of small lipophilic molecules, in addition to their application for the sustained release of sensitive drugs like peptides and proteins. Compared to “standard” polyesters such as PLGA, diblock copolymers avoid initial lag times and nonlinear “unpredictable” drug release profiles.

Synthetic derivatives of cyclic adenosine monophosphate, such as halogenated or other more hydrophobic analogs, are widely used compounds, to investigate diverse signal transduction pathways of eukaryotic cells. This inspired us to develop cyclic nucleotides, which exhibit chemical structures composed of brominated 7-deazaadenines and the phosphorylated ribosugar. The synthesized 8-bromo- and 7-bromo-7-deazaadenosine-3′,5′-cyclic monophosphates rank among the most potent activators of cyclic nucleotide-regulated ion channels as well as cAMP-dependent protein kinase. Moreover, these substances bind tightly to exchange proteins directly activated by cAMP.

Constraint-based modeling (CBM) is increasingly used to analyze the metabolism of complex microbial communities involved in ecology, biomedicine, and various biotechnological processes. While CBM is an established framework for studying the metabolism of single species with linear stoichiometric models, CBM of communities with balanced growth is more complicated, not only due to the larger size of the multi-species metabolic network but also because of the bilinear nature of the resulting community models. Moreover, the solution space of these community models often contains biologically unrealistic solutions, which, even with model linearization and under application of certain objective functions, cannot easily be excluded. Here we present RedCom, a new approach to build reduced community models in which the metabolisms of the participating organisms are represented by net conversions computed from the respective single-species networks. By discarding (single-species) net conversions that violate a minimality criterion in the exchange fluxes, it is ensured that unrealistic solutions in the community model are excluded where a species altruistically synthesizes large amounts of byproducts (instead of biomass) to fulfill the requirements of other species. We employed the RedCom approach for modeling communities of up to nine organisms involved in typical degradation steps of anaerobic digestion in biogas plants. Compared to full (bilinear and linearized) community models, we found that the reduced community models obtained with RedCom are not only much smaller but allow, also in the largest model with nine species, extensive calculations required to fully characterize the solution space and to reveal key properties of communities with maximum methane yield and production rates. Furthermore, the predictive power of the reduced community models is significantly larger because they predict much smaller ranges of feasible community compositions and exchange fluxes still being consistent with measurements obtained from enrichment cultures. For an enrichment culture for growth on ethanol, we also used metaproteomic data to further constrain the solution space of the community models. Both model and proteomic data indicated a dominance of acetoclastic methanogens (Methanosarcinales) and Desulfovibrionales being the least abundant group in this microbial community., Author summary Microbial communities are involved in many fundamental processes in nature, health and biotechnology. The elucidation of interdependencies between the involved players of microbial communities and how the interactions shape the composition, behavior and characteristic features of the consortium has become an important branch of microbiology research. Many communities are based on the exchange of metabolites between the species and constraint-based metabolic modeling has become an important approach for a formal description and quantitative analysis of these metabolic dependencies. However, the complexity of the models rises quickly with a growing number of organisms and the space of predicted feasible behaviors often includes unrealistic solutions. Here we present RedCom, a new approach to build reduced stoichiometric models of balanced microbial communities based on net conversions of the single-species models. We demonstrate the applicability of our RedCom approach by modeling communities of up to nine organisms involved in degradation steps of anaerobic digestion in biogas plants. As one of the first studies in this field, we compare simulation results from the community models with experimental data of laboratory-scale biogas reactors for growth on ethanol and glucose-cellulose media. The results also demonstrate a higher predictive power of the RedCom vs. the full models.