Interaction proteomics of synapse protein complexes

The brain integrates complex types of information, and executes a wide range of physiological and behavioral processes. Trillions of tiny organelles, the synapses, are central to neuronal communication and information processing in the brain. Synaptic transmission involves an intricate network of synaptic proteins that forms the molecular machinery underlying transmitter release, activation, and modulation of transmitter receptors and signal transduction cascades. These processes are dynamically regulated and underlie neuroplasticity, crucial to learning and memory formation. In recent years, interaction proteomics has increasingly been used to elucidate the constituents of synaptic protein complexes. Unlike classic hypothesis-based assays, interaction proteomics detects both known and novel interactors without bias. In this trend article, we focus on the technical aspects of recent proteomics to identify synapse protein complexes, and the complementary methods used to verify the protein–protein interaction. Moreover, we discuss the experimental feasibility of performing global analysis of the synapse protein interactome.     

Development of a microfluidic confocal fluorescence detection system for the hyphenation of nano-LC to on-line biochemical assays

One way to profile complex mixtures for receptor affinity is to couple liquid chromatography (LC) on-line to biochemical detection (BCD). A drawback of this hyphenated screening approach is the relatively high consumption of sample, receptor protein and (fluorescently labeled) tracer ligand. Here, we worked toward minimization of sample and reagent consumption, by coupling nano-LC on-line to a light-emitting diode (LED) based capillary confocal fluorescence detection system capable of on-line BCD with low-flow rates. In this fluorescence detection system, a capillary with an extended light path (bubble cell) was used as a detection cell in order to enhance sensitivity. The technology was applied to a fluorescent enhancement bioassay for the acetylcholine binding protein, a structural analog of the extracellular ligand-binding domain of neuronal nicotinic acetylcholine receptors. In the miniaturized setup, the sensitive and low void volume LED-induced confocal fluorescence detection system operated in flow injection analysis mode allowing the measurement of IC₅₀ values, which were comparable with those measured by a conventional plate reader bioassay. The current setup uses 50 nL as injection volume with a carrier flow rate of 400 nL/min. Finally, coupling of the detection system to gradient reversed-phase nano-LC allowed analysis of mixtures in order to identify the bioactive compounds present by injecting 10 nL of each mixture.     

Effects of Ultraviolet A-1 Radiation on Calcineurin Activity and Cytokine Production in (Skin) Cell Cultures†

Calcineurin (Cn) is the target of immunosuppressive drugs used for maintenance therapy of transplant patients. UV radiation is also known to be immunosuppressive and, like the Cn inhibitors, UV has been shown to positively influence various inflammatory skin diseases. Recently, Cn activity has been demonstrated in skin and skin cell cultures. In the present study we have investigated the effects of UV(A-1) irradiation on Cn activity in skin. In total skin we found a significant reduction in Cn activity after exposure to 450 kJ m⁻² of UVA-1 (340–400 nm). In repeated experiments cultures of fibroblasts and keratinocytes also showed dose-dependent and selective reduction in Cn activity after UVA-1 irradiation. UVB irradiation caused a decrease in the Cn activity of one of two fibroblast cultures and was ineffective in keratinocytes. In Jurkat cells and PBMC UVA-1 reduced Cn activity and also the production of cytokines such as interleukin (IL)-2, γ-interferon, IL-4 and IL-10 that are controlled by the Ca²⁺–Cn pathway. These results indicate that UV(A-1) irradiation may lead to inactivation of Cn in the skin and thus suppress the skin immune system in a similar fashion to the Cn inhibitors.     

Abscheidung von Kohlendioxid: Perspektiven für neue Materialien

Der stark ansteigende Kohlendioxidgehalt in der Atmosphäre ist eines der drängendsten Umweltprobleme unserer Zeit. Eine Option zur Verringerung anthropogener CO₂‐Emissionen ist die Abscheidung und Speicherung von Kohlendioxid (Carbon Capture and Storage, CCS) an Punktquellen wie Kraftwerken. Durch diese Sequestrierung steigt allerdings der Energiebedarf der Kraftwerke um 25–40 %. Wir berichten hier über die Technologien zur Abscheidung, die zur Verringerung der CO₂‐Emissionen wahrscheinlich am besten geeignet sind. Dazu zählen Postcombustion‐Verfahren, bei denen die Abscheidung nach der Verbrennung stattfindet (vor allem die CO₂/N₂‐Separation), Precombustion‐Verfahren, bei denen CO₂/H₂‐Gemische eingesetzt werden, und die Konditionierung von Erdgas (CO₂/CH₄). Der Schlüssel zu deutlichen Fortschritten sind bessere Trennmittel zur Separation. Wir werden hier aktuelle Entwicklungen und neuartige Konzepte zur CO₂‐Abtrennung durch Lösungsmittel‐Absorption, chemische und physikalische Adsorption und Membranen schildern und besonders auf Fortschritte auf dem wachsenden Gebiet Metall‐organischer Gerüste eingehen.     

Nanoporous Materials Can Tune the Critical Point of a Pure Substance

Molecular simulations and NMR relaxometry experiments demonstrate that pure benzene or xylene confined in isoreticular metal–organic frameworks (IRMOFs) exhibit true vapor–liquid phase equilibria where the effective critical point may be reduced by tuning the structure of the MOF. Our results are consistent with vapor and liquid phases extending over many MOF unit cells. These results are counterintuitive since the MOF pore diameters are approximately the same length scale as the adsorbate molecules. As applications of these materials in catalysis, separations, and gas storage rely on the ability to tune the properties of adsorbed molecules, we anticipate that the ability to systematically control the critical point, thereby preparing spatially inhomogeneous local adsorbate densities, could add a new design tool for MOF applications.     

Influence of a TiCl4 post-treatment on nanocrystalline TiO2 films in dye-sensitized solar cells

In this study, the influence of the TiCl(4) post-treatment on nanocrystalline TiO(2) films as electrodes in dye-sensitized solar cells is investigated and compared to nontreated films. As a result of this post-treatment cell efficiencies are improved, due to higher photocurrents. On a microscopic scale TiO(2) particle growth on the order of 1 nm is observed. Despite a corresponding decrease of BET surface area, more dye is adsorbed onto the oxide surface. Although it seems trivial to match this finding with the improved photocurrent, this performance improvement cannot be attributed to higher dye adsorption only. This follows from comparison between incident photon to current conversion efficiency (IPCE) and light absorption characteristics. Since the charge transport properties of the TiO(2) films are already more than sufficient without treatment, the increase in short circuit current density J(SC) cannot be related to improvements in charge transport either. Transient photocurrent measurements indicate a shift in the conduction band edge of the TiO(2) upon TiCl(4) treatment. It is concluded that the main contribution to enhanced current originates from this shift in conduction band edge, resulting in improved charge injection into the TiO(2).     

The effects of freeze/thaw periods and drying methods on isotopic and elemental carbon and nitrogen in marine organisms, raising questions on sample preparation

Stable isotopes are an increasingly important tool in trophic linkage ecological studies. In studies of large marine animals, isotopic sampling is often given secondary priority to sampling for diversity and biomass aspects. Consequently, isotopic samples are frequently collected subsequent to repeated freezing and thawing of animals, and the results of these studies are often based on the assumption that this pre-treatment does not affect the isotopic values. Our study tested this assumption and examined the difference between oven- and freeze-drying on isotopic values and elemental carbon-to-nitrogen (C:N) ratios. The values for δ(15)N and δ(13)C, percentage nitrogen and carbon, and the C:N ratios were determined from the tissues of six marine species, including invertebrates and fish, as (1) fresh samples, (2) samples thawed once, and (3) samples thawed twice. The drying method, thawing treatment and their interaction did significantly affect the δ(15)N and δ(13)C isotope values for all species. Oven-dried samples had slightly higher δ(13)C and δ(15)N values than freeze-dried samples, although not significant in most instances. For most species, oven-drying produced lower carbon and nitrogen percentage than freeze-drying for samples that had been thawed once, but the C:N ratio was unaffected by the drying method. Repeated freezing and thawing did not affect the isotope values, but it did decrease the percentage carbon and nitrogen for both desiccation methods. We recommend drying samples from fresh wherever possible, and careful choice of desiccation method in light of the fact that most lipid models are based on oven-dried samples and oven-drying could cause enrichment of (15)N or (13)C through evaporation of volatile compounds richer in lighter isotopes such as some lipids. Finally, we recommend that further studies on the specific effects of freezing and desiccation on elasmobranchs is needed. Overall we recommend the use of freeze-drying when possible and to use the samples from freshly caught organisms.     

Linearization of column responses by correlation chromatography. Separation improvement and reduction of undesired adsorption effects

Summary Experiments and simulations prove that correlation chromatography can greatly reduce the disadvantage of a non-linear response of the chromatographic column. A factor that has been accepted as being an important source of error in correlation or multiplex chromatography, has been shown not to be. Separations affected, improve dramatically when correlation chromatography is used, and a substantial amount of correlation noise only arises when there is a large difference in separation between a conventional chromatogram and a correlogram.A model has been developed for simulating these nonlinearities. It is shown that, especially for correlation chromatography, the simulation results match the practical measurements very well.     

Structure of Mg2.56V1.12W0.88O8 and vibrational raman spectra of Mg2.5VWO8 and Mg2.5VMoO8

Mg(2.56)V(1.12)W(0.88)O(8) crystals were grown from a MgO/V(2)O(5)/WO(3) melt. X-ray single-crystal diffraction studies revealed that it is orthorhombic with space group Pnma, a = 5.0658(5) A, b = 10.333(1) A, c = 17.421(2) A, Z = 6, and is isostructural with Mg(2.5)VMoO(8). Raman spectra are reported, and the assignment of the Raman bands is made by comparing the metal-oxygen vibrations of VO(4)/WO(4) tetrahedra in Mg(2.5)VWO(8) with the metal-oxygen vibrations of VO(4)/MoO(4) tetrahedra in Mg(2.5)VMoO(8). The stretching vibrations appearing at 1016 and 1035 cm(-)(1) are assigned to Mo=O and W=O double bonds, respectively, associated with the Mg(2+) cation vacancies.