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SummaryPrime editing is a highly versatile genome editing technology that holds great potential for treating genetic diseases1, 2. Whilein vivoprime editing has recently been conducted in the brain, liver, heart, and retina3–6, application of this technology to modulate neural circuits in the brain has not been reported yet. Here, we employ adeno-associated viral vectors to deliver optimized intein-split prime editors into the brain of mice. Delivery into newborn pups via intracerebroventricular injection resulted in up to 44.0% editing at theDnmt1locus in the cortex (on average 34.8±9.8% after 6 months). In addition, we obtained up to 28.1% editing at theAdrb1locus in the cortex (on average 14.7±11.6% after 6 months). The introducedAdrb1A187Vmutation is a naturally occurring variant of the β1-adrenergic receptor, which has previously been linked to increased activity and natural short sleep7. Similarly, we observed an increase in the activity and exploratory behavior of treated animals. This study demonstrates the potential of prime editing for treating genetic diseases in the central nervous system and for reprogramming molecular pathways that modulate animal behavior.

Zinc is the second most prevalent metal element present in living organisms, and control of its concentration is pivotal to physiology. The amount of zinc available to the cell cytoplasm is regulated by the activity of members of the SLC39 family, the ZIP proteins. Selectivity of ZIP transporters has been the focus of earlier studies which provided a biochemical and structural basis for the selectivity for zinc over other metals such as copper, iron, and manganese. However, several previous studies have shown how certain ZIP proteins exhibit higher selectivity for metal elements other than zinc. Sequence similarities suggest an evolutionary basis for the elemental selectivity within the ZIP family. Here, by engineering HEK293 cells to overexpress ZIP proteins, we have studied the selectivity of two phylogenetic clades of ZIP proteins, that is ZIP8/ZIP14 (previously known to be iron and manganese transporters) and ZIP5/ZIP10. By incubating ZIP over-expressing cells in presence of several divalent metals, we found that ZIP5 and ZIP10 are high affinity copper transporters with greater selectivity over other elements, revealing a novel substrate signature for the ZIP5/ZIP10 clade.

Faecal sterols and stable isotopes (δ13C and δ15N) from bulk organic matter (OM) were analysed in three sedimentary cores collected in two subtropical bays located in the South Atlantic to evaluate historical trends in the sewage input and to track possible changes in the bulk isotopic composition of OM in recent decades. The values of δ13C and δ15N ranged from −27.4 to −25.0‰ and from 0.5 to 3.9‰, respectively, without a clear trend in the variation over the whole period covered by sediment cores and with no conclusive interpretation of a specific range value typically related to the sewage input for these areas. The maximum coprostanol concentration was 0.19 μg g−1 in the upper 4 cm of one core, which was not considered contaminated by evaluation of the sterols diagnostic ratios. Even at low levels, the coprostanol concentrations followed variations in urban and economical regional development. Baseline values for faecal sterols (in average between 0.03 and 0.05 μg g−1), which may represent a previous non-impacted environment scenarios, were calculated for use in comparative perspectives for future evaluations of the sewage input and contamination.

The solute carrier (SLC) superfamily represents the biggest family of transporters with important roles in health and disease. Despite being attractive and druggable targets, the majority of SLCs remains understudied. One major hurdle in research on SLCs is the lack of tools, such as cell-based assays to investigate their biological role and for drug discovery. Another challenge is the disperse and anecdotal information on assay strategies that are suitable for SLCs. This review provides a comprehensive overview of state-of-the-art cellular assay technologies for SLC research and discusses relevant SLC characteristics enabling the choice of an optimal assay technology. The Innovative Medicines Initiative consortium RESOLUTE intends to accelerate research on SLCs by providing the scientific community with high-quality reagents, assay technologies and data sets, and to ultimately unlock SLCs for drug discovery.

Art. 2 EUV stellt die Europaische Union auf ein Wertefundament, das sich insbesondere aus Menschenrechten, Demokratie und Solidaritat zusammensetzt. Doch die dort propagierten Werte werden gegenwartig einer Ausnahmezustands- und Krisenrhetorik untergeordnet; an ihre Stelle werden Zweckmasigkeiten gesetzt. Menschenrechte, Demokratie und Solidaritat werden in Zeiten tatsachlicher oder vermeintlicher Krisen derart auser Acht gelassen, dass tatsachlich von einer Suspendierung der Werte des Art. 2 EUV zu sprechen ist. Verschiedene Formen dieser Suspendierung zeigen sich exemplarisch am sogenannten Hotspot-Konzept an den Ausengrenzen der Union, wo grundrechtliche Garantien systematisch verletzt werden, am zunehmenden Ausweichen auf informelle, exekutiv-zentrierte Verfahren, durch die demokratische Sicherungen umgangen werden, sowie am volligen Verzicht auf eine gemeinsame Lastenaufteilung, die dem Solidaritatsgedanken Rechnung tragen wurde. Halt man an der Relevanz europaischer Grundwerte fest, so ist ...

Este trabalho apresenta o modelamento de dois novos Nós Lógicos para a norma IEC 61850. Estes modelos foram implementados no monitoramento de dois transformadores de transmissão e distribuição para a criação de um sistema de gestão destes ativos, cuja topologia foi elaborada com base na norma IEC 61850. Este trabalho contribui para a revisão do capítulo da norma que versa sobre monitoramento e supervisão de transformadores (IEC 61850-7-4) e inaugura modelos de Diagnostico e Expectativa de sua Vida Útil. A implementação foi realizada na área piloto do projeto (subestação Pilarzinho), de forma experimental e, avaliados e validados em campo, através da utilização de uma técnica não invasiva de avaliação da degradação do sistema de isolamento dos transformadores, a chamada Espectroscopia Dielétrica por Corrente de Polarização e Despolarização (PDC) e a Medida de Condutividade do Óleo Isolante (LCM). Foi implementado também o processo de secagem da parte ativa baseado em filtros moleculares e que consiste na remoção seletiva das moléculas de água do óleo isolante provocando um desequilíbrio entre a umidade dos materiais isolantes.

Sedimentary sterols and linear alkylbenzenes associated with allochthonous organic matter (AOM) inputs were studied in surface sediments along the Tubarão riverbed, South Brazil. These markers were analysed in terms of concentrations, diagnostic ratios and by using multivariate analyses to identify the main organic matter sources. It was necessary to integrate all these factors to distinguish the sources and determine sewage contamination. Phytosterols predominated over faecal sterols, but the contributions of livestock waste along the river (determined in 50% of the sites) were confirmed by the fingerprint analysis. Raw sewage contamination was verified at one site, according to the increased levels of sewage molecular markers and confirmed by the multivariate analyses and diagnostic ratios calibrated to this region. A possible synergistic effect between inorganic nanoparticles from coal mine waste and organic contaminants related to AOM input was suggested and should not be ignored since both activities severely contribute to the environmental changes in much of this fluvial-estuarine gradient from the South Atlantic.

Gut vernetzt gut verwaltet. Wer mit Fragen der strategischen Steuerung, des Datenschutzes und des Verwaltungshandelns vor dem Hintergrund der Digitalisierung befasst ist, findet in diesem Band topaktuelle Antworten. Auf dem neuesten Rechtsstand – inkl. DSGVO und der Erklärung von Tallinn. Integrativ und ebenenübergreifend werden gleichermaßen die EU-, Bundes-, Landes- und Kommunalebene behandelt. Gut lesbar eignet sich der Band als Einstiegslektüre und zur Vertiefung. Mit einem Geleitwort der Staatsministerin für Digitalisierung, Dorothee Bär.

Faecal sterols and stable isotopes (δ

The advantage of the new generation IV iron preparations ferric carboxymaltose (FCM), ferumoxytol (FMX), and iron isomaltoside 1000 (IIM) is that they can be administered in relatively high doses in a short period of time. We investigated the physico-chemical properties of these preparations and compared them with those of the older preparations iron sucrose (IS), sodium ferric gluconate (SFG), and low molecular weight iron dextran (LMWID). Mossbauer spectroscopy, X-ray diffraction, and Fe K-edge X-ray absorption near edge structure spectroscopy indicated akaganeite structures (β-FeOOH) for the cores of FCM, IIM and IS, and a maghemite (γ-Fe2O3) structure for that of FMX. Nuclear magnetic resonance studies confirmed the structure of the carbohydrate of FMX as a reduced, carboxymethylated, low molecular weight dextran, and that of IIM as a reduced Dextran 1000. Polarography yielded significantly different fingerprints of the investigated compounds. Reductive degradation kinetics of FMX was faster than that of FCM and IIM, which is in contrast to the high stability of FMX towards acid degradation. The labile iron content, i.e. the amount of iron that is only weakly bound in the polynuclear iron core, was assessed by a qualitative test that confirmed decreasing labile iron contents in the order SFG ≈ IS > LMWID ≥ FMX ≈ IIM ≈ FCM. The presented data are a step forward in the characterization of these non-biological complex drugs, which is a prerequisite to understand their cellular uptake mechanisms and the relationship between the structure and physiological safety as well as efficacy of these complexes.

Intravenous iron preparations are typically classified as non-dextran-based or dextran/dextran-based complexes. The carbohydrate shell for each of these preparations is unique and is key in determining the various physicochemical properties, the metabolic pathway, and the immunogenicity of the iron-carbohydrate complex. As intravenous dextran can cause severe, antibody-mediated dextran-induced anaphylactic reactions (DIAR), the purpose of this study was to explore the potential of various intravenous iron preparations, non-dextran-based or dextran/dextran-based, to induce these reactions. An IgG-isotype mouse monoclonal anti-dextran antibody (5E7H3) and an enzyme-linked immunosorbent assay (ELISA) were developed to investigate the dextran antigenicity of low molecular weight iron dextran, ferumoxytol, iron isomaltoside 1000, ferric gluconate, iron sucrose and ferric carboxymaltose, as well as isomaltoside 1000, the isolated carbohydrate component of iron isomaltoside 1000. Low molecular weight iron dextran, as well as dextran-based ferumoxytol and iron isomaltoside 1000, reacted with 5E7H3, whereas ferric carboxymaltose, iron sucrose, sodium ferric gluconate, and isolated isomaltoside 1000 did not. Consistent results were obtained with reverse single radial immunodiffusion assay. The results strongly support the hypothesis that, while the carbohydrate alone (isomaltoside 1000) does not form immune complexes with anti-dextran antibodies, iron isomaltoside 1000 complex reacts with anti-dextran antibodies by forming multivalent immune complexes. Moreover, non-dextran based preparations, such as iron sucrose and ferric carboxymaltose, do not react with anti-dextran antibodies. This assay allows to assess the theoretical possibility of a substance to induce antibody-mediated DIARs. Nevertheless, as this is only one possible mechanism that may cause a hypersensitivity reaction, a broader set of assays will be required to get an understanding of the mechanisms that may lead to intravenous iron-induced hypersensitivity reactions.

During the operation of electrical equipments the compound 2-furfuraldehyde (2-FAL) and other related furanic substances are produced from the insulating paper degradation and can be detected in the mineral oil by high performance liquid chromatography (HPLC). Their concentration increase can be used to estimate the quality of the paper insulation inside the equipments. In this work colorimetric methodologies for quantification of these furanic degradation products in oil filled electrical equipment were evaluated. Two analytical methods were studied using aniline and modified Winkler’s reactions. For this purpose used oil samples and 2-FAL standard oil solutions were analyzed and the results for both methods were compared with HPLC analysis. The obtained results for 2-FAL standard oil solutions were in accordance with both methodologies. The results obtained for used mineral oil showed that the analysis of furanic compounds by the colorimetric methods may induce to false positive results due to the interference of oil oxidation by-products in the quantitative analysis.

Chitosan (Ch) and N-carboxymethylchitosan (N-CMCh) films were prepared by the casting method at concentrations of 1% and 2% of polymer, with or without plasticizer: polyethylene glycol (PEG-400) and glycerol (G), at 15% (w/w). The influence of composition on mechanical properties, water vapour transmission rate (WVTR), water saturation, and aqueous dissolution of the films was analysed. The thermal stability of the mixture (polymer:plasticizer, 1:1) was evaluated by thermogravimetric analysis (TGA). In general, all the properties were affected by the plasticizers. The plasticized films showed lower strength and a higher percentage of elongation (%E), in the following order: G > PEG-400 > unplasticized film. The total WVTR increased with Ch concentration, with a different WVTR profile for Ch and N-CMCh. While the PEG-400 addition did not significantly modify the WVTR profile of films, the glycerol enhanced the transport of water vapour through both polymers. The plasticizer addition increased the time of water film saturation, in the following order: G > PEG-400 > unplasticized film; this was more pronounced in the N-CMCh films, probably due to the formation of hydrogen bonds. The solubility of the films was also affected by their composition. Copyright © 2006 Society of Chemical Industry

Chitosan (degree of deacetylation of 90.2%) and N-carboxymethylchitosan (N-CMCh) (degree of substitution of 18.5%) were analyzed using thermogravimetric analysis in order to determine their thermal stability. Also, their films were evaluated using scanning electron microscopy (SEM) and mechanical and dynamic mechanical analysis (DMA). Both polymers showed a thermal degradation peak at Tm ∼ 250 °C, with Tonset and weight loss of 175 °C and 62% and 190 °C and 35% for chitosan and N-CMCh, respectively. N-CMCh showed a second thermal degradation peak at Tm = 600 °C, with an additional weight loss of 25%. Kinetic thermal analysis showed a slower process of degradation at 100 °C for N-CMCh compared with chitosan, and an activation energy 13 times higher for the former, confirming the higher stability of N-CMCh. Analysis of chitosan and N-CMCh films showed that the latter support a high tension, with lower elasticity, and, as revealed by DMA, N-CMCh has a more compact film structure, with a crossing arrangement of N-CMCh fibers, as compared with the chitosan films which were determined from SEM analysis to have fibers in one direction only. Copyright © 2006 Society of Chemical Industry

This article concerns the in situ compatibilization of immiscible isotatic polypropylene/styrene–butadiene–styrene triblock copolymer blends (i-PP/SBS) by use of a reactive mixture. For this purpose, maleated PP (PP–MAH) and SBS (SBS–MAH) were used as functionalized polymers and 4,4′-diaminediphenylmethane was used as a coupling agent between maleated polymers, resulting in a graft copolymer. Binary blends of i-PP/SBS, nonreactive ternary blends of i-PP/PP–MAH/SBS, and reactive ternary blends of i-PP/PP–MAH/SBS–MAH with varying diamine/anhydride molar ratios were prepared. The mechanical properties of the blends were determined by tensile and impact-resistance tests. The optimum improvement in the mechanical properties was found when the diamine/anhydride molar ratio in the ternary reactive blends was 0.5/1. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 87: 516–522, 2003

Blends of maleated polypropylene with maleic anhydride (PP–MAH) and maleated styrene–butadiene–styrene triblock copolymer with maleic anhydride (SBS–MAH) were compatibilized with 4,4′-diaminediphenylmethane, at various concentrations, as the coupling agent for the functionalized polymers. Their properties were compared with those of the corresponding blends of PP–MAH and styrene–butadiene–styrene triblock copolymer. The blends containing 15 and 50 wt % elastomers were prepared in a mixer at 190°C and 55 rpm. Torque measurements and solubility tests suggested a graft copolymerization during the melt blending of PP–MAH and SBS–MAH. Infrared spectroscopy was used to characterize the graft copolymer formed during the melt blending of the maleated polymers. The blends presented a morphology of a disperse elastomer phase in a continuous PP–MAH matrix. The size of the elastomeric domains decreased as the diamine and anhydride molar ratio increased. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 02: 366–371, 2002

The morphology and mechanical properties of blends of isotactic polypropylene (i-PP) and a maleated butadiene–styrene–butadiene triblock copolymer with maleic anhydride (SBS–MAH), i-PP/SBS–MAH, were investigated and compared with those of corresponding blends of i-PP and a styrene–butadiene–styrene triblock copolymer (SBS), i-PP/SBS. The blends were prepared in a mixer at 190°C with 5–25 wt % elastomer compositions. The blends exhibited a two-phase morphology with a disperse elastomer phase in the continuous i-PP matrix. The size and shape of the elastomer particles and the mechanical properties of the blends depended on the nature and concentration of the elastomer. The i-PP/SBS–MAH blends showed an impact strength superior to that of the corresponding i-PP/SBS blends. These results were attributed to the formation of the graft copolymer during the melt blending, which was verified by Fourier transform infrared. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 359–365, 2002

This article concerns the in situ compatibilization of immiscible isotactic polypropylene/butadiene-styrene-butadiene triblock copolymer blends (i-PP/SBS) by means of a reactive mixture. For this purpose, maleated PP (PP-MAH) and SBS (SBS-MAH) were used as functionalized polymers and 4,4′-diaminediphenylmethane was used as a coupling agent between maleated polymers, resulting in a graft copolymer. Binary blends i-PP/SBS, nonreactive ternary blends i-PP/PP-MAH/SBS, and reactive ternary blends i-PP/PP-MAH/SBS-MAH with varying diamine and anhydride molar ratios were prepared. Torque measurements suggest a graft copolymerization during the melt blending for ternary reactive blends, but the extension of the grafting does not vary with the diamine and anhydride molar ratio, but with the elastomer concentration. The morphology of the blends was investigated by scanning electron microscopy. The morphology of binary and ternary nonreactive blends is similar, exhibiting elastomer domains disperse in the i-PP matrix, whose sizes increase with elastomer concentration. On the other hand, the elastomer domain size in the ternary reactive blends is practically independent of the blends composition and of the diamine and anhydride molar ratio. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 85: 847–855, 2002

The bulk modification of SBS rubber with maleic anhydride in a mixing chamber of a Haake rheomixer was studied. The effect of temperature, maleic anhydride, and benzoyl peroxide concentrations on the grafting efficiency was evaluated. High grafting efficiency was achieved when the ratio of peroxide and maleic anhydride concentration was high. On the other hand, on this condition high insoluble fraction was generated. The addition of a diamine, 4,4′-diaminediphenylmethane to the reaction mixture minimizes the amount of insoluble polymer. However, the grafted MAH content also decreases. The graft copolymer was characterized by infrared spectroscopy and the grafting extension was determined by titration. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 2953–2960, 2002; DOI 10.1002/app.10355

AIMS Hyperphosphatemia in advanced chronic kidney disease (CKD) necessitates the use of phosphate binders. This in vitro study assessed phosphate binding and Fe release properties of the novel iron-based phosphate binder PA21. MATERIALS AND METHODS Phosphate adsorption and Fe release were assessed under conditions simulating administration of PA21 on an empty stomach and full stomach across a pH range to which PA21 would be exposed during passage through the gastrointestinal (GI) tract. RESULTS PA21 showed a robust phosphate binding capacity over the entire physiologically relevant pH range. The high binding capacity at low pH indicates that phosphate binding could begin in the stomach. Under the current experimental setting, the maximal bound phosphate to Fe ratio was 0.47 mmol P/mmol Fe. The largest amount of Fe release was observed at the lowest pH without phosphate and was much lower in the presence of phosphate. These results are in line with the formation of iron phosphate at low pH, as indicated by X-ray photoelectron spectroscopy and thermodynamic calculations. Fe release was minimal (≤ 0.35%) across pH 2.5 - 8.5. CONCLUSIONS These studies demonstrate that PA21 has potent phosphate binding capacity and low iron release over a physiologically relevant pH range in the GI tract. These features indicate PA21 could be an effective alternative phosphate binder for CKD patients.

An isocratic chromatographic method for the simultaneous determination of 10 benzodiazepines is presented. The selectivity of the assay was optimized by variation of stationary phase, temperature, as well as ionic strength, composition and pH of the mobile phase and the dependence of the detector response on the applied potential was investigated. The best results with respect to resolution at moderate retention times were obtained with a mixture of 0.02 mol/l phosphate buffer (pH 6) and acetonitrile in a volume ratio of 55:45 (v/v) on a LiChrospher-100 RP-8ec column (150x4.6 mm I.D.). Considerable improvement of selectivity was achieved if the column temperature was kept constant at 12 degrees C. Two detection modes were applied, UV detection at 250 nm, inserted upstream to the electrochemical detector, and reductive electrochemical detection at the hanging mercury drop electrode at -1.4 V (vs. Ag/AgCl), which proved to be especially sensitive in case of nitrophenyl-containing benzodiazepine species. The elaborated assay showed to be linear up to at least 2 mg/l for each compound. Detection limits generally were in the range of 6.5-123 ng/ml (130 pg-2.46 ng on-column, using a 20-microl loop) with relative standard deviations between 1.1 and 8.6% depending on the actual benzodiazepine investigated.

Deux villes réputées créatives sont comparées grâce à une approche inductive fondée sur des études de cas. Une grille d’analyse spécifiquement constituée pour cette étude met en lumière le rôle moteur d’un Underground créatif à Montréal alors que les acteurs institutionnels de l’Upperground battent la cadence à Munich. Cette comparaison nous amène à distinguer la dynamique qui anime chacune des deux villes : créativité à Montréal, innovation à Munich. Les succès des deux villes amènent leur cherté croissante, mettent en évidence le caractère cyclique des deux parcours et font surgir d’autres acteurs encore ignorés dans la littérature., Two cities with a reputation for creativity are compared using an inductive, case-study-based approach. An analysis grid specifically developed for this study highlights the leading role played by a creative Underground in Montreal while the institutional actors of the Upperground set the pace in Munich. This comparison leads us to distinguish the dynamics that drive each of the two cities: creativity in Montreal, innovation in Munich. The success of both cities comes with increasing living costs, highlights the cyclical nature of the two paths, and brings to the forefront other actors that the literature had ignored until now., El estudio compara dos ciudades consideradas creativas utilizando un enfoque inductivo basado en estudios de casos. Una tabla analítica diseñada específicamente para este estudio destaca el papel impulsor de los actores del Underground creativo en Montreal, mientras que los actores institucionales del Upperground imponen el ritmo en Munich. Esta comparación de actores permite distinguir la dinámica que impulsa a cada una de las dos ciudades: la creatividad en Montreal y la innovación en Munich. El éxito de ambas ciudades trae consigo un aumento de la carestía, resalta el carácter cíclico de ambas trayectorias y destaca a otros actores aún ausentes en la literatura.