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961.
Chemical Approach to Biological Safety: Molecular‐Level Control of an Integrated Zinc Finger Nuclease
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Dr. Masamitsu N. Asaka Dr. Kohsuke Kato Zita Fábián Prof. Dr. Chris Oostenbrink Dr. Hans E. M. Christensen Prof. Dr. Kyosuke Nagata Dr. Béla Gyurcsik 《Chembiochem : a European journal of chemical biology》2018,19(1):66-75
Application of artificial nucleases (ANs) in genome editing is still hindered by their cytotoxicity related to off‐target cleavages. This problem can be targeted by regulation of the nuclease domain. Here, we provide an experimental survey of computationally designed integrated zinc finger nucleases, constructed by linking the inactivated catalytic centre and the allosteric activator sequence of the colicin E7 nuclease domain to the two opposite termini of a zinc finger array. DNA specificity and metal binding were confirmed by electrophoretic mobility shift assays, synchrotron radiation circular dichroism spectroscopy, and nano‐electrospray ionisation mass spectrometry. In situ intramolecular activation of the nuclease domain was observed, resulting in specific cleavage of DNA with moderate activity. This study represents a new approach to AN design through integrated nucleases consisting of three (regulator, DNA‐binding, and nuclease) units, rather than simple chimera. The optimisation of such ANs could lead to safe gene editing enzymes. 相似文献
962.
DNA Primer Extension with Cyclopropenylated 7‐Deaza‐2′‐deoxyadenosine and Efficient Bioorthogonal Labeling in Vitro and in Living Cells
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Dr. Damian Ploschik Dr. Franziska Rönicke Hanna Beike Dr. Ralf Strasser Prof. Dr. Hans‐Achim Wagenknecht 《Chembiochem : a European journal of chemical biology》2018,19(18):1949-1953
A deoxyadenosine triphosphate (dATP) analogue for DNA labeling was synthesized with the 1‐methylcyclopropene (1MCP) group at the 7‐position of 7‐deaza‐2′‐deoxyadenosine and applied for primer extension experiments. The real‐time kinetic data reveals that this 1MCP‐modified dATP analogue is incorporated into DNA much faster than that of the similarly 1MCP‐modified deoxyuridine triphosphate (dUTP) analogue. The postsynthetic fluorescent labeling of these oligonucleotides works efficiently according to PAGE analysis, and can be applied for immobilization of a functional antibody on a surface. Site‐specific labeling at two different positions in DNA was achieved and the bioorthogonality of the postsynthetic fluorescent labeling was demonstrated in living HeLa cells. 相似文献
963.
Synthetic Indolactam V Analogues as Inhibitors of PAR2‐Induced Calcium Mobilization in Triple‐Negative Breast Cancer Cells
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Dr. Jan Stein Dr. Sonja Stahn Dr. Jörg‐M. Neudörfl Julia Sperlich Prof. Dr. Hans‐Günther Schmalz Prof. Dr. Nicole Teusch 《ChemMedChem》2018,13(2):147-154
Human proteinase‐activated receptor 2 (PAR2), a transmembrane G‐protein‐coupled receptor (GPCR), is an attractive target for a novel anticancer therapy, as it plays a critical role in cell migration and invasion. Selective PAR2 inhibitors therefore have potential as anti‐metastatic drugs. Knowing that the natural product teleocidin A2 is able to inhibit PAR2 in tumor cells, the goal of the present study was to elaborate structure–activity relationships and to identify potent PAR2 inhibitors with lower activity against the adverse target, protein kinase C (PKC). For this purpose, an efficient gram‐scale total synthesis of indolactam V (i.e., the parent structure of all teleocidins) was developed, and a library of derivatives was prepared. Some compounds were indeed found to exhibit high potency as PAR2 inhibitors at low nanomolar concentrations with improved selectivity (relative to teleocidin A2). The pseudopeptidic fragment bridging the C3 and C4 positions of the indole core proved to be essential for target binding, whereas activity and target selectivity depends on the substituents at N1 or C7. This study revealed novel derivatives that show high efficacy in PAR2 antagonism combined with increased selectivity. 相似文献
964.
Synthesis,Characterization, and Initial Biological Evaluation of [99mTc]Tc‐Tricarbonyl‐labeled DPA‐α‐MSH Peptide Derivatives for Potential Melanoma Imaging
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Dr. Feng Gao Dr. Wiebke Sihver Dr. Ralf Bergmann Dr. Birgit Belter Dr. Cristina Bolzati Dr. Nicola Salvarese Prof. Dr. Jörg Steinbach Prof. Dr. Jens Pietzsch Dr. Hans‐Jürgen Pietzsch 《ChemMedChem》2018,13(11):1146-1158
α‐Melanocyte stimulating hormone (α‐MSH) derivatives target the melanocortin‐1 receptor (MC1R) specifically and selectively. In this study, the α‐MSH‐derived peptide NAP‐NS1 (Nle‐Asp‐His‐d ‐Phe‐Arg‐Trp‐Gly‐NH2) with and without linkers was conjugated with 5‐(bis(pyridin‐2‐ylmethyl)amino)pentanoic acid (DPA‐COOH) and labeled with [99mTc]Tc‐tricarbonyl by two methods. With the one‐pot method the labeling was faster than with the two‐pot method, while obtaining similarly high yields. Negligible trans‐chelation and high stability in physiological solutions was determined for the [99mTc]Tc‐tricarbonyl–peptide conjugates. Coupling an ethylene glycol (EG)‐based linker increased the hydrophilicity. The peptide derivatives displayed high binding affinity in murine B16F10 melanoma cells as well as in human MeWo and TXM13 melanoma cell homogenates. Preliminary in vivo studies with one of the [99mTc]Tc‐tricarbonyl–peptide conjugates showed good stability in blood and both renal and hepatobiliary excretion. Biodistribution was performed on healthy rats to gain initial insight into the potential relevance of the 99mTc‐labeled peptides for in vivo imaging. 相似文献
965.
Jadran Vrabec Martin Bernreuther Hans‐Joachim Bungartz Wei‐Lin Chen Wilfried Cordes Robin Fingerhut Colin W. Glass Jürgen Gmehling René Hamburger Manfred Heilig Matthias Heinen Martin T. Horsch Chieh‐Ming Hsieh Marco Hülsmann Philip Jäger Peter Klein Sandra Knauer Thorsten Köddermann Andreas Köster Kai Langenbach Shiang‐Tai Lin Philipp Neumann Jürgen Rarey Dirk Reith Gábor Rutkai Michael Schappals Martin Schenk Andre Schedemann Mandes Schönherr Steffen Seckler Simon Stephan Katrin Stöbener Nikola Tchipev Amer Wafai Stephan Werth Hans Hasse 《化学,工程师,技术》2018,90(3):295-306
This article outlines advances in molecular modeling and simulation using massively parallel high‐performance computers (HPC). In the SkaSim project, partners from the HPC community collaborated with users from science and industry. The aim was to optimize the prediction of thermodynamic property data in terms of efficiency, quality and reliability using HPC methods. In this context, various topics were dealt with: atomistic simulation of homogeneous gas bubble formation, surface tension of classical fluids and ionic liquids, multicriteria optimization of molecular models, the development of the molecular simulation codes ls1 mardyn and ms2, atomistic simulation of gas separation processes, molecular membrane structure generators, transport resistors and the evaluation of predictive property data models based on specific mixture types. 相似文献
966.
The image analysis of dispersion layers in a vertical separator is realized by the development of an evaluation routine. The influence on the dispersion layer of the volume flow and hydrostatic pressure is investigated in continuous and discontinuous settling tests. A direct relationship between volumetric flow / hydrostatic pressure and dispersion layer height can be determined. The observed properties of the dispersion layer are explained by the underlying coalescence and sedimentation processes. The modeling of the dispersion layer is analyzed using four different model approaches. 相似文献
967.
Olga S. Kostromytska Cesar Rodriguez-Saona Hans T. Alborn Albrecht M. Koppenhöfer 《Journal of chemical ecology》2018,44(6):580-590
The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short cut turfgrass. Annual bluegrass, Poa annua L., is the most preferred and suitable host for ABW oviposition, larval survival and development. We investigated the involvement of grass volatiles in ABW host plant preference under laboratory and field conditions. First, ovipositional and feeding preferences of ABW adults were studied in a sensory deprivation experiment. Clear evidence of involvement of olfaction in host recognition by ABW was demonstrated. Poa annua was preferred for oviposition over three bentgrasses, Agrostis spp., but weevils with blocked antennae did not exhibit significant preferences. ABW behavioral responses to volatiles emitted by Agrostis spp. and P. annua were examined in Y-tube olfactometer assays. Poa annua was attractive to ABW females and preferred to Agrostis spp. cultivars in Y-tube assays. Headspace volatiles emitted by P. annua and four cultivars of Agrostis stolonifera L. and two each of A. capillaris L. and A. canina L. were extracted, identified and compared. No P. annua specific volatiles were found, but Agrostis spp. tended to have larger quantities of terpenoids than P. annua. (Z)-3-hexenyl acetate, phenyl ethyl alcohol and their combination were the most attractive compounds to ABW females in laboratory Y-tube assays. The combination of these compounds as a trap bait in field experiments attracted adults during the spring migration, but was ineffective once the adults were on the short-mown turfgrass. Hence, their usefulness for monitoring weevil populations needs further investigation. 相似文献
968.
969.
The aim of the study is to investigate if more competition leads to lower rents on the housing market. Data about the rent level for similar apartments in 30 cities in Sweden were available. Three hypotheses were formulated: (H1) Increased ‘internal’ competition, measured by the market share of the municipal housing company (that dominates the market and is price-leader according to the Swedish system of rent regulation), leads to lower rents. (H2) Increased ‘external’ competition measured by the price level on the market for single-family owner occupied housing, leads to lower rents. (H3) Lower capital expenditure in the municipal housing company leads to lower rents. The statistical analysis showed a strong correlation between the rent level and the level of external competition, but no relation was found for the level of internal competition and the level of capital expenditure. A possible conclusion is that policies that make it easier for households to leave the rental market are important for increasing the pressure on the firms in the rental sector and reducing rents. 相似文献
970.
Chlorination of drinking water in the presence of bromide and dissolved organic carbon (DOC) leads to the formation of brominated and chlorinated disinfection by-products (DBP). The concentration of bromide ions in the raw water is a significant factor in the speciation of DBP formed, and causes shifts in trihalomethane (THM) formation from chlorinated to brominated species. Drinking water treatment techniques that remove organic contaminants without affecting bromide ion concentrations cause increases in the brominated THM. For the present study, three water supplies containing different DOC and ambient bromide concentrations were filtered through biologically assisted granular activated carbon (BGAC). Similar to adsorption and coagulation treatment, this treatment does not remove bromide from drinking water; also, THMFP (trihalomethane formation potential) analysis indicated that the chlorinated effluent contained higher concentrations of brominated THM in comparison to the influent. Although BGAC may increase the brominated THM, which may be more toxic than the chlorinated THM, the overall reduction of THMFP by DOC removal far exceeds this negative change, thereby producing a much less toxic finished drinking water. This work is part of a study to make high DOC surface waters on the Canadian prairie safe and palatable for small volume users (individuals or small communities). 相似文献