582 research outputs found

    The photoelectron spectra of the diazanaphthalenes

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    The high-resolution He 584 Å photoelectron spectra of ten diazanaphthalenes are presented. The ordering of the π orbitals and the nitrogen “lone-pair” orbitals is discussed. Several semi-empirical quantum-chemical calculation methods have been screened against the experimental evidence

    Perfluoro effect in the photoelectron spectra of quinoline and isoquinoline

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    The high-resolution He 584Aophotoelectron spectra of heptafluoroquinoline and heptafluoroisoquinoline are compared with those of the parent compounds. Shifts in π ionisation potentials, due to the fluorine substitution, can be described with an inductive and a combined inductive-conjugative Hu¨ckel model

    Photoelectron spectra of fluorine substituted diazanaphthalenes: “Even cases”

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    The high resolution He 584 Ă… photoelectron spectra of three diazanaphthalenes and some of their fluorine derivatives are presented. The qualitative model that is used frequently in the discussion of lone-pair level splittings is examined

    Photoelectron spectra of some fluorine substituted diazanaphthalenes

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    The high resolution He 584 Å photoelectron spectra of fluorine substituted 1,2-diaza-, 1,3-diaza-, 1,4-diaza- and 2,3-diaza-naphthalene are presented. By means of fluorine substitution the analysis of the photoelectron spectra of the parent compounds can be made more definite. Unexpected shifts of the nitrogen “lone-pair” bands can be explained within the through-space and through-bond interaction model. From this explanation one can deduce that fluorine substitution can give experimental evidence about the symmetry character of the “lone-pair” molecular orbitals

    Photoelectron spectra of fluorine-substituted diazabenzenes

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    The high resolution He 584 Å photoelectron spectra of ten fluorine-substituted diazabenzenes are presented. By means of fluorine substitution the analysis of the photoelectron spectra of the parent compounds can be made more definite. “Lone-pair” ionization potentials are shown to correlate well with data calculated by a modified and iterative version of the extended Hückel method

    The Plant Propagation Algorithm on Timetables: First Results

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    One Stochastic HillClimber and two implementations of the Plant Propagation Algorithm (PPA-1 and PPA-2) are applied to an instance of the University Course Timetabling Problem from the Univer- sity of Amsterdam. After completing 10 runs of 200,000 objective function evaluations each, results show that PPA-1 outperforms the HillClimber, but PPA-2 makes the best timetables

    Flow focusing through gels as a tool to generate 3D concentration profiles in hydrogel-filled microfluidic chips

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    Laminar flow patterning is an iconic microfluidic technology used to deliver chemicals to specific regions on a two-dimensional surface with high spatial fidelity. Here we present a novel extension of this technology using Darcy flow within a three-dimensional (3D) hydrogel. Our test device is a simple 3-inlet microfluidic channel, totally filled with collagen, a cured biological hydrogel, where the concentration profiles of solutes are manipulated via the inlet pressures. This method allows solutes to be delivered with 50 micron accuracy within the gel, as we evidence by controlling concentration profiles of 40 kDa and 1 kDa fluorescent polysaccharide dyes. Furthermore, we design and test a 3D-printed version of our device with an extra two inlets for control of the vertical position of the concentration profile, demonstrating that this method is easily extensible to control of the concentration profile in 3D

    Unscheduled DNA synthesis at sites of local UV-induced DNA damage to quantify global genome nucleotide excision repair activity in human cells

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    Nucleotide excision repair (NER) removes a wide variety of structurally unrelated lesions from the genome, including UV-induced photolesions such as 6-4 pyrimidine-pyrimidone photoproducts (6-4PPs) and cyclobutane pyrimidine dimers (CPDs). NER removes lesions by excising a short stretch of single-stranded DNA containing the damaged DNA, leaving a single-stranded gap that is resynthesized in a process called unscheduled DNA synthesis (UDS). Measuring UDS after UV irradiation in non-dividing cells provides a measure of the overall NER activity, of which approximately 90% is carried out by the global genome repair (GGR) sub pathway. Here, we present a protocol for the microscopy-based analysis and quantification of UDS as a measurement for GGR activity. Following local UV-C irradiation, serum-starved human cells are supplemented with the thymidine analogue 5-ethynyl-2'-deoxyuridine (EdU), which is incorporated into repair patches following NER-dependent dual incision. The incorporated nucleotide analogue is coupled to a fluorophore using Click-iT chemistry, followed by immunodetection of CPD photolesions to simultaneously visualize both signals by fluorescence microscopy. Accompanying this protocol is a custom-built ImageJ plug-in to analyze and quantify UDS signals at sites of CPDmarked local damage. The local UDS assay enables an effective and sensitive fluorescence-based quantification of GGR activity in single cells with application in basic research to better understand the regulatory mechanism in NER, as well as in diagnostics to characterize fibroblasts from individuals with NER-deficiency disorder.Genome Instability and Cance
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