Aerobic intramolecular oxidative amination of alkenes catalyzed by NHC-coordinated palladium complexes

[reaction: see text] Palladium(II) complexes bearing a single N-heterocyclic carbene ligand serve as effective catalysts for the aerobic oxidative cyclization of alkenes with pendant sulfonamides. The use of carboxylic acid cocatalysts (AcOH and PhCO(2)H) often leads to significant improvements in catalyst stability and product yield and enables catalytic turnover to be achieved with air, rather than pure oxygen gas, as the source of O(2).     

Fluorescence-quenching-based homogeneous caspase-3 activity assay using photon upconversion

Caspase proteases are key mediators in apoptosis and thus of great interest in pharmaceutical industry. Enzyme-activity assays are commonly employed in the screening of protease inhibitors that are potential drug candidates. Conventional homogeneous fluorescence-based assays are susceptible to autofluorescence originating from biological material. This background autofluorescence can be eliminated by using upconverting phosphors (UCPs) that emit visible light upon excitation at near-infrared. In the assay energy was transferred from a UCP-donor to a conventional fluorophore acceptor that resided at one end of a caspase-3-specific substrate peptide. Attached to the other end was a quencher molecule that was used to attenuate the acceptor emission through intramolecular energy transfer in an intact peptide. In non-inhibitory conditions the enzyme reaction separated the fluorophore from the quencher and the emission of the fluorophore was recovered. The method was applied for the detection and characterization of a known caspase-3 inhibitor Z-DEVD-FMK, and the assay gave IC₅₀ values of approximately 13 nM for this inhibitor. We have demonstrated the applicability of UCPs on a fluorescence-quenching-based homogeneous enzyme-activity assay for the detection of caspase-3 inhibitors. The use of near-infrared excitable UCPs enables inexpensive instrumentation and total elimination of autofluorescence, while the use of an internally quenched substrate molecule diminishes the background resulting from radiatively excited acceptor molecules. The reduction of autofluorescence and radiative background result in high signal-to-background ratios (ratios of approximately 100 were obtained). By further utilizing assay miniaturization and signal enhancement in a white microtitration plate, a significant reduction in the reagent consumption can be achieved rendering the assay applicable for high-throughput screening.     

The role of stimulus cross-splicing in an event-related potentials study. Misleading formant transitions hinder automatic phonological processing

The mental organization of linguistic knowledge and its involvement in speech processing can be investigated using the mismatch negativity (MMN) component of the auditory event-related potential. A contradiction arises, however, between the technical need for strict control of acoustic stimulus properties and the quest for naturalness and acoustic variability of the stimuli. Here, two methods of preparing speech stimulus material were compared. Focussing on the automatic processing of a phonotactic restriction in German, two corresponding sets of various vowel-fricative syllables were used as stimuli. The former syllables were naturally spoken while the latter ones were created by means of cross-splicing. Phonetically, natural and spliced syllables differed with respect to the appropriateness of coarticulatory information about the forthcoming fricative within the vowels. Spliced syllables containing clearly misleading phonetic information were found to elicit larger N2 responses compared to their natural counterparts. Furthermore, MMN results found for the natural syllables could not be replicated with these spliced stimuli. These findings indicate that the automatic processing of the stimuli was considerably affected by the stimulus preparation method. Thus, in spite of its unquestioned benefits for MMN experiments, the splicing technique may lead to interference effects on the linguistic factors under investigation.     

Z-spectroscopy with Alternating-Phase Irradiation

Magnetization transfer (MT) MRI and Z-spectroscopy are tools to study both water-macromolecule interactions and pH-sensitive exchange dynamics between water and the protons of mobile chemical groups within these macromolecules. Both rely on saturation of frequencies offset from water and observation of the on-resonance water signal. In this work, an RF saturation method called Z-spectroscopy with Alternating-Phase Irradiation (ZAPI) is introduced. Based on the T(2)-selectivity of the irradiation pulse, ZAPI can be used to separate the different contributions to a Z-spectrum, as well as to study the T(2) distribution of the macromolecules contributing to the MT signal. ZAPI can be run at resonance for water and with low power, thus minimizing problems with specific absorption rate (SAR) limits in clinical applications. In this paper, physical and practical aspects of ZAPI are discussed and the sequence is applied in vitro to sample systems and in vivo to rat head to demonstrate the method.     

RELATIONSHIP BETWEEN LIGHT-INDUCED STATE I/II-TRANSITIONS and Mg2+-EFFECT ON FLUORESCENCE, TESTED IN SYNCHRONOUS CULTURES OF Scenedesmus obliquus

Abstract— Light induced transition of the photosynthetic apparatus from state II to state I by preillumination with PS I light in vivo and the addition of cations to chloroplasts or chloroplast particles prepared in low-salt buffer have rather similar effects on the fluorescence properties at room temperature and at 77 K. Using synchronous cultures of Scenedesmus obliquus, we were able to study both phenomena in parallel in different developmental stages under well defined conditions. Both phenomena show an inverse course during ontogenetic development of S. obliquus. This result suggests that Mg²⁺ ions and PS I light don't necessarily act in the same way. Hence, changes in the ionic environment cannot be the unique basis of light induced state changes within the photosynthetic apparatus in vivo.     

Antagonistic blue- and red-light regulation of cab-gene expression during photosynthetic adaptation in Scenedesmus obliquus.

During adaptation of the photosynthetic apparatus of the green alga Scenedesmus obliquus to various light qualities, the accumulation of chlorophylls and pigment-protein complexes (with specific consideration of chlorophyll a/b-binding (Cab) proteins) and cab-gene expression were determined. The fluence rate dependences for chlorophyll accumulation and cab-gene expression were very different. Very low fluence rates of violet (404 nm), blue (461 nm) and red (650 nm) light below the photosynthetic threshold, i.e. between 10(-3) and 10(-1) mumol m-2 s-1, inhibited all of these reactions in cells grown under heterotrophic conditions. At elevated fluence rates (above 1 mumol m-2 s-1), red light retained its negative regulation, whereas blue light stimulated pigment accumulation. Under autotrophic conditions the pattern was more complex, because chlorophyll accumulation was unaffected by light below the photosynthetic threshold. However, the expression of cab-genes was inhibited by red light but stimulated by blue light. Cells adapted to fluence rates, which ensured photosynthetic energy supply (above 1 mumol m-2 s-1), showed an increase in chlorophyll accumulation, blue light being more effective than red light. The results confirm and extend our previous discovery of two antagonistically acting photoreceptors in Scenedesmus which mediate and coordinate the complex functional and structural changes associated with photosynthetic adaptation. One of these receptor pigments is a blue-light receptor with positive action; the other is a violet-red-light receptor which can operate far below the photosynthetic threshold and exerts a negative regulation.