Substrate binding to vanadate-dependent bromoperoxidase from Ascophyllum nodosum: a vanadium K-edge XAS approach

The EXAFS region of vanadium K-edge XAS spectra of native vanadate-dependent bromoperoxidase (isoenzyme I) from Ascophyllum nodosum in the presence of the substrate bromide can be fitted to three shells (at 1.62, 1.73-1.78 and 1.99-2.07 A) in the first coordination sphere of vanadium plus two more distant shells at 4.1A, possibly corresponding to bromide, and 4.7 A due to light scatterers stemming from the protein pocket. Bromide does not directly bind to the vanadium centre. The XANES and the EXAFS features for the enzyme are essentially reproduced by model complexes of the general composition [VO(H(2)O)(n)(ONO)] (n= 1 or 2) where ONO is the dianion of a Schiff base from bromosalicylaldehydes (Brsal; with the Br substituent in the position 3, 4, 5 or 6) and amino acids. The 3-Brsal derivatives exhibit an outer sphere shell at 3.8 A, which is traced back to intermolecular contacts. The data obtained from EXAFS are compared to those obtained from single crystal X-ray diffraction of [VO(H(2)O)(2)(4-Brsal-gly)] and [VO(H(2)O)(2)(6-Brsal-gly)] (gly = glycinate). In the complex [VOBr(2)(ONO)']] ((ONO)' is the Schiff base from o-anisole and o-hydroxyaniline), the V-Br distance is 2.44 A.     

Bioactivation of self-immolative dendritic prodrugs by catalytic antibody 38C2

Self-immolative dendrimers have recently been developed and introduced as a potential platform for a multi-prodrug. These unique structural dendrimers can release all of their tail units, through a self-immolative chain fragmentation, which is initiated by a single cleavage at the dendrimer's core. Incorporation of drug molecules as the tail units and an enzyme substrate as the trigger can generate a multi-prodrug unit that will be activated with a single enzymatic cleavage. We have synthesized the first generation of dendritic prodrugs with doxorubicin and camptothecin as tail units and a retro-aldol retro-Michael focal trigger, which can be cleaved by catalytic antibody 38C2. The bioactivation of the dendritic prodrugs was evaluated in cell-growth inhibition assay with the Molt-3 leukemia cell line in the presence and the absence of antibody 38C2. The dendritic unit was applied as a platform for a heterodimeric prodrug, which achieved a remarkable increase in toxicity with its bioactivation.     

Detecting exposure-dependent changes of liver detoxification capacity of children in a formerly high-loaded industrial area with a 15N-labelled diagnostic tool

The effect of remediation activities in formerly high-loaded industrial areas is measured conventionally in terms of decreased toxic loads. It is more difficult to investigate the effect on physiological parameters of the people living there. A non-invasive 15N-based organ function test, the [15N]methacetin urine test, was adapted to environmental-medical purposes for the detection of small deviations from the norm-range hepatic detoxification capacity. Parallel to the measurement of region-specific external exposure (especially by tri- and tetrachloro ethylene) and of the corresponding internal load (urinary trichloro acetic acid), the hepatic mono oxygenation capacity was determined. The study cohort consisted of 35 kindergarten children who were 3.3+/-0.5 years old at the beginning of the study. They lived in a high-polluted industrial village (n = 23) and in a non-polluted control village close by (n = 12), respectively. Children living in the polluted area were exposed a 2.3 times higher load and showed detoxification capacities 6% lower than the children in the control area. Improvement of the environmental situation led to exposure data decreasing by 70%, which is also reflected in smaller internal load and improved liver detoxification of the children in the polluted area. Then, between loaded and control area the mean hepatic detoxification was no longer significantly different (0.6%). The study shows that chronic low-dose exposure can disturb a hepatic function and that thereafter the capacity is recovered quickly in children. The stable isotope-based test is suited to characterize a health effect of multicomponent exposure by bioeffect monitoring within the framework of screening and testing the remediation efficiency.     

Quantitative high-resolution on-line NMR spectroscopy in reaction and process monitoring

On-line nuclear magnetic resonance spectroscopy (on-line NMR) is a powerful technique for reaction and process monitoring. Different set-ups for direct coupling of reaction and separation equipment with on-line NMR spectroscopy are described. NMR spectroscopy can be used to obtain both qualitative and quantitative information from complex reacting multicomponent mixtures for equilibrium or reaction kinetic studies. Commercial NMR probes can be used at pressures up to 35 MPa and temperatures up to 400 K. Applications are presented for studies of equilibria and kinetics of complex formaldehyde-containing mixtures as well as homogeneously and heterogeneously catalyzed esterification kinetics. Direct coupling of a thin-film evaporator is described as an example for the benefits of on-line NMR spectroscopy in process monitoring.     

Chemistry in microstructured reactors

The application of microstructured reactors in the chemical process industry has gained significant importance in recent years. Companies that offer not only microstructured reactors, but also entire chemical process plants and services relating to them, are already in existence. In addition, many institutes and universities are active within this field, and process-engineering-oriented reviews and a specialized book are available. Microstructured systems can be applied with particular success in the investigation of highly exothermic and fast reactions. Often the presence of temperature-induced side reactions can be significantly reduced through isothermal operations. Although microstructured reaction techniques have been shown to optimize many synthetic procedures, they have not yet received the attention they deserve in organic chemistry. For this reason, this Review aims to address this by providing an overview of the chemistry in microstructured reactors, grouped into liquid-phase, gas-phase, and gas-liquid reactions.     

Slopes of vector bundles on projective curves and applications to tight closure problems

We study different notions of slope of a vector bundle over a smooth projective curve with respect to ampleness and affineness in order to apply this to tight closure problems. This method gives new degree estimates from above and from below for the tight closure of a homogeneous -primary ideal in a two-dimensional normal standard-graded algebra in terms of the minimal and the maximal slope of the sheaf of relations for some ideal generators. If moreover this sheaf of relations is semistable, then both degree estimates coincide and we get a vanishing type theorem.

     

Absorption and scattering imaging of tissue with steady-state second-differential spectral-analysis tomography

A novel approach to reconstructing both the absorption and the scattering properties of a turbid medium simultaneously from steady-state broadband spectral measurements is presented that utilizes second-differential fitting to the water spectrum to estimate the optical path length in tissue. Theoretical and experimental evidence is provided to demonstrate the robust accuracy of the spectroscopy approach and reconstructed absorption images. The steady-state broadband CCD system has the potential to provide accurate chromophore imaging without the technological complexity of time- or frequency-domain systems.