Detection of DNA-bound advanced glycation end-products by immunoaffinity chromatography coupled to HPLC-diode array detection

Sugars and sugar degradation products are formed during food processing, but also endogenously in vivo. In vitro, nucleosides and DNA react readily with these carbonyl compounds during the formation of the two diastereomers of N(2)-carboxyethyl-2'-deoxyguanosine (CEdG(A,B)), leading to a loss of DNA integrity. Only little is known about DNA glycation in vivo and about the influence of nutrition on CEdG formation. In this study, we developed a sensitive method to analyze DNA glycation by HPLC. For this purpose, immunoaffinity chromatography (IAC) using a polyclonal antibody against N(2)-carboxyethylguanine (CEguanine) was coupled to HPLC-DAD. In some samples, peak identity was confirmed by LC-MS/MS. The recovery of CEguanine from the IAC columns was 52.5% +/- 3.6 (n = 4). Thus, it was possible for the first time to detect CEdG(A,B), N(2)-carboxyethylguanosine (CEG(A,B)), and CEguanine in 11 human urine samples. However, due to imprecision of IAC, valid quantification of the adducts could not be achieved. Furthermore, CEdG was also detected in the DNA of cultured human smooth muscle cells (SMCs) and bovine aorta endothelium cells (BAECs). In BAECs, CEdG(A,B) were found by HPLC-DAD and LC-MS/MS after immunoaffinity purification, whereas in SMCs DNA-advanced glycation end-products were only detected with the more sensitive LC-MS/MS method.     

Influence of the Isolation Method on the Technofunctional Properties of Protein Isolates from Lupinus angustifolius L

The technofunctional properties of 2 protein isolates from Lupinus angustifolius L. Vitabor isolated by different procedures were investigated. The lupin protein isolate prepared by aqueous alkaline extraction with subsequent isoelectric precipitation (ILP) showed a significantly higher degree of protein denaturation and lower denaturation temperatures than the one obtained by aqueous salt‐induced extraction followed by dilutive precipitation (MLP) as determined by differential scanning calorimetry. Rheological investigations revealed higher firmness and a viscoelastic solid‐like behavior of ILP, in contrast to MLP that showed viscoelastic, liquid‐like properties. Protein solubility of MLP was higher compared to ILP and solubility minima were slightly different for both lupin protein isolates. The protein isolates exhibited different technofunctional properties with ILP showing higher water binding capacity, lower oil binding capacity and lower emulsifying capacity than MLP. This reflects the different putative application of both lupin protein isolates as food ingredients, for example for ILP as a moisture enhancer and for MLP as a “natural” emulsifier in mixed food systems.     

Development and performance of a parsimonious model to estimate temperature in sewer networks

This paper presents a model (inspired by another model) to calculate water temperature in free-surface flow with two main innovations: the convective heat transfer occurs only at the wetted perimeter of pipes, and the model was integrated to commercial software used for hydraulic calculations in drainage systems. Given these innovations, we could reduce the number of modeling input data to calculate the temperature of water and soil in the radial and tangential directions along the pipes, with the advantages of using industry-standard software. To test the performance of the model, it was firstly calibrated in two sets of experiments (to calibrate the hydraulic and the thermal parameters separately), and benchmarked with a third controlled discharge against the case model. The results indicate that in unsteady-state situations the parsimonious model can be twice as accurate as the underlying model because the parsimonious model considers the hydraulic influence of sewer infrastructure.     

Serial 1H-MRS in relapsing-remitting multiple sclerosis: effects of interferon-beta therapy on absolute metabolite concentrations

To assess the applicability of magnetic resonance spectroscopy (MRS) for long-term follow-up of neurological diseases a longitudinal 1H-MRS study at 3 T was carried out on ten patients having relapsing-remitting multiple sclerosis (MS) who, after baseline examination, received interferon-β (IFN) lb. At 8–20 examinations within up to 34 months absolute concentrations ofN-acetylaspartate (NAA), total creatine (tG), and choline-containing compounds (tCho) were determined in a large non-enhancing lesion and contralateral normal appearing white matter (NAWM). MR spectra were analyzed using a novel time domain-frequency domain method including non-parametric background characterization. For comparison at baseline, ten healthy controls were examined. The concentrations of tCho and tCr were found to be higher in MS brain than in control brain. Besides a non-significantly lower NAA concentration in lesions there were no concentration differences between lesions and NAWM. Over the follow-up period the measured metabolite concentrations exhibited a high variability. Most concentrations remained within this scatter, and statistical tests revealed significant fluctuations in the levels of metabolites in one case only. This stability of the metabolite concentrations over time might result from IFN therapy as for the spontaneous course of relapsing-remitting MS decreasing metabolite (NAA/tCr) ratios have been reported. The results further suggest that future treatment trials intending to use metabolite concentrations as a secondary outcome indicator use even longer observation periods and, besides group analysis of large cohorts, investigate the time behavior of selected single cases. The biochemical abnormalities found in NAWM emphasize the importance of analyzing both lesion and NAWM.     

Parameter estimation of human nerve C-fibers using matched filtering and multiple hypothesis tracking

We describe how multiple-target tracking may be used to estimate conduction velocity changes and recovery constants of human nerve C-fibers. These parameters discriminate different types of C-fibers and pursuing this may promote new insights into differential properties of nerve fiber membranes. Action potentials (APs) were recorded from C-fibers in the peroneal nerve of awake human subjects. The APs were detected by a matched filter constituting a maximum-likelihood constant false-alarm rate detector. Using the multiple-hypothesis tracking method and Kalman filtering, the detected APs (targets) in each trace (scan) were associated to individual nerve fibers (tracks) by their typical conduction latencies in response to electrical stimulation. The measurements were one-dimensional (range only) and the APs were spaced in time with intersecting trajectories. In general, the AP amplitude of each C-fiber differed for different fibers. Amplitude estimation was therefore incorporated into the tracking algorithm to improve the performance. The target trajectory was modeled as an exponential decay with three unknowns. These parameters were estimated iteratively by applying the simplex method on the parameters that enter nonlinearly and the least squares method on the parameters that enter linearly.     

Management decision support using long-term market simulation

In recent years Agent-based Computational Economics (ACE) has become an increasingly important method in market simulation. After liberalization of many former governmental owned or controlled industries the used operations research models are not longer sufficient to simulate market behavior due to individual action and increasing competition. Agent-based simulation appears to be an alternative approach considering also individual behavior and competition. Some short-term simulation approaches have shown promising results for the simulation in the domain of electricity markets. Picking up the desire for a long-term oriented simulation, this paper presents a basic agent-based model considering the investment decision within long-term planning of electricity markets. Additionally, regulatory agents are introduced as a third side in the market simulation to represent governmental decisions. This results in the definition of three types of agents representing electricity generating companies, consumers and governmental instances.     

Interfacial Modification of Polystyrene‐block‐polybutadiene‐block‐polystyrene/Magnesium Hydroxide Composites, 1

Composites based on polystyrene‐block‐polybutadiene‐block‐polystyrene (SBS triblock thermoplastic elastomer) and magnesium hydroxide (Mg(OH)₂) (5–60 wt.‐%) have been prepared by twin screw extrusion. Interfacial modifiers included dispersants, i.e., isostearic acid, oleic acid, stearic acid; and coupling agents, i.e., maleanised polybutadiene and vinyltriethoxysilane. In each case, approximately one monolayer of treatment was used. A dual bore motor driven extrusion rheometer was used for assessment shear and elongation flow behavior (Cogswell's method) over a shear rate range of 100 s^?1 to 5 000 s^?1. Untreated filler and filler treated with coupling agents gave composites that become increasingly pseudoplastic as filler level increased. Fatty acid structure was shown to have some influence over the level of melt viscosity reduction normally associated with such treatments; stearic acid gave the most pronounced reduction in melt viscosity possibly due to the tightly packed monolayer. Elongational flow properties, determined using Cogswell's method, indicated significant chain extension/branching of the bulk matrix when high levels of untreated filler were present and long range filler‐matrix interaction in composites modified with maleanised polybutadiene.

Elongational viscosity versus extensional stress (obtained by Cogswell's method) for SBS blended with filler surface treatments (□) unfilled matrix, and unfilled matrix plus (?) Hⁱst and (?) MPBD.