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In this study, we investigated galacturonic (GalAc)- and mannuronic (ManAc) acids as novel targeting ligands for receptor-mediated gene delivery. GalAc and ManAc were coupled to either polyethyleneimine (PEI) or PEIpolyethyleneglycol (PEG). Furthermore, lactobionic acid (LacAc), which comprises a GalAc-related carbohydrate ring, was coupled to each of the polymers through its open-chain gluconic acid moiety. The molar mass distributions of the polymers were characterized by analytical ultracentrifugation and size exclusion chromatography. PEI-conjugatepDNA complexes were transfected into HepG2-, HeLa-, and 16HBE14o--cells. Gene expression mediated by GalAc- and LacAc-functionalized PEI-conjugates was lower than for PEI. In contrast, gene expression mediated by ManAc-functionalized PEI-conjugates was up to three orders of magnitude higher than for the other tested PEI-conjugates, in particular for negatively charged gene vectors at low N/P ratios, independent of the cell line. Pre-incubation of cells with an excess of ManAc before transfection significantly inhibited transfection rates only for ManAc-functionalized PEI-conjugates. Coupling of methyl-α-D-mannuronic acid to PEI resulted in significantly lower transfection rates than for ManAc-PEI based complexes. Together with fluorescence microscopy images of fluorescein-labelled ManAc-functionalized dextrans and FACS analyses of cells, these results demonstrate that receptor-mediated endocytosis of ManAcPEI-conjugatepDNA complexes via ManAc-specific receptors was involved in gene transfer. In conclusion, ManAc-modification of PEI-polymers represents a novel strategy for receptor-mediated gene delivery which could be promising for in vivo application.
The objective of this paper is to demonstrate a new simulation technique which allows fast and automatic generation of temperature fields as input for subsequent thermomechanical welding simulation. The basic idea is to decompose the process model into an empirical part based on neural networks and a phenomenological part that describes the physical phenomena. The strength of this composite modelling approach is the automatic calibration of mathematical models against experimental data without the need for manual interference by an experienced user. As an example for typical applications in laser beam and GMA-laser hybrid welding, it is shown that even 3D heat conduction models of a low complexity can approximate measured temperature fields with a sufficient accuracy. In general, any derivation of model fitting parameters from the real process adds uncertainties to the simulation independent of the complexity of the underlying phenomenological model. The modelling technique presented hybridises empirical and phenomenological models. It reduces the model uncertainties by exploiting additional information which keeps normally hidden in the data measured when the model calibration is performed against few experimental data sets. In contrast, here the optimal model parameter set corresponding to a given process parameter is computed by means of an empirical submodel based on relatively large set of experimental data. The approach allows making a contribution to an efficient compensation of modelling inaccuracies and lack of knowledge about thermophysical material properties or boundary conditions. Two illustrating examples are provided.
The objective of this paper is to demonstrate a new simulation technique which allows the fast and automatic generation to temperature fields based on a combination of empirical and phenomenological modelling techniques. The automatic calibration of the phenomenological model is performed by a multi-variable global optimisation routine which yields the optimal fit between simulated and experimental weld charcteristics without the need for initial model parameters. For exemplary welding processes it is shown that linear 3D heat conduction models can approximate measured temperature fields with a high accuracy. The modelling approach presented comprises the automatic calibration against multiple experiments which permits simulating the temperature field for unknown process parameters. The validation of this composite simulation model is performed for exemplary welding processes and includes the prediction of the fusion line in the cross section and the corresponding thermal cycles.
Fast temperature field generation for welding simulation and reduction of experimental effort
(2009)
The quality of welding processes is governed by the occurring induced distortions yielding an
increase in production costs due to necessary reworking. Especially for more complex specimens
it is difficult to evaluate the optimal configuration of welding sequences in order to
minimise the distortion. Even experienced welding operators can solve this task only by trial
and error which is time and cost consuming.
In modern engineering the application of welding simulation is already known to be able to
analyse the heat effects of welding virtually. However, the welding process is governed by
complex physical interactions. Thus, recent weld thermal models are based on many simplifications.
The state of the art is to apply numerical methods in order to solve the transient heat
conduction equation. Therefore, it is not possible to use the real process parameters as input
for the mathematical model. The model parameters which allow calculating a temperature
field that is in best agreement with the experiments cannot be defined directly but inversely by
multiple simulations runs. In case of numerical simulation software based on finite discretisation
schemes this approach is very time consuming and requires expert users. The weld
thermal model contains an initial weakness which has to be adapted by finding an optimal set
of model parameters. This process of calibration is often done against few experiments. The
range of model validity is limited. An extension can be obtained by performing a calibration
against multiple experiments.
The focus of the paper is to show a combined modelling technique which provides an efficient
solution of the inverse heat conduction problem mentioned above. On the one hand the inverse
problem is solved by application of fast weld thermal models which are closed form
solutions of the heat conduction equation. In addition, a global optimisation algorithm allows
an automated calibration of the weld thermal model. This technique is able to provide a temperature
field automatically that fits the experimental one with high accuracy within minutes
on ordinary office computers. This fast paradigm permits confirming the application of welding
simulation in an industrial environment as automotive industry.
On the other hand, the initial model weakness is compensated by calibrating the model
against multiple experiments. The unknown relationship between model and process parameters
is approximated by a neural network. The validity of the model is increased successively
and enables to decrease experimental effort. For a test case it is shown, that this approach
yields accurate temperature fields within very short amount of time for unknown process parameters
as input data to the model contributing to the requirement to construct a substitute
system of the real welding process.
The objects of this investigation were sculptures taken from a ca. three hundred years old baroque epitaph of a church in Tonning, a town in Northern Germany. Around 1900 it was found in a disastrous state heavily damaged by wood-worm. At that time, the whole artwork was treated with the tar extract carbolineum as a remedy. Nowadays, this substance has been identified as carcinogenic, and its presence can be perceived by its stench and visually at certain spots on the surface where it has penetrated the covering paint.
A gold-painted sculpture of a massive wooden skull was interrogated with X-rays and fast neutrons to investigate the internal distribution of the carbolineum. The X-ray tomography, with its excellent spatial resolution revealed galleries left over from the worm infestation in the outer areas and cracks in the central region. The golden color coating appeared as a thick and dense layer. In comparison the tomography with fast neutrons, though being of lower resolution and yet unresolved artefacts revealed sections of slightly different densities in the bulk of the wood. These differences we attribute to the differences in the distribution of the impregnant in the wood, visible due to its higher hydrogen content making it less transparent for neutrons.
A European round robin test according to ISO 5725-2 was conceptually prepared, realised, and evaluated. The aim was to determine the inter-laboratory variability of the overall process for the ecotoxicological characterization of construction products in eluates and bioassays. To this end, two construction products BAM-G1 (granulate) and HSR-2 (roof sealing sheet), both made of EPDM polymers (rubber), were selected. The granular construction product was eluted in a one stage batch test, the planar product in the Dynamic Surface Leaching test (DSLT). A total of 17 laboratories from 5 countries participated in the round robin test: Germany (12), Austria (2), Belgium (1), Czech Republic (1) and France (1). A test battery of four standardised ecotoxicity tests with algae, daphnia, luminescent bacteria and zebrafish eggs was used. As toxicity measures, EC50 and LID values were calculated. All tests, except the fish egg test, were basically able to demonstrate toxic effects and the level of toxicity. The reproducibility of test results depended on the test specimens and the test organisms. Generally, the variability of the EC50 or LID values increased with the overall level of toxicity. For the very toxic BAM-G1 eluate a relative high variability of CV ¼ 73%e110% was observed for EC50 in all biotests, while for the less toxic HSR-2 eluate the reproducibility of EC50 varied with sensitivity: it was very good (CV ¼ 9.3%) for the daphnia test with the lowest sensitivity, followed by the algae test (CV ¼ 36.4%). The luminescent bacteria test, being the most sensitive bioassay for HSR-2 Eluate, showed the highest variability (CV ¼ 74.8%). When considering the complex overall process the reproducibility of bioassays with eluates from construction products was acceptable.