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Mouse nongenotoxic hepatocarcinogens phenobarbital (PB) and chlordane induce hepatomegaly characterized by hypertrophy and hyperplasia. Increased cell proliferation is implicated in the mechanism of tumor induction. The relevance of these tumors to human health is unclear. The xenoreceptors, constitutive androstane receptors (CARs), and pregnane X receptor (PXR) play key roles in these processes. Novel “humanized” and knockout models for both receptors were developed to investigate potential species differences in hepatomegaly. The effects of PB (80 mg/kg/4 days) and chlordane (10 mg/kg/4 days) were investigated in double humanized PXR and CAR (huPXR/huCAR), double knockout PXR and CAR (PXRKO/CARKO), and wild-type (WT) C57BL/6J mice. In WT mice, both compounds caused increased liver weight, hepatocellular hypertrophy, and cell proliferation. Both compounds caused alterations to a number of cell cycle genes consistent with induction of cell proliferation in WT mice. However, these gene expression changes did not occur in PXRKO/CARKO or huPXR/huCAR mice. Liver hypertrophy without hyperplasia was demonstrated in the huPXR/huCAR animals in response to both compounds. Induction of the CAR and PXR target genes, Cyp2b10 and Cyp3a11, was observed in both WT and huPXR/huCAR mouse lines following treatment with PB or chlordane. In the PXRKO/CARKO mice, neither liver growth nor induction of Cyp2b10 and Cyp3a11 was seen following PB or chlordane treatment, indicating that these effects are CAR/PXR dependent. These data suggest that the human receptors are able to support the chemically induced hypertrophic responses but not the hyperplastic (cell proliferation) responses. At this time, we cannot be certain that hCAR and hPXR when expressed in the mouse can function exactly as the genes do when they are expressed in human cells. However, all parameters investigated to date suggest that much of their functionality is maintained.
Increasing Scots pine (Pinus sylvestris L.) mortality has been recently observed in the dry inner valleys of the European Alps. Besides drought, infection with pine mistletoe (Viscum album ssp. austriacum) seems to play an important role in the mortality dynamics of Scots pines, but how mistletoes promote pine decline remains unclear. To verify whether pine mistletoe infection weakens the host via crown degradation, as observed for dwarf mistletoes, we studied the negative effects of pine mistletoe infestation on the photosynthetic tissues and branch growth of pairs of infested and non-infested branches. Pine mistletoe infection leads to crown degradation in its host by reducing the length, the radial increment, the ramification, the needle length and the number of needle years of the infested branches. This massive loss in photosynthetic tissue results in a reduction in primary production and a subsequent decrease in carbohydrate availability. The significant reduction in needle length due to mistletoe infection is an indication for a lower water and nutrient availability in infested branches. Thus, mistletoe infection might lead to a decrease in the availability of water and carbohydrates, the two most important growth factors, which are already shortened due to the chronic drought situation in the area. Therefore, pine mistletoe increases the risk of drought-induced mortality of its host when growing in a xeric environment.
C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli
(2010)
Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477.
This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40 °C and pH 7–11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40 °C and pH = 9.5 resulted in KM = 0.23 ± 0.01 mM and kcat = 167.5 ± 3.6 s⁻¹. MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease.
Bead-on-plate butt joints of 2.5 mm hot rolled DP600/DP600 and 1.2 mm cold rolled TRIP700/TRIP700 steel sheets were performed using 6 kW CO2 laser beam welding. The welding speed ranged from 1.5 to 3.0 and from 2.1 to 3.9 m/min in DP/DP and TRIP/TRIP steel weldments respectively. A top surface helium gas was used as a shielding gas at a flow rate of 20 l/min. Metallographic examinations and transverse tensile testing (DIN EN 895: 1995) were carried out to characterize the weldments. The formability of base metals and weldments were investigated by standard Erichsen test (DIN EN ISO 20482). It was found that the uniaxial plastic behavior of both DP600 and TRIP700 base metals was in agreement with Swift and modified Mecking–Kocks models respectively. In a perpendicular tensile test to the weld line, all specimens were fractured at the base metal however the strengths were somewhat higher than those of base metal. There was a significant reduction in formability caused by welding of both DP/DP and TRIP/TRIP steel weldments and the formability has been improved with the increase of the welding speed.
Shielding gas influences on laser weldability of tailored blanks of advanced automotive steels
(2010)
The effects of shielding gas types and flow rates on CO2 laser weldability of DP600/TRIP700 steel sheets were studied in this work. The evaluated shielding gases were helium (He), argon (Ar) and different mixtures of He and Ar. Weld penetration, tensile strength and formability (Erichsen test) of laser welds were found to be strongly dependent upon the shielding gas types. The ability of shielding gas in removing plasma plume and thus increasing weld penetration is believed to be closely related to ionization potential and atomic weight which determine the period of plasma formation and disappearance. It was found that the higher helium shielding gas flow rate, the deeper weld penetration and the lower weld width.
Patients after coarctation repair still have an increased risk of cardiovascular or cerebrovascular events. This has been explained by the persisting hypertension and alterations in the peripheral vessels. However, involvement of the central vessels such as the retinal arteries is virtually unknown. A total of 34 patients after coarctation repair (22 men and 12 women; 23 to 58 years old, age range 0 to 32 years at surgical repair) and 34 nonhypertensive controls underwent structural and functional retinal vessel analysis. Using structural analysis, the vessel diameters were measured. Using functional analysis, the endothelium-dependent vessel dilation in response to flicker light stimulation was assessed. In the patients after coarctation repair, the retinal arteriolar diameter was significantly reduced compared to that of the controls (median 182 μm, first to third quartile 171 to 197; vs 197 μm, first to third quartile 193 to 206; p <0.001). These findings were independent of the peripheral blood pressure and age at intervention. No differences were found for venules. The functional analysis findings were not different between the patients and controls (maximum dilation 3.5%, first to third quartile 2.1% to 4.5% vs 3.6%, first to third quartile 2.2% to 4.3%; p = 0.81), indicating preserved autoregulative mechanisms. In conclusion, the retinal artery diameter is reduced in patients after coarctation repair, independent of their current blood pressure level and age at intervention. As a structural marker of chronic vessel damage associated with past, current, or future hypertension, retinal arteriolar narrowing has been linked to stroke incidence. These results indicate an involvement of cerebral microcirculation in aortic coarctation, despite timely repair, and might contribute to explain the increased rate of cerebrovascular events in such patients.
Numerical solution of the heat equation with non-linear, time derivative-dependent source term
(2010)
The mathematical modeling of heat conduction with adsorption effects in coated metal structures yields the heat equation with piecewise smooth coefficients and a new kind of source term. This term is special, because it is non-linear and furthermore depends on a time derivative. In our approach we reformulated this as a new problem for the usual heat equation, without source term but with a new non-linear coefficient. We gave an existence and uniqueness proof for the weak solution of the reformulated problem. To obtain a numerical solution, we developed a semi-implicit and a fully implicit finite volume method. We compared these two methods theoretically as well as numerically. Finally, as practical application, we simulated the heat conduction in coated aluminum fibers with adsorption in the zeolite coating. Copyright © 2010 John Wiley & Sons, Ltd.
Oxorhenium(V) complexes [ReOX3(PPh3)2] (X = Cl, Br) react with phenylacetylene under formation of complexes with ylide-type ligands. Compounds of the compositions [ReOCl3(PPh3){C(Ph)C(H)(PPh3)}] (1), [ReOBr3(OPPh3){C(Ph)C(H)(PPh3)}] (2), and [ReOBr3(OPPh3){C(H)C(Ph)(PPh3)}] (3) were isolated and characterized by X-ray diffraction. They contain a ligand, which was formed by a nucleophilic attack of released PPh3 at coordinated phenylacetylene. The structures of the products show that there is no preferable position for this attack. Cleavage of the Re–C bond in 3 and dimerization of the organic ligand resulted in the formation of the [{(PPh3)(H)CC(Ph)}2]2+ cation, which crystallized as its [(ReOBr4)(OReO3)]2– salt.
AgTcO4 reacts with R3ECl compounds (E = C, Si, Ge, Sn, Pb; R = Me, iPr, tBu, Ph), tBu2SnCl2, or PhMgCl under formation of novel trioxotechnetium(VII) derivatives. The carbon and silicon derivatives readily undergo decomposition, which was proven by 99Tc NMR spectroscopy and the isolation of decomposition products such as [TcOCl3(THF)(OH2)]. Compounds [Ph3GeOTcO3], [(THF)Ph3SnOTcO3], [(O3TcO)SntBu2(OH)]2, and [(THF)4Mg(OTcO3)2] are more stable and were isolated in crystalline form and characterized by X-ray diffraction.
As the field strength and, therefore, the operational frequency in MRI is increased, the wavelength approaches the size of the human head/body, resulting in wave effects, which cause signal decreases and dropouts. Several multichannel approaches have been proposed to try to tackle these problems, including RF shimming, where each element in an array is driven by its own amplifier and modulated with a certain (constant) amplitude and phase relative to the other elements, and Transmit SENSE, where spatially tailored RF pulses are used. In this article, a relatively inexpensive and easy to use imaging scheme for 7 Tesla imaging is proposed to mitigate signal voids due to B1 field inhomogeneity. Two time-interleaved images are acquired using a different excitation mode for each. By forming virtual receive elements, both images are reconstructed together using GRAPPA to achieve a more homogeneous image, with only small SNR and SAR penalty in head and body imaging at 7 Tesla.
The objectives of the present work are to characterize the Gas Metal Arc Welding process of DP 600 sheet steel and to summarize the modelling techniques. The time-temperature evolution during the welding cycle was measured experimentally and modelled with the softwaretool SimWeld. To model the phase transformations during the welding cycle dilatometer tests were done to quantify the parameters for phase field modelling by MICRESS®. The important input parameters are interface mobility, nucleation density, etc. A contribution was made to include austenite to bainite transformation in MICRESS®. This is useful to predict the microstructure in the fast cooling segments. The phase transformation model is capable to predict the microstructure along the heating and cooling cycles of welding. Tensile tests have shown the evidence of failure at the heat affected zone, which has the ferrite-tempered martensite microstructure.
Epilepsy
(2010)
The CellDrum technology (The term 'CellDrum technology' includes a couple of slightly different technological setups for measuring lateral mechanical tension in various types of cell monolayers or 3D-tissue constructs) was designed to quantify the contraction rate and mechanical tension of self-exciting cardiac myocytes. Cells were grown either within flexible, circular collagen gels or as monolayer on top of respective 1-mum thin silicone membranes. Membrane and cells were bulged outwards by air pressure. This biaxial strain distribution is rather similar the beating, blood-filled heart. The setup allowed presetting the mechanical residual stress level externally by adjusting the centre deflection, thus, mimicking hypertension in vitro. Tension was measured as oscillating differential pressure change between chamber and environment. A 0.5-mm thick collagen-cardiac myocyte tissue construct induced after 2 days of culturing (initial cell density 2 x 10(4) cells/ml), a mechanical tension of 1.62 +/- 0.17 microN/mm(2). Mechanical load is an important growth regulator in the developing heart, and the orientation and alignment of cardiomyocytes is stress sensitive. Therefore, it was necessary to develop the CellDrum technology with its biaxial stress-strain distribution and defined mechanical boundary conditions. Cells were exposed to strain in two directions, radially and circumferentially, which is similar to biaxial loading in real heart tissues. Thus, from a biomechanical point of view, the system is preferable to previous setups based on uniaxial stretching.
Purpose:
MRI plays a leading diagnostic role in assessing the musculoskeletal (MSK) system and is well established for most questions at clinically used field strengths (up to 3 T). However, there are still limitations in imaging early stages of cartilage degeneration, very fine tendons and ligaments, or in locating nerve lesions, for example. 7 T MRI of the knee has already received increasing attention in the current published literature, but there is a strong need to develop new radiofrequency (RF) coils to assess more regions of the MSK system. In this work, an eight-channel transmit/receive RF array was built as a multipurpose coil for imaging some of the thus far neglected regions. An extensive coil characterization protocol and first in vivo results of the human wrist, shoulder, elbow, knee, and ankle imaged at 7 T will be presented.
Methods:
Eight surface loop coils with a dimension ofurn:x-wiley:00942405:media:mp7176:mp7176-math-0001 were machined from FR4 circuit board material. To facilitate easy positioning, two coil clusters, each with four loop elements, were combined to one RF transmit/receive array. An overlapped and shifted arrangement of the coil elements was chosen to reduce the mutual inductance between neighboring coils. A phantom made of body-simulating liquid was used for tuning and matching on the bench. Afterward, the S-parameters were verified on a human wrist, elbow, and shoulder. For safety validation, a detailed compliance test was performed including full wave simulations of the RF field distribution and the corresponding specific absorption rate (SAR) for all joints. In vivo images of four volunteers were assessed with gradient echo and spin echo sequences modified to obtain optimal image contrast, full anatomic coverage, and the highest spatial resolution within a reasonable acquisition time. The performance of the RF coil was additionally evaluated by in vivo B1 mapping.
Results:
A comparison of B1 per unit power, flip angle distribution, and anatomic images showed a fairly homogeneous excitation for the smaller joints (elbow, wrist, and ankle), while for the larger joints, the shoulder and especially the knee, B1 inhomogeneities and limited penetration depth were more pronounced. However, the greater part of the shoulder joint could be imaged.In vivo images rendered very fine anatomic details such as fascicles of the median nerve and the branching of the nerve bundles. High-resolution images of cartilage, labrum, and tendons could be acquired. Additionally, turbo spin echo (TSE) and inversion recovery sequences performed very well.
Conclusions:
This study demonstrates that the concept of two four-channel transmit/receive RF arrays can be used as a multipurpose coil for high-resolutionin vivo MR imaging of the musculoskeletal system at 7 T. Not only gradient echo but also typical clinical and SAR-intensive sequences such as STIR and TSE performed well. Imaging of small structures and peripheral nerves could in particular benefit from this technique.
Purpose: Image analysis by the retinal vessel analyzer (RVA) observes retinal vessels in their dynamic state online noninvasively along a chosen vessel segment. It has been found that high-frequency diameter changes in the retinal artery blood column along the vessel increase significantly in anamnestically healthy volunteers with increasing age and in patients with glaucoma during vascular dilation. This study was undertaken to investigate whether longitudinal sections of the retinal artery blood column are altered in systemic hypertension.
Methods: Retinal arteries of 15 untreated patients with essential arterial hypertension (age, 50.9 ± 11.9 years) and of 15 age-matched anamnestically healthy volunteers were examined by RVA. After baseline assessment, a monochromatic luminance flicker (530–600 nm; 12.5 Hz; 20 s) was applied to evoke retinal vasodilation. Differences in amplitude and frequency of spatial artery blood column diameter change along segments (longitudinal arterial profiles) of 1 mm in length were measured and analyzed using Fourier transformation.
Results: In the control group, average reduced power spectra (ARPS) of longitudinal arterial profiles did not differ when arteries changed from constriction to dilation. In the systemic hypertension group, ARPS during constriction, baseline, and restoration were identical and differed from ARPS during dilation (P < 0.05). Longitudinal arterial profiles in both groups showed significant dissimilitude at baseline and restoration (P < 0.05).
Conclusions: The retinal artery blood column demonstrates microstructural alterations in systemic hypertension and is less irregular along the vessel axis during vessel dilation. These microstructural changes may be an indication of alterations in vessel wall rigidity, vascular endothelial function, and smooth muscle cells in this disease, leading to impaired perfusion and regulation.
Various models have been proposed for the prediction of the necessary support pressure at the face of a shallow tunnel. To assess their quality, the collapse of a tunnel face was modelled with small-scale model tests at single gravity. The development of the failure mechanism and the support force at the face in dry sand were investigated. The observed displacement patterns show a negligible influence of overburden on the extent and evolution of the failure zone. The latter is significantly influenced, though, by the initial density of the sand: in dense sand a chimney-wedge-type collapse mechanism developed, which propagated towards the soil surface. Initially, loose sand did not show any discrete collapse mechanism. The necessary support force was neither influenced by the overburden nor the initial density. A comparison with quantitative predictions by several theoretical models showed that the measured necessary support pressure is overestimated by most of the models. Those by Vermeer/Ruse and Léca/Dormieux showed the best agreement to the measurements.