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Keywords
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.
Objective: As high-field cardiac MRI (CMR) becomes more widespread the propensity of ECG to interference from electromagnetic fields (EMF) and to magneto-hydrodynamic (MHD) effects increases and with it the motivation for a CMR triggering alternative. This study explores the suitability of acoustic cardiac triggering (ACT) for left ventricular (LV) function assessment in healthy subjects (n=14). Methods: Quantitative analysis of 2D CINE steady-state free precession (SSFP) images was conducted to compare ACT’s performance with vector ECG (VCG). Endocardial border sharpness (EBS) was examined paralleled by quantitative LV function assessment. Results: Unlike VCG, ACT provided signal traces free of interference from EMF or MHD effects. In the case of correct Rwave recognition, VCG-triggered 2D CINE SSFP was immune to cardiac motion effects—even at 3.0 T. However, VCG-triggered 2D SSFP CINE imaging was prone to cardiac motion and EBS degradation if R-wave misregistration occurred. ACT-triggered acquisitions yielded LV parameters (end-diastolic volume (EDV), endsystolic volume (ESV), stroke volume (SV), ejection fraction (EF) and left ventricular mass (LVM)) comparable with those derived fromVCG-triggered acquisitions (1.5 T: ESVVCG=(56± 17) ml, EDVVCG=(151±32)ml, LVMVCG=(97±27) g, SVVCG=(94± 19)ml, EFVCG=(63±5)% cf. ESVACT= (56±18) ml, EDVACT=(147±36) ml, LVMACT=(102±29) g, SVACT=(91± 22) ml, EFACT=(62±6)%; 3.0 T: ESVVCG=(55±21) ml, EDVVCG=(151±32) ml, LVMVCG=(101±27) g, SVVCG=(96±15) ml, EFVCG=(65±7)% cf. ESVACT=(54±20) ml, EDVACT=(146±35) ml, LVMACT= (101±30) g, SVACT=(92±17) ml, EFACT=(64±6)%). Conclusions: ACT’s intrinsic insensitivity to interference from electromagnetic fields renders
As high-field cardiac MRI (CMR) becomes more widespread the propensity of ECG to distortions and mistriggering increases and with it the motivation for a cardiac triggering alternative. Hence, this study explores the suitability of acoustic cardiac triggering (ACT) for left ventricular (LV) function assessment in healthy subjects at 1.5T and 3.0T.
Comparison of Intravenous Immunoglobulins for Naturally Occurring Autoantibodies against Amyloid-β
(2010)
Molke fällt im Rahmen der Käseherstellung allein in Deutschland in Mengen von über 11 Mio. Tonnen jährlich an. Dieses Nebenprodukt wurde trotz seines Reichtums an Milchzucker und Proteinen lange Zeit kaum industriell weiterverarbeitet und stellte ein bedeutendes Problem bei der Abwasserreinigung dar. Derzeit kommen meist kosten- und reinigungsintensive Membranfiltrationsverfahren bei der Auftrennung von Molke in ihre Hauptkomponenten Lactose und Molkenprotein zum Einsatz. Die Produkte finden vorwiegend in der Nahrungsmittelindustrie Anwendung als Süßungsmittel, Proteinzusatz oder Texturbildner. Die Mehrheit des Proteins wird dabei als Konzentrat bzw. Proteinpulver verarbeitet. Wegen der antibakteriellen, antiviralen und weiteren wertvollen physiologischen Eigenschaften der Molkeproteine stellt eine weitere Aufreinigung der einzelnen Molkeproteine für die pharmazeutische Industrie einen naheliegenden zusätzlichen Wertschöpfungsschritt dar. In Kooperation mit der Süd Chemie AG wurde damit begonnen, ein Verfahren zu entwickeln, das kostengünstige mineralische Adsorbentien verwendet. Bisher konnte die Abtrennung von Lactose von den Molkenproteinen aus verdünntem Molkekonzentrat in einem Verfahrensschritt ohne Vorbehandlung des Rohstoffs erfolgreich realisiert werden. Aktuelle Arbeiten beschäftigen sich mit der Verbesserung der Proteinbindekapazitätund chromatographischen Proteinauftrennung sowie dem Upscaling zum direkten Einsatz von Molkekonzentrat ohne Vorverdünnung.
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.
Objectives
Interest in cardiovascular magnetic resonance (CMR) at 7 T is motivated by the expected increase in spatial and temporal resolution, but the method is technically challenging. We examined the feasibility of cardiac chamber quantification at 7 T.
Methods
A stack of short axes covering the left ventricle was obtained in nine healthy male volunteers. At 1.5 T, steady-state free precession (SSFP) and fast gradient echo (FGRE) cine imaging with 7 mm slice thickness (STH) were used. At 7 T, FGRE with 7 mm and 4 mm STH were applied. End-diastolic volume, end-systolic volume, ejection fraction and mass were calculated.
Results
All 7 T examinations provided excellent blood/myocardium contrast for all slice directions. No significant difference was found regarding ejection fraction and cardiac volumes between SSFP at 1.5 T and FGRE at 7 T, while volumes obtained from FGRE at 1.5 T were underestimated. Cardiac mass derived from FGRE at 1.5 and 7 T was larger than obtained from SSFP at 1.5 T. Agreement of volumes and mass between SSFP at 1.5 T and FGRE improved for FGRE at 7 T when combined with an STH reduction to 4 mm.
Conclusions
This pilot study demonstrates that cardiac chamber quantification at 7 T using FGRE is feasible and agrees closely with SSFP at 1.5 T.
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.