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Optical coherence tomography : a potential tool to predict premature rupture of fetal membranes
(2013)
All cells generate contractile tension. This strain is crucial for mechanically controlling the cell shape, function and survival. In this study, the CellDrum technology quantifying cell's (the cellular) mechanical tension on a pico-scale was used to investigate the effect of lipopolysaccharide (LPS) on human aortic endothelial cell (HAoEC) tension. The LPS effect during gram-negative sepsis on endothelial cells is cell contraction causing endothelium permeability increase. The aim was to finding out whether recombinant activated protein C (rhAPC) would reverse the endothelial cell response in an in-vitro sepsis model. In this study, the established in-vitro sepsis model was confirmed by interleukin 6 (IL-6) levels at the proteomic and genomic levels by ELISA, real time-PCR and reactive oxygen species (ROS) activation by florescence staining. The thrombin cellular contraction effect on endothelial cells was used as a positive control when the CellDrum technology was applied. Additionally, the Ras homolog gene family, member A (RhoA) mRNA expression level was checked by real time-PCR to support contractile tension results. According to contractile tension results, the mechanical predominance of actin stress fibers was a reason of the increased endothelial contractile tension leading to enhanced endothelium contractility and thus permeability enhancement. The originality of this data supports firstly the basic measurement principles of the CellDrum technology and secondly that rhAPC has a beneficial effect on sepsis influenced cellular tension. The technology presented here is promising for future high-throughput cellular tension analysis that will help identify pathological contractile tension responses of cells and prove further cell in-vitro models.
Background
Minor changes in protein structure induced by small organic and inorganic molecules can result in significant metabolic effects. The effects can be even more profound if the molecular players are chemically active and present in the cell in considerable amounts. The aim of our study was to investigate effects of a nitric oxide donor (spermine NONOate), ATP and sodium/potassium environment on the dynamics of thermal unfolding of human hemoglobin (Hb). The effect of these molecules was examined by means of circular dichroism spectrometry (CD) in the temperature range between 25°C and 70°C. The alpha-helical content of buffered hemoglobin samples (0.1 mg/ml) was estimated via ellipticity change measurements at a heating rate of 1°C/min.
Results
Major results were:
1) spermine NONOate persistently decreased the hemoglobin unfolding temperature T u irrespectively of the Na + /K + environment,
2) ATP instead increased the unfolding temperature by 3°C in both sodium-based and potassium-based buffers and
3) mutual effects of ATP and NO were strongly influenced by particular buffer ionic compositions. Moreover, the presence of potassium facilitated a partial unfolding of alpha-helical structures even at room temperature.
Conclusion
The obtained data might shed more light on molecular mechanisms and biophysics involved in the regulation of protein activity by small solutes in the cell.
The importance of the availability of stored blood or blood cells, respectively, for urgent transfusion cannot be overestimated. Nowadays, blood storage becomes even more important since blood products are used for epidemiological studies, bio-technical research or banked for transfusion purposes. Thus blood samples must not only be processed, stored, and shipped to preserve their efficacy and safety, but also all parameters of storage must be recorded and reported for Quality Assurance. Therefore, blood banks and clinical research facilities are seeking more accurate, automated means for blood storage and blood processing.
Plant physiology and plant stress: Plant physiology will be much more important for human mankind because of yield and cultivation limits of crops determined by their resistance to stress. To assess and counteract various stress factors it is necessary to conduct plant research to gain information and results on plant physiology.
The demand of replacements for inoperable organs exceeds the amount of available organ transplants. Therefore, tissue engineering developed as a multidisciplinary field of research for autologous in-vitro organs. Such three dimensional tissue constructs request the application of a bioreactor. The UREPLACE bioreactor is used to grow cells on tubular collagen scaffolds OPTIMAIX Sponge 1 with a maximal length of 7 cm, in order to culture in vitro an adequate ureter replacement. With a rotating unit, (urothelial) cells can be placed homogeneously on the inner scaffold surface. Furthermore, a stimulation is combined with this bioreactor resulting in an orientation of muscle cells. These culturing methods request a precise control of several parameters and actuators. A combination of a LabBox and the suitable software LabVision is used to set and conduct parameters like rotation angles, velocities, pressures and other important cell culture values. The bioreactor was tested waterproof successfully. Furthermore, the temperature controlling was adjusted to 37 °C and the CO2 - concentration regulated to 5 %. Additionally, the pH step responses of several substances showed a perfect functioning of the designed flow chamber. All used software was tested and remained stable for several days.
Tests with palm tree leaves have just started yet and scan data are in the process to be analyzed. The final goal of future project for palm tree gender and species recognition will be to develop optical scanning technology to be applied to date palm tree leaves for in–situ screening purposes. Depending on the software used and the particular requirements of the users the technology potentially shall be able to identify palm tree diseases, palm tree gender, and species of young date palm trees by scanning leaves.