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Lipids in blood-brain barrier models in vitro I: Thin-layer chromatography and high-performance liquid chromatography for the analysis of lipid classes and long-chain polyunsaturated fatty acids.

The objectives of this study were to optimize a sensitive high-performance liquid chromatography (HPLC) method for fatty acid (FA) analysis for the quantification of polyunsaturated FAs (PUFAs) in cell lipid extracts and to analyze the lipid and FA patterns of three cell lines used in blood-brain barrier (BBB) models: RBE4, ECV304, and C6. Thin-layer chromatographic analysis revealed differences in the phosphatidylcholine-phosphatidylethanolamine (PC:PE) ratios and the triglyceride (TG) content. The PC:PE ratio was <1 for RBE4 cells but >1 for ECV304 and C6 cells. ECV304 cells displayed up to 9% TG depending on culture time, whereas the other cell lines contained about 1% TG. The percentages of docosahexaenoic acid were 9.4 +/- 1.7% of the unsaturated FAs in RBE4 cells (n = 5; 4 d in culture; 9.9% after 10 d), 8.1 +/- 2.0% in ECV304 cells (n = 11; 10 to 14 d), and 6.7 +/- 0.6% in C6 cells (n = 6; 10 to 14 d) and were close to the published values for rat brain microvascular endothelium. The percentage of arachidonic acid (C20:4) was about half that in vivo. ECV304 cells contained the highest fraction of C20:4, 17.8 +/- 2.2%; RBE4 cells contained 11.6 +/- 2.4%; and C6 cells 15.8 +/- 1.9%. It is concluded that a sensitive HPLC method for FAs is now optimized for the analysis of long-chain PUFAs. The results provide a useful framework for studies on the effects of lipid modulation and give reference information for the development of further BBB models.

Animals↗

Lipids in blood-brain barrier models in vitro II: Influence of glial cells on lipid classes and lipid fatty acids.

Lipids of brain tissue and brain microvascular endothelial cells contain high proportions of long-chain polyunsaturated fatty acids (long PUFAs). The blood-brain barrier (BBB) is formed by the brain endothelial cells under the inductive influence of brain cells, especially perivascular glia, and coculture of endothelial cells and glial cells has been used to examine this induction. The objective of this study was to investigate whether C6 glioma cells are able to influence the lipid composition and shift the fatty acid (FA) patterns of the BBB model cell lines RBE4 and ECV304 toward the in vivo situation. Lipid classes of the three cell lines were analyzed by thin-layer chromatography and lipid FA patterns by high-performance liquid chromatography. Only ECV304 cells showed altered lipid composition in coculture with C6 cells. The fractions of triglycerides and cholesteryl esters (depending on the support filter) were about twice as high in coculture as when the cells were grown alone. Triglyceride fractions reached 13 to 15% of total lipids in coculture. The three cell lines showed an increase in the percentage of long PUFAs with respect to unsaturated FAs, mainly because of an increase in the percentages of arachidonic acid, all cis-7,10,13,16-docosatetraenoic acid, and all cis-7,10,13,16,19-docosapentaenoic acid. It is concluded that glioma C6 cells are able to induce a more in vivo-like FA pattern in BBB cell culture models. However, changes were not significant for the individual PUFAs, and their levels did not reach in vivo values.

Animals↗

Reconstruction of deforming aortas in two-photon autofluorescence image sequences.

Information loss may occur frequently in the imaging of living tissues by using two-photon fluorescence microscopy due to the intensive deformation of the tissue. A landmark-based optical flow interpolation scheme is proposed for image reconstruction of living aorta walls in two-photon autofluorescence image sequences. Landmarks are extracted and evaluated by an active contour-based aorta model, and are aligned and reconstructed by use of a hierarchical algorithm. The accuracy of the calculation of optical flow is improved by applying landmark-based image warping. Experimental results show that the proposed scheme outperforms commonly used optical flow interpolation techniques for the reconstruction of intensively deforming tissues.

Aorta↗

Influence of perfusion depth on laser Doppler flow measurements with large source-detector spacing.

Laser Doppler flowmetry with a large source-detector spacing has been applied to measure blood perfusion in the deeper regions of tissue. The influence of the depth of perfusion on the Doppler spectrum for the large source-detector spacing is likely to be different from that for the conventional laser Doppler instruments with small source-detector spacing. In this study, the light propagation in a tissue model with a blood perfusion layer is predicted by the Monte Carlo simulation to discuss the influence of the depth of perfusion, blood volume, and source-detector spacing on the spectrum of the Doppler signal detected with large source-detector spacing. The influence of the depth of perfusion on the Doppler spectrum for the large source-detector spacing is different from that for the conventional laser Doppler instruments with small source-detector spacing, although the influence of source-detector spacing and blood volume on the Doppler spectrum for large source-detector spacing is almost the same as that for the conventional laser Doppler instruments. The influence of the depth of the perfusion on the Doppler spectrum depends on the path length that the detected light travels at different depths.

Laser-Doppler Flowmetry↗

Transillumination optical tomography of tissue-engineered blood vessels: a Monte Carlo simulation.

A Monte Carlo technique has been developed to simulate the transillumination laser computed tomography of tissue-engineered blood vessels. The blood vessel was modeled as a single cylinder layer mounted on a tubular mandrel. Sequences of images were acquired while rotating the mandrel. The tomographic image was reconstructed by applying a standard Radon transform. Angular discrimination was applied to simulate a spatial filter, which was used to reject multiply scattered photons. The simulation results indicated that the scattering effect can be overcome with angular discrimination because of the thin tissue thickness. However, any refractive-index mismatch among the tissue, the surrounding media, and the mandrel could produce significant distortions in the reconstructed image.

Animals↗

Cardiac arrhythmias and left ventricular function in respiratory failure from chronic obstructive pulmonary disease.

In 22 patients with COPD, we studied the relationship between left ventricular function and cardiac arrhythmias. Ventricular arrhythmias were detected on a 24-h ECG recorded at the beginning of the observation period and after a stable improvement of RF. Left ventricular function was evaluated by equilibrium-gated radionuclide angiocardiography measuring LVEF, PER and PFR. We found a significant decrease in the arrhythmia score after improvement of RF; LVEF and PFR were slightly depressed in six and nine patients, respectively. A "step-up" multiple regression analysis revealed a significant inverse correlation between PFR and ventricular arrhythmias during worsened RF, whereas LVEF, arterial blood gases and clinical data were not significantly predictive variables. Thus, a depressed left ventricular diastolic performance seems to be a predictive factor for arrhythmias during RF from COPD. The poor definition of the statistical model suggests that other presently unknown factors contribute to the genesis of ventricular arrhythmias.

Arrhythmias, Cardiac↗

Effects of injection site on the accuracy of thermal washout right ventricular ejection fraction measurements in clinical and model investigations.

The purpose of this investigation was to improve the accuracy of measurement of thermal right ventricular ejection fraction (RVEF) using the modified Swan-Ganz catheter. Three serial ejection fractions (EFs) (EF1, 2, 3) and the mean were calculated, based on Holt's theory. RVEFs were compared between right ventricular (RV) and atrial (RA) injection in ten intensive care unit (ICU) patients using a modified catheter having RV and RA orifices (15 cm and 30 cm from the distal end, respectively), and paired duplicate (two patients) or triplicate (eight patients) measurements were performed. To determine what factors interfere with RVEF, a model heart (with diastolic volume of 150 ml) was constructed, in which model injection of cold water to the direct inflow tract (RA), to the direct mixing chamber (RV), or through the catheter running in the inflow tract were compared. When EFs were compared between RV and RA injection, those for the former were greater (RV vs RA in EF1 and EFmean: 0.46 +/- 0.15 vs 0.23 +/- 0.11 in EF1, and 0.45 +/- 0.13 vs 0.28 +/- 0.11 in EFmean, mean +/- SD, p less than 0.01). When the serial EFs were compared in each injection type, in the RV injection EF3 was the smallest as was EF1 in the RA injection. The same phenomenon was observed in the model as in the patients, and moreover when cold water was injected in RA through a catheter running through the circuit, EFs were greatly underestimated (EF1 = 0.29 +/- 0.02 at preset EF = 0.4). We conclude that these phenomena were caused by sluggish movement of the cold indicator from RA to RV when injected into RA, and by interference with the cooled cardiac chamber and catheter. Consequently, the first or second EFs obtained from RV injection might be closest to the actual values because of the least interference with those factors.

Adult↗

Heart rate asymmetry by Poincaré plots of RR intervals.

The Poincaré plot is a widely used method for visualizing and calculating heart rate variability and for investigating the oscillatory nature of heart action. We show that the Poincaré plot produced using physiological data for RR intervals is asymmetric. This suggests that the processes of heart rate acceleration (shortening of consecutive RR intervals) and deceleration (prolongation of successive RR intervals) might be asymmetric. To investigate this phenomenon, we define descriptors quantifying the heart rate asymmetry and present the results of a study involving 5-min ECG recordings of 50 healthy subjects in which, despite of the shortness of the recordings, the asymmetry is clearly visible.

Adult↗

[LOGNORMAL-NLSQ-technique. Evaluation of a new mathematical method for determining blood volume].

This paper describes the investigation of a new mathematical method of calculating blood volume. The new method determines the blood volume by calculating the product of the mean circulation transit time. The mean transit time is calculated from the body transport function. To examine the accuracy of the LOGNORMAL-NLSQ technique, 45 concentration time curves were measured in an in vitro recirculation model with variable clearance. The calculated volume was 4% smaller than the actual volume. This may be attributed to the functional dead space within the model, and is tolerable for clinical situations. The LOGNORMAL-NLSQ technique might acquire considerable importance in future, especially since it provides accurate results very quickly.

Blood Flow Velocity↗

Optimizing deconvolution techniques by the application of the Münchhausen meta algorithm.

A deconvolution applied to disturbed data often gives poor results, due to fundamental difficulties associated with ill-posed problems. Many numerical and theoretical methods have been invented to circumvent this phenomenon. Their performance varies, depending on the given problem and data. The main aim of this paper is to provide a decision rule for choosing a method for deconvolution and application of this method to the same data. We have called this meta-algorithm Münchhausen. In this paper we introduce and describe for the first time the basic principle of artificial disturbance of the data in the set-up of deconvolution. We demonstrate some interesting features of the random procedure Münchhausen, such as the non parametric set-up, robustness to disturbance of the data and last but not least good performance.

Algorithms↗

Comparison of macroscopic models of excitation and force propagation in the heart.

Computer aided simulations of the heart provide knowledge of phenomena, which are commonly neither visible nor measurable with current techniques. This knowledge can be applied e.g. in cardiologic diagnosis and therapy. A variety of models was created to reconstruct cardiac processes, e.g. electrical propagation and force development. In this work different macroscopic models were compared, i.e. models based on excitation-diffusion equations and cellular automata. The comparison was carried out concerning reconstruct-ability of cardiac phenomena, mathematical and biophysical foundation as well as computational expense. Particularly, the reconstruct-ability of electromechanic feedback mechanisms was examined. Perspectives for further developments and improvements of models were given.

Animals↗

[Simulation of coupling behavior of heart rhythms for study of phase synchronization].

Biological systems exhibit numerous rhythmic processes. In the human, rhythms produced by e.g. the cardiac and respiratory systems show signs of synchronisation, reflecting a physiological coupling of the underlying oscillating processes. When examining such rhythms for evidence of coupling, it is difficult to distinguish between epochs of coincidental (numerical) and real (physiological) synchronisation. The objective of this study was to simulate model rhythms in order to examine the conditions under which coupling may be recognised and the influence of noise on this recognition. Different model rhythms with adjustable noise were implemented. The synchronisation between pairs of such rhythms was examined with the help of synchrograms. Functions were used to simulate coupling. The results of the simulation were compared to real fetal-maternal cardiac rhythms. The simulation showed that heart rate variability may be an important factor in distinguishing between coincidental and real synchronisation.

Computer Simulation↗

Observer design for haemodynamics in patients undergoing cardiac surgery.

During extracorporeal circulation, many important circulatory parameters are unknown, being inaccessible for measuring probes, as e.g. the perfusion of the brain. An observer system, which estimates such patient variables continuously throughout the operation, can extend the information basis for the decisions of the perfusionist regarding the control of the heart-lung machine and thus contribute to adjust this operation procedure to the actual patient situation. The observer design is based on a mathematical model of the human circulatory system. Beside the classical Luenberger observer design, a rule-based approach has been tested, which is also based on the structure of a Luenberger observer, however instead of an observer matrix a correction algorithm has been used in the feedback loop. A prototype of this system for animal experimental and clinical evaluation has been realised.

Algorithms↗