A novel method for serum lipoprotein profiling using high performance capillary isotachophoresis

A new capillary isotachophoresis (cITP) method for lipoprotein profiling with superior lipoprotein coverage compared to previous methods has been developed, resolving twice as many lipoprotein species (18 peaks/fractions) in serum or plasma in less than 9.5 min. For this, a novel mixture of 24 spacers, including amino acids, dipeptides and sulfonic acids, was developed and fine-tuned, using predictive software (PeakMaster) and testing of spiked serum samples. Lipoprotein peaks were identified by serum-spiking with reference lipoproteins. Compatibility with common lipophilic stains for selective lipoprotein detection with either UV/Vis or laser-induced fluorescence was demonstrated. A special new capillary with a neutral coating (combining water-compatible OV1701-OH deactivation and methylation) was used for the first time for electrodriven separations, allowing very stable separations in a pH 8.8–9.4 gradient system, being functional for more than 100 injections. Excellent reproducibility was achieved, with coefficients of variation lower than 2.6% for absolute migration times. Comparison was performed with human plasma samples analyzed by NMR, leading to similar results with cITP after multivariate statistics, regarding group-clustering and lipoprotein species correlation. The new cITP method was applied to the analysis of serum samples from a LDL receptor knock-out mice model fed either a normal diet or a western-type diet. Differences in the lipoprotein levels and in the sublipoprotein types were detected, showing a shift to more atherogenic particles due to the high cholesterol diet. In summary, this novel method will allow more detailed and informative profiling of lipoprotein particle subtypes for cardiovascular disease research.     

Predictive values instead of normal ranges: less data, more information

Diagnostic strategies can have various goals at two levels: to facilitate the diagnostic process on the cognitive level, and to serve considerations on the level of the doctor–patient relationship. Requests for laboratory tests could be intended to exclude a disease or to affirm the presence of disease. Thirdly, tactical motives to smoothen the negotiations between doctor and patient probably seem to be important as well. These three intentions differ in prior probability, should lead to different sets of tests, and to different interpretations. Even the cut-off points should differ. This leads to three different decision strategies, both at requesting, as at interpreting the results. Following this line of thought, post-test probabilities are more suitable than normal ranges. Excluding strategy: this is the most prevalent. However, the disadvantage of an excluding strategy (prior 1–5%) is a false-positive result. A positive test result should lead to follow-up by wait and see or by repeated testing. More extensive testing usually is not a very sensible strategy. In practice, physicians simply ignore slightly abnormal values. Mentally they put the cut-off points for normality more broader. The number of tests is small. Confirmative strategy: the disadvantage of a confirmative intention (prior 10–30%) is a false-negative result. Follow-up without testing, repeated testing, or even accepting marginal normal results as abnormal is a proper strategy. The number of tests is moderate to high. Tactical strategy: the tactical intention strategy to reassure the patient – or avoid referrals – could lead to ignoring all slightly positive test results by choosing a higher cut-off point. Actually, considering the usual insignificant diagnostic gain when testing for tactical reasons, all test results are clinically insignificant, unsuspected outliers excluded. Here, a very limited set of tests should be chosen. The laboratory test is the currency in mutual trading medical expectations and relationship considerations between doctor and patient. The number of tests is minimal. If the physician chooses a strategy, a limited range of prior probability is chosen. Then a possibly computerized algorithm produces a “Value (posterior probability)” as test result, replacing “Value (normal ranges)”. Thus one number less on the lab form, yielding more significant information.Presented at the 10th Conference Quality in the Spotlight, March 2005, Antwerp, Belgium     

Steroid oxidoreductase activity of alcohol dehydrogenases from horse, rat, and human liver.

Alcohol dehydrogenase from horse (isoenzyme SS and ES, but not EE), rat and human liver were found to catalyze the NAD-dependent oxidation of 3beta-hydroxy groups in 5alpha- and 5beta-steroids of the C19, C21, and C24 series. The enzymes from horse and rat liver were more active on 5beta-than on 5alpha-steroids. This difference was most marked with the enzyme from rat liver, especially with 3beta-hydroxyandrostan-17-ones and 3beta-hydroxypregnan-20-ones as substrates. The Km of isoenzyme ES from horse liver was lower for 3beta-hydroxy-5alpha-cholanoic acid (0.4 muM) than for 3beta-hydroxy-5beta-cholanoic acid (0.9 muM). 3alpha-Hydroxysteroids were not substrates for the enzymes from horse and rat liver. Human liver alcohol dehydrogenase had low affinity for 3beta-hydroxy-5alpha (and 5beta)-cholanoic acids, but oxidation could be clearly demonstrated by gas chromatographic analysis of the products.     

The fresh-salt water interface in a semi-pervious aquifer

The paper determines the location of the steady state interface between fresh and saltwater in a plane coastal aquifer. The lower boundary is totally impervious while the upper one is impervious below land and semi-pervious below the sea allowing an outflow through this part of the upper boundary. The model equations reduce to two boundary value problems, one valid in x < 0 and the other in x > 0 here x is measured along the aquifer with the origin at the coast. In each region unknown boundaries have to be determined as part of the solution using boundary and continuity conditions. Two cases are presented using the Dupuit approximation. One where the solution can be written down in terms of elementary functions and the other in which we have to use a phase space analysis.     

A Mathematical Model for Hysteretic Two-Phase Flow in Porous Media

We develop a mathematical model for hysteretic two-phase flow (of oil and water) in waterwet porous media. To account for relative permeability hysteresis, an irreversible trapping-coalescence process is described. According to this process, oil ganglia are created (during imbibition) and released (during drainage) at different rates, leading to history-dependent saturations of trapped and connected oil. As a result, the relative permeability to oil, modelled as a unique function of the connected oil saturation, is subject to saturation history. A saturation history is reflected by history parameters, that is by both the saturation state (of connected and trapped oil) at the most recent flow reversal and the most recent water saturation at which the flow was a primary drainage. Disregarding capillary diffusion, the flow is described by a hyperbolic equation with the connected oil saturation as unknown. This equation contains functional relationships which depend on the flow mode (drainage or imbibition) and the history parameters. The solution consists of continuous waves (expansion waves and constant states), shock waves (possibly connecting different modes) and stationary discontinuities (connecting different saturation histories). The entropy condition for travelling waves is generalized to include admissible shock waves which coincide with flow reversals. It turns out that saturation history generally has a strong influence on both the type and the speed of the waves from which the solution is constructed.     

Travelling waves in the transport of reactive solutes through porous media: Adsorption and binary ion exchange — Part 2

We study travelling wave solutions for the model developed in Part 1 of this paper. We develop and discuss a condition characterizing their existence. The possibility of finiteness is investigated. We consider the convergence to various limit cases and point out their different qualitative behaviour. Numerical examples are discussed.This work was supported by the EC project Filtration and Nonlinear Diffusion Processes (Contract No. SC1-0019-C(TT)).     

Analysis of Wet Pressing of Paper: The Three-Phase Model. Part II: Compressible Air Case

In this study we consider a model of wet pressing of paper. We use the techniques and results from the first part of this paper, where a simplified model is studied in details. The model is, using suitable transformation, rewritten in the standard parabolic-hyperbolic form. Numerical solution for typical example is given and the effects of plastic deformations of paper are investigated. Finally, the model is employed to adres the problem of choosing an optimal pressing regime.