Highly sensitive and rapid determination of tacrolimus in peripheral blood mononuclear cells by liquid chromatography–tandem mass spectrometry

After solid organ transplantation, tacrolimus is given to prevent rejection. Therapeutic drug monitoring is used to reach target concentrations of tacrolimus in whole blood. Because the site of action of tacrolimus is the lymphocyte, and tacrolimus binds ~80% to erythrocytes, the intracellular tacrolimus concentration in lymphocytes is possibly more relevant. For this purpose, we aimed to develop, improve and validate a UPLC–MS/MS method to measure tacrolimus concentrations in isolated peripheral blood mononuclear cells (PBMCs). PBMCs were isolated using a Ficoll separation technique, followed by a washing step using red blood cell lysis. A cell suspension of 50 μL containing 1 million PBMCs was used in combination with MagSiMUS‐TDM^PREP. To each sample we added 30 μL lysis buffer, 20 μL reconstitution buffer containing ¹³C²H₄‐tacrolimus as internal standard, 40 μL MagSiMUS‐TDM^PREP Type I Particle Mix and 175 μL Organic Precipitation Reagent VI for methanol‐based protein precipitation. A 10 μL aliquot of the supernatant was injected into the UPLC–MS/MS system. The method was validated, resulting in high sensitivity and specificity. The method was linear (r² = 0.997) over the range 5.0–1250 pg/1 × 10⁶ PBMCs. The inaccuracy was <5% and the imprecision was <15%. The washing steps following Ficoll isolation could be performed at either room temperature or on ice, with no effect of the temperature on the results. A method for the analysis of tacrolimus concentrations in PBMCs was developed and successfully validated. Further research will be performed to investigate the correlation between concentrations in PBMCs and clinical outcome.     

Water Transport in Bio-based Porous Materials: A Model of Local Kinetics of Sorption—Application to Three Hemp Concretes

Transport in Porous Media - The classic models describing the hygric mass transfers inside porous materials seem unsuitable in the case of bio-based materials. They are based on the assumption of...     

Nonlinear analysis of the pharmacological conversion of sustained atrial fibrillation in conscious goats by the class Ic drug cibenzoline

Methods from nonlinear dynamics were applied to test the hypothesis that the dynamics of sustained atrial fibrillation (AF) is modified by the class Ic drug cibenzoline during pharmacological conversion. The experiments were performed in conscious goats in which sustained AF was induced by continuous maintenance of AF via programmed electrical stimulation. Data were collected from electrophysiological experiments in five goats to terminate sustained AF by continuous infusion of cibenzoline. Sets of five unipolar epicardial electrograms of one minute duration were recorded from the left and right atrial free wall during sustained AF (control), and at three episodes during infusion of cibenzoline, when the mean AF interval had been prolonged to 25%, 50% and 85% with respect to control. Ventricular far-field potentials were removed from atrial electrograms by a coherent averaging procedure. Using the Grassberger-Procaccia method, the dynamics of the local atrial electrograms was investigated by estimating the (coarse-grained) correlation dimension and correlation entropy from the correlation integral. The results were related to a recently proposed classification (types I-III) of AF based on the degree of complexity of atrial activation patterns. The coarse-grained correlation dimension D(cg) and entropy K(cg) indicated that sustained AF corresponded to type II. During drug administration the coarse-grained parameters were not significantly different from control. Scaling regions in the correlation integral were observed after infusion of cibenzoline (3 out of 5 goats) suggesting that the drug introduced low-dimensional features (type I) in the dynamics of AF (correlation dimension D ranging from 2.8 to 4.4 and correlation entropy K from 1.6 to 6.2 nats/s). Sinus rhythm recorded shortly after cardioversion was very regular (D<2 and K<3 nats/s). The hypothesis that the electrograms during AF and sinus rhythm were generated by a static transformation of a linear Gaussian random process was rejected using a test for time reversibility. The nonlinear analysis revealed that cibenzoline does not significantly alter the dynamics of sustained AF during pharmacological conversion other than a slowing down of the atrial activation and a somewhat increasing global organization of the atrial activation pattern. The sudden change in the dynamical behavior at cardioversion suggests a mechanism that is reminiscent of a bifurcation. (c) 1997 American Institute of Physics.     

The Structure of the Algebra of Observables in the Intermediate Situation of the ∈-Model

We look at the action of the spin-1/2 operatorsof quantum mechanics on the state of an entity in aphysical way, and use this as a guideline to define theoperators of the intermediate situations of a general spin-1/2 measurement model called the-model. Then we test the possible linearity ofthe operators so constructed.     

Rhodium-catalyzed intramolecular conjugate addition of vinylstannanes to dihydro-4-pyridones: a simple method for stereoselective construction of 1-azabicyclic alkaloids

A straightforward route for the stereoselective synthesis of unsaturated quinolizidines and related higher homologs has been developed. Our results revealed that the rhodium complex [RhCl(cod)]₂ is an effective catalyst for the intramolecular conjugate addition of vinylstannanes to the dihydropyridones leading to the corresponding 1-azabicyclo[m.n.0]alkenes in a moderate to good yield and an excellent diastereoselectivity under neutral conditions at room temperature. This methodology has been successfully applied to the stereoselective synthesis of the racemic lasubine I.     

Self-consistent field modeling of adsorption from polymer/surfactant mixtures

We report on the development of a self-consistent field model that describes the competitive adsorption of nonionic alkyl-(ethylene oxide) surfactants and nonionic polymer poly(ethylene oxide) (PEO) from aqueous solutions onto silica. The model explicitly describes the response to the pH and the ionic strength. On an inorganic oxide surface such as silica, the dissociation of the surface depends on the pH. However, salt ions can screen charges on the surface, and hence, the number of dissociated groups also depends on the ionic strength. Furthermore, the solvent quality for the EO groups is a function of the ionic strength. Using our model, we can compute bulk parameters such as the average size of the polymer coil and the surfactant CMC. We can make predictions on the adsorption behavior of either polymers or surfactants, and we have made adsorption isotherms, i.e., calculated the relationship between the surface excess and its corresponding bulk concentration. When we add both polymer and surfactant to our mixture, we can find a surfactant concentration (or, more precisely, a surfactant chemical potential) below which only the polymer will adsorb and above which only the surfactant will adsorb. The corresponding surfactant concentration is called the CSAC. In a first-order approximation, the surfactant chemical potential has the CMC as its upper bound. We can find conditions for which CMC < CSAC . This implies that the chemical potential that the surfactant needs to adsorb is higher than its maximum chemical potential, and hence, the surfactant will not adsorb. One of the main goals of our model is to understand the experimental data from one of our previous articles. We managed to explain most, but unfortunately not all, of the experimental trends. At the end of the article we discuss the possibilities for improving the model.     

Structure elucidation and 1H/13C NMR spectral assignments of four trabectedin related compounds

This article presents the structure elucidation of four new compounds, formed during the hemisynthetic preparation of trabectedin, an anti-tumor natural product from Ecteinascidia turbinata. We report herein on the use of UV, MS and NMR spectroscopic data along with (1)H and (13)C spectral assignments obtained by means of 1D and 2D homo- and heteronuclear NMR techniques.     

Off-line pyrolysis and compound-specific stable carbon isotope analysis of lignin moieties: a new method for determining the fate of lignin residues in soil

Off-line pyrolysis was used to liberate lignin moieties from dung and soil and, after trimethylsilylation, the delta(13)C values of these derivatives were determined by gas chromatography-combustion-isotope ratio mass spectrometry. Initial delta(13)C values determined for 4-vinylphenol, syringol, 4-vinylguaiacol, 4-acetylsyringol, 4-vinylsyringol, 4-(2-Z-propenyl)syringol, 4-(2-E-propenyl)syringol and 4-(2-propenone)syringol pyrolysis products of the lignin polyphenol structure from C(4) (delta(13)C(bulk) = -12.6%) and C(3) (delta(13)C(bulk) = -30.1 per thousand) dung confirmed the robust and reproducible nature of the off-line preparation technique. C(4) dung was used as a treatment in a randomised field experiment to assess the short-term sequestration of dung carbon in managed grasslands. Since lignin was on average 3.5 per thousand depleted in (13)C compared with bulk dung delta(13)C values, this may have resulted in an under-estimation of dung C incorporation based on bulk delta(13)C values. Therefore, an investigation of the compound-specific delta(13)C values of dung-derived lignin moieties extracted from soils sampled up to 372 days was undertaken. Delta(13)C values between lignin moieties extracted from treated and untreated soils showed that dung-derived lignin was not especially resistant to degradation and suggested that individual moieties of the lignin macromolecule must: (i) move into soil, (ii) be degraded, or (iii) be transformed diagenetically at different rates. This adds to a gathering body of evidence that lignin is not particularly stable in soils, which has considerable significance for the perceived role of different biochemical components in the cycling of C in soils.