Method development for the quantitation of ABT-578 in rabbit artery tissue by 96-well liquid-liquid extraction and liquid chromatography/tandem mass spectrometric detection

Quantitative determination of drug concentrations in tissue samples can provide critical information for drug metabolism, kinetics, and toxicity evaluations. For analysis of tissue samples using liquid chromatography/tandem mass spectrometric (LC/MS/MS) detection, homogenization is a critical step in achieving good assay performance. Assay performance can be closely evaluated by spiking the drug directly into tissue samples prior to homogenization. It is especially important to include this assay evaluation for the analysis of artery tissue samples because artery tissue is very elastic, making it quite a challenge to develop an effective procedure for homogenization. An LC/MS/MS assay in 96-well format using liquid-liquid extraction was developed for analyzing ABT-578 in rabbit artery samples. Tissue quality control samples were prepared by spiking ABT-578 stock solutions directly into the tissue before homogenization. The usage of the tissue control samples gives a thorough evaluation of the sample preparation process that includes both homogenization and sample extraction. A 20% blood in saline solution was used as a homogenization solution. Calibration standards were made by spiking ABT-578 into rabbit whole blood. Blood quality control samples were also prepared by spiking ABT-578 into rabbit whole blood. These blood QC samples were used to confirm the validity of the calibration curve. A lower limit of quantitation of 0.050 ng/mL was achieved. The linear dynamic range of blood standards was from 0.050-30.3 ng/mL with the correlation coefficient (r) ranging from 0.9969-0.9996. Overall %CV was between 1.3 and 7.0%, and analytical recovery was between 98.2 and 105.8% for blood QC samples. The %CVs for tissue QC samples were between 6.7 and 13.0%, and analytical recovery after correction was between 93.5 and 114.3%.     

Electron binding energies of aqueous alkali and halide ions: EUV photoelectron spectroscopy of liquid solutions and combined ab initio and molecular dynamics calculations

Photoelectron spectroscopy combined with the liquid microjet technique enables the direct probing of the electronic structure of aqueous solutions. We report measured and calculated lowest vertical electron binding energies of aqueous alkali cations and halide anions. In some cases, ejection from deeper electronic levels of the solute could be observed. Electron binding energies of a given aqueous ion are found to be independent of the counterion and the salt concentration. The experimental results are complemented by ab initio calculations, at the MP2 and CCSD(T) level, of the ionization energies of these prototype ions in the aqueous phase. The solvent effect was accounted for in the electronic structure calculations in two ways. An explicit inclusion of discrete water molecules using a set of snapshots from an equilibrium classical molecular dynamics simulations and a fractional charge representation of solvent molecules give good results for halide ions. The electron binding energies of alkali cations computed with this approach tend to be overestimated. On the other hand, the polarizable continuum model, which strictly provides adiabatic binding energies, performs well for the alkali cations but fails for the halides. Photon energies in the experiment were in the EUV region (typically 100 eV) for which the technique is probing the top layers of the liquid sample. Hence, the reported energies of aqueous ions are closely connected with both structures and chemical reactivity at the liquid interface, for example, in atmospheric aerosol particles, as well as fundamental bulk solvation properties.     

The role of palladium catalyst and base in stereoselective transformations of (E)-2-chlorovinylsulfides

Stereoselective transformations of 2-chlorovinylsulfides in the presence of soluble (t-BuOK) or insoluble (solid KOH or Cs₂CO₃/18-crown-6) base and palladium catalyst (dppb)Pd(OAc)₂ have been studied. Depending on the substrate or catalytic system, the reaction leads to the formation of (E)-1,2-bis[aryl(or arylmethyl)thio]ethenes and/or (E)-1,4-bis[aryl(or arylmethyl)thio]-1-buten-3-ynes in yields of up to 93%.     

Sampling unfolding intermediates in calmodulin by single-molecule spectroscopy

We used single-pair fluorescence resonance energy transfer (spFRET) measurements to characterize denatured and partially denatured states of the multidomain calcium signaling protein calmodulin (CaM) in both its apo and Ca(2+)-bound forms. The results demonstrate the existence of an unfolding intermediate. A CaM mutant (CaM-T34C-T110C) was doubly labeled with fluorescent probes AlexaFlour 488 and Texas Red at opposing globular domains. Single-molecule distributions of the distance between fluorophores were obtained by spFRET at varying levels of the denaturant urea. Multiple conformational states of CaM were observed, and the amplitude of each conformation was dependent on urea concentration, with the amplitude of an extended conformation increasing upon denaturation. The distributions at intermediate urea concentrations could not be adequately described as a combination of native and denatured conformations, showing that CaM does not denature via a two-state process and demonstrating that at least one intermediate is present. The intermediate conformations formed upon addition of urea were different for Ca(2+)-CaM and apoCaM. An increase in the amplitude of a compact conformation in CaM was observed for apoCaM but not for Ca(2+)-CAM upon the addition of urea. The changes in the single-molecule distributions of CaM upon denaturation can be described by either a range of intermediate structures or by the presence of a single unfolding intermediate that grows in amplitude upon denaturation. A model for stepwise unfolding of CaM is suggested in which the domains of CaM unfold sequentially.     

Molecular structure of compounds containing a 2,2′-bithienyl fragment (review)

The results of X-ray crystallographic investigations of compounds containing a 2,2-bithienyl fragment are summarized and analyzed. The structural characteristics of unsubstituted bithiophene, 5-monosubstituted, 5,5-, 3,3-, 4,4-, and 3,4-disubstituted, and more highly substituted bithiophenes and also the structure of -conjugated oligothiophene derivatives are considered.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 6, pp. 725–760, June, 2000.     

Novel Stereoselective Phase Transfer Catalytic Synthesis and Some Applications of ( E )-2-Chlorovinylthioarenes and Hetarenes

A novel two-step method for the preparation of ( E )-2-chlorovinylthioarenes (or hetarenes) from thiols and 1,1,2-trichloroethane in the phase transfer catalytic systems solid K 2 CO 3 /solid KI/18-crown-6/xylene and solid KOH/18-crown-6/toluene has been developed. ( E )-2-chlorovinylthioarenes were isolated in yields up to 98%. Utilization of ( E )-2-chlorovinylthioarenes in the Heck and Stille reactions has been shown.     

Long-chain alkyl sulfonate micelle fission: a molecular dynamics study

In this study, we investigate micelle fission of long-chain alkyl sulfonate molecules using atomistic scale simulation. GROMACS software code with the united atom force field was applied. 0.5-μs parallel molecular dynamics simulation study was conducted for a surfactant/water system consisting of 192 sodium pentadecyl sulfonate and 40,553 water molecules. The large preassembled micelle was ruptured at Krafft above T?=?323-K temperature, and we track two ellipsoid-like micelles over the course of the production run. To estimate the micelle shape, we determined the principal moments of inertia and the eccentricity, which proved that the micelles have a pronounced prolate spheroid shape, which agrees well with our previous experimental data. The mechanism of micelle fission was explored in detail. The aggregation number, ionization degree, and other parameters obtained from simulation were consistent with existing experimental finding. The determined parameters in addition to simple visual inspection of trajectories revealed monomer-micelle exchange—with the estimated relaxation time τ ₁?=?10^??9s. We assume that the exchange process is conditioned by the unequal size of micelles leading to adjustment of aggregation number.     

Vinylation of pyridylcarboxamides with vinyltrimethoxysilane

A convenient method has been developed for the direct N-vinylation of pyridylcarboxamides with vinyltrimethoxysilane in the presence of copper(II) acetate and tetrabutylammonium fluoride. The yield of N-vinylamides depends on the nature of the starting amide and the solvent. The molecular structure of the products was confirmed by X-ray structural analysis.     

On data-guided optimal control simulation of human motion

This work investigates the combination of optical motion capturing data with optimal control simulations of human motion, which can be important in a wide range of applications in the professional as well as the private sector, ranging from health and ergonomics over human-machine-interaction to sports and games [1–3]. There are methodically very different approaches to include optical measurement data in the simulation of human motion, see e.g. [4–6]. Two different approaches to combine data and simulation are investigated in this work. Either we use a soft constraints approach, where the difference of simulated and measured marker positions is part of the objective function (1), or we formulate an hard constraints approach with nonlinear constraints that set an upper bound on this difference (2), while the objective function is purely physiologically motivated. (© 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim)