Rapid quantification of HIV protease inhibitors in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry

HIV protease inhibitors are important antiretroviral drugs which have substantially reduced the morbidity and mortality associated with HIV-1 infection. Recent data have shown relationships between plasma concentrations of the protease inhibitors and clinical response, which makes therapeutic drug monitoring valuable. We have developed and validated an assay, using liquid chromatography coupled with electrospray tandem mass spectrometry (LC/MS/MS), for the routine quantification of the six licensed protease inhibitors (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir) and the pharmacologically active nelfinavir metabolite M8 in plasma. The sample pretreatment consisted of protein precipitation with a mixture of methanol and acetronitrile using only 100 microl of plasma. Chromatographic separation was performed on an Inertsil ODS3 column (50 x 2.0 mm i.d., particle size 5 microm), with a quick stepwise gradient using an acetate buffer (pH 5) and methanol, at a flow rate of 0.5 ml min(-1). The analytical run time was 5.5 min. The use of a 96-well plate autosampler allowed batch sizes up to 150 patient samples. The triple-quadrupole mass spectrometer was operated in the positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over the concentration ranges 0.01-10 microg ml(-1) for indinavir and saquinavir, 0.1-10 microg ml(-1) for amprenavir, 0.05-10 microg ml(-1) for nelfinavir and ritonavir, 0.1-20 microg ml(-1) for lopinavir and 0.01-5 microg ml(-1) for M8. Saquinavir-d(5) and indinavir-d(6) were used as internal standards. The coefficients of variation were always <10% for both intra-day and inter-day precisions for each compound. Mean accuracies were also between the designated limits (+/-15%). The validated concentration ranges proved to be adequate in daily practice. This robust and fast LC/MS/MS assay is now successfully applied for routine therapeutic drug monitoring and pharmacokinetic studies in our hospital.     

含有陡峭势阱和凹凸非线性项的Kirchhoff型问题的多重正解*

在这篇文章中, 作者研究涉及凹凸非线性项的Kirchhoff型问题-(a + b ∫R3|▽u|²dx) Δu + λV (x)u = μf(x)|u|^q?2u + |u|^p?2u, x ∈ R³,u ∈ H¹(R³),其中a,b > 0 是常数, λ, μ > 0 是参数, 1 < q < 2, 4 < p < 6 且 V 是一个非负连续位势. 在f(x) 和 V 的合适条件下,此问题正解的存在性和集中性能够通过Nehari 流形和Ekeland 变分原理得到.     

Mode selective directional couplers for overmoded waveguide systems

Two types of directional couplers for transverse electric (TE) modes are described: short and multihole couplers, respectively. They selectively pick one mode out of a mode mixture in an overmoded circular waveguide system. Unwanted modes are either statistically kept at low level or are suppressed by destructive interference in the coupling waveguide. Mode selectivity and directivity in multihole couplers oscillate up and down with an increasing number of holes, finally reaching a minimum of approximately 20 dB, unless there are competing modes with rational fractions of the beat wavelength. A multihole coupler for the TE₀₂ mode (28 GHz, 63.4 mm waveguide diameter, 41 holes) and a length of 1.6 m shows a calculated directivity of 68 dB and suppresses the unwanted modes TE₀₁ with 34 dB (24 dB), TE₂₂ with 37 dB (45 dB), and further modes TE_m (<5, m<6) with 17 dB to 34 dB in forward direction (figures in parentheses are for unwanted modes propagating in backward direction).A short directional coupler for the TE₀₁ mode (28 GHz, 63.4 mm waveguide diameter) with 16 holes and a length of 230 mm shows a directivity of 55 to 100 dB between 27.9 and 28.1 GHz, suppressing the TE₀₂ mode with 35 to 80 dB, the TE₀₃ mode with 30 to 65 dB, and the TE₂₂ mode with 30 to 70 dB.     

Detection of single nucleotide polymorphisms using electrospray ionization mass spectrometry: validation of a one-well assay and quantitative pooling studies

Single nucleotide polymorphisms (SNPs) are currently being mapped and databased at a remarkable pace, providing a viable means for understanding disease susceptibility, differential drug response and human evolution. Consequently, there is an increasing demand for SNP genotyping technologies that are simple, rapid, cost effective and readily amenable to automation for high-throughput analyses. In this study, we improved the Survivor Assay, a SNP detection method based on electrospray ionization mass spectrometry (ESI-MS), with several developments. One improvement is the development of a one-well assay, requiring no off-line purification of the polymerase chain reaction product, achieved by simple addition of reagent solution into a single well. Another is the on-line separation of magnesium and dideoxynucleotides using an in-house made monolithic metal chelating column, eliminating any off-line sample preparation prior to mass spectrometric analysis. Here the Survivor Assay is extended from a proof-of-principle concept to a validated method by genotyping six SNPs from five different regions of human genomic DNA in 55 individual samples with 100% accuracy. This improved Survivor Assay eliminates the tedious and time-consuming steps of sample preparation, minimizes sample handing and offers a high-throughput analysis of SNPs by ESI-MS. The current combined preparation and analysis time is 2 min per sample. The simplicity of this method has potential for full automation and parallel chromatography and, thus, reduced analysis time. In addition, we have adapted the Survivor Assay for quantitative SNP analysis in pooled DNA samples. The capabilities and sensitivity of this approach were evaluated. We demonstrate that an allele occurring at a frequency of 2% can consistently be quantitated.     

Lipschitzian inverse functions,directional derivatives,and applications inC 1,1 optimization

The paper shows that Thibault's limit sets allow an iff-characterization of local Lipschitzian invertibility in finite dimension. We consider these sets as directional derivatives and extend the calculus in a way that can be used to clarify whether critical points are strongly stable inC ^1,1 optimization problems.Many fruitful discussions with colleagues D. Klatte and K. Tammer as well as with H. Th. Jongen and F. Nozicka have influenced the present investigations in a very constructive manner. For the original papers concerning the sets f(x; u), the author is indebted to Prof. L. Thibault.     

Analysis of the anisotropy of group velocity error due to spatial finite difference schemes from the solution of the 2D linear Euler equations

Numerical differencing schemes are subject to dispersive and dissipative errors, which in one dimension, are functions of a wavenumber. When these schemes are applied in two or three dimensions, the errors become functions of both wavenumber and the direction of the wave. For the Euler equations, the direction of flow and flow velocity are also important. Spectral analysis was used to predict the error in magnitude and direction of the group velocity of vorticity–entropy and acoustic waves in the solution of the linearised Euler equations in a two‐dimensional Cartesian space. The anisotropy in these errors, for three schemes, were studied as a function of the wavenumber, wave direction, mean flow direction and mean flow Mach number. Numerical experiments were run to provide confirmation of the developed theory. Copyright © 2012 John Wiley & Sons, Ltd.     

Characteristic behaviour of plasma fluctuations inside a plasma bubble in presence of a magnetic field due to the formation of a potential well

Experiments were carried out to observe the effect of a magnetic field and grid biasing voltage in presence of a plasma bubble in a magnetized, filamentary discharge plasma system. A spherical mesh grid of 80% optical transparency was negatively biased and introduced into the plasma for creating a plasma bubble. Diagnostics via an electrical Langmuir probe and a hot emissive probe were extensively used for scanning the plasma bubble. Plasma floating potential fluctuations were measured at three different positions of the plasma bubble. The instability in the pattern showed the dynamic transition from periodic to chaotic for increasing magnetic fields. Time scale analysis using continuous wavelet transform was carried out to identify the presence of non‐linearity from the contour plots. The mechanisms of the low‐frequency instabilities along with the transition to chaos could be qualitatively explained. Non‐linear techniques such as fast Fourier transform, phase space plot, and recurrence plot were used to explore the dynamics of the system appearing during plasma fluctuations. In order to demonstrate the observed chaotic phenomena in this study, characteristics of chaos such as the Lyapunov exponent were obtained from experimental time series data. The experimentally observed potential structure is confirmed with numerical analysis based on fluid hydrodynamics.     

Directional Self‐Assembly of Fluorinated Star Block Polymer Thin Films Using Mixed Solvent Vapor Annealing

We demonstrate the directional alignment of perpendicular‐lamellae domains in fluorinated three‐armed star block polymer (BP) thin films using solvent vapor annealing with shear stress. The control of orientation and alignment was accomplished without any substrate surface modification. Additionally, three‐armed star poly(methyl methacrylate‐block‐styrene) [PMMA‐PS] and poly(octafluoropentyl methacrylate‐block‐styrene) were compared to their linear analogues to examine the impact of fluorine content and star architecture on self‐assembled BP feature sizes and interdomain density profiles. X‐ray reflectometry results indicated that the star BP molecular architecture increased the effective polymer segregation strength and could possibly facilitate reduced polymer domain spacings, which are useful in next‐generation nanolithographic applications. © 2019 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2019 , 57, 1663–1672     

Effects of built-in electric field on donor binding energy in InGaN/ZnSnN2 quantum well structures

In_xGa${}_{1?x}$N/ZnSnN₂ quantum well structures are studied in terms of a binding energy of a donor atom. 1s and $2p_{\pm }$ impurity states are considered. The Schrödinger's and Poisson's equations are solved self-consistently. A hydrogenic type wave function to represent each impurity state is assumed. The calculations include band-bending in the potential energy profile introduced by the built-in electric field existing along the structures. The binding energy and the energy of the transition between the impurity states are represented as a function of the quantum well width, the donor position, and the indium concentration. An external magnetic field up to 10 T is included into the calculations to compute the Zeeman splitting. The maximum value of the transition energy is around 30 meV (nearly 7.3 THz) which occurs in a 15-Å In_0.3Ga_0.7N/ZnSnN₂ quantum well. Being strong, the built-in electric field makes the transition energy drop quickly with the decreasing well width. For the same reason, the energy curves are found to be highly asymmetric function of the donor position around the well center. Compared to the bulk value, the transition energy in the quantum well structures enhances nearly two-fold.     

Ground state solutions for Klein–Gordon–Maxwell system with steep potential well

In this paper, we study the following Klein–Gordon–Maxwell system $\left\{\begin{array}{c}-\Delta u+(\lambda a\left(x\right)+1)u-(2\omega +\varphi )\varphi u=f(x,u),\mathrm{in}{\mathbb{R}}^3,\hfill \\ -\Delta \varphi =-(\omega +\varphi )u^2,\mathrm{in}{\mathbb{R}}^3.\hfill \end{array}\right.$Using variational methods, we obtain the existence of ground state solutions under some appropriate assumptions on $a$ and $f$.