An automated, sheathless capillary electrophoresis-mass spectrometry platform for discovery of biomarkers in human serum

A capillary electrophoresis-mass spectrometry (CE-MS) method has been developed to perform routine, automated analysis of low-molecular-weight peptides in human serum. The method incorporates transient isotachophoresis for in-line preconcentration and a sheathless electrospray interface. To evaluate the performance of the method and demonstrate the utility of the approach, an experiment was designed in which peptides were added to sera from individuals at each of two different concentrations, artificially creating two groups of samples. The CE-MS data from the serum samples were divided into separate training and test sets. A pattern-recognition/feature-selection algorithm based on support vector machines was used to select the mass-to-charge (m/z) values from the training set data that distinguished the two groups of samples from each other. The added peptides were identified correctly as the distinguishing features, and pattern recognition based on these peptides was used to assign each sample in the independent test set to its respective group. A twofold difference in peptide concentration could be detected with statistical significance (p-value < 0.0001). The accuracy of the assignment was 95%, demonstrating the utility of this technique for the discovery of patterns of biomarkers in serum.     

Novel polyisoprenylated benzophenone derivatives from Garcinia paucinervis

Four novel polyisoprenylated benzophenone derivatives, paucinones A-D (1-4), were isolated from the leaves of the plant Garcinia paucinervis. Paucinones A-C (1-3) contained an unexpected cyclohexane-spiro-tetrahydrofuran moiety. A 1-methylene-3,3-dimethylcyclohexane group never reported before was found in the structure of paucinone D (4). The structures of these compounds were elucidated with spectroscopic evidences. The relative stereochemistries of 1-4 were determined by NOESY correlations. These compounds showed significant cytotoxicities against HeLa cells.     

Laser-induced breakdown detection combined with asymmetrical flow field-flow fractionation: application to iron oxi/hydroxide colloid characterization

The combination of asymmetrical flow field-flow fractionation (AsFlFFF) with the laser-induced breakdown detection (LIBD) is presented as a powerful tool for the determination of colloid size distribution at trace particle concentrations. Detection limits (D1) of 1, 4, and 20 microg/L have been determined for a mixture of polystyrene reference particles with 20, 50, and 100 nm in size, respectively. This corresponds to injected masses of 1, 4, and 20 pg, which is lower than found in a previous study with the symmetrical FlFFF (SyFlFFF). The improvement is mainly due to the lower colloid background discharged from the AsFlFFF channel. The combined method of AsFlFFF-LIBD is then applied to the analysis of iron oxi/hydroxide colloids being considered as potential carriers for the radionuclide migration from a nuclear waste repository. Our LIBD arrangement is less sensitive for iron colloid detection as compared to reference polystyrene particles which results in a detection limit of approximately 240 microg/L FeOOH for the AsFlFFF-LIBD analysis. This is superior to the detection via UV-Vis absorbance and comparable to ICP-MS detection. Size information (mean size 11-18 nm) for different iron oxi/hydroxide colloids supplied by the present method is comparable to that obtained by sequential ultrafiltration and dynamic light scattering. A combined on-line ICP-MS detection is used to gain insight into the colloid-borne main and trace elements.     

Lipid-lowering effects of ethyl 2-phenacyl-3-aryl-1H-pyrrole- 4-carboxylates in rodents

A series of substituted 2-phenacyl-3-phenyl-1H-pyrrole-4-carboxylates were prepared from substituted acetophenones in 6 steps. The final condensations between a chloroenal and an aminoketone were carried out under neutral conditions in parallel to yield the series listed below. Selected pyrrole derivatives proved to be potent hypolipidemic agents lowering serum triglyceride concentrations in CF-1 male mice after 14 days of I.P. administration. One agent orally lowered serum cholesterol in Sprague-Dawley male rats at 2mg/kg/day after 14 days. The agents demonstrated a lowering of mouse serum LDL- cholesterol levels and selected compounds showed an elevation of serum HDL-cholesterol levels. The cholesterol concentrations in the liver were raised while the cholesterol and triglyceride contents of the aorta were significantly lowered by the selected trisubstituted pyrrole.     

Versatile site-specific conjugation of small molecules to siRNA using click chemistry

We have previously demonstrated that conjugation of small molecule ligands to small interfering RNAs (siRNAs) and anti-microRNAs results in functional siRNAs and antagomirs in vivo. Here we report on the development of an efficient chemical strategy to make oligoribonucleotide-ligand conjugates using the copper-catalyzed azide-alkyne cycloaddition (CuAAC) or click reaction. Three click reaction approaches were evaluated for their feasibility and suitability for high-throughput synthesis: the CuAAC reaction at the monomer level prior to oligonucleotide synthesis, the solution-phase postsynthetic "click conjugation", and the "click conjugation" on an immobilized and completely protected alkyne-oligonucleotide scaffold. Nucleosides bearing 5'-alkyne moieties were used for conjugation to the 5'-end of the oligonucleotide. Previously described 2'- and 3'-O-propargylated nucleosides were prepared to introduce the alkyne moiety to the 3' and 5' termini and to the internal positions of the scaffold. Azido-functionalized ligands bearing lipophilic long chain alkyls, cholesterol, oligoamine, and carbohydrate were utilized to study the effect of physicochemical characteristics of the incoming azide on click conjugation to the alkyne-oligonucleotide scaffold in solution and on immobilized solid support. We found that microwave-assisted click conjugation of azido-functionalized ligands to a fully protected solid-support bound alkyne-oligonucleotide prior to deprotection was the most efficient "click conjugation" strategy for site-specific, high-throughput oligonucleotide conjugate synthesis tested. The siRNA conjugates synthesized using this approach effectively silenced expression of a luciferase gene in a stably transformed HeLa cell line.     

Use of modifiers in on-line and off-line supercritical fluid extraction

For many applications using supercritical fluid extraction (SFE), modifiers may be required.This paper will present some findings regarding the use of various modifiers including methanol, hexane, acetone, chloroform, dichloromethane, toluene, and tributylphosphate, in on-line and off-line SFE with cryogenic adsorbent trapping. The specific applications involved the extractions of petroleum hydrocarbons and pesticides from naturally incurred soils.     

Simulating Withdrawal Behaviors in Work Organizations: An Example of a Virtual Society

This article discusses computational modeling as a research discipline increasingly accessible to researchers in the social and behavioral sciences, a method necessary to allow us to address several important questions. Construction and use of virtual societies in which complex behavioral processes may be simulated in exquisite detail is possible because of the capacity and speed of desk top computers combined with requisite theoretical and methodological developments. Modeling has strengths orthogonal and complementary to those of the experimental tradition and correlational analyses of individual differences. These strengths can be exploited in many areas of psychology. Behavioral processes within a segment of a virtual society can be simulated with extreme levels of realism using overarching theoretical and empirical framework; segments can be combined to provide more macro statements. Variables that represent forces impinging on individuals from the macro-environment; variables that characterize relatively more micro-environments such as a work organization or the family or a classroom; and individual-level variables that summarize attitudinal, cognitive, and value states or traits can be included easily in simulations. Researchers can decompose and evaluate the interplay of multi-level causal forces as they interact within different theoretical models of how individuals enact different, interrelated behaviors. Temporal dynamics of behavioral processes, not well addressed by traditional research disciplines, can be exploited in modeling to permit the realistic study of dynamical, nonlinear systems. These points are illustrated by results from a program designed to simulate the process by which individuals withdraw from aversive work situations.     

A note on the interlacing of zeros and orthogonality

Let be a sequence of monic polynomials with deg(t_n)=n such that, for each n∈N, the zeros of t_n are real and simple and t_n and t_n+1 have no common zeros. We discuss the connection between the orthogonality of the sequence, the positivity of a certain ratio, and the interlacing of the zeros of t_n and t_n+1 for n≥1, n∈N.     

Noncompliant oligopolistic firms and marketable pollution permits: Statics and dynamics

In this paper, we consider the modeling, analysis, and computation of solutions to both static and dynamic models of multiproduct, multipollutant noncompliant oligopolistic firms who engage in a market for pollution permits. In the case of the static model, we utilize variational inequality theory for the formulation of the governing equilibrium conditions as well as the qualitative analysis of the equilibrium pattern, including sensitivity analysis. We then propose a dynamic model, using the theory of projected dynamical systems, whose set of stationary points coincides with the set of solutions to the variational inequality problem. We propose an algorithm, which is a discretization in time of the dynamic adjustment process, and provide convergence results using the stability analysis results that are also provided herein. Finally, we apply the algorithm to several numerical examples to compute the profit-maximized quantities of the oligopolistic firms' products and the quantities of emissions, along with the equilibrium allocation of licenses and their prices, as well as the possible noncompliant overflows and underflows. This is the first time that these methodologies have been utilized in conjunction to study a problem drawn from environmental policy modeling and analysis.