A switch-on fluorescence assay for bacterial β-lactamases with amyloid fibrils as fluorescence enhancer and visual tool

Herein is described the development of a novel switch-on fluorescence assay for detecting β-lactamases. The fluorescence assay comprises two components: solid beads coated with a β-lactam antibiotic, which is linked to an environment-sensitive fluorophore (dansylaminothiophenol, DTA), and amyloid fibrils of hen lysozyme (acting as fluorescence enhancer and visual tool). In the presence of the clinically significant TEM-1 β-lactamase, the DTA-antibiotic complex on the solid beads is hydrolyzed, thus releasing the DTA dye into solution. The DTA dye is only weakly fluorescent in solution but gives strong green fluorescence upon binding to lysozyme fibrils. These strongly fluorescent DTA-bound fibrils can be easily visualized by the naked eye upon illumination of the sample with a simple UV lamp. The fluorescence assay can detect TEM-1 at low concentration (0.01 nM). In contrast, no observable fluorescence appears when the fluorescence assay is performed on samples without the TEM-1 β-lactamase.     

Studies of wall painting fragments from Kaiping Diaolou by SEM/EDX, micro Raman and FT-IR spectroscopy

The multi-technique analytical approach has proved to be a very effective tool for the analysis of artwork, as demonstrated by various studies. In this work, four micro-analysis methods were used to analyze the wall painting fragments in Kaiping Diaolou, a world cultural heritage enlisted in 2007. Field-emission scanning electron microscopy (FE-SEM), energy dispersive X-ray micro-analysis (EDX), combined with micro Raman and Fourier transform infrared (FT-IR) spectroscopy, provided a vast amount of information concerning the raw materials present in the pigments, organic binder, plasters and mortars of the wall painting. Four types of pigments (goethite, lazurite, chromium green and calcite) were identified on the surface layer of the wall paintings. The substrate under the pigment layer was found to be composed of cubic-like calcite (CaCO₃), micro-rod bundle-shaped syngenite (K₂Ca(SO₄)₂·H₂O), gypsum (CaSO₄·2H₂O) and silica (SiO₂). The organic binder can be attributed to animal glue (such as egg) and drying oil by micro FT-IR spectroscopy. These analysis results can provide important information for the conservation and restoration of the Kaiping Diaolou.     

Chromatography-free Wittig reactions using a bifunctional polymeric reagent

The first example of a polystyrene bearing two distinct reagent groups has been prepared. This phosphine and amine functionalized material was used in one-pot Wittig reactions with an aldehyde and either an α-halo-ester, -ketone, or -amide. Due to the heterogeneous nature of the polymer, the desired alkene product of these reactions could be isolated in excellent yield in essentially pure form after only filtration and solvent removal.     

Selection of aptamers for signal transduction proteins by capillary electrophoresis

High‐affinity aptamers for important signal transduction proteins, i.e. Cdc42‐GTP, p21‐activated kinase1 (PAK1) and MRCK (myotonic dystrophy kinase‐related Cdc42‐binding kinase) α were successfully selected in the low micro‐ to nanomolar range using non‐systematic evolution of ligands by exponential enrichment (SELEX) with at least three orders of magnitude enhancement from their respective bulk affinity of naïve DNA library. In the non‐SELEX procedure, CE was used as a highly efficient affinity method to select aptamers for the desired molecular target through a process that involved repetitive steps of partitioning, known as non‐equilibrium CE of equilibrium mixtures with no PCR amplification between successive steps. Various non‐SELEX conditions including the type, concentration and pH of the run buffer were optimized. Other considerations such as salt composition of selection buffer, protein concentration and sample injection size were also studied for high stringency during selection. After identifying the best enriched aptamer pool, randomly selected clones from the aptamer pool were sequenced to obtain the individual DNA sequences. The dissociation constants (K_d) of these sequences were in the low micromolar to nanomolar range, indicating high affinity to the respective proteins. The best binders were also subjected to sequence alignment to generate a phylogenetic tree. No significant consensus region based on approximately 50 sequences for each protein was observed, suggesting the high efficiency of non‐SELEX for the selection of numerous unique sequences with high selectivity.     

Photo-activated pheophorbide a inhibits the growth of prostate cancer cells

Pheophorbide a (PhA) was identified as a photosensitizer to exert cytotoxicity on tumor cells. However, the efficacy of this compound on the treatment of prostate cancer remains unknown. The aim of this study was to evaluate the photodynamic effect of PhA on prostate cancer cells. Cellular uptake of PhA and cell viability after photo-activation was studied in LNCaP prostate cancer cells. The corresponding production of reactive oxygen species within cells was determined after photodynamic therapy (PDT). Our results showed that the uptake of PhA into LNCaP cells was in a time-dependent manner and the cytotoxicity of PhA-PDT was photosensitizer dose- and light dose-dependent. The intracellular reactive oxygen species was remarkably induced after PDT treatment, which was responsible for the inhibition effect on prostate cancer cells. This is the first report to evaluate the photodynamic effect of PhA on prostate cancer. Our findings demonstrate that PhA-PDT may be a potentially promising treatment for localized prostate cancer, which can be a therapeutic option after the failures of radiotherapy and hormone therapy.     

A grid based particle method for solving partial differential equations on evolving surfaces and modeling high order geometrical motion

We develop numerical methods for solving partial differential equations (PDE) defined on an evolving interface represented by the grid based particle method (GBPM) recently proposed in [S. Leung, H.K. Zhao, A grid based particle method for moving interface problems, J. Comput. Phys. 228 (2009) 7706–7728]. In particular, we develop implicit time discretization methods for the advection–diffusion equation where the time step is restricted solely by the advection part of the equation. We also generalize the GBPM to solve high order geometrical flows including surface diffusion and Willmore-type flows. The resulting algorithm can be easily implemented since the method is based on meshless particles quasi-uniformly sampled on the interface. Furthermore, without any computational mesh or triangulation defined on the interface, we do not require remeshing or reparametrization in the case of highly distorted motion or when there are topological changes. As an interesting application, we study locally inextensible flows governed by energy minimization. We introduce tension force via a Lagrange multiplier determined by the solution to a Helmholtz equation defined on the evolving interface. Extensive numerical examples are also given to demonstrate the efficiency of the proposed approach.     

Transport properties of Pr0.7Ca0.3MnO3/Nb:SrTiO3 heterojunctions

Current–voltage (J–V) characteristics of epitaxial hetero-junctions composed of Pr_0.7Ca_0.3MnO₃ and Nb:SrTiO₃ were studied under forward and reversed bias conditions. Detailed analysis showed that the J–V characteristics of these heterojunctions can be well-fitted by the thermally-assisted tunnelling model. While the dielectric constant of Nb:SrTiO₃ extracted under the forward bias was about one order of magnitude smaller than that of bulk SrTiO₃, the value obtained under reverse bias was very close to that of the bulk SrTiO₃. The result can be explained by the existence of an interface layer on the Nb:SrTiO₃ substrate with a smaller effective dielectric constant. The current finding suggested that the properties of interface layer should be taken into account in order to accurately simulate the J–V characteristics of such heterojunctions.     

Finite quotients of the automorphism group of a free group

The automorphism group AutF_n of a free group F_n of rank n acts on the product of n copies of a group G by substituting n elements of G into the words defining an automorphism of the free group. This gives rise to an antihomomorphism from AutF_nto a permutation group. We determine this antihomomorphic image of AutF_n when G is the semidirect product Z_p x Z_q     

Minimizing maximum lateness of jobs in inventory constrained scheduling

This paper focuses on a machine scheduling problem having applications in truck scheduling at transshipment terminals. Jobs increase and decrease, respectively, the level of a central inventory. Naturally, jobs decreasing the inventory level can be processed only if the level of the inventory is high enough not to drop below zero. We consider the problem to find a schedule for jobs such that the maximum lateness among all jobs is minimized. We develop properties of optimal solutions, lower bounds, and heuristic methods in order to find upper bounds. These are incorporated in four branch and bound algorithms that are based on fixing sequences of jobs in forward or backward direction in two different types of representations. By means of a computational study, we compare these approaches with each other in order to show their efficiency.     

Group and individual heterogeneity in a stochastic frontier model: Container terminal operators

Container ports are a major component of international trade and the global supply chain. Hence, the improvement of port efficiency can have a significant impact on the wider maritime economy. This paper deconstructs a representation in the existing literature that neglects the heterogeneity of individual and group-specific terminal operators. In its place, we present a hierarchical model to make a connection between efficiency and terminal operator group characteristics. The paper develops a stochastic frontier model that controls not only individual heterogeneity but also group-specific variations. The model decomposes the total stochastic derivation from the frontier into inefficiency, individual heterogeneity, group-specific variations, and noise components, with the estimation being performed using Markov chain Monte Carlo simulations. The validity of the model is tested with a panel of container terminal operator data from 1997-2004. Our findings show that terminal operator groups are important in promoting terminal efficiency at the global level, and that the operators with stevedore backgrounds show a higher efficiency than carriers.