Applications of aptamers in cancer cell biology

Identifying cells associated with specific disease states is critically important for the early detection and diagnosis of cancer. To facilitate this task, molecular probes, which bind biomarkers that are either specifically or differentially expressed in diseased cells relative to healthy cells, provide a simple and effective method. This review focuses on the use of DNA aptamers as molecular probes for cancer cells. These aptamers are created by means of the cell-based Systematic Evolution of Ligands by EXponential enrichment (SELEX) process, which uses whole disease cells as targets. We describe at length the steps of the cell-SELEX process and discuss several applications for the aptamers, including profiling leukemia patient samples and discovering cell-surface cancer biomarkers. We conclude with a discussion of an aptamer-conjugated nanoparticle enrichment and detection scheme.     

Real-time quantitative imaging of submolecular layers

Using a recent optical contrast method, real-time and quantitative imaging of submolecular layers was performed with the help of a simple optical microscope. The measuring technique is exposed and documented by three examples. In particular, it allowed label-free detection of peptide-antibody binding interactions with 50 pg/mm2 sensitivity while keeping full optical lateral resolution.     

聚多巴胺表面改性技术在分离科学领域中的应用

聚多巴胺作为新型仿生材料,具有制备过程简单、环保和适用面广（可用于各种类型基质表面改性）等优点,已被广泛应用于化学、生物医学、药学、传感器和电池制造等领域。在分离科学领域,聚多巴胺不仅可用于制备色谱固定相,也可用于制备新型的富集材料。该文对聚多巴胺的形成机理研究现状进行了简单介绍,主要综述了近年来聚多巴胺在色谱分离和富集领域的应用,包括毛细管电泳/电色谱、液相色谱、分子印迹固相萃取、分散固相微萃取和固相微萃取等技术领域。     

Applications of the deferred standard technique in process gas chromatography

Three applications of the deferred standard (DS) technique in on-line gas chromatographic analysis with a rotary injection valve are reported. It is demonstrated how both absolute and relative measurements are affected by several deviations of the experimental conditions, such as atmospheric pressure, sample injection during either flow or stopped-flow conditions and instrumental settings (carrier gas flow-rate, column oven temperature, detector bridge current) of the analytical system. It is shown that the relative DS concept provides an excellent means of improving the reliability of the analytical equipment. Under standard conditions, accurate results were obtained with a relative standard deviation < 1%. Significant deviations in the instrumental settings affected the accuracy of the final analytical results by less than 4%. It is further shown how the DS technique can be applied to obtain early information on maintenance requirements of the system. A diagnostic table is included that relates observed changes with time of the signals, obtained with a thermal conductivity detector, to possible maintenance requirements.     

Direct submolecular scale imaging of mesoscale molecular order in supported dipalmitoylphosphatidylcholine bilayers

Supported dipalmitoylphosphatidylcholine (DPPC) bilayers are widely used membrane systems in biophysical and biochemical studies. Previously, short-range positional and orientational order of lipid headgroups of supported DPPC bilayers was observed at room temperature using low deflection noise frequency modulation atomic force microscopy (FM-AFM). While this ordering was supported by X-ray diffraction studies, it conflicted with diffusion coefficient measurements of gel-phase bilayers determined from fluorescence photobleaching experiments. In this work, we have directly imaged mica-supported DPPC bilayers with submolecular resolution over scan ranges up to 146 nm using low deflection noise FM-AFM. Both orientational and positional molecular ordering were observed in the mesoscale, indicative of crystalline order. We discuss these results in relation to previous biophysical studies and propose that the mica support induces mesoscopic crystalline order of the DPPC bilayer at room temperature. This study also demonstrates the recent advance in the scan range of submolecular scale AFM imaging.     

Applications of laboratory robotics in spectrophotometric sample preparation and experimental optimization

Optimal experimental conditions for spectrophotometric solution-preparation procedures for determinations of iron(III) and phosphate were determined by a completely robotic application of the SIMPLEX algorithm, factorial design methods, and response surface fitting. The overall reliability of the approach, the precision observed and its significance are discussed.     

A new optimization strategy for gaseous phase sampling by an internally cooled solid-phase microextraction technique

This study describes a new optimization strategy for internally cooled solid-phase microextraction based on a multivariate approach. The coating temperature was changed in an extraction while manipulating the extraction times to improve the extraction of compounds with different volatilities. Polycyclic aromatic hydrocarbons (PAHs) and phthalic acid esters (PEs) and adipate were used as model compounds in this study. The optimization strategy was in two steps: (1) multivariate optimization of extraction time and initial coating temperature and (2) multivariate optimization of total extraction time and the time required to cool the coating to a lower temperature as determined in step 1. The observed analytical response in relation to the coating temperature was found to be dependent on the analyte volatility and size. The optimized extraction condition for PEs was 23 min extraction while maintaining the coating at 140°C, followed by 7 min of cooling the coating at 10°C. For the PAHs the coating temperature was maintained at 60°C for the first 20 min and at 5°C in the last 20 min of extraction. Comparisons have been made between the proposed optimized conditions with the conventional internally cooled fiber approach and the results thoroughly discussed. The proposed optimization strategy was found to be more effective for all the analytes, especially for the semi-volatiles, compared to the conventional method.     

A hybrid technique of orthonormality constrained orbital optimization in SCF calculations

A recently proposed orthonormality constrained orbital optimization technique is operationally modified further by coupling it to a gradient biased method, namely the steepest descent procedure of McWeeny. The hybrid technique developed in this way is shown to have better convergence properties in closed and unrestricted open-shell calculations. The technique can be adapted to MCSCF procedures as well. The important role played by "orbital symmetries" in the operation of the method is analysed. Similarities and differences of the present method with the orthogonal gradient method are pointed out. Possible avenues of circumventing convergence difficulty that one may encounter in pathological cases, particularly in ab initio calculations involving extended basis set, are suggested.     

Applications of Padé approximants of type II in partitioning technique

Padé approximants of type II have been applied to solve equations of the type F(z) = 0. The method is compared with the first-order iteration procedure, the Aitken-Samuelson formula, and the Newton-Raphson tangential method. As a test example the partitioning technique in its most simple form is applied to the Hamiltonian of a rigid symmetric-top molecule in a static electric field. The proposed algorithm is found to be superior to the first-order procedure and the Aitken-Samuleson formula, and at least as effective as the Newton-Raphson method.     

The optimization of an ICP injection technique and the application to the direct analysis of small-volume serum samples

The optimization and the analytical properties of an injection technique for the analysis of small-volume samples (50–200 μl) by inductively coupled plasma atomic emission spectroscopy (ICP-AES) are described. Samples are injected into a small funnel connected to a concentric glass nebulizer. A 3 kW argon/nitrogen ICP with power stabilization is used as excitation source. When operating the nebulizer at an argon pressure of 5 bar, relative detection limits for calcium, copper, iron, magnesium and zinc (0.2–50 $\text{ng}\text{ml}$) are a factor of 2 to 10 higher when compared with ICP methods using continuous nebulization. However, the full power of detection of the injection method is obtained at a 50 μl sampling volume. Matrix effects caused by sodium salt concentrations of 5 $\text{g}\text{l}$ are lower than 10%. Relative standard deviations s_r,(I_X) ranged from 0.03 to 0.07. The method was applied to the sequential determination of trace elements (copper, iron and magnesium) in human serum samples after a 1 + 4 dilution with Herrmann solution. The accuracy of the method is illustrated by the analysis results for calcium, copper, iron, magnesium and zinc in a series of test serum samples.     

Applications of imaged capillary isoelectric focussing technique in development of biopharmaceutical glycoprotein-based products

CE-based methods have increasingly been applied to the analysis of a variety of different type proteins. One of those techniques is imaged capillary isoelectric focusing (icIEF), a method that has been used extensively in the field of protein-based drug development as a tool for product identification, stability monitoring, and characterization. It offers many advantages over the traditional labor-intensive IEF slab gel method and even standard cIEF with on-line detection technologies with regard to method development, reproducibility, robustness, and speed. Here, specific examples are provided for biopharmaceutical glycoprotein products such as mAbs, erythropoietin (EPO), and recombinant Fc-fusion proteins, though the technique can be adapted for many other therapeutic proteins. Applications of iCIEF using a Convergent Bioscience instrument (Toronto, Canada) with whole-field imaging technology are presented and discussed. These include a quick method to establish an identity test for many protein-based products, product release, and stability evaluation of glycoproteins with respect to charge heterogeneity under accelerated temperature stress, different pH conditions, and in different formulations. Finally, characterization of glycoproteins using this iCIEF technology is discussed with respect to biosimilar development, clone selection, and antigen binding. The data presented provide a "taste' of what icIEF method can do to support the development of biopharmaceutical glycoprotein products from early clone screening for better product candidates to characterization of the final commercial products.     

Two-dimensional Raman study of the submolecular, electric field-induced reorientation of a nematic liquid crystal

Two-dimensional Raman scattering is presented as a technique for the monitoring of electric field-induced, submolecular reorientation in liquid crystals. The motions of the flexible part and the rigid core of 4-pentyl-(4-cyanophenyl)cyclohexane (PCH5) are independently monitored in response to both step and oscillatory electric fields. Step voltage experiments show that the flexible group reorients before the rigid core. Also, oscillatory electric field experiments demonstrate that the flexible and rigid groups reorient asynchronously. In fact, at periodicities that are shorter than the bulk reorientation times, it is observed that the reorientation of the flexible part is amplified, while the motion of the rigid core is inhibited. The data suggest that the flexible group possesses a small, local dielectric anisotropy that can couple with the electric field to induce an independent, cooperative reorientation when the mobility of the rigid core is restricted.     

Atomic force microscope images of the surfaces of aspirin crystals at submolecular resolution.

The atomic force microscope has been developed and used to image arrays of molecules at the (001) and (100) faces of aspirin crystals in water and in air. Lattice spacings composed of methyl groups and the part of the phenyl groups on the surface of the (001) in water, are consistent with X-ray diffraction data. The surface of (100) face which shows most perfect cleavage in bulk, is more difficult to image. This initial success in imaging at drug crystal surfaces clarified the different structural behavior at the submolecular level for three crystal faces, and the close relationship to the differences in the dissolution velocity.     

Applications of the optical trapping technique to analyze chemical reactions in single emulsion particles

The applicability of the single beam gradient force optical trap and the optical levitation technique to investigate dynamic processes in microparticles is examined. The device allows to follow chemical reactions, like the emulsion polymerization and the ester hydrolysis reaction, in single emulsion droplets. Furthermore the usability of optical traps to investigate living cells is demonstrated.     

Applications of the optical trapping technique to analyze chemical reactions in single emulsion particles

The applicability of the single beam gradient force optical trap and the optical levitation technique to investigate dynamic processes in microparticles is examined. The device allows to follow chemical reactions, like the emulsion polymerization and the ester hydrolysis reaction, in single emulsion droplets. Furthermore the usability of optical traps to investigate living cells is demonstrated.     

Shielding optimization in an in vivo activation analysis setup

A neutron irradiation setup for in vivo activation analysis was characterized to optimize its spectrum measurements. In order to have a reliable spectrum, some shields and filters were defined to eliminate as much background possible. Since there were different types of background sources, different states were studied and necessary shields were employed to optimize the spectrum. To use a neutron shield, boron carbide layers (3 cm) around 3 × 3 in. sodium iodine detectors were employed. To prevent γ-ray background, bismuth (2 cm) is used under the detectors.