Quantitative in vivo microsampling for pharmacokinetic studies based on an integrated solid-phase microextraction system

An integrated microsampling approach based on solid-phase microextraction (SPME) was developed to provide a complete solution to highly efficient and accurate pharmacokinetic studies. The microsampling system included SPME probes that are made of poly(ethylene glycol) (PEG) and C18-bonded silica, a fast and efficient sampling strategy with accurate kinetic calibration, and a high-throughput desorption device based on a modified 96-well plate. The sampling system greatly improved the quantitative capability of SPME in two ways. First, the use of the C18-bonded silica/PEG fibers minimized the competition effect from analogues of the target analytes in a complicated sample matrix such as blood or plasma samples, which is a common problem associated with solid coating SPME fibers for quantitative analysis. Moreover, the C18-bonded silica/PEG fibers provide high sensitivity and a large dynamic range that covers the possible sample concentration range during diazepam administration and elimination. Second, the kinetic calibration method offers more accurate quantitation than the calibration curve method for in vivo SPME, because it compensates for convection and matrix effects during sampling. Therefore, it is especially suitable as a fast sampling technique for pre-equilibrium SPME. Furthermore, with the high-throughput desorption device, the integrated system offers compactness and high efficiency. Its feasibility for in vivo sampling was demonstrated by monitoring diazepam pharmacokinetics and validated by conventional chemical assays and equilibrium SPME. In addition, we propose a simple method to determine the apparent distribution constant between an SPME fiber and a blood matix (Kfs) and the distribution constant between an SPME fiber and a pure PBS buffer sample matrix (Kfb). As a result, both total and free concentrations of the drug and its metabolites can be detected simultaneously. Accordingly, the binding constants to the blood matrix can be obtained, which are of special significance for clinical diagnosis and drug discovery.     

Dynamic kinetic capillary isoelectric focusing: a powerful tool for studying protein-DNA interactions

A new method called dynamic kinetic capillary isoelectric focusing (DK-CIEF) is presented for the study of protein-DNA interactions. The method is based on CIEF with laser-induced fluorescence-whole column imaging detection in which protein-DNA complexes are separated with spatial resolution while dissociations of the complexes are dynamically monitored using a CCD camera with temporal resolution. This method allows for the discrimination of different complexes and the measurement of the individual dissociation rate constants.     

Cysteine-specific, covalent anchoring of transition organometallic complexes to the protein papain from Carica papaya

Site-directed and covalent introduction of various transition metal-organic entities to the active site of the cysteine endoproteinase, papain, was achieved by treatment of this enzyme with a series of organometallic maleimide derivatives specially designed for the purpose. Kinetic studies made it clear that time-dependent irreversible inactivation of papain occurred in the presence of these organometallic maleimides as a result of Michael addition of the sulfhydryl of Cys25. The rate and mechanism of inactivation were highly dependent on the structure of the organometallic entity attached to the maleimide group. Combined ESI-MS and IR analysis indicated that all the resulting papain adducts contained one organometallic moiety per protein molecule. This confirmed that chemospecific introduction of the metal complexes was indeed achieved. Thus, three novel reagents for heavy-atom derivatization of protein crystals, which include ruthenium, rhenium and tungsten, are now available for the introduction of electron-dense scatterers for phasing of X-ray crystallographic data.     

Kapazit?t der verschiedenen Teile des Verdauungsapparates des Rothirsches (Cervus elaphus L.)

Zusammenfassung An drei Hirschen wurden die Gewichte und Volumina des Verdauungstraktes festgestellt. Die absolute und relative Kaparität, das Verhältnis der Kapazität zum Körpergewicht sowie die Länge und Gewichte der einzelnen Verdauungsabschnitte und deren Inhalt sind in Tabellen angegeben.

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Summary In case of three red deer weights and volumes of the digestive tract were stated. Absolute and relative capacity, the relation of capacity to body weights as well as length and weights of different digestive sections and their contents have been stated in tables.

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Résumé Sur trios cerfs on déterminait les poids et les volumes de l'appareil de la digestion. La capacité absolue et relative, la rélation de la capacité au poids du corps, ainsi que la longueur et les poids des diffrentes sections digestives et leur contenu sont présentés en tableaux.

     

A technique to compare polythiophene solid-state dye sensitized TiO2 solar cells to liquid junction devices

In this communication, we report on a technique to fabricate solid-state polythiophene-based dye sensitized solar cells (DSSCs) that can be directly compared to analogous liquid junction devices. The device configuration is based on non-porous TiO₂ thin films and one of the three undoped polythiophene hole conductors: poly[3-(11 diethylphosphorylundecyl) thiophene], P3PUT, poly(4-undecyl-2,2′-bithiophene), P4UBT, or poly(3-undecyl-2,2′-bithiophene), P3UBT. These polymers were spin coated and cast from organic solutions onto the TiO₂ films. The dense TiO₂ thin films (ca. 30 nm) were deposited on conductive glass via facile spray pyrolysis and sol–gel techniques. After that, cis-(SCN)₂ Bis(2,2′ bipyridyl-4,4′-dicarboxylate) ruthenium(II) (a.k.a. Ru N3 dye) was adsorbed on the TiO₂ surface, and the polythiophenes were utilized as hole conductors in a simplified solar cell geometry. The results were compared to the control DSSC device made with dense TiO₂ and a liquid electrolyte, or 2,2′,7,7′-tetrakis(N,N-di-p-methoxyphenylamine)-9,9′-spirobifluorene (a.k.a. Spiro-MeOTAD). The polythiophenes exhibited bandgaps in the range 1.9–2.0 eV, and HOMO energy levels of approximately 5 eV (vs. vacuum). The P3PUT DSSC device exhibited an AM1.5 V_OC=0.8 V, a J_SC=0.1 mA/cm², as well as an IPCE=0.5–1%. The AM1.5 short-circuit photocurrents and quantum efficiencies for DSSCs made with the polythiophenes, the Spiro-MeOTAD and the standard liquid electrolyte (I⁻/I₃⁻) were found to be identical within the limits of experimental uncertainty and reproducibility. Our results indicate that a solid-state replacement to the liquid junction is not necessarily limited by the fundamental aspect of hole transfer, one of the three fundamental aspects that must be met for an efficient DSSC. Rather than suggest that P3UBT or P4UBT could be used to create efficient “organic solar cells” with the exclusion of the Ru dye, we suggest that transparent thiophene compounds could be attractive candidates for high-surface area solid-state DSSCs, and that the technique presented can be applied to other hole conductors. It can allow a verification of one of the things necessary for the DSSC, so that parallel studies using high-surface area materials can proceed with confidence.     

Functional Validation of cas9/GuideRNA Constructs for Site-Directed Mutagenesis of Triticale ABA8′OH1 loci

Cas endonuclease-mediated genome editing provides a long-awaited molecular biological approach to the modification of predefined genomic target sequences in living organisms. Although cas9/guide (g)RNA constructs are straightforward to assemble and can be customized to target virtually any site in the plant genome, the implementation of this technology can be cumbersome, especially in species like triticale that are difficult to transform, for which only limited genome information is available and/or which carry comparatively large genomes. To cope with these challenges, we have pre-validated cas9/gRNA constructs (1) by frameshift restitution of a reporter gene co-introduced by ballistic DNA transfer to barley epidermis cells, and (2) via transfection in triticale protoplasts followed by either a T7E1-based cleavage assay or by deep-sequencing of target-specific PCR amplicons. For exemplification, we addressed the triticale ABA 8′-HYDROXYLASE 1 gene, one of the putative determinants of pre-harvest sprouting of grains. We further show that in-del induction frequency in triticale can be increased by TREX2 nuclease activity, which holds true for both well- and poorly performing gRNAs. The presented results constitute a sound basis for the targeted induction of heritable modifications in triticale genes.     

Neural network for predicting the performance of credit card accounts

This paper reports the interim results of an experimental project using neural networks as a decision support tool for credit card risk assessment within a major bank. Two prototype neural network systems have been developed: one which emulates the decisions of the current risk assessment system, and another which attempts to predict the performance of credit card accounts based on the accounts historical data. This paper focuses on the development of the neural network model for credit card account performance prediction. The study has shown that such a tool can help in discovering the potential problems with credit card applicants at the very early stage of the credit account life cycle.     

Measurements of Water-Air Flow Phenomena in a Chamber with a Rotating Shaft

The paper concerns the phenomena of the fluid film that occurs in the bearing chamber of an aircraft engine.The geometry of the system includes two concentric cylinders,between which there is a mixture of oil and air.The fluid circulates in a closed circuit.The rotary movement of the inner cylinder causes creation of the fluid film on the walls of the chamber.The article proposes a measurement method that is allowing observation of this fluid film and,in particular,analysis of the movement of the air bubbles occurring in the film.An own model of the bearing chamber with transparent walls was constructed for the research.For the investigation,water was used instead of oil.Observation of the behaviour of the flow was possible thanks to video recordings made with the use of a fast-capture camera.The results presented in the paper include velocity magnitudes of the air bubbles in the fluid film in dependence on the rotational speed of the shaft and water volume fraction and with a range from 0.37 to 0.91 m/s.The results presented in this article can be used for the bearing chamber numerical models validation.     

The Influence of Process Parameters on the Low-Cycle Fatigue Properties of 316L Steel Parts Produced by Powder Bed Fusion

Metallurgical and Materials Transactions A - In this paper, the influence of the additive manufacturing (AM), powder bed fusion (PBF) process parameters on the low-cycle fatigue (LCF) properties of...