Pehrsonpersson3294

The developed HLAS-SPCE combines the use of a new sample preparation procedure for volatiles with, as far as we know, the first determination of formaldehyde (as the derivative product, DDL) on unmodified SPCEs, offering a promising alternative for the determination of formaldehyde in WBPs and other samples.A novel method is successfully tested for non-covalent imprinting. Conditions are used which practically exclude the formation of prepolymerization complexes. The template is cholesterol, and no so-called functional monomer is used. The polymers contain only an acrylic diester crosslinker. The porogen isopropanol prevents even hydrogen bonding between the template and the monomer in the prepolymerization solution. Despite of these apparently very disadvantageous conditions, appreciable imprinting factors for cholesterol and imprinted selectivity against some other steroids are observed, similar to other cholesterol MIPs with proven analytical usefulness.Hydrogen peroxide (H2O2), an important member of the family of reactive oxygen species (ROS), has a significant impact on cell signal transduction, energy conversion and immune responses of living organisms. Therefore, accurate detection of the content of H2O2 in living cells is of vital importance. In this paper, we report on the synthesis of a novel colorimetric and near-infrared fluorescent probe HAA, a heterocyclic aromatic amine with acetyl group for the specific detection of both exogenous and endogenous H2O2 in living cells. Our results show that the probe not only possesses high specificity and sensitivity, but also has advantages of low cytotoxicity and good biocompatibility. Theoretical computations elucidated the luminescence and quenching mechanism of HAA in the absence and presence of H2O2. In addition, HAA was applied to the determination of H2O2 in human serum and the imaging of endogenous H2O2 in living cells, during which it demonstrated excellent performance and good potential for future bioanalysis applications.Enzyme-linked immunoassay (ELISA) is highly specific and selective towards target molecules and is convenient for on-site detection. However, in many cases, lack of high sensitivity makes it hard to reveal a significant colorimetric signal for detecting a trace amount of target molecules. Thus, analytical instruments are required for detection, which limits the application of ELISA for on-site detection. In the present study, a highly sensitive and naked-eyed detectable colorimetric biosensor for chloramphenicol (CAP) was prepared by incorporating ELISA onto surfaces of microporous and nanofibrous membranes. The high specific surface areas of the nanofibers significantly increased the number of antibodies covalently linked onto the fiber surfaces and binding capacity of the sensor with antigens present in a sample. With such an integration, the sensitivity of the ELISA sensor was dramatically increased, and a trace number of targets could reveal a naked-eye detectable color. The immunoassay sensor exhibited a significant naked-eye distinguishable color to chloramphenicol (CAP) at 0.3 ng/mL. The successful design and fabrication of the nanofibrous membrane immunoassay sensor provide new paths towards the development of on-site inspection sensors without the assistance from any instrument.Profiling blood-brain barrier permeability of bioactive molecule is an important issue in early drug development, being a part of the optimization process of a compound's physicochemical properties, and hence pharmacokinetic profile. The study aimed to develop and optimize a new in vitro method for assessment of the compound's brain penetration. The tool is proposed as an alternative to the PAMPA-BBB (Parallel Artificial Membrane Permeability Assay for Blood-Brain Barrier) and based on a capillary electrochromatography (CEC) technique. It utilizes liposomes as structural substitutes of biological membranes, which are used as a capillary inner wall coating material. Following optimization of analysis conditions, migration times for a set of 25 reference drugs (mainly non-ionized in pH 7.4) were examined in a liposome coated capillary. On that basis, the retention factor (log k) was determined for each reference drug. Obtained log k values and experimentally received reference permeability parameters log BB (in vivo data) and log Pe (PAMPA-BBB data) were compared with one another. Correlation coefficients were calculated, giving comparable results for CEC log k/log BB and analogical PAMPA-BBB log Pe/log BB analyses. Approximate ranges of log k for the central nervous system (CNS) permeable (CNS(+)) and non-permeable (CNS(-)) drugs were established.Peroxynitrite (ONOO-), a highly reactive oxygen species generated by the reaction of nitric oxide and superoxide radical anion, is involved in numerous physiological and pathological processes in the human body. To identify important pathogenic mechanisms, it is crucial to develop a reliable tool for detecting peroxynitrite in living systems. In the present study, a new difluoroboron β-diketonate-based fluorescent probe for detecting exogenous and endogenous peroxynitrite in living systems was designed. The red emitting fluorophore can be synthesized in a simple three-step procedure. This probe reacts quickly and selectively with peroxynitrite and its detection limit is determined to be as low as 19.8 nM. It allows for clear imaging of peroxynitrite in RAW 264.7 cells and was successfully applied to visualize changes of intracellular peroxynitrite induced by reactive oxygen species inhibitors. This designed probe is an effective tool for investigating the physiological and pathological role of peroxynitrite in living cells.After the Fukushima Dai-ichi nuclear power plant disaster, the demand for a rapid method for the detection of environmental radioactivity increased drastically. Since the development of extraction chromatography using resins, analytical methods have advanced significantly in terms of simplicity and required labor. Herein, a home-made automated separation system that is applicable radio-extraction chromatographic separation techniques is reported. A simple, rapid, and high-throughput method was developed using this home-made automated separation system to analyze radiostrontium in seawater in emergency and routine situations. For emergency situations, radiostrontium in seawater is pre-concentrated on a cation exchange resin and consecutively purified using the Sr-resin. Fifty minutes are required for the purification of 90Sr in four samples (100 ml). The minimum detectable activity (MDA) for 90Sr is 0.2 Bq kg-1 at 100 min counting, with a recovery of 70% and counting efficiency of 95% in the scintillation mode. For routine monitoring, 90Y that is in equilibrium with 90Sr is first separated from the sample matrix using DGA. Treatment of 30 L of each seawater sample requires ~2 h. The MDA for this method is 0.3 mBq kg-1 at 400 min counting with a recovery of 70% and counting efficiency of 67% in the Cerenkov mode. Bafilomycin A1 By employing the developed method, the measured 90Sr in seawater collected from the coastal area of Korea is 0.92 ± 0.18 mBq kg-1, which is comparable to that reported previously. The measurements were obtained using a liquid scintillation counter, and the entire separation process was performed by employing the home-made separation system.β-Galactosidase (β-gal) is an important biomarker for primary ovarian cancers. Developing noninvasive bioimaging probes for studying the activity of β-gal is highly desirable for cancer diagnosis. Herein, a turn-on near-infrared (NIR) fluorescent probe， 2-((6-(((2S, 3R, 4S, 5R, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran -2-yl)oxy)-2,3-dihydro-1H-xanthen-4-yl)methylene)malononitrile named DXM-βgal, was rationally designed based on enzymatic reaction for the detection of β-gal activity both in vitro and in vivo. Upon incubating with β-gal, DXM-βgal displayed a significant fluorescence enhancement at 640 nm, accompanying by a color change of solution color from red to purple. DXM-βgal exhibited high selectivity and sensitively to β-gal with low limit of detection (2.92 × 10-4 U mL-1). Besides, based on its advantages of long-wavelength emission and excellent biocompatibility, DXM-βgal was successfully applied to imaging β-gal in living cells and zebrafish. Given these prominent properties, we believe that DXM-βgal will be a potential tool for investigating β-gal activity in biomedical research.Rapid quantification of pathogenic Salmonella Typhimurium (S. Typhimurium) and total bacteria in eggs is highly desired for food safety control. However, the complexity of egg matrix presents a significant challenge for sensitive detection of bacteria. In this study, a sample pretreatment protocol, including dilution, fat dissolution, protein degradation, filtration, and washing was developed to circumvent this challenge. A laboratory-built nano-flow cytometer (nFCM) that is hundreds of fold more sensitive than the conventional flow cytometer was employed to analyze individual bacteria upon nucleic acid and immunofluorescent staining. Eggs spiked with pathogenic S. Typhimurium and harmless Escherichia coli K12 (E. coli K12) were used as the model system to optimize the sample pretreatment protocol. S. Typhimurium and total bacteria in eggs can be quantified without cultural enrichment, and the whole process of sample pretreatment, staining, and instrument analysis can be accomplished within 1.5 h. The bacterial recovery rate upon sample pretreatment, detection limit, and dynamic range for S. Typhimurium in eggs were 92%, 2 × 103 cells/mL, and from 2 × 103 to 4 × 108 cells/mL, respectively. The as-developed approach can specifically distinguish S. Typhimurium from other bacteria and successful application to bacterial detection in eggs freshly purchased from supermarket and spoiled eggs upon inappropriate storage was demonstrated.In this study, a real-time target-recycled enzyme-free amplification strategy-based test (Trefas test) was developed for rapid, simple, isothermal, and highly sensitive microRNA (miRNA) detection. The Trefas relies on rationally designed sequence-specific hairpins (HPs, HP1 and HP2) and the strand displacement process completely free of environment-susceptible enzymes, enhancing the stability and reproducibility of the test. In the absence of target miRNA, the HP2, modified with a fluorophore and a quencher, maintains stem-loop structure so that the fluorescent signal is quenched. However, in the presence of target miRNA, the target miRNA is repeatedly used to trigger continuous HP1-HP2 hybridizations, restoring fluorescence due to the opening of HP2. The developed miR-21 real-time Trefas test exhibited a broad linear dynamic range of 1 pM to 1 μM and a detection limit of 0.58 pM for miR-21 detection in vitro. In particular, the high specificity of the developed miR-21 real-time Trefas test was prominently exhibited by discriminating single base differences in miRNA sequences. Finally, the expression level of miR-21 in the cell lines and clinical tissues was evaluated by the developed miR-21 real-time Trefas test, and the detection results were highly consistent with the results obtained by stem-loop RT-PCR. In summary, our developed test exhibited great potential for further application in biomedical research and early clinical diagnosis.