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BACKGROUND: As emergency ultrasound use explodes around the world, it is important to realize the path its development has taken and learn from trials and tribulations of early practitioners in the field.METHODS: Approaches to education, scanning, documentation, and organization are also described.RESULTS: Machines have reduced in price and once purchased further material costs are low. Staffing costs in terms of training, etc have yet to be assessed, but indications from elsewhere are that these are low. Length of stay in the emergency department dramatically decreases, thus increasing patient satisfaction while maintaining an even higher standard of care.CONCLUSION: Emergency screening ultrasound is now a nationally accepted tool for the rapid assessment of the emergency patient.

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Solid-supported membranes immobilized on gold electrodes were used to detect and characterize the spontaneously inserting anion-selective protein channel (Clavibacter anion channel, CAC) present in the culture fluid of Clavibacter michiganense ssp. nebraskense. Three different membrane systems varying in the composition of the first chemisorbed monolayer were investigated by means of impedance spectroscopy. Conductance changes of the immobilized lipid membranes were sensitively detected after adding the culture fluid of the bacteria to the solid-supported membranes, indicating that the relative change in conductance is largest if the lipid layer is attached to the surface via a flexible lipid anchor. Variation in the d.c. potential revealed that CAC exhibits a voltage dependence in these tethered membranes which can be described by an exponential function in accordance with previous results obtained from patchclamp measurements and impedance analysis. The addition of an inhibitor that selectively blocks anion channels abolished the channel conductance almost completely, indicating that the increased conductivity can be attributed to the specific insertion of the CAC. A linear dependence of the channel conductance on the chloride concentration was found, which was modulated by the charges of the second lipid monolayer. The results demonstrate that tethered lipid membranes on gold surfaces in conjunction with impedance spectroscopy allows one to monitor and characterize water-soluble spontaneously inserting channels, providing an effective means to probe for bacterial toxins.

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The quantification of small molecules in aqueous solution by surface bound supramolecular host molecules is an important goal in the research field of chemo- and biosensor development. In this paper we present an attempt to quantify the interaction of different charged guest molecules with chemisorbed monothiolated ß-cyclodextrin monolayers by means of impedance spectroscopy in the presence of the redox couple [Fe(CN)(6)](3)(-)/[Fe(CN)(6)](4)(-). Self-assembled monolayers of mercaptopropane-N-mono-6-deoxy-ß-cyclodextrin amide (MPA-CD) on gold surfaces were formed with coverage of 99-100%. The inclusion of charged guest molecules was detected by monitoring the changes in the charge-transfer resistance, which is sensitive to the surface charge density in terms of repulsion or attraction of the redox active ions. Adsorption of positively charged 1-adamantanamino hydrochloride (1-ADHC) led to a considerable increase in the charge-transfer resistance, whereas the inclusion of both negatively charged 1-adamantanecarboxylic acid (1-ADC) and 2-(p-toluidinyl)naphthalene-6-sulfonate (2,6-TNS) caused a decrease. Applying the Frumkin correction to obtain the surface charge density and the Gouy-Chapman-Stern theory to account for the electrochemical double layer, we were able to quantify the binding of the charged guest molecules in terms of binding isotherms. The isotherms display a distinct two step adsorption process probably owing to the presence of two energetically different binding sites on the surface. Complete reversibility of the binding process of the guest molecules could be demonstrated by the addition of ß-cyclodextrin in solution, which allowed the reuse of the functionalized surfaces.

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Overexpression of P-glycoprotein in tumor cells can represent a severe drawback for cancer chemotherapy. P-glycoprotein acts as an efflux transporter for a variety of chemotherapeutic agents. It is encoded by multidrug resistance (mdr) genes of the subfamily 1 in humans (MDR1) and rodents (mdr1a and 1b). Because mdr1 gene expression is inducible in cultured rat hepatocytes and in rat liver with chemical carcinogens such as 2-acetylaminofluorene or aflatoxin B1, which form DNA-binding electrophiles during their metabolism, we investigated whether the DNA-damaging chemotherapeutic drug mitoxantrone may induce multidrug resistance in rodents and in hepatocytes in primary culture. In H4IIE rat hepatoma cells stably transfected with a luciferase construct containing the rat mdr1b promoter, mitoxantrone caused a concentration-dependent increase in promoter activity. Mdr1 gene expression in cultured rat hepatocytes was enhanced at mitoxantrone concentrations greater than or equal to 0.1 microM and in mouse hepatocytes at 5 microM. In hepatocytes from both species, a correlation was found between mdr1 induction and the inhibition of protein synthesis. In vivo, mitoxantrone was a very powerful inducer of mdr1 gene expression in rat liver and small intestine. In rat kidney, induction of mRNA was lower, and a marginal effect was seen in lung. In contrast with rats, no significant induction of mdr1 gene expression was obtained in mouse liver. Probably as a consequence of inhibition of protein synthesis, mitoxantrone did not lead to a pronounced elevation of P-glycoprotein levels in rat liver and kidney.

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P-Glycoprotein the multidrug resistance (mdr) efflux transporter is encoded by class 1 mdr genes (mdr1) in humans and rodent species. In rat liver and in rat hepatocytes in primary culture, expression of mdr1 genes can be induced with the carcinogenic aromatic amine 2-acetylaminofluorene (2-AAF). As a consequence, increased P-glycoprotein levels led to an accelerated efflux of vinblastine from the hepatocytes and to resistance towards vinblastine-mediated cytotoxicity. N-Hydroxylation, an obligatory initial step in the activation of 2-AAF into electrophilic DNA-binding metabolites is catalyzed predominantly by cytochrome P450 (CYP)1A2, an isozyme present in normal rat liver. In rat hepatocytes in primary culture, mdr1 induction with 2-AAF could be inhibited by addition of the CYP1A-inhibitor alpha-naphthoflavone, indicating the requirement for metabolic conversion of 2-AAF to act as an inducer of mdr1 gene expression. Both N-hydroxy-2-AAF and the mutagenic 2-AAF derivative N-acetoxy-2-AAF (AAAF) were more potent than 2-AAF as mdr1 inducers. mdr1 induction also decreased when deacetylation of AAAF, which strongly accelerates its conversion into a mutagen, was inhibited with paraoxon. Furthermore, rat liver epithelial cells stably transfected with mouse CYP1A2 showed inducibility of mdr1 gene expression with 2-AAF, whereas the parental cell line, which is devoid of CYP1A2 activity, did not. These findings indicate that electrophilic metabolites formed during 2-AAF or AAAF metabolism are responsible for mdr1 induction in rat hepatocytes. The increased mdr1 gene expression may reflect an adaptive cellular response to electrophiles which includes enhanced synthesis of P-glycoprotein aimed to protect the cell from further damage.

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