Solid-phase microextraction and gas chromatography-mass spectrometry for the determination of polycyclic aromatic hydrocarbons in environmental solid matrices.

A solid-phase microextraction (SPME) and gas chromatography-mass spectrometry method for determining polycyclic aromatic hydrocarbons (PAHs) in environmental solid matrices is developed. Investigated matrices include seaweed (Undaria pinnatifida and Himanthalia elongata), humic substances (isolated from a wetland out-flow and purchased from Aldrich), and soil. Optimal conditions for a good SPME efficiency of 16 hydrocarbon compounds are obtained using a 100- micro m polydimethylsiloxane fiber directly immersed in aqueous carrier medium. The method is remarkable for presenting short extraction times and considerably high sensitivities. The SPME results obtained by using internal calibration give the total analyte concentration based on the identical partitioning behavior of native and spiked pollutants. The detection limits range from 0.001 to 0.1 mg of PAH per kilogram of dry matrix. SPME external calibration provides information regarding freely dissolved analytes. The detection limits range from 0.001 to 0.05 micro g of PAH per liter of carrier medium. The SPME with external calibration procedure can be applied to measure free concentrations of a target compound spiked into a carrier medium and onto a matrix. Based on a comparison of results obtained for the two samples, the partitioning of the target analyte between the matrix and the carrier medium is calculated.

Visualization of agonist-induced internalization of histamine H2 receptors.

Histamine H2 receptors were tagged at the N-terminus with the eight amino acid Flag epitope to allow the immunological identification of the receptor peptide with the monoclonal anti-Flag M2 antibody. The introduction of the epitope did not modify the binding of several H2 ligands to the H2 receptor, nor the ability of histamine to stimulate the H2 receptor mediated cAMP production in HEK-293 cells. Western blots revealed a major protein band of 57 +/- 1 kDa, whereas a second band of 31 +/- 1 kDa was probably the result of a proteolytic breakdown of the 57 kDa band. Immunofluorescence measurements of stably transfected HEK-293 cells revealed the presence of anti-Flag-immunoreactivity in the plasma membrane. This immunoreactivity completely disappeared after a one hour treatment with histamine. The receptor internalization was reversible and blocked by the endocytosis inhibitor phenylarsine oxide. Forskolin did not induce H2 receptor internalization, indicating that histamine causes H2 receptor internalization via a cAMP-independent pathway.

[Radial and proximo-distal distribution of hydrolytic enzymes in the small intestine at rest and during digestion]. / Radial'noe i proksimo-distal'noe raspredelenie gidroliticheskikh fermentov v tonkoi kishke v pokoe i pri pishchevarenii.

Alpha-amylase was present in considerable amount in the chyme in rats both at rest and during digestion, in the latter case its level being increased. Activity of intrinsic intestinal enzymes dominated in the homogenate of the mucosa. Transition from fasting to feeding and digestion increased the level of alkaline phosphatase and dipeptidase activities in the homogenate of the mucosa. The data obtained suggest the final stages of biopolymers' hydrolysis to be localized in various enterocyte structures. The surface mucus, owing to absorbed pancreatic enzymes, binding proteins and other types of physiologically active molecules, is assumed to play the role of a specific molecular filter and take part in the initial stages of digestion.