Importance of the Arg-Gly-Asp triplet in human thrombin for maintenance of structure and function

Site-directed mutagenesis was employed to assess the importance of the Arg-Gly-Asp triplet that comprises residues 197 to 199 in the B-chain of thrombin. Properties of the R197E and the D199E variants were compared with those of zeta-thrombin and the inactive S205A variant wherein the active site Ser is replaced by Ala. Relative to zeta-thrombin, the R197E thrombin variant under the assay conditions used exhibits 26% activity toward a small chromogenic substrate, 13% activity in the activation of protein C in the presence of thrombomodulin, < 3% activity in processing fibrinogen, and 1% activity in inducing platelet activation. Thus, the substrate specificity of thrombin was altered by the R197-->E replacement. The D199E variant was essentially inactive. It exhibited only 0.02% of the activity of thrombin toward the chromogenic substrate and its reactivity toward the active site-directed alkylating agent D-Phe-Pro-Arg-CH2Cl was 10,000-fold lower than that of thrombin. Like the inactive S205A thrombin variant, the D199E variant antagonized the interactions of thrombin with hirudin and thrombomodulin, but was a less effective antagonist. The dependence of the antagonism of the thrombin-thrombomodulin interaction on the concentration of D199E thrombin variant provided evidence suggesting the presence of two or more domains in thrombin that independently interact with their counterparts in thrombomodulin. Although the S205A thrombin variant antagonized the action of thrombin on platelets no such activity could be demonstrated for the D199E variant in the concentration range studied (< 800 nm). Comparison of the circular dichroism spectra of zeta-thrombin, the D199E, R197E, and S205A variants indicated that subtle differences in conformation exist between the D199E variant and the other thrombins. These differences in conformation might well account for the altered behavior of the D199E variant with respect to its interactions toward thrombomodulin, hirudin, and platelets.     

Has interleukin 2 modified the outcome of metastatic kidney cancer?

Mesenteric Cystic Lymphangiomas is a rare benign tumor. The first case was described after an autopsy in 1507 by Benivieni, an anatomist from Florence. In 1880 by Tillaux. Since then many cases have been described. None of these cases is associated with a non specific necrosis due to Bauhins' valvula compression; in the following lines we are going to describe this exceptional case.     

Echolocation — From range to outline segments

A ring of ultrasonic sensors mounted on a mobile robot is used to map a room. The sensors are modelled with an arc model: the object causing the reflection lies on an arc with radius equal to the range and arc angle equal to twice the beam angle. By fusing sensing with motion, a surface is displayed as a sequence of arcs. In this paper, the algorithm for obtaining outline segments from the arcs is presented.     

Double coupling Edman chemistry for high-sensitivity automated protein sequencing

Capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) is a promising new method for the analysis of protein sequencing products. It gives 10 zmol (1 zmol = 10(-21) mol) limits of detection (3 sigma) for fluorescein thiohydantoin (FTH) amino acids. We have developed a separation for the (FTH)-amino acid products generated from 18 of the 20 coded amino acids. The extremely low volume requirement associated with CE-LIF makes it incompatible with commercial sequencers. For this reason, we have also been developing a miniaturized sequencer that can be more easily coupled to our detection system. Both the CE-LIF system and the miniaturized sequencer are described.     

Rheological Property and Stress Development during Drying of Tape-Cast Ceramic Layers

Rheological property and stress development of tape-cast ceramic layers derived from nonaqueous alumina (A1₂O₃)-poly(vinyl butyral) (PVB) suspensions were observed during drying. Casting suspensions exhibited strong shear-thinning behavior, with a low shear Newtonian plateau apparent viscosity >10² Pa^.s. The apparent suspension viscosity displayed a power-law dependence on the A1₂O₃ volume fraction during the initial stage of drying (<30% solvent loss). Stress development, measured by a cantilever deflection method, and parallel weight loss measurements were performed during the drying of tape-cast layers and pure binder coatings. Maximum drying stresses (σ_max) of 1.37-0.77 MPa were observed for plasticized tapes cast at gap heights of 150-400 μm. In contrast, nonplasticized tapes of similar thickness displayed a more gradual stress increase, with σ_max values approximately an order of magnitude higher than their plasticized counterparts. The stress histories of the corresponding binder coatings were quite similar to the tape-cast layers, albeit slightly lower σ _max values were observed. Stresses decayed beyond σ_max with a logarithmic time dependence to an almost constant value of 0.2-0.4 MPa for the plasticized tapes. Based on these observations, process methodologies have been offered to minimize stress development and retention in tape-cast ceramic layers     

The importance of glossaries

Standards cannot be set, specifications cannot be written uniformly and unambiguously, and methods cannot be described succinctly, without an agreed terminology. Producing a glossary is a painstaking process and the fundamental principles involved are described. Some difficulties and misunderstandings are identified.