ABSTRACT
In this paper we present some initial resolution measurements of an improved magnetic-electrostatic detector objective lens for a low-voltage scanning electron microscope. The electron optical design of the lens was already proposed by the authors [G. Knell, E. Plies, Nucl. Instr. & Meth. A 427 (1999) 99]. The magnetic circuit of this lens has a radially arranged pole-piece gap. Thus, the specimen is immersed in a strong magnetic field of 106 mT (working distance: 1 mm, primary electron energy: 200 eV). The electrostatic field strength of our optimized lens variant amounts to a moderate value of 100 V/mm for a working distance of 1 mm. At a final beam energy of 1 keV a resolution of 3 nm, at 260 eV a resolution of 5 nm was obtained.
ABSTRACT
5-Oxo-L-prolinase (5-OPase) catalyses the hydrolysis of 5-oxo-L-proline to glutamate with concomitant stoichiometric cleavage of ATP to ADP, a reaction which is known to be part of the gamma-glutamyl cycle-an interrelated series of reactions involved in the synthesis and metabolism of glutathione. As recent studies indicate, this cyclic pathway plays a crucial role in the regulation of amino acid transport. Apparently, the intermediate product 5-oxo-L-proline functions as a second messenger molecule that upregulates the activity of certain amino acid transport systems. Thus, the degradation of 5-oxo-L-proline by 5-OPase leads to the downregulation of this stimulus. In this study, a new sensitive fluorimetric assay for 5-OPase activity was established which is based on the derivatization of glutamate with o-phthaldialdehyde in the presence of thiols and subsequent separation of the products by HPLC. The method is suitable for the screening of chromatography fractions as well as for the determination of the kinetic parameters Km and Vmax of purified 5-OPase. Additionally, it can be used for the measurement of enzyme activity in crude cell extracts and evaluation of tissue distribution.
