Equity is out of fashion? An essay on autonomy and health policy in the individualized society

The 67-kDa laminin receptor (67LR) is a nonintegrin cell surface receptor that mediates high-affinity interactions between cells and laminin. Overexpression of this protein in tumor cells has been related to tumor invasion and metastasis. Thus far, only a full-length gene encoding a 37-kDa precursor protein (37LRP) has been isolated. The finding that the cDNA for the 37LRP is virtually identical to a cDNA encoding the ribosomal protein p40 has suggested that 37LRP is actually a component of the translational machinery, with no laminin-binding activity. On the other hand, a peptide of 20 amino acids deduced from the sequence of 37LR/p40 was shown to exhibit high laminin-binding activity. The evolutionary relationship between 23 sequences of 37LRP/p40 proteins was analyzed. This phylogenetic analysis indicated that all of the protein sequences derive from orthologous genes and that the 37LRP is indeed a ribosomal protein that acquired the novel function of laminin receptor during evolution. The evolutionary analysis of the sequence identified as the laminin-binding site in the human protein suggested that the acquisition of the laminin-binding capability is linked to the palindromic sequence LMWWML, which appeared during evolution concomitantly with laminin.     

High-performance liquid chromatographic determination of erdosteine and its optical active metabolite utilizing a fluorescent chiral tagging reagent, R-(-)-4-(N,N-dimethylamiosulfonyl)-7-(3-aminopyrrolidin-1-yl)-2 ,1,3-benzoxadiazole

Chiral separation of racemic M1 metabolized from erdosteine was investigated by reversed-phase chromatography. The sensitive determination of M1 and erdosteine with UV detection was difficult because of their low absorptivity in the effective wavelength region. To improve the sensitivity and separatability, one thiol and two carboxyl groups in the M1 structure were labelled with DBD-F and R-(-)-DBD-APy, respectively. Non-fluorescent DBD-F quantitatively reacted with thiol in M1 at room temperature for 30 min in borate buffer (pH 9.3) to produce the fluorescent derivative. On the other hand, the labelling of two carboxyls was carried out with a chiral fluorescent reagent, R-(-)-DBD-APy, in acetonitrile containing DPPA. The derivatives corresponding to a pair of the enantiomers were completely separated with water-acetonitrile containing 0.1% TFA as the mobile phase by an ODS column. Erdosteine with a carboxyl group was also labelled with R-(-)-DBD-APy and separated together with M1 derivatives. The detection limits (S/N=3) of erdosteine and M1 were 0.37 and 0.22 pmol, respectively. The proposed derivatization and separation methods were applied to simultaneous determination of racemic M1 and erdosteine in rat plasma after administration of erdosteine. The amounts of both enantiomers of M1 were essentially the same in oral and intravenous administrations. In contrast, total amounts (reduced-form and oxidized-form) of S-(-)-M1 in rat plasma were higher than those of R(+)-M1 in both administrations.     

Development of systems reliability analysis code SECOM-2 for seismic PSA

An integrated code system SECOM-2, developed at the Japan Atomic Energy Research Institute (JAERI), has the following functions for systems reliability analysis in seismic probabilistic safety assessments (PSAs): (1) calculation of component failure probability, (2) extraction of minimal cut sets (MCSs) from a given fault tree (FT), (3) calculation of frequencies of accident sequences and core damage, (4) importance analysis with several measures with consideration of unique parameters of seismic PSAs, (5) sensitivity analysis, and (6) uncertainty analysis. This paper summarizes the special features of SECOM-2 to perform the analyses mentioned above. At JAERI, using an integrated FT which represents seismically induced core damage due to all initiating events as a system model to calculate core damage frequency of a nuclear power plant, SECOM-2 can calculate conditional point estimate probabilities of system failures, losses of safety functions, and core damage as a function of earthquake motions. The point estimate is computed by a method which gives an exact numerical solution using the Boolean arithmetic model method. As for consideration of correlation of component failure, which has been an important issue in seismic PSAs, a new technique based on direct FT quantification by a Monte Carlo simulation is being added to SECOM-2. Adding this technique, the core damage frequency can be calculated not only with the upper bound approximation based on MCSs but also with a near exact solution taking into account the correlation among all components. This paper also presents the preliminary results of a seismic PSA of a generic BWR plant in Japan performed at JAERI to demonstrate the functions of the SECOM-2 code.     

Formulary review of the carbapenems: comparison of imipenem/cilastatin and meropenem

The duplicative mutation of an Ala-Val-Arg sequence at positions 208 to 210 in the loop structure of Enterobacter cloacae class C beta-lactamase caused substrate specificity extension to oxyimino beta-lactam antibiotics and this chromosomal mutation provided bacterial cells with high resistance to the beta-lactams (M. Nukaga et al, 1995, J. Biol. Chem. 270, 5729-5735). In order to confirm the universality of this phenomenon among other class C beta-lactamases, the duplicative mutation was applied to a class C beta-lactamase of Citrobacter freundii, which has 74% homology to the E. cloacae beta-lactamase amino acid sequence. The counterpart sequence to the Ala-Val-Arg of the E. cloacae enzyme in C. freundii beta-lactamase was identified to be Pro-Val-His. A Pro-Val-His sequence was inserted just after the native Pro-Val-His sequence at positions 208 to 210 in the C. freundii beta-lactamase. The resulting mutant of C. freundii beta-lactamase obtained a striking characteristic that we expected, showing substrate specificity extension to oxyimino beta-lactams. Nearly the same result was obtained with the insertion of an Ala-Val-Arg sequence after the native Pro-Val-His sequence. These results indicate that structural modification of this locus commonly induces modification of the substrate specificity to unfavorable substrates for many chromosomal class C beta-lactamases produced by gram-negative bacteria.     

Prodrug esters of the indolocarbazole CEP-751 (KT-6587)

Prodrug esters of the indolocarbazole CEP-751 (KT-6587) were prepared with the goal of identifying water soluble, stable but cleavable forms for intravenous dosing. A dipeptide proform Lys-beta-Ala (16, CEP-2563/KT-8391) was identified for advancement to clinical trials.     

Investigation of the action patterns of pectinmethylesterase isoforms through kinetic analyses and NMR spectroscopy. Implications In cell wall expansion

Well characterized pectin samples were incubated with cell wall-bound and -solubilized pure isoforms of pectinmethylesterase from mung bean hypocotyls (Vigna radiata). Both enzyme activity and average product structure were determined at intervals along the deesterification pathway at pH 5.6 and 7.6. The latter analyses were performed by 13C NMR spectroscopy, and the degree of esterification was probed by both 13C NMR and potentiometric measurements. A dichotomy was observed in the behavior of the alpha and gamma isoforms when compared with that of the beta isoenzyme. Ideal blockwise deesterification mechanisms reproduced the experimental average structures (methylester distribution) throughout the course of the reaction. In the case of the alpha and gamma isoforms, a single chain mechanism associated with a free carboxyl group at the second nearest neighbor position could be postulated at pH 5.6, whereas some multiple attack character was required to reproduce the data at pH 7.6. Several mechanisms that differed from the preceding ones were compatible with the data for the beta isoform at the two pH values. Both the nature of the polysaccharides produced in these reactions and the role of pectinmethylesterase in the cell wall-stiffening process along the growth gradient are discussed.