In vitro evaluation of 5-arylidene-2-thioxo-4-thiazolidinones active as aldose reductase inhibitors

2-Thioxo-4-thiazolidinone derivatives were evaluated as aldose reductase inhibitors (ARIs) and most of them exhibited good or excellent in vitro efficacy. Out of the tested compounds, most N-unsubstituted analogues were found to possess inhibitory effects at low micromolar doses and two of them exhibited higher potency than sorbinil, used as a reference drug. The insertion of an acetic chain on N-3 of the thiazolidinone scaffold led to analogues with submicromolar affinity for ALR2 and IC₅₀ values very similar to that of epalrestat, the only ARI currently used in therapy.     

Reduction in magnetic resonance imaging T2 burden of disease in patients with relapsing-remitting multiple sclerosis: analysis of 48-week data from the EVIDENCE (EVidence of Interferon Dose-response: European North American Comparative Efficacy) study

Background  

The EVIDENCE (EVidence of Interferon Dose-response: European North American Comparative Efficacy) study was an international, randomized, open-label, assessor-blinded, parallel-group study assessing the efficacy and tolerability of interferon (IFN) beta-1a, 44 mcg subcutaneously (sc) three times weekly (tiw), and IFN beta-1a, 30 mcg intramuscularly (im) once weekly (qw), in patients with relapsing-remitting multiple sclerosis (RRMS). The aim of this analysis was to assess whether reductions in T2 burden of disease (BOD) were greater for patients receiving IFN beta-1a, 44 mcg sc tiw, than for those treated with IFN beta-1a, 30 mcg im qw, and to assess the impact of neutralizing antibodies (NAbs).     

Alpha-5-phosphoribosyl-1-pyrophosphate-independent salvage of purines in cultured Chinese hamster lung fibroblasts

A variant clone of cultured chinese hamster lung fibroblasts (V79), selected for resistance to 8-azaguanine (V79 azagrst), although lacking hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8), is able to convert hypoxanthine into IMP via purine-nucleoside phosphorylase (EC 2.4.2.1) and nucleoside kinase. In addition to the phosphoribosylation pathway, we also present evidence for the occurrence of a kinase-mediated pathway of recovery of hypoxanthine in the wild-type cells. The lower rate of formation of IMP in the V79 azagrst cells, apparently correlated with the phosphorylation of the nucleoside, suggests possible differences in the catalytic and/or regulatory properties of nucleoside kinase in the two cell lines. This fact might be of particular relevance in evaluating the mechanisms of resistance to purine analogs displayed by several cell types.     

CCK-Containing paraneurons in human adenomatous prostate

The presence of CCK-containing neuroendocrine cells in human adenomatous prostates, and the colocalization of CCK together with serotonin in the same cell, have been demonstrated by means of an immunohistochemical technique and by a double labeling immunofluorescent staining. CCK-containing neuroendocrine cells had a focal distribution in the prostates and sometimes showed dendrite-like cytoplasmic processes. The major part of CCK (96.55%) colocalized with serotonin.CCK probably stimulates muscle contraction and endocrine/exocrine secretions in the urogenital tract.     

pH influence on enzymic activity: the involvement of two active ionized forms of either substrate or enzyme in the reaction.

The interpretation of graphs of pKm as a function of pH by the theory of Dixon (Dixon, 1953) is based on the assumption that each component of the reaction system is active in a particular ionic state and completely inactive in other ionic states. In this paper we analyse the case in which two ionic forms of either the substrate or the enzyme are active to different degrees. After definition of reaction system of this type, a generalized equation is derived connecting the reaction rate to the total substrate concentration. The pH dependence of Km deduced from this equation is discussed and compared with that deriving from the theory of Dixon. The present model predicts that the pKm dependence on pH has the form of a “wave” with two inflexion points, thus providing an alternative interpretation of such behaviour to that afforded by Dixon's formulation.     

Some kinetic aspects of the mechanism of hydrolysis of phosphoric acid esters by nonspecific acid phosphatase from Schizosaccharomyces pombe.

1. The kinetics of the hydrolysis of nitrophenylphosphate by nonspecific acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2.) from Schizosaccharomices pombe was studied. 2. The kinetic parameters, Km and V, were determined as well as the inhibition constants, K1, for the inhibitors, phosphate and fluoride, as a function of pH. 3. The results, interpreted according to the theories of Dixon and Waley indicated the presence of three ionizable groups on the enzyme itself and one on the enzyme-substrate complex. 4. A model of the hydrolysis of phosphoric acid monoesters by the S. pombe acid phosphatase is proposed based on the ionization state of the reactants and on the results of the inhibition by the competitive inhibitors.     

Chaperone-like activity of alpha-crystallin toward aldose reductase oxidatively stressed by copper ion

The protective action of alpha-crystallin against copper-induced protein stress is studied using bovine lens aldose reductase (ALR2) as protein model. The oxidative inactivation of ALR2 induced by CuCl2 at the stoichiometric Cu2+/ALR2 ratio of 2/1 [I. Cecconi, M. Moroni, P.G. Vilardo, M. Dal Monte, P. Borella, G. Rastelli, L. Costantino, D. Garland, D. Carper, J.M. Petrash, A. Del Corso, U. Mura, Biochemistry 37 (1998) 14167-14174] is accompanied by protein aggregation phenomena when the metal ion concentration is increased (Cu2+/ALR2>3). Protein oxidation precedes protein precipitation. Both inactivation and precipitation of ALR2 are prevented by alpha-crystallin in a concentration-dependent manner. The rationale for the stabilization of ALR2 exerted by alpha-crystallin at low metal concentration is given on the basis of the ability of alpha-crystallin to chelate copper. However, the overall protective action exerted by alpha-crystallin at higher copper concentration may be explained invoking the contribution of the special features of alpha-crystallin to easily interact with target proteins undergoing structural rearrangement.     

Reproduction Strategy of the Deep-sea Hermit Crabs Pagurus alatus and Pagurus excavatus of the Central-Western Mediterranean Sea

Some aspects of the reproductive biology of two hermit crabs Pagurus alatus (Fabricius, 1775) and P. excavatus (Herbst, 1791) were studied. Specimens were collected monthly from April 2000 to March 2001 on the continental shelf and the upper-middle slope off the Southwestern coasts of Sardinia, in the central-western Mediterranean Sea. Crab size (minimum and maximum shield length) was 1.7 and 9.5 mm for 1150 females of P. alatus; 4.0 and 10.4 mm for 347 females of P. excavatus, respectively. Females of P. alatus with ripe ovaries were only found in February-April and ovigerous females were observed throughout the year except in March, with the highest incidence in summer. Females of P. excavatus with fully developed ovaries were collected during all months of the year reaching a peak from April to June. Ovigerous females of P. excavatus occurred throughout the year and the main spawning period occurred in March and April. Fecundity of both species was calculated to assess seasonal variation of reproductive intensity and was positively correlated with the size of the individuals. Monthly fecundity did not reveal significant differences in P. alatus ovigerous females, whereas P. excavatus exhibited significant differences in monthly fecundity depending on the time of year. Difference in fecundity among P. alatus and P. excavatus can be related to the size of the ovigerous females and species of hermit crab, but their different reproductive strategies suggest that they are specifically related to different environmental conditions.     

In situ stability of uridylyl removing-uridylyltransferase of Escherichia coli

The difference in sensitivity of UR/UT toward Lubrol WX permeabilization treatment of stationary phase E. coli cells is not uniquely related to nitrogen availability during cellular growth. The sensitivity of UR/UT to detergent treatment appears to be related to differences in the balance between fermentative and oxidative glucose metabolism. The possible occurrence of a third cycle in the glutamine synthetase regulatory cascade mechanism is considered.