Synthesis of glutaminyl adenylate analogues that are inhibitors of glutaminyl-tRNA synthetase.

Glutaminol adenylate 5 is a competitive inhibitor of glutaminyl-tRNA synthetase with respect to glutamine (Ki = 280 nM) and to ATP (Ki = 860 nM). The corresponding methyl phosphate ester 4 is a weaker inhibitor (Ki approximately 10 microM) with respect to glutamine.

Acute phase proteins and plasma lipoproteins during antimony treatment in infantile visceral leishmaniasis.

The purpose of this study was to determine the effects of antimony treatment on the altered lipoprotein system of four young Tunisians with visceral Leishmaniasis and to further investigate possible relationships between acute phase proteins (C-reactive protein and apolipoprotein SAA)-known to be increased in this disease-and lipid or apolipoprotein (apo) parameters measured before and throughout the treatment. A marked improvement of all investigated parameters was already observed after the first cure: the lecithin-cholesterol acyltransferase activity was the only parameter remaining deficient even at the end of the second cure. Statistical analysis reveals a significant inverse relationship between changes in C-reactive protein and changes in the plasma ratio of esterified to total cholesterol (r = -0.78; p < 0.001) as well as in plasma apoA-I concentration (r = -0.64; p < 0.01). Because no such correlation could be observed with apoSAA, results of this study suggest that the major mechanism of lowering the plasma levels of apoA-I and the plasma-cholesteryl content could be closely related to the appearance of large amounts of C-reactive protein in plasma or more likely to the process inducing the hepatic synthesis of this acute phase protein.

Alterations in lipoprotein density classes in infantile visceral leishmaniasis: presence of apolipoprotein SAA.

This study describes the alterations in the plasma lipoproteins from nine young Tunisian children with active visceral Leishmaniasis. The plasma lipid profile from affected patients was characterized by a marked hypertriglyceridaemia associated with reduced levels of total and high density lipoprotein (HDL)-cholesterol and a significant increase in the plasma ratio of unesterified to total cholesterol. Quantitative determination of plasma apolipoproteins revealed significantly decreased levels of all measured apolipoproteins, especially of apolipoproteins A-I and A-II, with the exception of apolipoprotein E, the levels of which were markedly increased. Moreover, at least two isoforms of the apolipoprotein serum amyloid A (SAA), an acute phase protein, were detected in all patients' plasma using two-dimensional electrophoresis. Immunochemical evidence was presented that apolipoproteins E and SAA, although both primarily associated with apolipoprotein A- (A-I and A-II) as well as with apolipoprotein B-containing lipoproteins, could occur as LP-E and LP-SAA subspecies, devoid of apolipoproteins A and B. However, it should be pointed out that LP-SAA particles were found in HDL2 from only two patients whereas the abnormal LP-E particles were detected in LDL and HDL2 from all investigated patients. The polydispersity and heterogeneity of patients' HDL3 were assessed by electron microscopy. It was further suggested that the profound changes in the lipoprotein metabolism of these young patients may be due to the increased hepatic synthesis of apolipoprotein SAA and/or to their altered immune function during active visceral Leishmaniasis.

In vivo metabolism of apolipoproteins A-IV and A-I associated with high density lipoprotein in normolipidemic subjects.

The kinetics of apolipoprotein A-IV associated with high density lipoproteins (HDL) of plasma from fasting human subjects was followed for 15 days in five healthy normolipidemic volunteers. Purified apoA-IV and apoA-I were radioiodinated, respectively, with 125I and 131I, incubated in vitro with normal HDL, isolated at density 1.250 g/ml, and finally reinjected intravenously as HDL-125I-labeled apoA-IV and HDL-131I-labeled apoA-I. Blood samples were withdrawn at regular intervals for 15 days, and 24-h urine samples were collected. More than 93% (93.5 +/- 0.9%) of apoA-IV was recovered in apoA-I-containing lipoprotein particles after affinity chromatography on an anti-apoA-I column and 69.7 +/- 4.8% was bound to apoA-II in apoA-I:A-II particles separated on an anti-apoA-II column. 125I-labeled apoA-IV showed a much faster decay than 131I-labeled apoA-I for the first 5 days and thereafter the curves became parallel. Urinary/plasma ratios (U/P) for the 125I-labeled parallel. Urinary/plasma ratios (U/P) for the 125I-labeled apoA-IV were much higher than those for 131I-labeled apoA-I for the first days, but the U/P curves became parallel for the last 7 days, suggesting heterogeneity of apoA-IV metabolism. A heterogeneous multicompartmental model was constructed to describe the metabolism of lipoprotein particles containing apoA-IV and apoA-I and to calculate the kinetic parameters, fitting simultaneously all plasma and urine data for both tracers.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein C-II and C-III metabolism in a kindred of familial hypobetalipoproteinemia.

Three affected members of a kindred with asymptomatic hypobetalipoproteinemia (HBL) showed low levels of triglycerides, low-density lipoprotein (LDL)-cholesterol, and apolipoproteins (apo) B, C-II, and C-III. Turnover of iodine-labeled apo C-II and apo C-III associated in vitro to plasma lipoproteins was studied after intravenous injection. Radioactivity in plasma and lipoproteins (95% recovered in high-density lipoprotein [HDL] density range) and in 24-hour urine samples was observed for 16 days. A parallelism of the slowest slopes of plasma decay curves was observed between apo C-II and apo C-III, indicating a partial common catabolic route. Urine/plasma radioactivity ratio (U/P) varied with time, suggesting heterogeneity of metabolic pathways. A new compartmental model using the SAAM program was built, not only fitting simultaneously plasma and urine data, but also taking into account the partial common metabolism of lipoprotein particles (LP) containing apo C-II and apo C-III. The low apo C-II and C-III plasma concentrations observed in HBL compared with normal resulted from both an increased catabolism and a reduced synthesis, these changes being more marked for apo C-III. The modifications in the rate constants of the different pathways calculated from the new model are in favor of an increased direct removal of particles following the fast pathway, likely in the very-low-density lipoprotein (VLDL) density range.

Plasma lipids, lipoproteins and apolipoproteins in two kindreds of hypobetalipoproteinemia.

Plasma lipids and apolipoproteins were quantified in two kindreds of hypobetalipoproteinemia. All affected members were asymptomatic but showed a decrease of 75% in apolipoprotein B and of 69% in LDL-cholesterol. There were no major changes in apo A-I and A-II but all affected family members had reduced levels of apo C-II (by 58%) and C-III (by 59%) without significant decrease in apo C-I and no specific decrease of apo C-III1. Apolipoprotein E is decreased in SDS-PAGE. The plasma level and phenotype of Lp(a) are not affected by HBL, suggesting that a catabolic rather than a synthetic mechanism is responsible for the disease. As shown by density gradient ultracentrifugation, HDL2 particles that contain essentially apolipoprotein A-I, cholesterol and phospholipids represent in affected subjects the major part of HDL. Due to the net reduction of apolipoprotein B-containing particles (VLDL and LDL) as acceptors of lipids in HBL, there is an accumulation of large particles rich in cholesteryl esters.

Characterization of anti-apolipoprotein A-I monoclonal antibodies and their use in the measurement of apolipoprotein A-I by a two-site enzyme immunoassay.

Six monoclonal antibodies raised against pure apo A-I or high density lipoprotein were characterized for epitope specificity by enzyme and radioimmunoassays, immunodiffusion and affinity chromatography. The six antibodies were classified into three groups according to the region of apo A-I they reacted with. The antibody VI10H, from group II, appeared to recognize a region fully exposed on native HDL-apo A-I, whereas group I comprised antibodies specific for a partially masked region. Group III comprised only one antibody. Use of the non-ionic detergent Tween 20 in the immunoassays permitted antibodies from the three groups to react with their respective epitope on native HDL-apo A-I. An antibody from group I (V4F) was chosen as the first antibody and VI10H, the antibody showing the highest affinity, was chosen for the anti-A-I-peroxidase conjugate in a two-site enzyme immunoassay.

Apolipoproteins C-II and C-III metabolism in hypertriglyceridemic patients. Effect of a drastic triglyceride reduction by combined diet restriction and fenofibrate administration.

Four hypertriglyceridemic patients, who had received an equilibrated high calorie diet and no lipid lowering drug for 1 month, were injected intravenously with 125I-apo C-II and 131I-apo C-III labeled homologous lipoproteins. Plasma and urine radioactivity, lipid and apolipoprotein levels were followed at regular intervals for 15 days. At the end of this first kinetic study the patients were advised to adhere for 1 month to a more restricted diet, limited in fat, and were given additionally 300 mg fenofibrate daily. After this treatment, a new kinetic study involving intravenous injection (similar to the first one) was performed. The protocols of both studies were identical. Treatment (diet plus drug) (1) reduced total cholesterol by 26 +/- 8%, triglycerides by 56 +/- 15%, apo C-II by 36 +/- 14%, and apo C-III by 48 +/- 10%; (2) modified the distribution of radioactivity between lipoproteins proportionally to the change in their mass ratio (decrease in VLDL and increase in HDL); (3) changed the kinetics of both apoproteins by rising the fractional removal rate, shortening residence time and decreasing the synthesis rate of both apolipoproteins C-II and C-III. The treatment was, however, unable to reduce the synthesis rate of apo C-III to normal, suggesting a major role of the apoprotein overproduction in the triggering of hypertriglyceridemia.

Modifications of plasma lipids, lipoproteins and apolipoproteins in advanced cancer patients treated with recombinant interleukin-2 and autologous lymphokine-activated killer cells.

Intravenous injection of recombinant interleukin-2 (r-Met-hu-IL-2(Ala-125] and LAK cells induced dramatic changes of lipoproteins in 12 patients with advanced cancer. After r-IL-2 injection (1) total cholesterol was reduced by 47% as a mean, LDL-cholesterol by 62%, HDL-cholesterol by 77%; (2) the triglyceride/cholesterol ratio was greatly increased (352%); (3) apolipoproteins B, A-I and A-II showed a mean reduction of 26%, 55% and 51%, respectively; and (4) very low density lipoproteins relatively increased, and HDL were separated into two definite fractions (I and II). LAK cell administration accentuated all the above effects and in most patients, HDL-fraction I almost completely disappeared. An action on hepatic synthesis of acute phase proteins is pointed out by the increase in C-reactive protein and apolipoprotein S concentrations contrasting with an unexpected reduction of fibrinogen. Surprisingly the drastic changes caused by treatment were quickly and completely reversible.

Non-competitive enzyme-linked immunosorbent assay for human apolipoprotein SAA or S.

Purified antibodies against human apolipoprotein S were used to measure apo S levels in human plasma by a sandwich ELISA technique. The method developed was highly sensitive and specific. A reliable calibration of a plasma standard was obtained by reassociating the purified protein with plasma HDL. A significant correlation (r = 0.69) was found between plasma apo S and CRP values. This correlation coefficient reached 0.97 when the plasma samples were selected according to additional clinical and laboratory criteria.

Comparison of different sandwich enzyme immunoassays for the quantitation of human apolipoproteins A-I and A-II.

Variants of a non-competitive enzyme immunoassay for human apolipoproteins A-I and A-II are described. The method developed is highly sensitive and highly specific. There is a good correlation between the reference technique (electroimmunoassay) and both apolipoprotein A-I and A-II assays (r=0.92). For the apolipoprotein A-I assay, seven simple treatments were studied. Two of these treatments (plasma plus Tween 20 and serum plus apo A-II) greatly increased the precision and the accuracy of the results. For the apo A-II measurement, no treatment of the samples is necessary, but plasma should be preferentially used rather than serum.