L.N. Castilho

Reactive oxygen species generation in peripheral blood monocytes and oxidized LDL are increased in hyperlipidemic patients

OBJECTIVES Experimental and in vitro evidences have established that reactive oxygen species (ROS) generated by vascular wall cells play a key role in atherogenesis. Here, we evaluated the rate of ROS generation by resting peripheral monocytes in naive hyperlipidemic subjects. DESIGN AND METHODS Primary hypercholesterolemic, combined hyperlipidemic, and normolipidemic individuals were studied. ROS generation and the mitochondrial electrical transmembrane potential were estimated by flow cytometry. Plasma oxidized (ox) LDL levels and lipid profile were measured by ELISA and enzymatic colorimetric methods. RESULTS Both hyperlipidemic groups presented significantly higher rates of monocyte ROS generation and elevated plasma levels of ox-LDL. Combined hyperlipidemic subjects presented increased levels of small dense LDL and insulin. Significant positive correlations between monocyte ROS generation and ox-LDL concentrations were found in pooled data. CONCLUSIONS These data provide evidence that ROS production by circulating monocytes from hyperlipidemic subjects may contribute to the systemic oxidative stress and possibly to atherogenesis.

RHD Allelic Identification Among D−Brazilian Blood Donors as a Routine Test Using Pools of DNA

RHD alleles leading to a reduced expression of D antigen of the red blood cell (RBC) surface may be erroneously typed as D− by serology and may cause anti‐D immunizations when transfused to recipients.

Opposite lipemic response of Wistar rats and C57BL/6 mice to dietary glucose or fructose supplementation

The metabolic effects of carbohydrate supplementation in mice have not been extensively studied. In rats, glucose- and fructose-rich diets induce hypertriacylglycerolemia. In the present study, we compared the metabolic responses to two monosaccharide supplementations in two murine models. Adult male Wistar rats (N = 80) and C57BL/6 mice (N = 60), after 3 weeks on a standardized diet, were submitted to dietary supplementation by gavage with glucose (G) or fructose (F) solutions (500 g/L), 8 g/kg body weight for 21 days. Glycemia was significantly higher in rats after fructose treatment (F: 7.9 vs 9.3 mM) and in mice (G: 6.5 vs 10 and F: 6.6 vs 8.9 mM) after both carbohydrate treatments. Triacylglycerolemia increased significantly 1.5 times in rats after G or F supplementation. Total cholesterol did not change with G treatment in rats, but did decrease after F supplementation (1.5 vs 1.4 mM, P < 0.05). Both supplementations in rats induced insulin resistance, as suggested by the higher Homeostasis Model Assessment Index. In contrast, mice showed significant decreases in triacylglycerol (G: 1.8 vs 1.4 and F: 1.9 vs 1.4 mM, P < 0.01) and total cholesterol levels (G and F: 2.7 vs 2.5 mM, P < 0.05) after both monosaccharide supplementations. Wistar rats and C57BL/6 mice, although belonging to the same family (Muridae), presented opposite responses to glucose and fructose supplementation regarding serum triacylglycerol, free fatty acids, and insulin levels after monosaccharide treatment. Thus, while Wistar rats developed features of plurimetabolic syndrome, C57BL/6 mice presented changes in serum biochemical profile considered to be healthier for the cardiovascular system.

Oxidation of LDL enhances the cholesteryl ester transfer protein (CETP)-mediated cholesteryl ester transfer rate to HDL, bringing on a diminished net transfer of cholesteryl ester from HDL to oxidized LDL

Cholesteryl ester transfer protein (CETP) plays a controversial role in atherogenesis by contributing to the net transfer of high density lipoprotein (HDL) cholesteryl ester (CE) to the liver via apolipoprotein-B-containing lipoproteins (apoB-LP). We evaluated in vitro the CETP-mediated bidirectional transfer of CE from HDL to the chemically modified pro-atherogenic low density lipoprotein (LDL) particles. Acetylated or oxidized (ox) LDL, either unlabeled or [3H]-CE labeled, were incubated with [14C]-CE-HDL in the presence of the lipoprotein-deficient plasma fraction (d>1.21 g/ml) as the source of CETP. The amount of radioactive CE transferred was determined after dextran sulfate/MgCl(2) precipitation of LDL. The results showed a 1.4-2.8-fold lower HDL-CE transfer to acetylated LDL while no effect was observed on the CE transfer to oxidized LDL. However, the reverse transfer rate of [3H]CE-LDL to HDL was 1.4-3.6 times greater when LDL was oxidized than when it was intact. Overall, HDL(2) was better than HDL(3) as donor of CE to native LDL, probably reflecting the relatively greater CE content of HDL(2). Oxidation of LDL enhanced the CETP-mediated cholesteryl ester transfer rate to HDL, bringing on a reduced net transfer rate of cholesteryl ester from HDL to ox LDL. This may diminish the oxLDL particles atherogenic effect.

Biological Rhythms of Biochemical Serum Parameters in a Brazilian Population: a Three‐Year Study

A marked decrease in analytical and post‐analytical variability has been achieved in clinical laboratories by the use of automated analytical systems. Current studies are now focused on the origin of pre‐analytical variability, such as that due to intra‐individual differences and biological rhythms. The objective of this work was to evaluate the occurrence of biological rhythms in several biochemical serum parameters in a Brazilian population. A retrospective study (1996 to 1998) was carried out to collect the test results within the reference intervals of adults, from 21 to 50 yr of age (average age of 36 yr) attending the outpatient clinics of the Teaching Hospital at the University of Campinas, São Paulo, Brazil. The reference sample was 52.9% male and 47.1% female and encompassed 15,036 calcium, 7,478 phosphorus, 53,641 urea, 58,315 creatinine and 6,433 uric acid determinations (140,903 in total). Significant annual rhythms were detected in serum calcium (p≤0.001), with maximum and minimum values in fall and spring, and in serum creatinine (p≤0.002), with maximum and minimum values in summer and winter. The other parameters did not present significant annual rhythmicity. The seasonal rhythms present in the serum concentrations of calcium and creatinine observed in this large population study, although of small amplitude, should be considered a component of the pre‐analytical variation of these clinical laboratory tests.

Hyperlipidemic pancreatitis in a primigravida adolescent

Acute pancreatitis complicating pregnancy occurs very rare ly. Its incide nce has been estimated at between one in 1000 and one in 3000 (1± 3). Chole lithiasis is the most commonly associate d cause (17 to 90% of the cases) followe d by hyperlipidemia. Other cause s are : acute hepatic steatosis, preeclampsia, hypercalcemia, hype rparathyroidism, infections and drugs like hydrochlorothiazid e, furosemide , sulfurs, metronidazole , corticoste roids, tetracycline s, estrogens, and methyldopa (2, 4 ± 6). Gestational pancreatitis carries a signi® cant risk of mortality for both mother and fetus: up to 37% and up to 13.5% respective ly (3, 6, 7). The important elements in diagnosis are : 1) clinical symptoms; 2) high serum amylase and lipase which are not indispensable for the diagnosis, principally in hyperlipidemic patients, who can have normal amylasemia; 3) compatible ultrasonography; 4) computerized tomography (only at the end of pregnancy) (2± 4, 5). Early diagnosis, adequate observation, and control of the mother’ s and fetus’ s vitality reduce complications such as pseudocysts, abcesses, and maternal and fetal mortality rates (3). The patients should be carefully informed about postpartum contraceptive s, particularly the avoidance of synthe tic estrogens in cases of hyperlipide mia (3). It is very important to search for the cause of the pancreatitis in order to treat and prevent new acute episodes. We report a case of hypertriglyceridemia-induce d pancreatitis occurring during pregnancy in a primigravida adole scent.

Reversible flow of cholesteryl ester between high-density lipoproteins and triacylglycerol-rich particles is modulated by the fatty acid composition and concentration of triacylglycerols

We determined the influence of fasting (FAST) and feeding (FED) on cholesteryl ester (CE) flow between high-density lipoproteins (HDL) and plasma apoB-lipoprotein and triacylglycerol (TG)-rich emulsions (EM) prepared with TG-fatty acids (FAs). TG-FAs of varying chain lengths and degrees of unsaturation were tested in the presence of a plasma fraction at d > 1.21 g/mL as the source of CE transfer protein. The transfer of CE from HDL to FED was greater than to FAST TG-rich acceptor lipoproteins, 18% and 14%, respectively. However, percent CE transfer from HDL to apoB-containing lipoproteins was similar for FED and FAST HDL. The CE transfer from HDL to EM depended on the EM TG-FA chain length. Furthermore, the chain length of the monounsaturated TG-containing EM showed a significant positive correlation of the CE transfer from HDL to EM (r = 0.81, P < 0.0001) and a negative correlation from EM to HDL (r = -041, P = 0.0088). Regarding the degree of EM TG-FAs unsaturation, among EMs containing C18, the CE transfer was lower from HDL to C18:2 compared to C18:1 and C18:3, 17.7%, 20.7%, and 20%, respectively. However, the CE transfer from EMs to HDL was higher to C18:2 than to C18:1 and C18:3, 83.7%, 51.2%, and 46.3%, respectively. Thus, the EM FA composition was found to be the rate-limiting factor regulating the transfer of CE from HDL. Consequently, the net transfer of CE between HDL and TG-rich particles depends on the specific arrangement of the TG acyl chains in the lipoprotein particle core.

Determinants of serum lipoprotein(a) concentration in normolipidaemic individuals without clinical atherosclerosis

Background: Lipoprotein(a) (Lp(a)) is an independent risk factor for coronary artery diseases. The main objective of this work was to evaluate the determinants of plasma Lp(a) concentrations in normolipidaemic individuals. Methods: Immunonephelometric quantification of Lp(a) was made in 177 volunteers. A multivariate analysis was employed to verify the influence of clinical and biochemical parameters on plasma Lp(a) concentration. Results: The serum Lp(a) concentration in this population ranged from 0.7 to 40 nmol/L. The Lp(a) predictors were: sex (female), HDL2 triglyceride (negative) and LDL-cholesterol (positive). Conclusions: The modulation of plasma Lp(a) concentration in this study points to pro-atherogenic lipoprotein associations.

Phospholipid transfer protein activity in two cholestatic patients.

CONTEXT Plasma phospholipid transfer protein mediates the transfer of phospholipids from triglyceride-rich lipoproteins, very low density lipoproteins and low density lipoproteins to high density lipoproteins, a process that is also efficient between high density lipoprotein particles. It promotes a net movement of phospholipids, thereby generating small lipid-poor apolipoprotein AI that contains particles and subfractions that are good acceptors for cell cholesterol efflux. CASE REPORT We measured the activity of plasma phospholipid transfer protein in two cholestatic patients, assuming that changes in activity would occur in serum that was positive for lipoprotein X. Both patients presented severe hypercholesterolemia, high levels of low density lipoprotein cholesterol and, in one case, low levels of high density lipoprotein cholesterol and high levels of phospholipid serum. The phospholipid transfer activity was close to the lower limit of the reference interval. To our knowledge, this is the first time such results have been presented. We propose that phospholipid transfer protein activity becomes reduced under cholestasis conditions because of changes in the chemical composition of high density lipoproteins, such as an increase in phospholipids content. Also, lipoprotein X, which is rich in phospholipids, could compete with high density lipoproteins as a substrate for phospholipid transfer protein.