apoB Ratio Research Articles

ApoB-100 carrying lipoprotein, but not apoB-48, is the major subset of proatherogenic remnant-like lipoprotein particles detected in plasma of sudden cardiac death cases

We have previously reported that plasma levels of remnant-like lipoprotein particles (RLP) significantly increased in sudden cardiac death cases with and without coronary atherosclerosis. In this study we have elucidated the major subset of proatherogenic RLP, containing both apoB-48 and apoB-100-carrying remnants, in plasma of SCD and control death cases. One hundred and sixty seven Japanese cases of sudden cardiac death and 78 cases of control death underwent autopsy within 12 h after death were studied. Heart weight was 9.2% higher in SCD cases than controls ( P < 0.05). Moreover 57.5% or 96/167 of the cases had more than grade (2+) coronary atherosclerosis versus 21.8% or 17 of 78 controls ( P < 0.01). Approximately 2/3 of the cases had full stomach, reflecting the postprandial state at the time of death. Plasma TC, TG, VLDL-C, LDL-C were significantly elevated ( P < 0.001) together with RLP-C ( P < 0.01), RLP-TG ( P < 0.005) in SCD cases. Plasma RLP-apoB-100 levels were significantly elevated in SCD ( P < −0.001), but apoB-48 levels were not. The median ratio of apoB-100/apoB-48 in RLP was 7.1 in SCD. The median RLP-TG/RLP-C ratio was 4.7, which suggested a large VLDL size. When apoB-48 and apoB-100 in RLP were divided into two groups, above and below the median level, respectively, apoB-48 inversely correlated with RLP-C ( P < 0.05) and RLP-TG ( P < 0.01), while apoB-100 in RLP positively correlated with RLP-C ( P < 0.01) in SCD cases. In conclusion, these results indicated that apoB-100 carrying lipoproteins, not apoB-48 carrying lipoproteins, were the major subset of RLP associated with sudden cardiac death in the postprandial state, regardless to the severity of coronary atherosclerosis.

Low-density lipoprotein apolipoprotein B100 turnover in hypopituitary patients with GH deficiency: a stable isotope study

Epidemiological studies suggest that hypopituitary patients have an increased risk for cardiovascular mortality. The dyslipidaemia associated with this condition is often characterised by an increase in total cholesterol (TC) and low-density lipoprotein (LDL) cholesterol (LDL-C) and may contribute to these findings. The underlying mechanisms are not fully elucidated. LDL apolipoprotein B (apoB) production rate and metabolic clearance rate were measured in seven patients with hypopituitarism (including GH deficiency) under stable conventional replacement therapy (three males and four females; age 40-16.1 years; body mass index 29.0-6.1 kg/m(2) (means +/- s.d.)) and seven age-, gender- and body mass index-matched control subjects with an infusion of 1-(13)C-leucine. Fasting lipid profile and lipid composition of LDL were also measured. Fasting TC, triglycerides (TG), high-density lipoprotein-C, LDL-C and free fatty acid concentrations were not different between hypopituitary patients and control subjects. LDL-TG (P < 0.006) and LDL-TG/LDL apoB ratio (P < 0.02) were significantly increased in hypopituitary patients. LDL apoB pool size was not statistically different between patients and control subjects. In the hypopituitary patients, LDL apoB metabolic clearance rate (P < 0.05) and LDL apoB production rate (P < 0.02) were lower than in the control subjects. The present results suggest that LDL apoB turnover and LDL composition is altered in hypopituitary patients. Whether these findings explain the increased risk for cardiovascular disease in hypopituitary patients remains to be established.

Delayed In Vivo Catabolism of Intermediate-Density Lipoprotein and Low-Density Lipoprotein in Hemodialysis Patients as Potential Cause of Premature Atherosclerosis

Premature cardiovascular disease is the leading cause of death in patients with end-stage renal disease treated by hemodialysis (HD). Low-density lipoprotein (LDL) levels are not generally increased in HD patients, but their LDL metabolism is still poorly understood. We therefore investigated the in vivo metabolism of apoB-containing lipoproteins in two different ethnic populations of HD patients and controls. We performed stable isotope kinetic studies using a primed constant infusion of deuterated leucine in 12 HD patients and 13 healthy controls. Tracer/tracee ratio of apoB was determined by means of gas chromatography/mass spectrometry, and the modeling program SAAMII was used to estimate the fractional catabolic rate (FCR) of apoB. Mean LDL-apoB plasma concentrations were almost identical in both groups (HD: 95+/-30 mg/dL, controls: 91+/-40 mg/dL), whereas LDL-apoB FCR was 50% lower in HD patients as compared with controls (0.22+/-0.12 days(-1) versus 0.46+/-0.20 days(-1), P=0.001) with concomitantly decreased production rates of LDL. Compared with controls, intermediate-density lipoprotein (IDL)-apoB FCR was 65% lower (2.87+/-1.02 days(-1) versus 8.89+/-4.94 days(-1), P=0.014), accompanied by 1.5-fold higher IDL-apoB levels in HD. Very low-density lipoprotein metabolism was similar in both study groups. In vivo catabolism of LDL and IDL is severely impaired in HD patients but misleadingly masked by normal plasma cholesterol levels. The resulting markedly prolonged residence times of both IDL and LDL particles might thus significantly contribute to the well-documented high risk for premature cardiovascular disease in HD patients.

Oxidized Phospholipids, Lp(a) Lipoprotein, and Coronary Artery Disease

Lp(a) lipoprotein binds proinflammatory oxidized phospholipids. We investigated whether levels of oxidized low-density lipoprotein (LDL) measured with use of monoclonal antibody E06 reflect the presence and extent of obstructive coronary artery disease, defined as a stenosis of more than 50 percent of the luminal diameter. Levels of oxidized LDL and Lp(a) lipoprotein were measured in a total of 504 patients immediately before coronary angiography. Levels of oxidized LDL are reported as the oxidized phospholipid content per particle of apolipoprotein B-100 (oxidized phospholipid:apo B-100 ratio). Measurements of the oxidized phospholipid:apo B-100 ratio and Lp(a) lipoprotein levels were skewed toward lower values, and the values for the oxidized phospholipid:apo B-100 ratio correlated strongly with those for Lp(a) lipoprotein (r=0.83, P<0.001). In the entire cohort, the oxidized phospholipid:apo B-100 ratio and Lp(a) lipoprotein levels showed a strong and graded association with the presence and extent of coronary artery disease (i.e., the number of vessels with a stenosis of more than 50 percent of the luminal diameter) (P<0.001). Among patients 60 years of age or younger, those in the highest quartiles for the oxidized phospholipid:apo B-100 ratio and Lp(a) lipoprotein levels had odds ratios for coronary artery disease of 3.12 (P<0.001) and 3.64 (P<0.001), respectively, as compared with patients in the lowest quartile. The combined effect of hypercholesterolemia and being in the highest quartiles of the oxidized phospholipid:apo B-100 ratio (odds ratio, 16.8; P<0.001) and Lp(a) lipoprotein levels (odds ratio, 14.2; P<0.001) significantly increased the probability of coronary artery disease among patients 60 years of age or younger. In the entire study group, the association of the oxidized phospholipid:apo B-100 ratio with obstructive coronary artery disease was independent of all clinical and lipid measures except one, Lp(a) lipoprotein. However, among patients 60 years of age or younger, the oxidized phospholipid:apo B-100 ratio remained an independent predictor of coronary artery disease. Circulating levels of oxidized LDL are strongly associated with angiographically documented coronary artery disease, particularly in patients 60 years of age or younger. These data suggest that the atherogenicity of Lp(a) lipoprotein may be mediated in part by associated proinflammatory oxidized phospholipids.

Overproduction of VLDL 1 Driven by Hyperglycemia Is a Dominant Feature of Diabetic Dyslipidemia

We sought to compare the synthesis and metabolism of VLDL1 and VLDL2 in patients with type 2 diabetes mellitus (DM2) and nondiabetic subjects. We used a novel multicompartmental model to simultaneously determine the kinetics of apolipoprotein (apo) B and triglyceride (TG) in VLDL1 and VLDL2 after a bolus injection of [2H3]leucine and [2H5]glycerol and to follow the catabolism and transfer of the lipoprotein particles. Our results show that the overproduction of VLDL particles in DM2 is explained by enhanced secretion of VLDL1 apoB and TG. Direct production of VLDL2 apoB and TG was not influenced by diabetes per se. The production rates of VLDL1 apoB and TG were closely related, as were the corresponding pool sizes. VLDL1 and VLDL2 compositions did not differ in subjects with DM2 and controls, and the TG to apoB ratio of newly synthesized particles was very similar in the 2 groups. Plasma glucose, insulin, and free fatty acids together explained 55% of the variation in VLDL1 TG production rate. Insulin resistance and DM2 are associated with excess hepatic production of VLDL1 particles similar in size and composition to those in nondiabetic subjects. We propose that hyperglycemia is the driving force that aggravates overproduction of VLDL1 in DM2.

Effect of antiatherogenic l-aspartate and l-glutamate on serum lipoproteins cholesterol and apolipoproteins A-1 and B in rabbits fed with high cholesterol diet

It has been shown that aspartate and glutamate inhibit mononuclear cell adhesion to the endothelium and formation of foam cells in the intima of thoracic aorta in cholesterol-fed rabbits. The purpose of the present study was to investigate whether a high cholesterol diet supplemented with aspartate and glutamate may alter lipoproteins cholesterol and apolipoproteins A-1 and B levels in rabbits. Serum total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglycerides (TG), apolipoprotein A-1 (apoA-1), apolipoprotein B (apoB), atherogenic index (AI) and apoA-1/apoB ratio were determined in 17 male New Zealand white rabbits fed a cholesterol plus corn oil diet (control group) or the same diet supplemented with aspartate and glutamate (Asp+Glu group) for 4 weeks. Both diets were found to increase TC, LDL-C, apoB and AI, while apoA-1/apoB ratio was decreased compared to baseline values. TG did not seem to be affected in the 4 weeks time in both groups. There was a significant increase of HDL-C in Asp+Glu group and a marked decrease of apoA-1 in control group during the study. Oral administration of aspartate and glutamate has been shown to inhibit fatty streak initiation in cholesterol-fed rabbits. The two amino acids did not have any effect on serum TC, LDL-C, TG and apoB concentrations. However, they increased HDL-C and maintained apoA-1 levels. Their antiatherogenic effect probably may be explained by different mechanisms than these related to the atherogenic lipids lowering, and it is possible to involve HDL-C and apoA-1.

Assessment of cysteine synthesis in very low–birth weight neonates using a [ 13C 6]glucose tracer

Background/Purpose Cysteine is an amino acid necessary for the synthesis of all proteins, the antioxidant glutathione, and the neuromodulator taurine. Whether cysteine is an essential amino acid for premature neonates remains controversial. Using a [ 13C 6]glucose precursor in very-low–birth weight (VLBW) premature neonates, we measured the 13C content of cysteine in hepatically derived apolipoprotein (apo) B-100 and in the plasma to determine whether cysteine synthesis occurs and to relate minimum synthetic capacity to neonatal maturity. Methods Twelve VLBW premature neonates (birth weight, 907 ± 274 [SD] g; gestational age, 26.8 ± 2.4 weeks) were studied on day of life 7.8 ± 4.2 while on total parenteral nutrition (TPN) for 5.6 ± 4.5 days. A 4-hour intravenous infusion of [ 13C 6]glucose was administered. Blood samples were obtained immediately before and at the end of the infusion. Isotopic enrichment of cysteine was determined by gas chromatography/mass spectrometry. Analysis of variance, Student's t test, and linear regression were used for comparisons. Results The 13C isotope ratio of apo B-100–derived cysteine after the [ 13C 6]glucose infusion was significantly higher than baseline (18.57 ± 0.38 [SEM] vs 17.54 ± 0.25 mol%, P < .05). The 13C isotope ratio of plasma cysteine was also significantly higher than baseline (17.36 ± 0.25 vs 16.91 ± 0.16 mol%, P < .05). When expressed as a product/precursor ratio, the mole percent above baseline of [ 13C]apo B-100 cysteine/[ 13C 6]glucose correlated with birth weight ( r = 0.74, P < .01). Conclusions Very low–birth weight neonates are capable of cysteine synthesis as evidenced by incorporation of 13C label into hepatically derived apo B-100 cysteine and plasma cysteine from a glucose precursor. The minimum capacity for intrahepatic cysteine synthesis appears to be directly proportional to the maturity of the neonate and may impact the capabilities of VLBW neonates to counteract oxidative stresses such as bronchopulmonary dysplasia and necrotizing enterocolitis.

Apolipoprotein B-containing Lipoprotein Particle Assembly: LIPID CAPACITY OF THE NASCENT LIPOPROTEIN PARTICLE

We previously proposed that the N-terminal 1000-residue betaalpha(1) domain of apolipoprotein B (apoB) forms a bulk lipid pocket homologous to that of lamprey lipovitellin. In support of this "lipid pocket" hypothesis, we demonstrated that apoB:1000 (residues 1-1000) is secreted by a stable transformant of McA-RH7777 cells as a monodisperse particle with high density lipoprotein 3 (HDL(3)) density. In contrast, apoB:931 (residues 1-931), missing only 69 residues of the sequence homologous to lipovitellin, was secreted as a particle considerably more dense than HDL(3). In the present study we have determined the stoichiometry of the lipid component of the apoB:931 and apoB:1000 particles. The secreted [(3)H]glycerol-labeled apoB:1000 particles, isolated by nondenaturing gradient gel electrophoresis, contained 50 phospholipid (PL) and 11 triacylglycerol (TAG) molecules/particle. In contrast, apoB:931 particles contained only a few molecules of PL and were devoid of TAG. The unlabeled apoB:1000 particles, isolated by immunoaffinity chromatography, contained 56 PL, 8 TAG, and 7 cholesteryl ester molecules/particle. The surface to core lipid ratio of apoB:1000-containing particles was approximately 4:1 and was not affected by oleate supplementation. Although very small amounts of microsomal triglyceride transfer protein (MTP) were associated with apoB:1000 particles, it never approached a 1:1 molar ratio of MTP to apoB. These results support a model in which (i) the first 1000 amino acid residues of apoB are competent to complete the lipid pocket without a structural requirement for MTP; (ii) a portion, or perhaps all, of the amino acid residues between 931 and 1000 of apoB-100 are critical for the formation of a stable, bulk lipid-containing nascent lipoprotein particle, and (iii) the lipid pocket created by the first 1000 residues of apoB-100 is PL-rich, suggesting a small bilayer type organization and has a maximum capacity on the order of 50 molecules of phospholipid.

Nutritional assessment, health markers and lipoprotein profile in postmenopausal women belonging to a closed community.

To assess the dietary characteristics of a closed community and their relationship with several health markers and lipid and lipoprotein values in postmenopausal women. Energy and nutrient intake, serum lipids, lipoproteins, antioxidants, peroxides and low-density lipoprotein (LDL) peroxides in addition to several health markers were measured in a closed, postmenopausal female community consuming a diet without meat, meat products and alcoholic beverages. Departamento de Nutrición and Sección Departamental de Química Analítica and Escuela de Especialización de Análisis Clínicos, Universidad Complutense de Madrid, Spain and Lerma, Burgos, Spain. Cereals, vegetables, legumes and fruit, together with milk and eggs, constituted the most important ingredients of the diet consumed. Dietary carbohydrates contributed 42%en and lipids 46.4%en. The SFA/MUFA/PUFA ratio was 1/2/1 and the n-3/n-6 ratio 0.05 (SFA=saturated fatty acids, MUFA=monounsaturated fatty acids, PUFA=polyunsaturated fatty acids). The study community diet was monotonous and made for possible deficiencies of iron, magnesium, zinc, vitamin B(6) and vitamin D, in variable proportions. Routine biochemical and haematological normality markers indicate that only one woman presented hyperglycaemia and hyperuricaemia. Two women had haemoglobin levels <12 g/dl, but their mean corpuscular volume or mean corpuscular haemoglobin was normal. The prevalence of high cholesterol values (>6.21 mmol/l) was 42.8%, while that of high LDL-cholesterol levels (>3.88 mmol/l) was 35.7%, but none of the women displayed levels of high-density lipoprotein (HDL)-cholesterol <1.16 mmol/l, triglycerides >1.2 mmol/l or an LDL-cholesterol/HDL-cholesterol ratio>3. Only one woman had apolipoprotein (Apo) B levels >1.5 g/l, while most of the women presented Apo B values <1.2 g/l and an ApoA-1/ApoB ratio &<1.1. Plasma and LDL-peroxide levels, together with the tocopherol and carotene intakes, suggest a good antioxidant status in this population. The diet of the study group seems compatible with a healthy life-profile and permits a more-than-acceptable degree of cardiovascular disease protection. However, the consumption of certain nutrients should be improved.

Normal VLDL metabolism despite altered lipoprotein composition in type 1 diabetes mellitus.

Patients with type 1 diabetes are at increased risk of cardiovascular disease, which may be related to abnormal lipid metabolism. Secretion and clearance of VLDL apolipoprotein B100 (apoB) are important determinants of plasma lipid concentrations and are known to be influenced by hormones, including insulin and growth hormone. This study examined overnight VLDL apoB metabolism and VLDL composition in six lean patients with type 1 diabetes during euglycaemia (controlled by a varying insulin infusion) and in six age-, sex- and BMI-matched control subjects. VLDL apoB kinetics were determined using a primed constant 1-13C leucine infusion, and VLDL apoB enrichment was measured by gas-chromatography mass-spectrometry. Fasting lipid profile, IGF-I, IGFBP-3, overnight GH profiles and free insulin concentrations were also assessed. Fasting concentrations of triglycerides (TG), total cholesterol (TC), HDL-cholesterol (HDL-C) and LDL-cholesterol (LDL-C) were similar in both groups. The VLDL apoB secretion and metabolic clearance rates were not significantly different between the two groups, but the VLDL-TGNLDL apoB and the VLDL-CNLDL apoB ratios were significantly increased in those with diabetes (P < 0.02 and P < 0.03, respectively). Total IGF-I concentrations were similar between the two groups; however, the GH area under the curve and free insulin concentrations were increased in patients with type 1 diabetes (GH: diabetes: 94.8 +/- 15.1 vs. controls: 45.6 +/- 10-6, mU/L/h, P < 0.04; free insulin: diabetes: 78.4 +/- 5.0 vs. controls: 28.3 +/- 3.26, pmol/l, P < 0.001). IGFBP-3 concentrations were lower in diabetic patients (diabetes: 2,454.2 +/- 68.7 vs. controls: 3,219.4 +/- 76.4, ng/ml, P < 0.001). In the control group overnight GH secretion correlated negatively with fasting TC (P < 0.01) and LDL-C (P < 0.03) concentrations, whereas free insulin concentrations correlated positively with fasting TG concentrations (P < 0.009). No significant correlations were found in the patients with diabetes. This study suggests that in euglycaemic conditions patients with type 1 diabetes mellitus have normal VLDL apoB kinetics but altered VLDL composition. The altered VLDL composition may be associated with accelerated atherogenesis. We speculate that the disrupted hormonal balance and, in particular, the increased GH secretion might be responsible for the compositional changes of VLDL particles in type 1 diabetes mellitus.

A Deficiency of Microsomal Triglyceride Transfer Protein Reduces Apolipoprotein B Secretion

Microsomal triglyceride transfer protein (MTP) transfers lipids to apolipoprotein B (apoB) within the endoplasmic reticulum, a process that involves direct interactions between apoB and the large subunit of MTP. Recent studies with heterozygous MTP knockout mice have suggested that half-normal levels of MTP in the liver reduce apoB secretion. We hypothesized that reduced apoB secretion in the setting of half-normal MTP levels might be caused by a reduced MTP:apoB ratio in the endoplasmic reticulum, which would reduce the number of apoB-MTP interactions. If this hypothesis were true, half-normal levels of MTP might have little impact on lipoprotein secretion in the setting of half-normal levels of apoB synthesis (since the ratio of MTP to apoB would not be abnormally low) and might cause an exaggerated reduction in lipoprotein secretion in the setting of apoB overexpression (since the MTP:apoB ratio would be even lower). To test this hypothesis, we examined the effects of heterozygous MTP deficiency on apoB metabolism in the setting of normal levels of apoB synthesis, half-normal levels of apoB synthesis (heterozygous Apob deficiency), and increased levels of apoB synthesis (transgenic overexpression of human apoB). Contrary to our expectations, half-normal levels of MTP reduced the plasma apoB100 levels to the same extent ( approximately 25-35%) at each level of apoB synthesis. In addition, apoB secretion from primary hepatocytes was reduced to a comparable extent at each level of apoB synthesis. Thus, these results indicate that the concentration of MTP within the endoplasmic reticulum rather than the MTP:apoB ratio is the critical determinant of lipoprotein secretion. Finally, we found that heterozygosity for an apoB knockout mutation lowered plasma apoB100 levels more than heterozygosity for an MTP knockout allele. Consistent with that result, hepatic triglyceride accumulation was greater in heterozygous apoB knockout mice than in heterozygous MTP knockout mice.

Screening for hypercholesterolaemia in 10,000 neonates in a multi-ethnic population.

An immunoassay was developed for the detection of hypercholesterolaemia in dried blood spots collected from 6-day-old neonates. Blood spot samples (9,673) were subjected to immunoturbidimetric assay to determine the levels of apolipoprotein B (apoB), an index of plasma low density lipoprotein, and of apolipoprotein A-1 (apoA-1), an index of plasma high density lipoprotein. Infants with raised apoB or a reduced apoA-1/apoB ratio were recalled, retested and, if appropriate, referred to the Vascular Risk Clinic at King's College Hospital for medical management. A total of 189 were recalled for further testing; of whom 82 (45%) attended the recall clinic. A group of 16 families (24 individuals) had abnormal lipid profiles; of these, 7 families (14 individuals) had lipid profiles consistent with inherited hypercholesterolaemia. Neonatal screening for hypercholesterolaemia using blood spot apolipoprotein measurements is feasible but ethnic variations in disease prevalence must be considered in the design of a screening programme.

Caco-2 cells and human fetal colon: a comparative analysis of their lipid transport

Caco-2 cells and human colonic explants were compared for their ability to esterify lipid classes, synthesize apolipoproteins and assemble lipoproteins. Highly differentiated cells and colonic explants were incubated with [ 14C]oleic acid or [ 35S]methionine for 48 h. Caco-2 cells demonstrated a higher ability to incorporate [ 14C]oleic acid into cellular phospholipids (13-fold, P<0.005), triglycerides (28-fold, P<0.005) and cholesteryl ester (2-fold, P<0.01). However, their medium/cell lipid ratio was 11 times lower, indicating a limited capacity to export newly synthesized lipids. De novo synthesis of apo B-48 and apo B-100 was markedly increased (7%0 and 240%, respectively), whereas the biogenesis of apo A-I was decreased (60%) in Caco-2 cells. The calculated apo B-48/apo B-100 ratio was substantially diminished (107%), suggesting less efficient mRNA editing in Caco-2 cells. When lipoprotein distribution was examined, it displayed a prevalence of VLDL and LDL, accompanied along with a lower proportion of chylomicron and HDL. In addition, differences in lipoprotein composition were evidenced between colonic explants and Caco-2 cells. Therefore, our findings stress the variance in the magnitude of lipid, apolipoprotein and lipoprotein synthesis and secretion between the two intestinal models. This may be due to various factors, including the origin of Caco-2 cell line, i.e., colon carcinoma.

Divergent responses of the liver to increased delivery of glucose or fatty acids: implications for the pathogenesis of type IV hyperlipoproteinemia

This study examines the patterns of response of primary cultures of hamster hepatocytes to increased delivery of glucose or oleate. Increased glucose in the medium produced: (1) increased triglyceride in the cells and the medium; (2) no change in cholesterol ester in the cells or the medium; (3) no change in apo B100 secreted into the medium; (4) more apo B100 particles within the VLDL range with an increase in the VLDL triglyceride to apo B100 ratio. By contrast, increased oleate in the medium resulted in: (1) increased triglyceride in the cells and the medium; (2) increased cholesterol ester in the cells and the medium; and (3) increased apo B100 secreted into the medium. Important differences in the intracellular metabolism of triglyceride and cholesterol ester were also documented. Under all circumstances, there was substantially more radiolabelled triglyceride (overall eight times more) in the cell than in the medium, indicating that up to 90% of the newly synthesized triglyceride enters the cellular pool rather than being secreted with apo B100. By contrast, almost half of the newly synthesized cholesterol ester molecules were secreted with apo B100, pointing to an equal probability of entering the cell storage pool as opposed to being secreted. The data establish therefore two patterns of response of the liver to increased triglyceride synthesis depending on whether the substrate drive is glucose or oleate.

Effects of medium chain fatty acids (MCFA), myristic acid, and oleic acid on serum lipoproteins in healthy subjects

In this study we investigated the effects on lipoproteins of medium chain fatty acids (MCFA) and myristic acid relative to those of oleic acid. Thirty-seven women and 23 men consumed a 3-wk run-in diet enriched in oleic acid followed by a 6-wk test diet rich in MCFA (n = 21), myristic (n = 20), or oleic acid (n = 19). Experimental fats were incorporated into solid foods. Total fat intake was 40 En% fat. The dietary compositions were the same except for 10 En%, which was provided by MCFA, myristic, or oleic acids, respectively. With the myristic acid diet, low density lipoprotein (LDL) cholesterol was 0.37 mmol/L higher compared with the oleic acid diet (P = 0.0064 for difference in changes). The MCFA diet increased LDL cholesterol, though not significantly, with 0.23 mmol/L relative to the oleic acid diet (P = 0.0752). Compared with the oleic acid diet, HDL cholesterol concentrations increased with the myristic acid diet by 0.10 mmol/L (P = 0.0273) but not with the MCFA diet. The MCFA diet slightly elevated triacylglycerol concentrations, but responses did not significantly differ between the diets. The MCFA diet significantly decreased the apoA-I to apoB ratio compared with both other diets (P < 0.02). We conclude that MCFA raise LDL cholesterol concentrations slightly and affect the apoA-I to apoB ratio unfavorably compared with oleic acid. Myristic acid is hypercholesterolemic, although less than predicted earlier, and raises both LDL and HDL cholesterol concentrations compared with oleic acid.

MUCOSAL AND HEPATIC PROTEIN SYNTHESIS MEASURED SIMULTANEOUSLY WITH INTRAGASTRIC AND INTRAVENOUS STABLE ISOTOPIC TRACERS IN FED PIGLETS

The pool of amino acids used for intestinal mucosal and hepatic protein synthesis can derive from the diet, from intracellular proteolysis and from arterial amino acids. The relative contributions of these sources to mucosal and hepatic protein were quantified in piglets (N=4; 7.4 kg body weight), previously implanted with gastric (IG), portal (PORT), jugular (IV) and arterial (ART) catheters. After an overnight fast, the animals were fed hourly meals of a milk-based diet and were infused, for 6 h, with U13C-algal protein (IG) and 2H5-phenylalanine(PHE; IV). The tracer:tracee ratios of [U13C] and [2H5]PHE in ART and PORT blood, as well as in VLDL apolipoproteinB-100 (APOB), fibrinogen, hepatic and mucosal protein were measured by methane negative chemical ionization selected ion monitoring mass spectrometry. The table shows the mean ratios (± SD) of the isotopic enrichments of IV tracer to that of the IG tracer in ART, PORT and APOB-100 over the last h, and in plasma fibrinogen and hepatic protein at the end of the infusion. The ratio of the IV:IG tracers was (P<0.01) lower in PORT than in ART PHE. The tracer:tracee ratio of the IG tracer in APOB (4.98 ± 1.69 mol%) was higher (P<0.025) than in hepatic free PHE (1.92 ± 0.85). The IV:IG tracer ratio of APOB-100, fibrinogen and hepatic protein were all lower (P<0.05) than that of ART PHE, but the IV:IG ratio was higher in APOB than in fibrinogen and liver constitutive protein. The ratio of the two tracers in mucosal protein (0.27 ± 0.02) was 30% (P=0.05 by paired t-test) higher than their ratio in mucosal free PHE (0.21 ± 0.06). We conclude (i) that PHE for mucosal and hepatic protein synthesis derives from the extracellular input rather than from amino acids released by intracellular proteolysis and (ii) that ART PHE may be a preferential source of mucosal and PORT PHE a preferential source of hepatic protein synthesis.

Lipid, apolipoprotein, and lipoprotein synthesis and secretion during cellular differentiation in Caco-2 cells.

Although Caco-2 cells are frequently employed for the study of enterocyte lipid metabolism, variable results have been reported regarding their ability to synthesize and secrete lipids and apolipoproteins. The major goal of this investigation is to examine the capacity of Caco-2 cells to elaborate and secrete lipids, lipoproteins, and apolipoproteins at different degrees of morphological and functional differentiation. Cells were cultured in medium with 5% fetal bovine serum (FBS), on permeable polycarbonate filters from 2 to 30 d in the presence of 14C-oleate or 35S-methionine. Cellular differentiation, as assessed by morphology (light and electron microscopy), transepithelial resistance, free fatty acid flux, and sucrase activity, progressed steadily up to 20 d of culture. Caco-2 cells esterified oleic acid mainly into phospholipids, triglycerides (TG), and smaller amounts of cholesterol esters. Lipid synthesis began as early as 2 d, and TG secretion was enhanced with increased duration of culture. However, very low efficiency of lipid export was observed at all levels of differentiation, reaching a maximum of only 6% of intracellular lipids. VLDL and LDL were the dominant lipoproteins secreted, with HDL comprising < 20% of the total. VLDL secretion increased, while LDL decreased, whereas the lipid composition of lipoproteins varied little with increasing duration of culture. Apoprotein B and A-I synthesis and secretion increased markedly from 11 to 20 d of culture. The ratio of apo B-100/B-48 decreased between 11 and 30 d, consistent with enhanced apo B editing of more mature enterocytes. Taken together, our data suggest that from 20 d of culture, Caco-2 cells are morphologically and functionally mature, capable of lipid esterification, and lipoprotein and apolipoprotein synthesis. However, despite their functional and morphological similarities to mature enterocytes, Caco-2 cells have a very limited lipid export capacity.

The usefulness of measuring body fat deposition for detecting obesity and atherogenesity in Japanese school children.

Body fat deposition was measured in overweight and non-overweight children using a bioelectrical impedance method, and its relationship with serum lipids and apolipoproteins was investigated in 90 overweight children (over 120% of their ideal weight) and 241 non-overweight children aged 10-15 years in Niigata Prefecture. The results were as follows. Overweight boys had significantly higher levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), atherogenic index (AI), RLP-cholesterol (RLP-C), apoA1, apoA2, apoB, apoC2, apoC3, apoE and the ratio of apoB to apoA1 than non-overweight boys. Overweight girls had significantly higher levels of TC, LDL-C, AI, remnant-like lopoprotein cholesterol (RLP-C), apoA2, apoB, apoC2, apoC3, apoE and the ratio of apoB to apoA1 than non-overweight girls. It has been reported that of all children studied 2.1% had higher levels of RLP-C than its upper limit known for adults (12 mg/dL). Of the overweight children in the present study, 4.4% had a high level of RLP-C whereas only 1.2% of non-overweight children had a high RLP-C level. No difference in the lipoprotein levels was found between overweight and non-overweight children. In both boys and girls, relative weight, body fat, skinfold thickness and body mass index (BMI) were correlated with the lipoprotein levels. Non-overweight boys whose body fat was over 20% had significantly higher levels of TC, LDL-C, apoA2, apoB, apoC2, apoE and apoB/A1 than those whose body fat was less than 20%. It was concluded that the measurement of body fat deposition, together with relative weight, was useful for detecting obesity and atherogenesity in Japanese school children.

Lipoprotein analyses in patients with stable angina and acute coronary syndrome

We have performed lipid analyses by using a table-top ultracentrifuge based on the Hatch and Lees' method in 77 subjects (60 men, 17 women; mean age, 63 years) to clarify lipoprotein disorders in coronary artery disease. Sixty-four subjects had coronary artery disease and 13 normal subjects were controls. They were divided into the groups with stable angina pectoris and with unstable angina pectoris or acute myocardial infarction (acute coronary syndromes). In patients with coronary artery disease, there were no significant differences from control in age, body mass index, total cholesterol, however, HDL cholesterol was significantly lower than those in the controls. LDL cholesterol:LDL apoB ratio, which is thought to reflect the size of LDL in coronary artery disease, was significantly smaller than that in the controls; mean values were 1.2 in coronary artery disease and 1.4 in controls. There were no significant differences in those lipoprotein disorders between the patients with stable angina and those with acute coronary syndromes. Though these lipoprotein abnormalities would not play a trigger role in acute coronary syndromes, they are characteristic of the lipid profile of patients with coronary artery disease.

Genetic contributions to LDL-C, Apo-B and LDL-C/Apo-B ratio in a sample of Israeli offspring with a parental history of myocardial infarction.

One hundred and forty sibships consisting of 280 brothers and 256 sisters with a family history of myocardial infarction were investigated for the possible involvement of a major gene in the determination of LDL-C, Apo-B and LDL-C/ Apo-B ratio (as a surrogate for LDL subclasses). The mean ages were 29.5 years (range 15-48) and 29.2 years (range 15-47), for brothers and sisters, respectively, and values were initially adjusted for age effects through multiple regression analysis. Results from commingling analysis indicated that for LDL-C a single normal distribution fitted the data as well as a mixture of two distributions. For Apo-B, a mixture of two normal distributions fitted the data significantly better than a single distribution (chi 2 = 7.8, df = 2, p = 0.02). For LDL-C/ Apo-B ratio a mixture of three normal distributions fitted the data significantly better than two distributions (chi 2 = 9.2, df = 2, p = 0.01). A regression analysis applied to the logarithm of the sex- and age-adjusted mean and variance within sibship, showed no indication of a major gene involvement for LDL-C. For Apo-B and LDL-C/Apo-B ratio, there existed, however, significant linear relationships between the logarithmically transformed means and within sibship variances which support the involvement of major genes. In addition, the Bartlett test applied to the data of within-sibship variances also rejected the null hypothesis of multifactorial transmission for Apo-B and LDL-C/Apo-B ratio (p < 0.0001). Lastly, the results from sib-pair linkage analyses provided significantly positive evidence for linkage between ApoB levels and the Apo-B XbaI restriction site.