Alogliptin ameliorates postprandial lipemia and postprandial endothelial dysfunction in non- diabetic subjects: a preliminary report

Background

Osteoprotegerin is a member of the tumor necrosis factor-related family and inhibits RANK stimulation of osteoclast formation as a soluble decoy receptor. The goal of this study was to determine the relationship of serum osteoprotegerin with vascular calcification in patients with type 2 diabetes.

Methods

The subjects were 124 patients with type 2 diabetes mellitus, including 88 males and 36 females with a mean (± SD) age of 65.6 ± 8.2 years old. Serum levels of osteoprotegerin, osteocalcin, fibroblast growth factor 23 (FGF23), 25-hydroxyvitamin D3 and adiponectin were measured by ELISA. Vascular calcification in the cervical artery was examined by ultrasound sonography. The subjects were divided into 4 quartiles depending on serum osteoprotegerin levels.

Results

Vascular calcification was significantly higher in the 4th quartile and significantly lower in the 1st quartile of serum osteoprotegerin levels, compared to other quartiles. There were no differences in serum osteoprotegerin and vascular calcification among patients with different stages of diabetic nephropathy, but serum FGF23 levels were elevated in those with stage 4 diabetic nephropathy. Simple regression analysis showed that serum osteoprotegerin levels had significant positive correlations with age, systolic blood pressure and serum adiponectin levels, and significant negative correlations with BMI and serum 25-hydroxyvitamin D3.

Conclusions

These findings suggest that elevated serum osteoprotegerin may be involved in vascular calcification independently of progression of diabetic nephropathy in patients with type 2 diabetes.     

Protocol for the study of the metabolism of retinyl esters in plasma lipoproteins during postprandial lipemia.

An efficient protocol is described for the study of the kinetics of retinyl esters in whole plasma and several lipoprotein fractions following the consumption of an oral fat load containing vitamin A (retinol). To allow for a more complete characterization of the kinetics of retinyl esters in different lipoprotein fractions, a simplified two-step ultracentrifugation procedure is reported for the efficient and reproducible isolation of triglyceride-rich chylomicrons from nonfasting subjects, VLDL-sized lipoprotein particles, and the triglyceride-poor lipoprotein fraction. The present method for the determination of retinyl esters is based on the direct application of the lipid fraction onto a normal phase HPLC column without requiring the lipid extract to be desiccated and resolubilized. All of the commonly occurring esters of retinol elute as a single peak with a retention time of 1.6-1.8 min followed by retinyl acetate (serving as the internal standard) and retinol with retention times of 2-2.5 min and 5-5.5 min, respectively. With this system, a new sample can be processed every 10 min and a complete set of 60 samples from a typical oral fat load can analyzed in one working day with minimal technical interaction. By normalizing to the area under the internal standard to correct for variability in the injected volume, the coefficient of variability for the concentration retinyl esters within a single run is less than 5% and less than 10% between runs.     

Effect of exercise and medium-chain fatty acids on postprandial lipemia.

The purpose of this study was to evaluate the effect of medium-chain triglycerides (MCT) with and without exercise on postprandial lipemia (PPL). Subjects were 25 young men and women. Each subject performed three trials: 1) control (fat meal only, 1.5 g fat/kg) 2) MCT (substitution of MCT oil, 30% of fat calories), and 3) MCT + Ex (exercise 12 h before the MCT meal). Before each trial, the subject underwent consistent dietary preparation. Blood was collected on 2 separate days for baseline measurements of postheparin lipases and, in each trial, at 0 h (premeal), at 2, 4, 6, and 8 h after the fat meal for triglycerides and cholesterol ester transfer protein (CETP), and at 8 h for postheparin lipoprotein lipase (LPL) and hepatic lipase activities (HL). ANOVA indicated that the partial substitution of MCT oil to the fat meal did not affect the PPL response. However, the PPL was significantly lower after the MCT + Ex trial vs. the other trials. LPL activity was significantly elevated after all trials compared with baseline, whereas HL was lower in the MCT + Ex trial only. CETP mass was significantly lower at 4 and 8 h than 0 h during all trials but relatively higher in the MCT + Ex trial vs. the nonexercise trials. These results suggest that MCT does not affect the TG response to a fat meal. LPL and CETP are affected by a fat meal with or without exercise, but HL is affected only when exercise is included.     

Effects of low and moderate exercise intensity on postprandial lipemia and postheparin plasma lipoprotein lipase activity in physically active men.

This study was designed to assess differences in the intensity of exercise to attenuate postprandial lipemia (PPL). Thirteen healthy men (age 23.8 +/- 0.9 yr) participated in three random-ordered trials: in low-(25% peak oxygen consumption; Low) and moderate-intensity (65% peak oxygen consumption; Mod) exercise trials, which were completed 1 h before a high-fat meal (1.3 g fat/kg body mass), and a control (Con), fat meal only, trial. Venous blood samples were obtained before the fat meal, and at 2, 4, 6, 8, and 20 h after the fat meal. PPL in the Mod trial (267 +/- 50 mg.dl-1.8 h) was lower compared with that in either Con (439 +/- 81 mg.dl-1.8 h) or Low (403 +/- 91 mg.dl-1.8 h) trials (P < 0.05), whereas there was no difference in PPL between Con and Low trials (P > 0.05). High-density lipoprotein cholesterol (HDL-C) and HDL subtype 2 cholesterol were not different between or within trials (P > 0.05). Postprandial insulinemia was lower in the Mod trial (20.5 +/- 5.7 microIU.ml-1.8 h; P < 0.05), but not in the Low trial (31.4 +/- 4.7 microIU.ml-1.8 h), compared with that in the Con trial (34.9 +/- 5.0 microIU.ml-1.8 h). Postheparin lipoprotein lipase activity at 8 h was higher in the Low trial compared with that in either Con or Mod trials, whereas there were no differences between trials at 20 h. These results suggest that, when exercise is performed 1 h before a fat meal, only exercise of moderate but not of low intensity attenuates PPL and that this effect is not associated with changes in postheparin lipoprotein lipase activity.     

Effects of exercise and n-3 fatty acids on postprandial lipemia.

Because n-3 fatty acid ingestion and aerobic exercise each has been associated with diminished postprandial lipemia (PPL), the purpose of this study was to evaluate the effect of a combination of these two factors on PPL. Sedentary men underwent a standard dietary preparation, including a 12-h fast before each trial. Six subjects performed a control trial (fat meal, 100 g fat) and an n-3 fatty acid trial (fat meal after 3 wk of n-3 fatty acid supplementation at 4 g/day). In a parallel experiment, six different subjects underwent a control trial and n-3 fatty acid supplementation + 60 min of exercise before ingestion of the fat meal. Supplementation with n-3 fatty acid significantly decreased baseline triglyceride (TG) concentrations but did not significantly affect PPL. The combination of n-3 fatty acid and exercise had no effect on the postprandial TG response. The present study suggests that n-3 fatty acid supplementation lowers resting TG concentrations but inhibits the beneficial effect of aerobic exercise on the postprandial TG response.     

Obesity and postprandial lipemia in adolescents: risk factors for cardiovascular disease

In the last 50 years, obesity has become a global epidemic and is one of the main public health problems in many parts of the world. Adolescence is a critical period regarding weight control. The factors determining obesity include a complex group of interrelated biological, behavioral and environmental factors which reinforce each other. In children and adolescents, obesity is associated with premature cardiovascular diseases, diabetes mellitus type 2, acanthosis nigricans, respiratory and skeletal muscle problems, as well as psychological problems. The clinical manifestations of cardiovascular disease begin in middle age. Nevertheless, studies indicate that the atherosclerotic process begins to develop during childhood. Postprandial hyperlipemia is a physiological process that occurs several times a day after the complete absorption of a diet including lipids and has been suggested as a risk factor for coronary heart disease (CHD). New study areas include the effects of different fatty acids, lipid sources (endogenous and exogenous), and the effect ingesting alcoholic beverages during meals. Given the evidence that postprandial lipidemia is an independent risk factor for CHD, it is vital to establish normative values for children and adolescents such that more effective and efficient preventive and therapeutic measures can be adopted.     

Effects of glucose ingestion on postprandial lipemia and triglyceride clearance in humans

To determine whether the metabolism of diet-derived triglycerides (TG) is acutely regulated by the consumption of insulinogenic carbohydrates, we measured the effects of glucose ingestion on oral and intravenous fat tolerance, and on serum triglyceride concentrations obtained during duodenal fat perfusion. Postprandial lipemia was diminished by the ingestion of 50 g (148 +/- 121 mg.dl-1 x 7 h-1 vs 192 +/- 124 mg.dl-1 x 7 h-1, P less than 0.05) and 100 g (104 +/- 106 mg.dl-1 x 7 h-1 vs 171 +/- 104 mg.dl-1 x 7 h-1, P less than 0.05) glucose. Peak postprandial TG concentrations occurred later after meals containing glucose and fat than after meals containing fat alone. This effect could be reproduced when an iso-osmotic quantity of urea was substituted for glucose in the test meal. Starch ingestion had no discernible effect on postprandial lipemia. Intravenous fat tolerance was similar before (4.9 +/- 1.2%.min-1) and 2 h (4.4 +/- 1.3%.min-1) and 4 h (4.8 +/- 1.5%.min-1) after 50 g glucose ingestion. During duodenal fat perfusion, glucose ingestion caused a progressive decrease in plasma triglyceride concentrations. These data suggest that glucose ingestion diminishes postprandial lipemia in a dose-dependent manner, but that this effect is not due to increased clearance of triglyceride from the circulation. The hypotriglyceridemic effects of glucose appear to reflect delayed gastric emptying and decreased hepatic secretion of triglyceride.     

Impaired fat-induced thermogenesis in obese subjects: the NUGENOB study

Objectives: To study energy expenditure before and 3 hours after a high‐fat load in a large cohort of obese subjects (n = 701) and a lean reference group (n = 113). Research Methods and Procedures: Subjects from seven European countries underwent a 1‐day clinical study with a liquid test meal challenge containing 95% fat (energy content was 50% of estimated resting energy expenditure). Fasting and 3‐hour postprandial energy expenditures, as well as metabolites and hormones, were determined. Results: Obese subjects had a reduced postprandial energy expenditure after the high‐fat load, independent of body composition, age, sex, research center, and resting energy expenditure, whereas within the obese group, thermogenesis increased again with increasing BMI category. Additionally, insulin resistance, habitual physical activity, postprandial plasma triacylglycerols, and insulin were all independently positively related to the postprandial energy expenditure. Resting energy expenditure, adjusted for fat‐free mass, increased with degree of obesity, a difference that disappeared after adjustment for fat mass. Furthermore, insulin resistance, fasting plasma free fatty acids, and cortisol were positively associated, whereas fasting plasma leptin and insulin‐like growth factor‐1 were negatively associated, with resting energy expenditure. Discussion: The 3‐hour fat‐induced thermogenic response is reduced in obesity. It remains to be determined whether this blunted thermogenic response is a contributory factor or an adaptive response to the obese state.     

Increased postprandial homocysteinemia in a group of depressed patients

Summary Elevated tissue and serum concentrations of homocysteine (HCY) are associated with neuropsychiatric disorders as well as with premature occlusive vascular disease, as seen in homocystinuria. In order to study dietary-related modifications in plasma HCY, total HCY was assayed in the fasted state and 2 hr after meals in 12 depressed female patients aged 54 to 81 yr and in 12 female controls aged 50 to 85 yr. Fasting HCY was also studied in 4 patients with dementia. Postprandial HCY varied only slightly in the controls compared with their fasting values, whereas a significant increase was noted in the depressives. To study the influence of normal and low protein diets on this abnormality, fasting and postprandial HCY were investigated in 4 of the depressives after one week of a normal diet, after a week on a diet without meat, fish or eggs, and then again after return to a normal diet for one week. Persistence of the abnormal increase in postprandial HCY in 2 of these 4 patients while on the low-protein diet may have been due to an inherited defect in HCY metabolism. Folate deficiency can also cause hyperhomocysteinemia, and as folate supplements constantly lower HCY concentrations, nutritional counseling and folate therapy might prove helpful in the treatment of depression.     

Increased plasma level of endothelin with vasoconstriction after dextran infusion.

Our previous study demonstrated that volume expansion with dextran produced blood pressure elevation due to vasoconstriction 3 hours after the cessation of infusion. To examine whether endogenous endothelin contributes to this vasoconstriction, we measured plasma level of endothelin before, immediately after, and 3 hours after the administration of dextran. Plasma level of endothelin decreased immediately after the administration (from 1.5 +/- 0.3 pg/ml to 1.1 +/- 0.2 pg/ml, P less than 0.05), and increased 3 hours after the administration (2.1 +/- 0.3 pg/ml, P less than 0.05). However, the changes in the plasma level of endothelin did not significantly correlated with those in blood pressure or total peripheral resistance. Thus, vasoconstriction after dextran infusion was accompanied by an increase in the plasma level of endothelin, but further evaluation is needed for the direct role of this peptide in the vasoconstrictive blood pressure elevation.     

Effects of a 1,3-diacylglycerol oil-enriched diet on postprandial lipemia in people with insulin resistance

Postprandial hypertriglyceridemia is common in individuals with insulin resistance, and diets enriched in 1,3-diacylglycerol (DAG) may reduce postprandial plasma triglycerides (PPTGs). We enrolled 25 insulin-resistant, nondiabetic individuals in a double-blind, randomized crossover trial to test the acute and chronic effects of a DAG-enriched diet on PPTG. Participants received either DAG or triacylglycerol (TAG) oil, in food products, for 5 weeks. Fasting lipids, and two separate postprandial tests, one with DAG oil and one with TAG oil, were performed at the end of each 5 week diet period. We found no acute or chronic effects of DAG oil on PPTG. Thus, neither the DAG oil PPTG (h/mg/dl) on a chronic TAG diet [area under the curve (AUC) = 503 +/- 439] nor the TAG oil PPTG on a chronic DAG diet (AUC = 517 +/- 638) was different from the TAG oil PPTG on a chronic TAG diet (AUC = 565 +/- 362). Five weeks of a DAG-enriched diet had no acute or chronic effects on PPTG in insulin-resistant individuals. We suggest further studies to evaluate the effects of DAG on individuals with low and high TG levels.     

Changes in plasma high-density lipoprotein cholesterol concentration after weight reduction in grossly obese subjects.

Changes in serum lipoproteins associated with weight loss were assessed in 13 grossly obese (relative weight 183%) patients who had participated in an outpatient semi-starvation diet consisting of liquid protein and carbohydrate. At the follow-up examination an average of six and a half months after the start of refeeding the mean weight loss was 16.1 +/- 4.5 kg or 15% of initial body weight. Significant increases in high-density lipoprotein (HDL) cholesterol of 0.16 +/- 0.05 mmol/l (6 +/- 2 mg/100 ml) and decreases in triglycerides (0.8 +/- 0.23 mmol/l; 73 +/- 20 mg/100 ml) and fasting blood sugar (0.6 +/- 0.22 mmol/l; 11 +/- 4 mg/100 ml) were observed. Changes in HDL cholesterol correlated significantly with changes in weight (r = 0.667) and percentage change in weight. The intercept of the regression equation relating HDL cholesterol to percentage change in weight was -7.3, indicating that a change in HDL cholesterol greater than zero required a weight loss of at least 7.3% of body weight. Thus, weight loss can significantly increase HDL cholesterol concentrations but a considerable amount of weight must be lost to produce a significant increase in HDL cholesterol concentration.     

Increased resting-state functional connectivity in obese adolescents; a magnetoencephalographic pilot study

Background

Obesity is not only associated with metabolic abnormalities, but also with cognitive dysfunction and changes in the central nervous system. The present pilot study was carried out to investigate functional connectivity in obese and non-obese adolescents using magnetoencephalography (MEG).

Methodology/Principal Findings

Magnetoencephalographic recordings were performed in 11 obese (mean BMI 38.8±4.6 kg/m²) and 8 lean (mean BMI 21.0±1.5 kg/m²) female adolescents (age 12–19 years) during an eyes-closed resting-state condition. From these recordings, the synchronization likelihood (SL), a common method that estimates both linear and non-linear interdependencies between MEG signals, was calculated within and between brain regions, and within standard frequency bands (delta, theta, alpha1, alpha2, beta and gamma). The obese adolescents had increased synchronization in delta (0.5–4 Hz) and beta (13–30 Hz) frequency bands compared to lean controls (P _{delta total} = 0.001; P _{beta total} = 0.002).

Conclusions/Significance

This study identified increased resting-state functional connectivity in severe obese adolescents. Considering the importance of functional coupling between brain areas for cognitive functioning, the present findings strengthen the hypothesis that obesity may have a major impact on human brain function. The cause of the observed excessive synchronization is unknown, but might be related to disturbed motivational pathways, the recently demonstrated increase in white matter volume in obese subjects or altered metabolic processes like hyperinsulinemia. The question arises whether the changes in brain structure and communication are a dynamic process due to weight gain and whether these effects are reversible or not.     

Metabolic syndrome is not a risk factor for kidney dysfunction in obese non-diabetic subjects

Objective: To investigate whether insulin resistance (IR) and the metabolic syndrome (MS) are associated with kidney dysfunction in obese non‐diabetic (OND) subjects. Methods and Procedures: Three‐hundred and eighty (113M/267F; age = 41 ± 14 years) OND subjects (BMI ≥ 30 kg/m²; range = 43 ± 8 kg/m²) were studied. Anthropometric measures, blood pressure, fasting glucose, insulin, lipid profile, and serum creatinine were evaluated. Glomerular filtration rate (GFR) was estimated (e‐GFR) with the Modification of Diet in Renal Disease equation. Chronic kidney disease (CKD) was defined as e‐GFR <60 ml/min/1.73 m². Results: e‐GFR was associated with gender (being lower in women) (P = 0.001) and age (P < 0.0001). CKD was present in 32 subjects (8.4%), who were older (P < 0.0001) and more frequently affected by hypertension (P = 0.04) as compared to subjects without CKD. MS was present in 212 (55.8%) subjects. They were older (P< 0.001), had lower e‐GFR (P = 0.02) and were more frequently affected by CKD (odds ratio (OR), 95% confidence interval (CI) = 2.3, 1.1–5.1) than those without MS. However, differences in e‐GFR values and in the risk of CKD were no longer statistically significant after adjusting for age (P = 0.99 for e‐GFR and OR, 95% CI = 1.2, 0.5–2.8 for the risk of CKD, respectively). Homeostasis model assessment of IR (HOMA_IR) index was neither higher in subject with CKD (P = 0.1) nor inversely correlated with e‐GFR (r = 0.1, P = 0.1). Discussion: In OND individuals the risk of CKD is independent of the MS and related abnormalities. This suggests that these individuals are not susceptible to a further deleterious role on kidney function on the top of that played by obesity itself.     

Increased intracellular glycerophosphoinositol is a biochemical marker for transformation by membrane-associated and cytoplasmic oncogenes

Transformation of rodent fibroblasts by cytoplasmic (mos, raf) and membrane-associated (ras, src, met, trk), but not nuclear (myc, fos) oncogenes results specifically in a very significant elevation of intracellular levels of glycerophosphoinositol (GPI). This elevation is specifically associated with the transformed state of the cells and not merely with their active state of proliferation. The basal phospholipase A2 (PLA2) activity of the same cells is also significantly stimulated in vivo. Our results are consistent with the notion that the elevated levels of GPI result from deacylation of lysophosphatidylinositol released by the enhanced PLA2 activity. GPI is a water-soluble, easily detectable metabolite which may constitute a convenient biochemical marker for malignant transformation by this particular group of oncogenes.