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Biomedical subjects

S M Humphreys

Publications and source records attributed to S M Humphreys.

At least 55 records · Page 3Linked to original sources
PubMed1995

Human triacylglycerol-rich lipoprotein subfractions as substrates for lipoprotein lipase.

In order to test the hypothesis that lipoprotein lipase (LPL) acts preferentially on larger lipoprotein particles, we determined the susceptibility of triacylglycerol-rich lipoprotein (TRL) subfractions to hydrolysis by LPL in vitro. Chylomicrons (Sf > 400), very low density lipoproteins (VLDL)1 (Sf 60-400) and VLDL2 (Sf 20-60) were isolated from six subjects with a range of plasma-triacylglycerol (TAG) concentrations following an overnight fast and for up to 6 h after the consumption of a mixed meal (41% fat). The percent of TRL-TAG hydrolysed by LPL in subfractions isolated following overnight fast was VLDL1 > VLDL2 (46.8 +/- 10.2 vs. 25.9 +/- 7.4%, P = 0.006) and 3 h after the meal it was chylomicrons > VLDL1 > VLDL2 (81.0 +/- 12.6 vs. 52.8 +/- 10.2 vs. 27.7 +/- 6.2%, chylomicrons vs. VLDL1 and VLDL1 vs. VLDL2, both P < or = 0.005). The percent of VLDL1-TAG hydrolysed increased both within and between subjects as VLDL1-TAG concentrations increased. This relationship could be explained by the positive correlation observed between VLDL1-TAG and VLDL1-TAG:apolipoprotein B. In conclusion, increasing the size and TAG content of a lipoprotein particle increases its susceptibility to hydrolysis by LPL.

Adult↗
PubMed1995

Coordinated regulation of hormone-sensitive lipase and lipoprotein lipase in human adipose tissue in vivo: implications for the control of fat storage and fat mobilization.

The enzymes lipoprotein lipase (LPL, EC 3.1.1.34) and hormone-sensitive lipase (HSL, EC 3.1.1.3) apparently catalyze opposing functions in white adipose tissue: the former is concerned with fat storage, the latter with fat mobilization. We have studied their regulation in vivo in normal subjects in the postabsorptive state and after eating meals of different compositions, by measurement of arteriovenous concentration differences for triacylglycerol, non-esterified fatty acids and glycerol across a subcutaneous adipose depot. The two enzymes are regulated in a broadly reciprocal manner: in the overnight-fasted state, HSL is more active, but after a meal HSL is suppressed whilst LPL is activated. The movement of fatty acids in and out of adipose tissue appears to be driven by concentration gradients generated by regulation of these two enzymes, and also by activation, in the postprandial period, of the process of fatty acid esterification. The results show some interesting and perhaps unexpected features of metabolic regulation. Of the fatty acids generated by the action of LPL on circulating TAG, a large proportion is released directly into the venous plasma: close to 100% in the overnight-fasted state, and 50% or more at the peak of LPL action after a meal, making what appear reasonable assumptions. We suggest that this apparent 'inefficiency' of fat storage reflects the energetic cost of maintaining precise control over such a fundamental process. Although LPL is usually thought of as the enzyme regulating fat deposition, in fact the fatty acids and glycerol it releases from circulating TAG represent a substantial proportion of those released from adipose tissue, especially in the postprandial state. In addition, although HSL is considered the enzyme responsible for fat mobilization, suppression of its activity is essential to normal regulation of fat deposition. Thus, fat storage and fat mobilization during normal daily life are controlled by coordinated regulation of a number of enzymatic processes in white adipose tissue.

Adipose Tissue↗
PubMed1995

Impaired postprandial clearance of triacylglycerol-rich lipoproteins in adipose tissue in obese subjects.

Adipose tissue is an important site of clearance of circulating triacylglycerol (TAG), especially in the postprandial period. Postprandial lipemia is usually increased in obesity. We studied the extraction of TAG from plasma and TAG-rich lipoproteins (TRLs) in subcutaneous adipose tissue in 11 control and 8 obese subjects before and after a mixed meal. Clearance of plasma TAG and very low-density lipoprotein (VLDL)-TAG was decreased in the obese subjects after an overnight fast. After the meal, chylomicron-TAG extraction increased in the control group whereas VLDL-TAG clearance decreased; these changes were not seen in the obese group, in whom the VLDL particles appeared to be better able to compete with the chylomicrons for clearance by lipoprotein lipase. In the control subjects, removal of TAG from the TRL in the postprandial period was accompanied by a shift toward addition of cholesterol to the high-density lipoprotein (HDL) fraction; this was not observed in the obese subjects. We conclude that disturbed TRL-TAG clearance in adipose tissue is related both to the elevated plasma TAG concentrations and the depressed HDL-cholesterol concentrations typical of obesity.

Adipose Tissue↗
PubMed1995

Catecholamine interference with enzymatic determination of nonesterified fatty acids in two commercially available test kits.

We present evidence that catecholamines, which are commonly used to stimulate lipolysis in adipose tissue in vitro, interfere with the enzymatic determination of non-esterified fatty acid (NEFA) in two commercially available kits. Measurement of a 100 mumol/L standard with the Wako "NEFA C" test kit was 60% inhibited by 100 mumol/L norepinephrine and was completely inhibited by 100 mumol/L isoproterenol or by 1 mmol/L norepinephrine or epinephrine. Measurement with the Boehringer Mannheim "Free Fatty acids, Half-micro test" was completely inhibited by 100 mumol/L norepinephrine and was also affected by concentrations as low as 0.1 mumol/L. We propose that this effect is due to the catecholamines interfering with a step common to the two kits, the generation of hydrogen peroxide and oxidation of a chromagen; furthermore, this interference appears to be stoichiometric. We also give details of an alternative in-house method, which does not depend on the generation of hydrogen peroxide and is not affected by catecholamines.

Acyl-CoA Oxidase↗
PubMed1994

Forearm substrate utilization during exercise after a meal containing both fat and carbohydrate.

1. Whilst the provision of exogenous carbohydrate has been shown to be beneficial to endurance exercise performance, little attention has been paid to the possibility of dietary manipulation of the availability of fat. 2. Ten normal subjects were studied on two occasions: after an overnight fast (postabsorptive state) and after a meal containing 80 g of fat and 80 g of carbohydrate (fed state). Forearm substrate exchange was studied during 60 min of isometric forearm exercise (5 s contraction, 5 s relaxation). 3. In the fed state concentrations of plasma triacylglycerol (1510 +/- 150 versus 850 +/- 80 mumol/l, P < 0.01), blood ketone bodies (151 +/- 21 versus 80 +/- 10 mumol/l, P < 0.01) and plasma insulin (17 +/- 3 versus 7 +/- 1 m-units/l, P < 0.01) were elevated compared with the postabsorptive state; plasma glucose and non-esterified fatty acid concentrations were not significantly different. 4. Forearm blood flow and O2 consumption each increased about 6-fold during exercise, with no differences between the two nutritional states. The potential contribution of individual substrates to forearm O2 consumption (a calculation which is independent of blood flow) was assessed: for triacylglycerol it was significantly greater in the fed state (P < 0.01). The sum of the potential contributions of triacylglycerol, non-esterified fatty acids, glucose and ketone bodies to forearm O2 consumption was significantly greater, both before and during exercise, in the fed than in the postabsorptive state (P < 0.05), implying the sparing of endogenous (forearm) fuels. 5. These studies highlight the potential for manipulation of substrate supply during exercise by feeding meals containing both carbohydrate and fat.

Adult↗
PubMed1994

Fasting plasma triacylglycerol concentrations predict adverse changes in lipoprotein metabolism after a normal meal.

The changes in lipoprotein metabolism which follow the ingestion of a large fat load have been well described. The hypothesis was tested that similar changes in lipoprotein metabolism would occur after a relatively normal meal. Plasma and lipoprotein triacylglycerol, cholesterol and apolipoprotein concentrations were determined in twenty subjects (ten female) given a mixed meal containing approximately one-third of the daily intake of major nutrients in the typical Western diet. Fasting plasma triacylglycerol concentrations (range 0.38-2.70 mm/l) and the postprandial rise in plasma triacylglycerol varied considerably between subjects and were significantly associated (P < 0.01). The rise in plasma triacylglycerol corresponded to marked increases in the triacylglycerol concentration of the triacylglycerol-rich lipoproteins (TRL; chylomicrons and very-low-density lipoproteins). TRL cholesterol also increased after the meal. An increase in high-density-lipoprotein (HDL)-triacylglycerol following the meal was accompanied by a decrease in HDL-cholesterol concentration, presumably due to the action of the cholesteryl-ester transfer protein. The increases in HDL-triacylglycerol and in TRL-cholesterol were correlated with the postprandial rise in triacylglycerol in the TRL (P < 0.01). We conclude that potentially adverse changes occur in both triacylglycerol-rich and high-density lipoproteins following a typical mixed meal, as they do after large fat loads. The changes are exaggerated in those subjects with greater fasting plasma triacylglycerol concentrations.

Adult↗
PubMed1994

Immediate metabolic availability of dietary fat in combination with carbohydrate.

We tested the hypothesis that when fat is ingested in combination with carbohydrate, direct release of fatty acids into the plasma may occur. Eight normal subjects ingested two meals: high-fat (80 g fat, 80 g carbohydrate, and 18 g protein) and low-fat (< 1 g fat, otherwise closely matched). Over the subsequent 6 h, net fat oxidation was greater after the high-fat meal (20.7 vs 10.6 g, P < 0.01). Plasma nonesterified fatty acid (NEFA) concentrations were markedly suppressed after the low-fat meal, but relatively maintained after the high-fat meal (P < 0.01). The profile of plasma NEFAs changed after the high-fat meal, consistent with entry of meal-derived fatty acids into the NEFA pool. We suggest that after ingestion of combinations of carbohydrate and fat, the action of lipoprotein lipase on chylomicron-triacylglycerol leads to direct release of fatty acids into the plasma and increased fat oxidation.

Adult↗
PubMed1994

Regulation of fatty acid movement in human adipose tissue in the postabsorptive-to-postprandial transition.

There is net outward flow of fatty acids from adipose tissue in the fasted state but net inward flow and storage in the postprandial state. We investigated how this is regulated. Arteriovenous differences were measured across a subcutaneous adipose depot in six normal subjects before and for 5 h after a meal containing 80 g fat and 80 g carbohydrate. In five further experiments, insulin was infused at 40 mU.m-2.min-1 from 30 min after the meal, clamping the plasma glucose. Net transcapillary fatty acid flow changed from negative (outward flow from tissue to capillaries) in the postabsorptive state to consistently positive (net inward flow, implying fat storage) after the meal despite continued net efflux of fatty acids into venous blood. In the "clamped" experiments (with additional insulin), net fatty acid efflux in the venous blood was suppressed and positive transcapillary flux (storage) was more marked. Regulation of fatty acid flow appeared to depend on coordinated changes in hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) action and fatty acid esterification. Additional insulin caused no further suppression of HSL or activation of LPL but markedly stimulated fatty acid retention (presumed to represent esterification). In the absence of additional insulin, a high proportion of the fatty acids liberated by LPL are released into the venous plasma in both postabsorptive and postprandial states. We hypothesize that this "loss" of fatty acids is necessary to give precise control to the pathway of fat storage.

Absorption↗
PubMed1993

Mono-, di- and triacylglycerol concentrations in human plasma: effects of heparin injection and of a high-fat meal.

A sensitive method has been developed to measure specific mono-, di- and triacylglycerol concentrations in human plasma, using thin-layer chromatography and enzymatic assay. The levels of partial acylglycerols in human plasma from fasting subjects were lower than previous reports had suggested and amounted to less than 3% of the total acylglycerols. After heparin injection the plasma monoacylglycerol concentration increased markedly (P < 0.01) while the triacylglycerol concentration decreased significantly (P < 0.001). The plasma diacylglycerol concentration did not change significantly although it increased as a percentage of the total (P < 0.05); after heparin partial acylglycerols accounted for more than 10% of the total. After a high-fat meal the plasma concentrations of di- and triacylglycerol increased approximately two-fold (P < 0.005) but no significant change was observed for mono-acylglycerol. The percentage contribution of partial acylglycerols was unchanged (2.6% fasting, 2.4% postprandially).

Adult↗
PubMed1993

Periprandial regulation of lipid metabolism in insulin-treated diabetes mellitus.

We have examined the regulation of lipid and glucose metabolism in the postabsorptive and postprandial states in six subjects with insulin-treated diabetes mellitus, and compared them with eight nondiabetic subjects. Blood or plasma concentrations of metabolites and fluxes across forearm and subcutaneous adipose tissue were studied after an overnight fast and for 6 hours after a mixed meal (3.1 MJ, 41% from fat). In the postabsorptive state, regulation of lipid metabolism in the two groups appeared basically similar except that a wider spread of plasma (free) insulin concentrations in the diabetic group led to a wider range of values of plasma nonesterified fatty acid (NEFA) release from adipose tissue, plasma NEFA concentrations, and blood ketone body concentrations. Extraction of ketone bodies across adipose tissue was positively correlated with arterial concentration in both groups (as it was in the forearm), confirming the ability of human adipose tissue to utilize ketone bodies. A single subcutaneous injection of insulin before the meal in the diabetic group produced a plasma free-insulin profile that was blunted and prolonged compared with the postprandial response in the control group. Postprandial forearm glucose uptake followed very closely the plasma (free) insulin concentration. Postprandial suppression of NEFA release from adipose tissue was essentially normal in the diabetic group, and the normal postprandial decrease in plasma NEFA concentrations was reproduced extremely closely. Forearm and adipose tissue blood flow did not differ between the groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1993

Subcutaneous adipose tissue metabolism studied by local catheterization.

Adipose tissue is a metabolically active tissue which plays a key role in regulating the concentrations of circulating lipid fuels. In recent years, two new techniques have become available for the study of human adipose tissue metabolism in vivo: microdialysis, and catheterization of the venous drainage from the subcutaneous abdominal adipose tissue. Blood obtained by the latter method shows all the characteristics expected of adipose tissue drainage. The characteristics of this tissue are quite distinct from those of the superficial (mainly skin) or the deep (mainly muscle) tissues of the forearm. The depot studied appears to be typical of adipose tissue as a whole in terms of non-esterified fatty acid release. In comparison with the microdialysis technique, the arterio-venous difference technique allows easier quantification of substrate uptake and release, and allows the study of hydrophobic molecules (e.g. fatty acids, triacylglycerol). On the other hand, it does not allow the study of more than one depot, nor the local introduction of effectors of metabolism (e.g. adrenergic agents). The two techniques are clearly complementary.

Adipose Tissue↗
PubMed1992

Adipose tissue metabolism in obesity: lipase action in vivo before and after a mixed meal.

Physiological actions of insulin include suppression of fat mobilization from adipose tissue and activation of adipose tissue lipoprotein lipase. Here, we report measurements of adipose tissue hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) action in vivo in 10 normal and eight obese subjects, with the latter group having varying degrees of glucose intolerance. HSL and LPL actions (per gram of adipose tissue) were similar in the two groups, after an overnight fast. In the normal subjects, HSL action was suppressed after a meal (by 75% +/- 6% between 60 to 300 minutes, P less than .01), and the action of LPL was increased (clearance of circulating triacylglycerol [TAG] increased by 140% +/- 57% at 300 minutes, P less than .05). Despite hyperinsulinemia, these responses were blunted in the obese subjects (P less than .05 for each change being less than in normal group). The adipose tissue of the obese subjects showed continued nonesterified fatty acid (NEFA) release at a time when NEFA mobilization was completely suppressed in the normal group. Both impaired suppression of HSL and low fractional retention of fatty acids for reesterification within the adipose tissue contributed to this abnormal NEFA release. Impaired activation of LPL was associated with a greater absolute increase in plasma TAG concentration postprandially in the obese. In obese subjects, adipose tissue HSL and LPL fail to respond to immunoreactive insulin postprandially, which may be an important maladaptation in terms of lipoprotein metabolism and risk of coronary heart disease.

Adipose Tissue↗
PubMed1991

Glycerol and lactate uptake in human forearm.

Arteriovenous differences for lactate and glycerol reported across the human forearm are inconsistent in direction and magnitude. Such inconsistency could represent the effects of differing forearm compositions. The hypothesis was tested by examination of 37 studies of forearm arteriovenous differences for lactate, glycerol, glucose, and oxygen (only measured in 25 studies) in 23 normal subjects after overnight fast. In 16 studies, glycerol was taken up rather than released by the forearm, and in 12 of these the arteriovenous difference was greater than could be accounted for by analytical variation. The arteriovenous differences for glycerol and lactate were positively correlated (r = .44, P less than .01). The hypothesis that glycerol and lactate uptake might reflect a more "oxidative" forearm was not borne out, since neither glycerol nor lactate arteriovenous differences correlated with that for oxygen, although oxygen and glucose arteriovenous differences and fluxes were correlated (fluxes: r = .60, P less than .01). The arteriovenous difference for glycerol was positively related to body mass index, arguing against a variable contribution from fat. The hypothesis that the direction of glycerol and lactate exchange would reflect the forearm composition was not borne out by the analysis of repeated studies on the same individual, which showed that the variation within subjects was not significantly less than that between subjects. Therefore, we conclude that in approximately 40% of studies in normal subjects after an overnight fast, the forearm will show glycerol uptake, although we have been unable to identify any physiological reason for this phenomenon. Peripheral glycerol uptake has implications for studies in which glycerol release is taken as a measure of lipolysis.

Adult↗