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The aim of this study was to determine whether the final enzymes in the two biosynthetic pathways for platelet-activating factor (PAF) (the 'de novo' and the 'membrane remodelling' pathways) are present in mouse embryos, zygotes and oocytes. The enzymes are dithiothreitol-insensitive cytidinediphosphocholine: 1-O-alkyl-2-acetyl-sn-glycerol cholinephosphotransferase (cholinephosphotransferase) in the de novo pathway and acetyl-coenzyme A:1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine acetyltransferase (acetyltransferase) in the membrane remodelling pathway. Activity of both enzymes was detected in the unfertilized oocyte, the zygote and also in the preimplantation embryo (48, 72 and 96 h after the ovulatory injection of hCG). In both cases the activity was destroyed by boiling and increased linearly with incubation time and the concentration of embryo homogenate present, indicating that the reactions were catalysed by enzymes. The product of the reactions was confirmed as PAF using HPLC and structural analyses by enzymatic digestion. Cholinephosphotransferase required Mg2+ and was inhibited by Ca2+, while acetyltransferase required the presence of NaF (a phosphatase inhibitor). The activity of cholinephosphotransferase was similar in unfertilized oocytes and zygotes, and did not change significantly with advancing developmental stage in preimplantation embryos. Acetyltransferase had a significantly lower specific activity (0.078 +/- 0.044 fmol PAF per oocyte per min, mean +/- SEM) in unfertilized oocytes than in zygotes of corresponding age (0.358 +/- 0.097 fmol PAF per zygote per min) (P < 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Detection and preliminary characterization of two enzymes involved in biosynthesis of platelet-activating factor in mouse oocytes, zygotes and preimplantation embryos: dithiothreitol-insensitive cytidinediphosphocholine: 1-O-alkyl-2-acetyl-sn-glycerol cholinephosphotransferase and acetyl-coenzyme A:1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine acetyltransferase. 793 73

The purpose of this study was to examine the effect of one bout of low-intensity exercise on the lipemic response to a high-fat meal. Twelve (six women, six men) normolipidemic adults aged 25.8 +/- 1.2 years (mean +/- SEM) took part in two trials. In the exercise trial, subjects walked for 2 hours on a treadmill at 30.9% +/- 1.6% of maximal oxygen uptake (VO2 max) 15 hours before ingestion of the test meal. In the control trial, subjects rested the day before the test meal. After a 12-hour fast, blood samples were obtained by venous cannulation before ingestion and hourly after ingestion for 6 hours. Serum was analyzed for triacylglycerol (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and HDL2-C, apolipoproteins (apos) A-I and B, free fatty acids (FFA), free glycerol, glucose, and insulin. TG values were corrected for free glycerol. Fasting serum TG and peak TG concentrations were lower (Wilcoxon, P < .05) for the exercise trial than for the control trial (0.74 +/- 0.03 v 0.92 +/- 0.08 and 1.98 +/- 0.18 v 2.59 +/- 0.32 mmol.L-1, respectively). The total lipemic response (area under the TG/time curve, normalized to the 0-hour level) was 31% +/- 7% lower in the exercise trial (4.28 +/- 0.66 v 6.46 +/- 1.08 mmol.L-1.h, P < .01). No differences were found between trials in the other parameters. These results show that a single bout of low-intensity exercise reduces the extent of postprandial lipemia in normolipidemic young adults. One possible mechanism is enhanced lipoprotein lipase (LPL) activity in the exercised skeletal muscle.
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PMID:The effect of a single bout of brisk walking on postprandial lipemia in normolipidemic young adults. 802 6

This study investigated the percentage of carbohydrate utilization than can be accounted for by glucose ingested during exercise performed after the ingestion of the potent lipolysis inhibitor Acipimox. Six healthy male volunteers exercised for 3 h on a treadmill at about 45% of their maximal oxygen uptake, 75 min after having ingested 250 mg of Acipimox. After 15-min adaptation to exercise, they ingested either glucose dissolved in water, 50 g at time 0 min and 25 g at time 60 and 120 min (glucose, G) or sweetened water (control, C). Naturally labelled [13C]glucose was used to follow the conversion of the ingested glucose to expired-air CO2. Acipimox inhibited lipolysis in a similar manner in both experimental conditions. This was reflected by an almost complete suppression of the exercise-induced increase in plasma free fatty acid and glycerol and by an almost constant rate of lipid oxidation. Total carbohydrate oxidation evaluated by indirect calorimetry, was similar in both experimental conditions [C, 182, (SEM 21); G, 194 (SEM 16) g.3 h-1], as was lipid oxidation [C, 57 (SEM 6); G, 61 (SEM 3) g.3 h-1]. Exogenous glucose oxidation during exercise G, calculated by the changes in 13C:12C ratio of expired air CO2, averaged 66 (SEM 5) g.3 h-1 (19% of the total energy requirement). Consequently, endogenous carbohydrate utilization was significantly smaller after glucose than after placebo ingestion: 128 (SEM 18) versus 182 (SEM 21) g.3 h-1, respectively (P < 0.05). Symptoms of intense fatigue and leg cramps observed with intake of sweet placebo were absent with glucose ingestion.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Availability of glucose ingested during muscle exercise performed under acipimox-induced lipolysis blockade. 807 20

1. The effect of infusion of noradrenaline (0.42 mumol min-1 kg-1) on the exchange of nonesterified fatty acids, glycerol and other metabolites across subcutaneous abdominal adipose tissue was investigated in five healthy subjects using an arteriovenous catheterization technique and measurement of adipose tissue blood flow using the 133Xe clearance technique. At the same time, the net rate of fat oxidation in the whole body was assessed by indirect calorimetry, and the turnover of glycerol in the whole body and in subcutaneous adipose tissue was estimated using [5-2H]glycerol, which was administered as a primed constant infusion for 1 h before (basal turnover) noradrenaline administration and continued during the 1 h of noradrenaline infusion. 2. The noradrenaline infusion increased the plasma noradrenaline concentration from a basal value of 0.9 +/- 0.1 to 12.6 +/- 1.2 nmol/(mean +/- SEM) at 60 min. It also increased the arterialized concentration of glycerol by 50% (basal value 81 +/- 11 mumol/l-1) and that of plasma non-esterified fatty acids three-fold (basal value 357 +/- 86 mumol/l). 3. Noradrenaline increased the net release of glycerol by adipose tissue three-fold and that of non-esterified fatty acids three- to four-fold. Although the ratio of non-esterified fatty acid to glycerol release by adipose tissue increased in all subjects from a mean value of 2.7 in the basal period to 3.6 and 3.9 at 50 and 60 min of the noradrenaline infusion, respectively (P < 0.02), at no time point did the ratio differ significantly from 3.0.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effect of noradrenaline on glycerol turnover and lipolysis in the whole body and subcutaneous adipose tissue in humans in vivo. 814 28

A group of 17 children, 8.5-11 years old, performed a 60-min cycle exercise at 60% of maximal oxygen uptake (VO2max) 2 h after a standardized breakfast. They were 10 young boys (pubertal stage = 1) and 7 young girls (pubertal stage < or = 2) of similar VO2max (respective values were 48.5 ml min-1 kg-1, SEM 1.8; 42.1 ml min-1 kg-1, SEM 2.4). Blood samples of 5 ml were withdrawn by heparinized catheter, the subjects being in a supine position, 30 min before the test, then after 0, 15, 30 and 60 min of exercise and following 30 min recovery. Haematocrit was immediately measured. Thereafter plasma was analysed for glucose, non-esterified fatty acid, glycerol, catecholamine (noradrenaline, adrenaline), insulin and glucagon concentrations. This study showed two main results. First, the onset of exercise induced a significant glucose decrease (of about 11.4%) in all the children. Secondly, both the glycaemic and the hormonal responses were obviously different according to the sex. In boys only, the initial glucose drop was significantly correlated to the pre-exercise insulin values. Whatever the time, the glycaemic levels and the catecholamine responses were lower in girls than in boys, whereas the insulin values remained higher. However, none of these two hormonal parameters seemed to be really responsible for the lower glucose values in girls. On the one hand, the great individual variability of noradrenaline and adrenaline and differences in their relative intensity at the end of the exercise between boys and girls might contribute to the lower catecholamine levels in girls.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Glucoregulation and hormonal changes during prolonged exercise in boys and girls. 816 19

Normozoospermic semen samples (n = 82) were examined to investigate whether the degree of sperm tail swelling in hypo-osmotic medium (fructose and sodium citrate; 150 mOsm/l), and motility characteristics after a 15-min exposure to hypotonic saline (sodium chloride; 150 mOsm/l) could predict the cryosurvival rate (% post-thaw motility/% pre-freeze motility x 100%) of spermatozoa after cryopreservation by the liquid nitrogen vapour freezing method using the TEST-glycerol-egg yolk buffer. The CellSoft automated semen analyser was used to analyse sperm motility in pre-freeze and post-thaw semen samples, and after exposure to hypotonic saline. Sperm tail hypo-osmotic swelling and sperm motility in pre-freeze semen showed no significant correlations (P > 0.05) with the cryosurvival rate. There were significant correlations (P < 0.05) between the cryosurvival rate and the following sperm motility parameters in hypotonic saline: % motility (r = 0.2846), motility index (% motility x curvilinear velocity; r = 0.2809) and % decrease in motility index from the baseline value in semen (r = 0.3378). The % decrease in motility index after hypotonic saline treatment was significantly less (P < 0.05) in the normal (> or = 50% cryosurvival rate; mean +/- SEM 5.9 +/- 3.2%; n = 33) compared with the subnormal (< 50% cryosurvival rate; 27.3 +/- 4.8%; n = 49) cryosurvival groups. This parameter was also determined, by multivariate discriminant analysis, to be capable of classifying each pre-freeze semen sample into either cryosurvival group with 69.5% accuracy.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Human spermatozoal tail hypo-osmotic swelling test, motility characteristics in hypotonic saline, and survival of spermatozoa after cryopreservation. 831 66

The adenosine A1 receptor antagonist (FR113453) prevents glycerol- but not mercury-induced acute renal failure. To clarify this mechanism, adenosine concentration in the renal vein was measured serially. Plasma adenosine in the renal vein increased from the preinjection value of 120.6 +/- 15.4 (mean +/- SEM) pmol/ml to 426.9 +/- 107.5, 407.0 +/- 70.1 and 283.9 +/- 22.9 pmol/ml at 1, 5 and 60 min after intramuscular injection of 10 mg/kg of 50% glycerol into Sprague-Dawley rats. On the other hand, intramuscular vehicle (0.9% NaCl) injection and subcutaneous administration of 10 mg/kg of HgCl2 did not change or caused mild elevation of adenosine concentration in the renal vein. Furthermore, simultaneous blood collection from the carotid artery, renal vein and inferior vena cava revealed a greater increase in adenosine concentration in the inferior vena cava than in the artery or renal vein. These findings were not affected by the administration of FR113453 or vehicle (methylcellulose). The increase in adenosine in the inferior vena cava was derived from the release from the acutely damaged muscles due to glycerol injection. These findings suggest that the effect of adenosine A1 antagonist to prevent glycerol-induced acute renal failure is due to the inhibition of adenosine A1 receptor in the kidneys during the release of adenosine through the inferior vena cava. Therefore, the release of adenosine from the muscle and hemolysis plays an important role to induce acute renal failure in the glycerol-injected rat.
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PMID:Changes of adenosine levels in the carotid artery, renal vein and inferior vena cava after glycerol or mercury injection in the rat. 836 87

Several studies have reported that parenteral lipid emulsions containing medium-chain triglycerides (MCT) and structured lipids (SL) are better utilized than those containing long-chain triglycerides (LCT). The objective of this study was to test the hypothesis that parenteral LCT require more extensive modification via hydrolysis and reesterification (triglyceride-free fatty acid [TG-FFA] recycling) for effective utilization, whereas MCT and SL do not. As an index of TG-FFA cycling activity, we measured glycerol and palmitate kinetics in rats (204 to 243 g) fed parenterally one of three isocaloric (250 kcal/kg/d) isonitrogenous (1.5 g N/kg/d) diets with half of the nonprotein energy from glucose and the rest from either LCT, LCT plus MCT, or SL for 5 days. Two experiments were performed. On day 5, rats were given a 7-to 8-hour infusion of either 5H2 Glycerol and 1-14C Palmitate bound to albumin to measure palmitate and glycerol kinetics (experiment 1), or U-13C glucose to determine the proportion of endogenous glycerol production derived from glucose (experiment 2). Data are presented as means +/- SEM. Endogenous glycerol production was significantly higher with LCT (11.33 +/- 2.89 mmol/kg/h) than with SL (2.91 +/- 0.62 mmol/kg/h). The value for the physical mixture of LCT plus MCT (5.46 +/- 1.29 mmol/kg/h) fell midway between that for LCT and SL (P = NS). There were no significant differences in palmitate kinetics or oxidation. The increased glycerol production is due to the mobilization of endogenous triglyceride and is consistent with a higher rate of TG-FFA cycling being involved in the metabolism of LCT than of SL.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Glycerol kinetics with parenteral lipid emulsions (long-chain triglycerides, medium-chain triglycerides, and structured lipids) in rats. 851 May 20

Osmotic permeability characteristics and the effects of cryoprotectants are important determinants of recovery and function of spermatozoa after cryopreservation. The primary purpose of this study was to determine the osmotic permeability parameters of human spermatozoa in the presence of cryoprotectants. A series of experiments was done to: 1) validate the use of an electronic particle counter for determining both static and kinetic changes in sperm cell volume; 2) determine the permeability of the cells to various cryoprotectants; and 3) test the hypothesis that human sperm water permeability is affected by the presence of cryoprotectant solutes. The isosmotic volume of human sperm was 28.2 +/- 0.2 microns3 (mean +/- SEM), 29.0 +/- 0.3 microns3, and 28.2 +/- 0.4 microns3 at 22, 11, and 0 degrees C, respectively, measured at 285 mOsm/kg via an electronic particle counter. The osmotically inactive fraction of human sperm was determined from Boyle van't Hoff (BVH) plots of samples exposed to four different osmolalities (900, 600, 285, and 145 mOsm/kg). Over this range, cells behaved as linear osmometers with osmotically inactive cell percentages at 22, 11, and 0 degrees C of 50 +/- 1%, 41 +/- 2%, and 52 +/- 3%, respectively. Permeability of human sperm to water was determined from the kinetics of volume change in a hyposmotic solution (145 mOsm/kg) at the three experimental temperatures. The hydraulic conductivity (Lp) was 1.84 +/- 0.06 microns.min-1.atm-1, 1.45 +/- 0.04 microns.min-1.atm-1, and 1.14 +/- 0.07 microns.min-1.atm-1 at 22, 11, and 0 degrees C, respectively, yielding an Arrhenius activation energy (Ea) of 3.48 kcal/mol. These biophysical characteristics of human spermatozoa are consistent with findings in previous reports, validating the use of an electronic particle counter for determining osmotic permeability parameters of human sperm. This validated system was then used to investigate the permeability of human sperm to four different cryoprotectant solutes, i.e., glycerol (Gly), dimethylsulfoxide (DMSO), propylene glycol (PG), and ethylene glycol (EG), and their effects on water permeability. A preloaded, osmotically equilibrated cell suspension was returned to an isosmotic medium while cell volume was measured over time. A Kedem-Katchalsky model was used to determine the permeability of the cells to each solute and the resulting water permeability. The permeabilities of human sperm at 22 degrees C to Gly, DMSO, PG, and EG were 2.07 +/- 0.13 x 10(-3) cm/min, 0.80 +/- 0.02 x 10(-3) cm/min, 2.3 +/- 0.1 x 10(-3) cm/min, and 7.94 +/- 0.67 x 10(-3) cm/min, respectively. The resulting Lp values at 22 degrees C were reduced to 0.77 +/- 0.08 micron.min-1.atm-1, 0.84 +/- 0.07 micron.min-1.atm-1, 1.23 +/- 0.09 microns.min-1.atm-1, and 0.74 +/- 0.06 micron.min-1.atm-1, respectively. These data support the hypothesis that low-molecular-weight, nonionic cryoprotectant solutes affect (decrease) human sperm water permeability.
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PMID:Effect of cryoprotectant solutes on water permeability of human spermatozoa. 852 30

Renal vascular reactivity was studied in rats with acute renal failure (ARF) to investigate whether changes in sensitivity to the renal haemodynamic effects of adenosine can explain why adenosine plays a significant role in some but not all forms of ARF. Experiments involved rats with glycerol-induced ARF in which adenosine antagonists have been shown previously to have beneficial effects and rats with HgCl2-induced ARF which was not ameliorated by treatment with the selective A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine (0.1 mg/kg). Close renal arterial injections of adenosine (0.1-10 micrograms) or noradrenaline (0.003-0.1 microgram) produced falls in renal blood flow in rats with HgCl2-induced ARF which were not statistically different from controls. Adenosine evoked falls in renal blood flow in rats with glycerol-induced ARF which were significantly greater 16 and 48 h, but not 30 min after glycerol injection. The enhanced responsiveness to adenosine's renal constrictor effects was most pronounced 48 h following glycerol injection when, for example, a dose of 10 micrograms produced a fall of 60 +/- (SEM) 5% (n = 8) in renal blood flow in comparison to a fall of 27 +/- 5% (n = 8) in controls. By contrast to the renal vascular response to adenosine, the falls in renal blood flow induced by noradrenaline in rats 48 h following glycerol injection were not statistically different from the decreases in renal blood flow recorded in control animals.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Renal haemodynamic responses to adenosine in acute renal failure. 856 52


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