Sir Barry Albert Cross, C.B.E. 17 March 1925-27 April 1994.
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History, 20th Century↗
Biomedical subjects
R G Dyer
Publications and source records attributed to R G Dyer.
7-ketocholesterol, a specific indicator of lipoprotein oxidation, and malondialdehyde in non-insulin dependent diabetes and peripheral vascular disease.
Increased free radical-mediated lipoprotein oxidation may contribute to the increased prevalence of atherosclerosis in non-insulin dependent diabetes. We have determined levels of malondialdehyde (MDA) and 7-ketocholesterol, a specific indicator of free radical-mediated oxidation of lipoprotein cholesterol, in serum in very low density lipoprotein, intermediate density lipoprotein, low density lipoprotein (LDL) and high density lipoprotein fractions of serum separated by sequential flotation ultracentrifugation. Four groups of male subjects were studied: normal controls, diabetic patients with no evidence of microvascular complications or macrovascular disease, diabetic and non-diabetic patients with peripheral vascular disease (PVD). MDA was increased in vascular disease patients (diabetic 4.5 (3.7-5.8), non-diabetic 4.4 (3.2-5.7) mumol/l, median (2.5-97.5 percentiles)) than controls (3.6 (2.9-5.0) mumol/l) (P < 0.01), but was not increased in uncomplicated diabetic patients (3.8 (3.0-4.8) mumol/l). There were no significant differences in 7-ketocholesterol concentration in LDL, but calculated total 17-ketocholesterol was lower in non-diabetic vascular patients than controls (P < 0.01). Vitamin C concentration was reduced in diabetic and non-diabetic patients with vascular disease. No significant difference in concentration of vitamin E or A was found. In six normal subjects the concentration of MDA was low in lipoproteins separated by ultracentrifugation but high in the residue following lipoprotein fractionation (70-80% total serum MDA). In conclusion, the concentration of MDA by the thiobarbituric acid assay in untreated serum may not reflect free radical damage to lipoproteins. There was no evidence of increased lipoprotein oxidation using 7-ketocholesterol in NIDDM or PVD.
Adult↗
Simultaneous measurement of phytosterols (campesterol and beta-sitosterol) and 7-ketocholesterol in human lipoproteins by capillary column gas chromatography.
7-Ketocholesterol (a major cholesterol oxidation product) and phytosterols are important indicators of lipoprotein oxidation and lipoprotein metabolism respectively. We describe a simple, sensitive and reproducible method for the simultaneous measurement of these sterols in human lipoprotein samples by capillary column gas liquid chromatography. The method is suitable for clinical studies as small quantities of lipoprotein are required. Sterols are analysed after extraction from lipoprotein samples obtained by sequential flotation ultracentrifugation. The method involves briefly: extraction from lipoprotein samples using chloroform-methanol, saponification of sterol esters using cold potassium hydroxide, purification and derivatisation to trimethylsilyl ethers using BSTFA and 1% TMCS. Oxidation is prevented by drying under nitrogen and the use of powerful antioxidants. Separation is achieved using a DB-1 capillary column and a two-stage temperature ramp from 180-250 degrees C and detection using FID. The identity of sterols can be confirmed by GC-MS. Phytosterols and 7-ketocholesterol are present at low concentration in all the major lipoproteins. Using [3,4-13C]cholesterol and GC-MS we present evidence that cholesterol oxidation does not occur during the processing of lipoproteins using this technique.
Capillary Action↗
The effects of lipid lowering drugs on metabolic control and lipoprotein composition in type 2 diabetic patients with mild hyperlipidaemia.
Patients with Type 2 diabetes are at increased risk from macrovascular disease whether or not they are hyperlipidaemic. Several factors may contribute to this increased risk including abnormalities of lipoprotein composition. The aim of our study was to determine the effects of lipid lowering drugs on lipoprotein composition (lipoprotein fractions were separated by sequential flotation ultracentrifugation) and insulin sensitivity (measured by a modified Harano technique) in 44 patients with mild hyperlipidaemia. All patients had total cholesterol concentrations between 5.2 and 6.5 mmol l-1 and total triglyceride concentrations < 3.0 mmol l-1, and were randomized by minimization to receive treatment for 12 weeks with bezafibrate, acipimox, simvastatin or placebo. Total cholesterol concentrations were decreased by simvastatin, 5.7 +/- 0.4 to 3.7 +/- 0.6 mmol l-1 (p < 0.05), due mainly to reduced LDL-cholesterol levels (-1.25 mmol l-1; p < 0.05), and bezafibrate 5.7 +/- 0.6 to 4.6 +/- 0.4 mmol l-1 (p < 0.05). The LDL:HDL-cholesterol ratio was reduced in the simvastatin group 2.0 +/- 0.5 to 1.2 +/- 0.3 (p < 0.005). There was no effect of the drugs on glycated haemoglobin or insulin sensitivity. In conclusion bezafibrate and simvastatin improve the lipid profile in Type 2 diabetic patients without adversely affecting diabetic control.
Apolipoproteins B↗
Is the inhibitory action of estradiol on luteinizing hormone pulse frequency in anestrous ewes mediated by noradrenergic neurons in the preoptic area?
This study tested the hypothesis that estradiol inhibits luteinizing hormone (LH) pulse frequency in anestrous ewes by increasing the activity of an inhibitory noradrenergic (NE) system that acts in the ovine preoptic area (POA). The effects of estradiol on the release of NE and other neurotransmitters in the POA were determined using intracranial microdialysis. Microdialysis probes (5 mm membrane length) were inserted via chronic guide tubes into the POA. Ringer's solution was pumped through the probes at a rate of 2 microliters/min for 8 h, the alpha-adrenergic antagonist phenoxybenzamine (PBZ; 60 micrograms/ml of Ringer's solution) was then administered via the probe from hours 8 to 12, and Ringer's solution alone was given the last 4 h. The outflow from the dialysis probes was collected every 20 min from 2 to 16 h and concentrations of aminergic transmitters and gamma aminobutyric acid determined by high-performance liquid chromatography. Blood samples were collected every 10 min throughout the experiment and the LH pulse patterns determined. Dialysis was done in the same neural area twice in each ewe, without (ovariectomy only) and with estradiol treatment (ovariectomy and a 1.5-cm-long Silastic capsule filled with crystalline estradiol placed subcutaneously for 2 days); the order of dialysis was randomized. As expected, estradiol decreased the LH pulse frequency. This negative feedback action of estradiol was associated with a decrease in mean NE concentrations in dialysate samples and an increase in the intra-animal variability of NE. Estradiol had no effect on any other neurotransmitter measured.(ABSTRACT TRUNCATED AT 250 WORDS)
Anestrus↗
Analysis of brainstem A1 and A2 noradrenergic inputs to the preoptic area using microdialysis in the rat.
Noradrenergic inputs to the preoptic area (POA) are involved in regulating a variety of homeostatic functions. However, the accurate measurement of endogenous noradrenaline (NA) release in the POA has been difficult to achieve and consequently little has been done to characterise the different noradrenergic pathways. By combining the technique of intracranial microdialysis with tissue pre-loading of [3H]NA we have developed a sensitive index of NA release in the POA [8]. Using this method we have now examined and compared the effects of electrical stimulation of the brainstem A1 and A2 cell groups on NA release in the POA. Anaesthetised proestrus rats were implanted with microdialysis probes either unilaterally or bilaterally in the POA and stimulating electrodes positioned in either the A1 or A2 regions. Electrical stimulation (10 Hz, 10s on/off for 20 min) of the A1 region resulted in repeatable, calcium-dependent increases in radioactivity outflow from the ipsilateral POA (P < 0.01). A1-evoked release was twice as large as that observed after equivalent 10 Hz electrical stimulation of the A2 region (P < 0.05). In experiments using bilateral POA microdialysis and A1 stimulation, a significant increase in release from the contralateral POA, amounting to approximately 80% of that observed in the ipsilateral POA, was observed. Experiments involving the blockade of A1-stimulated release in the ipsilateral POA by perfusion with a calcium-free medium demonstrated that increases in radioactivity measured in the contralateral POA were not originating from the ipsilateral POA.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Traditional treatment of obesity: does it work?
Obesity is the most important nutritional disorder in the developed world, since up to 10% of the population are obese. The place of physical activity and diet in the aetiology of obesity is discussed. The traditional treatment of obesity includes change in lifestyle, nutritional education and modification and increase in exercise. These changes are important for long-term success. There are a number of other treatment options including anorectic drugs, the use of very low calorie diets and surgical techniques which may have some clinical role. For the extremely obese patient with established complications surgery may be the most appropriate intervention and may be life-saving. Most studies of traditional treatment have demonstrated limited success. The prevention of obesity is therefore of great importance. Large-scale studies have shown that it is possible to modify behaviour and cardiovascular risk factors. The prevention of obesity requires a coordinated approach with targeting of children and their carers. Governmental involvement and legislation is essential. The future holds the promise of more imaginative and coordinated therapies for obesity using the skills of physicians, nutritionists, exercise physiologists and psychologists. Different forms of treatment may be appropriate for different groups of obese patients.
Adipose Tissue↗
Characterization of tritiated noradrenaline release from the rat preoptic area with microdialysis in vivo.
Present techniques are unable to provide a sensitive and accurate index of noradrenergic activity in the rat preoptic area. In this study, we have examined the brainstem A1 noradrenergic input to the preoptic area using a new technique whereby [3H]noradrenaline is preloaded into the preoptic area and release of radioactivity from this region is measured subsequently using microdialysis in vivo. Electrical stimulation of the ipsilateral A1 area for 20 min at 5, 10, and 15 Hz evoked significant increases in dialysate radioactivity that were repeatable and frequency-dependent. After removal of calcium from the perfusion medium, basal release of radioactivity was markedly reduced and the effect of A1 stimulation abolished. Changing to a 100 mM K+ medium evoked an increase in the release of radioactivity that was sixfold greater than that seen after A1 stimulation. Separation of the dialysate with HPLC showed that 33% of the increase in measured radioactivity after A1 stimulation was directly attributable to [3H]noradrenaline and the remainder to the metabolites vanillylmandelic acid, 3,4-dihydroxymandelic acid, and 3,4-dihydroxyphenylglycol. In contrast, the increase in radioactivity after K+ depolarization was due almost completely to [3H]noradrenaline. Addition of 10 microM clonidine to the perfusion medium markedly reduced basal release of radioactivity, but had no effect on evoked release following A1 stimulation. Conversely, perfusion with 10 microM yohimbine had no effect on basal release, but significantly increased evoked release after A1 stimulation. These results now provide a characterization of noradrenergic activity in the preoptic area and indicate the importance of the A1 noradrenergic input to this region. The technique of measuring radioactivity with microdialysis after preloading with [3H]noradrenaline provides a relatively simple, sensitive index of noradrenergic activity in vivo with good temporal resolution.
Animals↗
Lipoprotein compositional abnormalities and insulin resistance in type II diabetic patients with mild hyperlipidemia.
Lipoprotein composition was determined using ultracentrifugation in 20 non-insulin-dependent (NIDDM) diabetic patients on diet only (D), 20 NIDDM patients on diet and sulfonylurea therapy (T), and 20 nondiabetic control subjects (C), all of whom had total plasma cholesterol concentrations < 6.5 mmol/L and total plasma triglyceride concentrations < 3.0 mmol/L. Although the groups were well matched for age, body mass index, total triglyceride levels, and total cholesterol concentrations, there were significant compositional abnormalities in the low-density lipoprotein (LDL) fractions of diabetic subjects. The LDL total lipid to apolipoprotein B weight ratio (representing the density distributions of LDL particles) was reduced in both diabetic groups: 3.75 +/- 0.3, 3.50 +/- 0.28, and 3.54 +/- 0.22 in C, D, and T groups, respectively (mean +/- SD; P < .05). This was associated with a significant shift in the hydrated density distributions of LDL in the diabetic groups, with the average peak densities being 1.0320 g/mL (in C), 1.0365 g/mL (in D), and 1.0380 g/mL (in T) (P < .05). The LDL particles were also smaller in the NIDDM patients: 21.1 +/- 0.7, 20.4 +/- 0.5, and 20.6 +/- 0.5 nm in C, D, and T groups, respectively (P < .05). When the NIDDM groups were analyzed together, the LDL peak density was found to correlate with both insulin resistance (measured by a modified Harano technique; r = 0.37, P < .015) and total triglyceride concentrations (r = 0.40, P < .01). The results show that diabetic patients have small, dense LDL particles, which may be related to insulin resistance, and that these occur with minimal elevations of total triglyceride concentrations. These potentially atherogenic changes may contribute to the increased coronary heart disease in diabetic patients with mild hyperlipidemia.
Aged↗
Opioid binding in the rostral hypothalamus is reduced following lesion of the ventral noradrenergic tract in female rats.
Experiments were undertaken to establish whether opioid receptors exert a direct presynaptic influence on noradrenergic (NA) terminals in the preoptic/anterior hypothalamus (PO/AH) of the female rat. Thus, opioid binding studies were performed in rats with lesions of the ventral NA tract (VNAT; the main NA projection to the hypothalamus) to assess whether a loss of NA terminals may also result in a decrease in opioid binding in the PO/AH. In the first experiment, unilateral electrolytic lesions of the VNAT caused a significant reduction in both the NA content and specific [3H]-diprenorphine binding to membrane homogenates in the ipsilateral PO/AH. In the second experiment bilateral 6-hydroxydopamine (6-OHDA)-induced lesions of the VNAT caused a significant reduction in NA levels in the PO/AH as well as significant decreases in the density of [3H]-diprenorphine binding to tissue sections of the PO/AH when compared to control animals. These results strongly suggest that the NA input to the PO/AH is regulated by endogenous opioid peptides, and provide an anatomical substrate to explain opioid-NA interactions in the control of gonadotrophin releasing hormone (GnRH) and gonadotrophin secretion.
Animals↗
Role of medial preoptic GABA neurones in regulating luteinising hormone secretion in the ovariectomised rat.
The role of GABA neurones in the medial preoptic area (MPOA) in regulating the activity of the luteinising hormone-releasing hormone (LHRH) neurones projecting to the median eminence was investigated in the conscious ovariectomised rat. Plasma luteinising hormone (LH) concentrations were measured while (1) endogenous GABA release from the MPOA was monitored with the technique of microdialysis, or (2) activity at the GABA receptor was modulated by local infusions into the MPOA. Microdialysis studies revealed a fluctuating level of GABA release in the MPOA which did not correlate with pulsatile LH secretion. Infusion of 10 microM GABA (n = 8) or bicuculline methiodide (BMI, n = 6) into the MPOA, at a rate of 1 microliter/30 min, significantly inhibited mean LH concentrations (P less than 0.05-0.001) and LH pulse frequency (P less than 0.05-0.001) compared with controls (n = 8). LH pulse amplitude was not significantly altered by infusion of GABA (P greater than 0.05) while too few pulses were found after BMI treatment to enable statistical analysis. Infusions of GABA into the ventral half of the MPOA had a more significant inhibitory effect upon LH secretion compared with dorsal infusions (P = 0.012). A similar relationship did not exist for BMI infusions. These results show that acute changes in preoptic GABA receptor occupancy result in disruption of pulsatile LH secretion in the ovariectomised rat. This suggests that GABA neurones provide a tonic input important for the functional integrity of the neural network controlling LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
The changing shape of obesity, Japan 1990.
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Diabetes Mellitus, Type 2↗
Effect on luteinizing hormone secretion of GABA receptor modulation in the medial preoptic area at the time of proestrous luteinizing hormone surge.
Using a bilateral medial preoptic area (MPOA) infusion system in conscious female rats we have investigated the role of the GABA system in this area on the proestrous luteinizing hormone (LH) surge. Fifteen-minute blood samples for LH estimation were taken throughout the afternoon of proestrus from female rats exhibiting 4-day oestrous cycles and implanted at least 2 weeks prior with cerebral guide cannulae. Between 15:00 and 17:00 h rats received an infusion (1 microliter/30 min) of artificial cerebrospinal fluid (n = 7), 10 microM GABA (n = 6) or 10 microM bicuculline methiodide (BMI, n = 6). Animals infused with GABA failed to exhibit an LH surge, while BMI-treated animals displayed an LH surge which was not significantly different to controls. These data show that on the afternoon of proestrus, there are no tonic modulatory actions of the GABA system, acting through the GABAA receptor, on neural elements controlling the LH surge in the MPOA. If, however, GABA levels are elevated in the MPOA at this time then the LH surge is blocked. In conjunction with data from correlative studies showing a decrease in endogenous GABA release prior to the LH surge, we suggest that this fall in activity is an essential component of the LH surge mechanism.
Animals↗
Endogenous release of gamma-aminobutyric acid from the medial preoptic area measured by microdialysis in the anaesthetised rat.
The characteristics of gamma-aminobutyric acid (GABA) release as monitored by microdialysis have been investigated in the chloral hydrate anaesthetised rat. The high outflow of GABA following insertion of the microdialysis probe (membrane 2 mm in length, 0.5 mm in diameter) into the medial preoptic area was found to decline to a stable baseline level after 2 h. After this time, perfusion with a medium containing 100 mM potassium ions evoked a 56-fold increase in GABA outflow. The addition of the calcium channel blocker verapamil (100 microM) to the perfusion medium induced significant 25 and 50% reductions in basal and potassium-stimulated GABA outflow, respectively. In the same animals, verapamil caused an 80% decrease in potassium-stimulated noradrenaline outflow. The glutamic acid decarboxylase inhibitors 3-mercaptopropionic acid and L-allylglycine added to the perfusion medium at a concentration of 10 mM reduced basal GABA release by approximately 50% with different time-courses of action. Ethanolamine-O-sulfate, a GABA-transaminase inhibitor, induced significant increases in basal GABA outflow 90 min after inclusion in the perfusion medium. These results demonstrate that microdialysis is a suitable technique with which to monitor extracellular levels of GABA and provide in vivo data on GABA release and degradation mechanisms.
4-Aminobutyrate Transaminase↗
Oestrogen modulation of excitatory A1 noradrenergic input to rat medial preoptic gamma aminobutyric acid neurones demonstrated by microdialysis.
The effect of A1 cell group electrical stimulation on the simultaneous release of endogenous noradrenaline (NA) and gamma-aminobutyric acid (GABA) from the medial preoptic area (MPOA) was monitored with microdialysis. In ovariectomised (OVX) rats a 15-min period of A1 stimulation induced an immediate increase in both NA and GABA release in the MPOA. Electrical stimulation lateral to the A1 region did not alter NA or GABA release. The addition of the alpha-adrenergic antagonist phenoxybenzamine to the perfusion medium resulted in a significant increase in basal NA levels, and electrical stimulation during this period further increased NA release while GABA levels were not significantly altered throughout. The effect of oestrogen on this pathway was examined in animals at a time of oestrogen-negative feedback on luteinising hormone (LH) secretion (OVX-EBn) and prior to the expected oestrogen-induced LH surge (OVX-EBp). Activation of A1 neurones in OVX-EBn rats resulted in NA and GABA increases in the MPOA similar to that observed with OVX rats. In OVX-EBp animals, basal GABA levels were found to be significantly higher compared with OVX rats but NA release induced by A1 stimulation had no effect on GABA levels. Depolarisation of the MPOA by increasing the potassium ion concentration of the perfusion medium evoked significantly greater GABA release from OVX-EBp rats compared with the OVX and OVX-EBn animals. Potassium-stimulated NA release was not significantly altered by oestrogen administration. These results demonstrate an excitatory alpha-adrenergic mediated noradrenergic input to GABA neurones in the MPOA.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Oestrogen and noradrenaline modulate endogenous GABA release from slices of the rat medial preoptic area.
Endogenous gamma-aminobutyric acid (GABA) release from the rat medial preoptic area (MPOA) was measured in an in vitro slice technique with sensitive HPLC analysis. Oestrogen is demonstrated to increase GABA activity in the ovariectomised, oestrogen-primed (OVX-EB) rat prior to the luteinising hormone (LH) surge compared with ovariectomised (OVX) animals. Noradrenaline (NA) at a concentration of 10 microM was found to significantly enhance GABA release in response to 30 mM potassium stimulation in both OVX and OVX-EB animals. A significantly greater response to NA was observed in the OVX-EB animal. No effect of NA on basal GABA release was detected. The effects of NA were blocked by the alpha-adrenergic receptor blocker phenoxybenzamine (PB). These data suggest that GABA activity is modulated both by oestrogens and noradrenergic-mediated input in the MPOA.
Animals↗
Opioid peptides inhibit noradrenergic transmission in the preoptic area to block LH secretion: evidence from neonatally androgenised rats.
A series of experiments are described which show (a) that the release of noradrenaline in the preoptic area of rats is regulated by a presynaptic opioid receptor and, (b) that changes in the opioid control of preoptic noradrenaline release are associated with the anovulatory condition found in adult female rats which have been exposed to androgen neonatally. The experiments support the hypothesis that opioid-noradrenergic interaction in the preoptic area is an important component in the neural mechanism regulating gonadotrophin secretion.
Action Potentials↗