Research in experimental medicine. Zeitschrift fur die gesamte experimentelle Medizin einschliesslich experimenteller Chirurgie最新文献

Antiretroviral therapy (ART) is frequently associated with metabolic alterations, including insulin resistance and diabetes mellitus. In this pilot study, we evaluated the effect of the PPARgamma activator troglitazone on ART-associated insulin resistance in HIV-infected patients with ART-associated diabetes mellitus. Six patients with protease inhibitor (PI)-associated diabetes mellitus, lipodystrophy and dyslipidemia were treated with troglitazone 400 mg q.d. for 3 months. Previous oral antidiabetics were discontinued prior to the study. At baseline and after 3 months, insulin sensitivity (intravenous insulin tolerance test), body composition (multifrequence bioelectrical impedance analysis) and fat distribution (CT scan quantification) were assessed. Glycaemic control (fasting and postprandial blood glucose, fructosamine, glycosylated haemoglobin) and serum lipid status were determined monthly. In four of the six patients, there was a clear improvement in insulin sensitivity, resulting in a reversal of insulin resistance in two of these patients. Overall, there was an increase in lean body mass and a decrease in total body fat. The volume of visceral adipose tissue decreased whilst the volume of subcutaneous adipose tissue increased. Total cholesterol, LDL and HDL cholesterol increased, and total triglycerides and VLDL-cholesterol decreased. No adverse effects such as hepatotoxicity were observed. Treatment with troglitazone 400 mg q.d. can ameliorate and in some cases even reverse ART-associated insulin resistance. Therefore, further studies including non-diabetic patients with ART-associated insulin resistance may be helpful in evaluating the long-term effects of thiazolidinediones on ART-associated insulin resistance and other metabolic complications, such as adipose maldistribution and dyslipidaemia.

In metastasis research it would be useful to determine the number of blood borne tumor cells which are released from a primary tumor into the blood circulation. One way to quantify the number of released tumor cells could be to take blood from a vessel which is located close to a primary tumor and is draining the tumor. The number and viability of tumor cells released into the blood stream at any given time could be measured in cancer patients, especially those known to bear a primary, hematogenous metastasizing tumor. Plating efficiency is a precise method for the quantitative determination of the number of colony-forming cells in an adherent cell population. We performed initial in vitro experiments using plating efficiency to count adherent tumor cells within whole human blood. Exploiting the difference in adherence properties of colon carcinoma cells and blood cells in standard cell culture medium, these initial investigations show that it is possible to determine the plating efficiency of colon carcinoma cells within fresh whole human blood.

Intracortical injection of iron salts causes seizures. Oxidation of lipids in neural membranes by reactive oxygen species is involved in the mechanism responsible for iron-induced seizures as a model of posttraumatic epilepsy. In this study, we examined the effect of trimetazidine (TMZ) and deferoxamine (DFO) on lipid peroxidation after cortical injection of 5 microliters of an aqueous solution containing 100 mM of ferric chloride (FeCl3) in rats. Animals were divided into four groups (n = 7 each) and treated as follows: group 1, saline injection into the cortex (control group); group 2, iron injection into the cortex (injury group); group 3, iron injection into the cortex plus TMZ; group 4, iron injection into the cortex plus DFO. The animals were killed 3 h after injections, and the levels of malondialdehyde (MDA), a lipid peroxidation product, and reduced glutathione (GSH) were measured. A significant elevation of MDA was observed in group 2 (P < 0.05). MDA levels were found to be lower in both the TMZ-treated (P < 0.05) and DFO-treated (P < 0.05) groups than in the injury group. Tissue GSH levels were significantly decreased in group 2 (P < 0.05). GSH levels were increased in the TMZ-treated (P < 0.05) and DFO-treated (P < 0.05) groups compared to the injury group. The results of our study suggest that lipid peroxidation is a critical event in iron-induced epilepsy and that treatment with TMZ and DFO is effective in preventing the formation of free radicals and reducing lipoperoxides in brain tissue.

An increase in oxidative stress may contribute to the development of oxidative protein damage in streptozotocin diabetic rats. In the present study, the influence of alpha-lipoic acid supplementation on plasma protein carbonyl, plasma thiol, and plasma lipid hydroperoxide levels was examined in order to characterize the relationship between the oxidative stress and the oxidative protein damage. Rats were randomly divided into three groups of equal body weight. Chronic hyperglycemia was induced by intravenous streptozotocin injection in both the group of male Wistar rats to be supplemented with alpha-lipoic acid and the group that was not to receive alpha-lipoic acid. Nondiabetic rats formed the control group and received a saline injection. In streptozotocin diabetic rats with and without alpha-lipoic acid supplementation, plasma carbonyl levels were significantly increased, while plasma thiol levels were significantly decreased compared with those of the control group. Plasma lipid hydroperoxide levels were significantly increased in diabetic rats without alpha-lipoic acid supplementation compared with those of the controls, but the lipid hydroperoxide levels in the alpha-lipoic acid supplemented group were no different from those of the controls. In streptozotocin-diabetic rats, oxidative stress was significantly decreased in the alpha-lipoic acid-supplemented group. The results of this study suggest that alpha-lipoic acid, by decreasing oxidative stress, may be effective in preventing oxidative protein damage, which may contribute to the development of diabetic complications.

3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors decrease mevalonate and subsequently cholesterol synthesis competitively. Mevalonate is also the precursor of ubiquinone. Ubiquinone is an important component of electron transport chain. We therefore investigated the effect of simvastatin on rat blood and tissue ATP concentrations and the lipid composition of red blood cell membranes after 4 weeks of therapy. Significant reductions in rat plasma cholesterol, triglyceride, and blood ATP concentrations were detected. Tissue ATP levels were not affected. Membrane phospholipids increased, while cholesterol and the cholesterol to phospholipid ratio decreased (P < 0.05). A positive correlation between the plasma cholesterol concentration and the cholesterol to phospholipid ratio was noted (P < 0.05, r = 0.851). Our results show that HMG-CoA reductase inhibitors change the composition and probably also the functions of cell membrane lipids and blood ATP concentration.

Weanling rats fed a methyl-deficient diet develop acute renal failure, the morphological features of which vary from focal tubular necrosis to widespread cortical necrosis. We and others have shown that coconut oil, rich in saturated fatty acids, has a renal protective effect in this experimental model. In the experiment we are reporting now, we studied which fatty acid is involved in the protection afforded by coconut oil by feeding five groups of methyl-deficient rats a mixture of corn oil and hydrogenated vegetable oil, C6-C8-C10 fatty acids, C12 fatty acid, C14 fatty acid and C16-C18 fatty acids. Five groups of rats receiving the same diets supplemented with choline chloride were used as controls. The group of methyl-deficient rats fed C14 fatty acid (myristic acid) showed a greater percentage of surviving animals and lower renal damage than the other groups of methyl-deficient rats, indicating that the protective effect of coconut oil found in previous experiments is due to its high content of myristic acid.

The effect of tirilazad mesylate (U-74006F), mannitol, and their combination was investigated on focal cerebral ischemia induced by permanent middle cerebral artery (MCA) occlusion in rabbits. Rabbits were divided into four groups receiving vehicle, U-74006F, mannitol, and U-74006F plus mannitol. Hematocrit (hct), glucose, mean arterial blood pressure (MABP), pH, PCO2, and PO2 were measured both before and after occlusion. Seventy-two hours following the permanent MCA occlusion, the neurological outcome was assessed and a quantitative neuropathologic examination was performed in all rabbits. The neurological outcome was better in the rabbits treated with U-74006F plus mannitol than in the other groups. The size of infarction of the affected hemisphere following MCA occlusion was 49.7% in the control group, 30.6% in the U-74006F group, 47.6% in the mannitol group, and 24.1% in the U-74006F plus mannitol group. There was a statistically significant reduction in infarct size in the U-74006F plus mannitol group compared with the other groups (P < 0.05). The ratio of ischemic neurons to total neurons in the cortex was smaller in the U-74006F plus mannitol group than in the other groups. The ratio of ischemic neurons to total neurons in the subcortex was significantly lower in the U-74006F plus mannitol group than in the other groups (P < 0.05). Our data provide evidence for the beneficial effects of both U-74006F and U-74006F plus mannitol in promoting neurological recovery and preservation of the ischemic area.

Based on the previously suggested hypothesis that the generation of free radicals leading to lipid peroxidation is involved in the genesis of vasospasm and vasculopathy following subarachnoid hemorrhage, the therapeutic effect of EGb 761 as an antioxidant on experimental vasospasm and vasculopathy was evaluated in a double hemorrhage dog model of chronic cerebral vasospasm. For this study 14 dogs were randomly assigned to two groups, a control and a Ginkgo biloba group. The control group was only administered saline in a volume equivalent to a dose of 100 mgEGb 761/kg while the treatment group was given 100 mg EGb 761/kg. The diameter of the basilar artery decreased from 1.95 +/- 0.16 mm at day 0 to 1.11 +/- 0.07 mm at day 8 in the control group, while in the treatment group the vessel diameter decreased from 2.01 +/- 0.17 mm at day 0 to 1.72 +/- 0.16 mm at day 8. These results correspond a decrease in vessel diameter of 15.1% in the treatment group and of 43.1% in the control group (P < 0.05). Histopathological studies of the specimens obtained from basilar arteries showed that pathological signs of proliferative vasculopathy, including narrowing of the vessel lumen, corrugation of the lamina elastica and subendothelial thickening, were present in all the animals in the control group, while they could not be demonstrated in the Ginkgo biloba group. These results suggest that Ginkgo biloba may have a protective effect against subarachnoid hemorrhage-induced vasospasm and vasculopathy as a result of antioxidants.

Recent investigations have shown that antihypertensive drug treatment leads to enhanced myocardial beta-adrenoceptor sensitivity. This study was therefore conducted to establish whether or not such hypersensitivity might trigger myocardial arrhythmia subsequent to adrenaline exposure. Adult male Wistar rats (n = 6 per group) were treated with either placebo (vehicle), metoprolol (2.40 mg.kg-1.day-1), timolol (0.075 mg.kg-1.day-1), verapamil (5.50 mg.kg-1.day-1) or enalapril (0.50 mg.kg-1.day-1) for 20 consecutive days. Hearts were excised and perfused ad modum Langendorff in the presence of an adrenaline gradient (0-300 nM) for 20 min with either 3.0 mM or 5.9 mM of potassium in the perfusion buffer. Adrenaline threshold concentration (ATC, nanomolar) at myocardial fibrillation was recorded, as well as tissue cAMP contents, beta-adrenoceptor number, G-protein levels and signalling effector enzyme activities. The main findings were: (1) ATC and cAMP levels were affected in hearts perfused with low-concentration potassium buffer only. In terms of ATC, the beneficial effect of each drug regimen appeared to be in the rank order of: placebo = enalapril > verapamil > timolol > metoprolol. There was an inverse correlation between ATC and myocardial cAMP contents at the start of fibrillation; (2) Subsequent to fibrillation, beta-adrenoceptor number, hormone-elicited adenylate cyclase activities and Gs alpha:Gi2 alpha-ratio were no different from preperfusion values; (3) Significant inverse correlations between beta 1-adrenoceptor numbers and ATC were observed. We conclude that alterations in beta-adrenoceptor number, G proteins and cAMP induced by antihypertensive drugs are predictive of the myocardial sensitivity to adrenaline in terms of time to continuous and irrevocable fibrillation.

The effect of moderate changes of peak aortic and end-diastolic pressure and of heart rate on the left ventricular relaxation of isolated working rat and guinea pig hearts was investigated by multivariate regression analysis. Each of these three independent variables was set to three different levels, yielding 27 sets of data by combination in each experiment. Relaxation was quantified by the maximum pressure fall velocity (min LVdP/dt) and by the time constant tau of left ventricular isovolumic pressure fall. tau was obtained by fitting that pressure curve to the three-parametric exponential regression model p(t) = P infinity + (P0-P infinity) exp (-t/tau) and to an extended four-parametric model p(t) = P infinity + (P0-P infinity) exp [(-t/(tau 0 + r tau t)]. The influence of the three independent variables on min LVdP/dt, tau, and r tau was checked by analysis of variance and quantified by standardized regression coefficients obtained by trivariate regression analysis. A positive dependence of min LVdP/dt on precedent maximum pressure and of the three-dimensionally (but not four-dimensionally) estimated tau on beat interval length are the only unequivocally significant effects of the investigated hemodynamic changes on the lusitropic parameters min LVdP/dt and tau. Principal species differences do not occur. It is concluded that considerable lusitropic effects seen especially in pharmacologic studies must be attributed to intrinsic effects of the substance rather than to hemodynamic changes caused by the substance if the former remain moderate. The four-dimensionally calculated tau is slightly more often found to be independent of the investigated hemodynamic parameters than the usual three-dimensional estimate. This may indicate higher reliability of the four-dimensional regression model of isovolumic pressure fall.