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Biomedical subjects

E Bertoli

Publications and source records attributed to E Bertoli.

At least 73 records · Page 4Linked to original sources
PubMed1990

Differential scanning calorimetry characterization of oxidized egg phosphatidylcholine liposomes.

Differential scanning calorimetry has been used to characterize liposomes made from mixtures of unoxidized and singlet oxygen oxidized egg phosphatidylcholine. Cooling scans reveal that trapped water decreases when the oxidized phosphatidylcholine content is increased in the liposomes. Liposomes made from mixtures containing more than 50% by weight of oxidized phosphatidylcholine do not show trapped water.

Calorimetry↗
PubMed1990

Interaction of the herbicide atrazine with model membranes. I: Physico-chemical studies on dipalmitoyl phosphatidylcholine liposomes.

Atrazine (2-chloro-4 ethylamino-6-(isopropylamino)-s-triazine) is one of the most widely used herbicides. Fourier transform infrared spectroscopy, differential scanning calorimetry and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and of its derivative 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) were used to study the interaction of atrazine with dipalmitoyl phosphatidylcholine liposomes used as a model for biological membranes. The results show that atrazine does not perturb the hydrophobic core of the lipid bilayer and suggest that the herbicide localizes near the glycerol backbone of the lipid.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1990

Interaction of the herbicide atrazine with model membranes. II: Effect of atrazine on fusion of phospholipid vesicles.

The effect of atrazine on Ca2+ induced fusion of cardiolipin(CL) and phosphatidylserine (PS) vesicles is studied by Tb3+/dipicolinic acid fluorescence and turbidity measurements. The interaction of herbicide with CL and PS membranes is studied by DPH fluorescence polarization. At low concentrations the pesticide partially inhibits fusion, especially in CL vesicles. Higher concentrations of atrazine decrease inhibition of fusion in CL, while fusion is slightly increased in PS. The Ca2(+)-induced increase of turbidity is not affected by atrazine in both PS and CL aggregation experiments. DPH polarization measurements show a perturbation only of the membrane hydrophobic core of PS, in presence of Ca2+. It is hypothesized that this biphasic effect shown by low and high atrazine concentrations on Ca2(+)-induced fusion of vesicles is due to a different localization of the pesticide in the membrane.

Atrazine↗
PubMed1990

Alterations of the respiratory burst of polymorphonuclear leukocytes from diabetic children. A chemiluminescence study.

The respiratory burst of polymorphonuclear leukocytes was investigated in 24 children with insulin dependent diabetes mellitus and 24 healthy controls. This oxygen dependent, membrane associated process generates a number of toxic oxygen metabolites which are implicated in the pathogenesis of endothelial damage. The activity of polymorphonuclear leukocytes was studied in terms of luminol amplified chemiluminescence. It was found that the resting luminol amplified chemiluminescence activity of isolated polymorphonuclear leukocytes from diabetic children was significantly higher than that of controls (342,000 +/- 174,000 cpm vs. 165,000 +/- 82,000 cpm, p less than 0.01). The addition of respiratory burst inhibitors caused a significant reduction of basal chemiluminescence (greater than 80%). When the ratio of phorbol myristate acetate stimulated activity to basal activity was calculated and used as an activation index, it was found to be significantly reduced in diabetics relative to controls (4.29 +/- 2.46 vs. 8.34 +/- 3.21, p less than 0.01). These observations suggest that increased release of toxic oxygen metabolites from polymorphonuclear leukocytes in diabetic subjects may play a role in the development of diabetic angiopathies.

Adolescent↗
PubMed1990

Altered cellular Ca2+ and Na+ transport in diabetes mellitus.

Platelet intracellular Ca2+ concentration ([Ca2+]i) and its response to stimuli (ADP and thrombin) were studied in 15 insulin-dependent and 22 non-insulin-dependent diabetes mellitus patients with the fluorescent probe Fura 2. The activity of Ca2(+)-ATPase and Na(+)-K(+)-ATPase, membrane fluidity, and cholesterol and phospholipid content were also determined in platelet membranes. Compared with control subjects, diabetic patients showed 1) increased platelet [Ca2+]i in the resting state, 2) higher Ca2+ levels after stimulation with thrombin and ADP, due entirely to increased resting concentrations, 3) reduced activity of Na(+)-K(+)-ATPase, 4) increased activity of Ca2(+)-ATPase, 5) higher fluidity of the platelet membrane, and 6) increased membrane concentration of total phospholipids. Na(+)-K(+)-ATPase activity was inversely related to platelet [Ca2+]i in each group studied, whereas Ca2(+)-ATPase activity was positively correlated with intracellular Ca2+ levels. The data obtained in diabetic subjects suggest an abnormality in Ca2+ and Na+ transport across the platelet membrane that might be responsible for the reported platelet hyperreactivity to stimuli in diabetes.

Adenosine Diphosphate↗
PubMed1990

Direct oxidation of glutamate by mitochondria from porcine adrenal cortex.

1. Mitochondria isolated from porcine adrenal cortex under State 3 conditions oxidized succinate with a rate of 47 +/- 4.48 na oxygen/min/mg/protein and with ADP:O ratio 0.98 +/- 0.09. In the presence of 15 microM deoxycorticosterone the rate of succinate oxidation was 36.8 +/- 3.08 na oxygen/min/mg/protein. 2. Under the same conditions the rate of glutamate oxidation was 22.8 +/- 2.21 and 16.8 +/- 0.65 na oxygen/min/mg/protein, respectively. ADP:O ratio was 1.45 +/- 0.14. 3. Introduction of trace amounts of malate into the mitochondria oxidizing glutamate only slightly increased the rate of O2 uptake. 4. The glutamate dehydrogenase activity in these mitochondria was 12.5 +/- 0.69 nmol/min/mg.

Adrenal Cortex↗
PubMed1990

Steady state fluorescence polarization and Fourier transform infrared spectroscopy studies on membranes of functionally senescent human erythrocytes.

Human erythrocytes were separated using Percoll/Albumin density gradient centrifugation into 5 subpopulations which represent cell subpopulations of different ages. The physico-chemical properties of the membranes and their lipid extracts, prepared from the 5 subpopulations, were studied by 1,6-diphenyl-1,3,5-hexatriene fluorescence polarization and by Fourier Transform Infrared Spectroscopy respectively. The fluorescence polarization data show a decrease of membrane fluidity in the youngest erythrocyte fraction and the infrared spectroscopy reveals a lower degree of unsaturation of the lipids from this fraction. Changes of membrane fluidity among the 5 erythrocyte subpopulations is assigned to the variations of the degree of unsaturations of lipids rather than to variations of cholesterol content.

Adult↗
PubMed1989

Permeability of oxidized phosphatidylcholine liposomes.

Permeability of liposomes made from mixtures of unoxidized and singlet oxygen oxidized phosphatidylcholine has been related to the degree of lipid oxidation expressed as hydroperoxide moiety content in the lipids. The effect of oxidation on the liposomes permeability has been studied by fluorometry using calcein as a fluorescent probe that undergoes self quenching when highly concentrated inside liposomes. The liposomes containing 73% and 5% of hydroperoxides retain respectively 64.5 and 96.3% of calcein with respect to that retained by the liposomes made from unoxidized phosphatidylcholine. The fluorescence data show a linear relationship between the liposome permeability and the oxidation degree of lipids.

Chromatography, High Pressure Liquid↗
PubMed1989

Steady-state fluorescence anisotropy and multifrequency phase fluorometry on oxidized phosphatidylcholine vesicles.

Multilamellar liposomes, from mixtures of unoxidized (control) and singlet oxygen oxidized phosphatidylcholine, were studied by steady-state fluorescence anisotropy and multifrequency phase fluorometry using 1,6-diphenyl-1,3,5-hexatriene (DPH) as fluorescent probe. Lifetime fluorescence decay of the DPH-labeled liposomes was analyzed either by a model of discrete exponential components and a model that assumes a continuous distribution of lifetime values. Increasing the oxidized phosphatidylcholine content in the liposomes, an increase of the membrane interior polarity and a decrease of membrane fluidity occurs which can be related to the hydroperoxide-lipids and double bonds conjugation, respectively.

Diphenylhexatriene↗
PubMed1989

Calcium and sodium transport in gestational hypertension.

The enzymatic activities of Na+/K+ ATPase and Ca2+ ATPase were determined on erythrocyte membranes from 9 normotensive and 9 gestational hypertensive pregnant women near term. A reduction in the activity of the Na+/K+ ATPase and a relative increase in the activity of the Ca2+ ATPase were found in the hypertensive patients, possibly due to a conformational alteration of erythrocyte membranes. This observation supports the possible role of the transmembrane cation transport in the pathogenesis of gestational hypertension.

Adult↗
PubMed1989

Modifications induced by diabetes on the physicochemical and functional properties of erythrocyte plasma membrane.

Increasing evidence suggests that in experimental diabetes an impairment in Na+,K+-ATPase activity plays a central role in the pathophysiology of diabetic complications, while only a few data are available with regard to human subjects. We studied the erythrocyte membrane Na+,K+-ATPase activity and membrane fluidity in insulin-dependent and non-insulin-dependent diabetic subjects. A significant decrease in the enzyme activity and in fluorescence polarization values was found in both groups compared with normal subjects. Neither Na+,K+-ATPase activity nor membrane fluidity was found to be related to metabolic control, assessed by means of fasting blood glucose levels and HbA1c. On the contrary, a significant correlation was observed between Na+,K+-ATPase activity and membrane fluidity in both insulin-dependent and non-insulin-dependent diabetic subjects. The present work provides evidence that a reduction in the Na+,K+-ATPase activity is present in the plasma membranes of insulin-dependent and non-insulin-dependent diabetics. Furthermore, it suggests that the change in enzyme activity might be related to modifications in membrane fluidity.

Adult↗
PubMed1989

Modifications of erythrocyte membrane fluidity of in vivo functionally aged human erythrocytes.

The process of red blood cell senescence in the blood stream results in many changes in their physical and biochemical properties. In this work we have studied the physico-chemical state of erythrocyte membranes prepared from 5 subpopulations of erythrocytes of different age by using the fluorescence technique. Membrane fluidity has been evaluated by the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and a further study of the fluorescence decay of this probe has been performed by multifrequency phase and modulation fluorometry. DPH fluorescence polarization is significantly increased in the membranes prepared from the youngest fraction of erythrocytes, indicating a decreased fluidity without any significant change in DPH fluorescence decay.

Erythrocyte Aging↗
PubMed1988

Increased membrane heterogeneity in stimulated human granulocytes.

TMA-DPH fluorescence decay in human PMN before and after stimulation with FMLP was studied using frequency domain fluorometry. Membrane heterogeneity was assessed by the width of the continuous distributions of lifetime values of Lorentzian shape used to describe the fluorescence decay. In non-stimulated granulocytes TMA-DPH fluorescence decay is characterized by two distributions of lifetime values centered at 6.5 and 1.0 ns and full width at half maximum of 0.3 and 1.2 ns, respectively. Within 15 min after stimulation, the center values of the two distribution components were 5.1 and 0.8 ns and the distribution width was 0.8 and 0.6 ns, respectively. These results indicate changes of membrane domain organization which can be ascribed to compositional changes and redistribution of membrane components.

Cell Membrane↗
PubMed1988

Changes of erythrocyte membrane fluidity associated with childhood obesity: a molecular study using fluorescence spectroscopy.

Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene has been used to investigate the physical state of erythrocyte membranes from 36 obese children in absence of endocrine and metabolic disorders involving alterations of blood lipid pattern. A decrease of membrane fluidity, as shown by an increase of fluorescence polarization, was evident in obese subjects in comparison with the controls. The study of membrane composition has revealed an increase of cholesterol content and of cholesterol to phospholipid molar ratio, both being positively correlated with the observed changes of membrane fluidity. Moreover a correlation of membrane fluidity is also observed with body-mass index and plasma cholesterol levels. The results are discussed in the light of the possible use of erythrocyte membrane as a model to detect early alterations in the exchanges between erythrocyte membrane and lipoproteins in obesity.

Adolescent↗
PubMed1988

Abnormal membrane cation transport in pregnancy-induced hypertension.

An abnormality of sodium handling has been suggested as one of the mechanisms responsible for the development of pregnancy-induced hypertension. We analysed the plasma and urinary concentrations, and the intraerythrocyte activities of Na and K, and the RBC membrane Na+/K+-ATPase activity of 77 hypertensive and 133 normal pregnant women. Umbilical cord blood of infants from 21 hypertensive and 28 control women was studied. The Na+/K+-ATPase activity was determined by measuring the inorganic phosphate released by incubation in a reaction medium in the presence and absence of K ions or ouabain. The intra-erythrocyte sodium and potassium activities were measured by ion-selective electrode analysis of the haemolysates, after washing the RBCs in 110 mmol/l MgCl2. We found a significant increase in intracellular sodium and a reduction in Na+/K+-ATPase activity in the hypertensive women in comparison with the control subjects during pregnancy. No difference was observed in early puerperium. Cord blood from infants of pregnancy-induced hypertensive women showed an increase in intracellular Na+ activity and a decrease in the erythrocyte membrane Na+/K+-ATPase activity in comparison with cord blood samples from control subjects. The observed abnormalities in the plasma membrane sodium transport may play a major role in the pathophysiology of pregnancy-induced hypertension.

Biological Transport↗
PubMed1988

Abnormal membrane fluidity and acetylcholinesterase activity in erythrocytes from insulin-dependent diabetic patients.

An alteration in the enzymatic properties of the erythrocyte membrane acetylcholinesterase (AchE) and Na+,K+-ATPase has been described in experimental diabetes mellitus. We studied erythrocyte membrane fluidity and AchE and Na+,K+-ATPase activities in 15 insulin-dependent diabetic patients and 11 normal subjects. Fluidity was assessed by fluorescence polarization, using 1,6-diphenyl-1,3,5-hexatriene as a probe, and AchE and Na+,K+-ATPase activities were measured enzymatically. We found a significant increase in the enzymatic activity of AchE and a change in its enzymatic properties in diabetic patients compared with those in normal subjects. AchE activity correlated inversely with membrane fluorescence polarization, which was decreased in the diabetic patients, indicating an increase in membrane fluidity. Na+,K+-ATPase activity was reduced in the diabetic patients and correlated positively with the fluorescence polarization values. We hypothesize that the abnormal dynamic properties of the erythrocyte membrane may play a major role in determining the described change in enzymatic activity.

Acetylcholinesterase↗
PubMed1988

Sodium/potassium-adenosine triphosphatase on erythrocyte ghosts from pregnant women and its relationship to pregnancy-induced hypertension.

The regulation of electrolytes has been proposed as a possible determinant in hypertensive conditions, including pregnancy-induced hypertension. We report a study of sodium/potassium-adenosine triphosphatase in erythrocyte membranes of 59 patients with pregnancy-induced hypertension and 48 normotensive pregnant controls. The kinetics of the enzyme were also investigated, and the enzyme activity related to different degrees of pathology. A marked reduction of the enzyme activity was found in hypertensive patients, compared with controls. This reduction was greater when hypertension was associated with worse prognostic signs, such as proteinuria. These features are in agreement with the increased sodium content and the increased vascular reactivity found in pregnancy-induced hypertension. The enzyme activity appears to be decreased by means of a conformational modification of the enzyme sites, a phenomenon related to the hypertensive condition.

Edema↗