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Biomedical subjects

G Curatola

Publications and source records attributed to G Curatola.

At least 91 records · Page 5Linked to original sources

Changes of lymphocyte membrane fluidity in rheumatoid arthritis: a fluorescence polarisation study.

Fluorescence polarisation of 1,6-diphenyl-1,3,5-hexatriene was used to study the lymphocyte membrane in rheumatoid arthritis. The increase of polarisation value in the patients (n = 27) compared with healthy controls (n = 32) suggests a decrease of membrane fluidity. Moreover, erythrocyte sedimentation rate (ESR) and plasma fibrinogen concentrations were positively correlated with lymphocyte fluorescence polarisation values (r = 0.66 and r = 0.76 respectively). The results suggest that the changes in lymphocyte membrane fluidity could be involved in the pathogenetic mechanism of rheumatoid arthritis.

Adult↗

Altered properties of platelet membrane in childhood obesity.

Using 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5,hexatriene (TMA-DPH) as fluorescent probe, we have studied platelet membrane fluidity and thrombin induced exocytosis in ten obese children and fifteen controls. Our results indicate a decrease of membrane fluidity, as shown by an increase of fluorescence anisotropy, in platelets from obese patients in comparison with the controls. Associated with the changes in membrane fluidity, platelets of obese subjects showed a decreased sensitivity to thrombin stimulation.

Adolescent↗

Erythrocyte membrane heterogeneity studied using 1,6-diphenyl-1,3,5-hexatriene fluorescence lifetime distribution.

The fluorescence decay of 1,6-diphenyl-1,3,5-hexatriene has been used to characterize the structural organization of erythrocyte membranes. At 37 degrees C a large fraction of the decay (0.96) is associated with a lifetime value of 11.31 ns, while a minor fraction has a short lifetime of 2.63 ns. The distribution analysis approach has shown that the 11 ns component can be described using a Lorentzian distribution function having a full width at half maximum of 0.27 ns. The width of this component is associated with the membrane structural organization since liposomes from erythrocyte total lipid extract exhibit a narrower width. Moreover the distribution width is sensitive to different treatments of erythrocyte membrane.

Diphenylhexatriene↗

Modification of lipid composition of erythrocyte membranes in chronic alcoholism.

The lipid composition and microviscosity of erythrocyte membranes have been studied in chronic alcoholics with macrocytosis and minimal liver dysfunction. The findings have been compared with those obtained in subjects with macrocytosis not related to ethanol abuse. In all instances, macrocytosis was accompanied by an increased microviscosity of erythrocyte membranes, together with an increase of the cholesterol/phospholipid and of saturated/unsaturated fatty acids ratio. Both microviscosity and saturated/unsaturated ratio were significantly higher in alcoholics compared to subjects with macrocytosis not related to ethanol abuse. These findings confirm that altered lipid composition and fluidity of erythrocyte membrane may be observed in chronic alcoholics with minimal liver dysfunction, possibly due to the "stabilising" properties of ethanol - among a number of drugs- on biological membranes.

Adult↗

Changes of membrane fluidity in erythrocytes of obese children: a spin label study.

Red blood cells depend on circulating lipoproteins for the turnover of membrane phospholipids. Thus abnormalities in lipid and lipoprotein metabolism are frequently associated with changes in composition and fluidity of erythrocyte membranes. Using the electron spin resonance technique we studied the physical state of erythrocyte membranes from 15 obese children with a body mass index ranging from 22.8 to 34 kg . m-2 and slight alterations of plasma lipids. The spin labels used, 5-doxylstearate and 16-doxylstearate, have shown an increase of the order parameter and of the correlation time, respectively. These observations indicate a decrease of fluidity at the surface and in the hydrophobic core of erythrocyte membranes from obese subjects. The results are statistically significant and they are discussed in relation to clinical aspects of childhood obesity. Studies on erythrocyte membrane molecular organization could represent a sensitive way to find out small metabolic alterations hardly detectable with compositional studies.

Adolescent↗

Changes of membrane fluidity in chemotactic peptide-stimulated polymorphonuclear leukocytes.

Although the phenomenon of stimulus-response coupling in polymorphonuclear leukocytes involves a series of membrane events the influence of stimulation on membrane fluidity is to clarify. In our experiments we have used 1-(4-trimethylaminophenyl) 6-phenyl-1,3,5-hexatriene and 1,6-diphenyl-1,3,5-hexatriene fluorescence polarization technique to evaluate membrane fluidity in living polymorphonuclear leukocytes after stimulation with N-formyl-methyonil-leucyl-phenylalanine peptide which has a well defined membrane receptor on the plasma membrane. We report that polymorphonuclear leukocytes stimulation increases 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene polarization, only when colcemid, a microtubule disrupting drug, is added to polymorphonuclear leukocytes. This can be viewed as an indirect evidence that microtubules are involved in the control of polymorphonuclear leukocytes membrane fluidity. On the contrary no changes have been observed with 1,6-diphenyl-1,3,5-hexatriene. This study indicates the potential use of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene to evaluate the involvement of plasma membrane physical state during intact cell activity.

Cell Membrane↗

The kinetic and structural changes of the mitochondrial F1-ATPase with temperature.

Mitochondrial F1-ATPase shows a break in the Arrhenius plot with an increase of the activation energy below 17 degrees C, this may imply that the F1-ATPase undergoes a conformational change at this temperature. Further, a structural change of the F1-ATPase is indicated by analysis of the intrinsic fluorescence at 307 nm between 33 and 11 degrees C and also by evaluation of the circular dichroism spectra of the enzyme at temperatures below and above the temperature corresponding to the discontinuity of the Arrhenius plot. It is therefore suggested that F1-ATPase exists in two temperature dependent conformational states to which different catalytic properties may be assigned.

Adenosine Triphosphate↗

Effect of chronic ethanol abuse on the physico-chemical properties of erythrocyte membranes in man.

Chronic alcoholics (greater than 150 g/day) showing minor serum and histological changes have been studied, compared to healthy non alcoholic subjects, and the following parameters have been considered: S. (Serum)cholesterol (CH), S.phospholipids (PH), S.folate level, and mean corpuscular volume (MCV). Erythrocyte ghosts have been studied for CH and PH content and membrane fluidity using diphenylhexatriene as a probe. All alcoholics showed decreased fluidity of red cell membrane with increased CH/PH ratio, even in patients showing normal MCV or minimal alterations of functional tests, suggesting that changes in red cell membrane fluidity represent an early sign of ethanol abuse. These likely reflect the diffuse interaction of ethanol with biological membranes. The administration of N5-Methyltetrahydrofolate produced an increase of membrane fluidity over the 3 weeks considered, associated with modest changes of MCV. The latter were delayed with respect to changes in fluidity. If changes of red cell membrane fluidity are a sensitive index of alcohol abuse, they could be a useful marker for detection and follow-up of chronic alcoholism.

Adult↗

S-100 protein-induced changes in the physical state of synaptosomal particulate fractions as monitored by spin labels.

This report documents changes in the physical state of synaptosomal particulate fractions (SYN) upon binding of S-100 protein, as monitored by spin labels. Studies were conducted on SYN labeled with either 5-doxylstearic acid or 16-doxylstearic acid, which probe the polar region and the hydrophobic core of the lipid bilayer, respectively. S-100 perturbs to some extent both the polar surface and the hydrophobic core of SYN in a time- and temperature-dependent manner. Ca2+ is essential for S-100 to perturb the membranes. K+ almost completely inhibits the S-100 perturbing effect if present in the incubation medium, but fails to reverse the S-100-induced changes if added after S-100 has interacted with SYN. At room temperature and below, the overall S-100 effect registered after about 30 min of association of the protein with SYN is an increase in the fluidity of both the surface and the interior of the membranes. Spectra registered at intervals at room temperature indicate that the S-100 perturbing effect on the membrane surface is practically monophasic, consisting of an increase in fluidity, while that on the membrane interior is multiphasic, consisting of a decrease in fluidity during the first 10 min of association, followed by an increase in fluidity during the subsequent 20 min and a return to starting values during the second 30 min of association. Around 37 degrees C, on the contrary, a decrease in fluidity is registered in both regions. The data suggest that S-100 induces a spatial rearrangement of membrane components (proteins) involved in the specific binding and/or partially penetrates into the lipid bilayer.

Animals↗

Age-related modification of dopaminergic and beta-Adrenergic receptor system: restoration to normal activity by modifying membrane fluidity with S-adenosylmethionine.

Microviscosity of membranes prepared from striata and cortex of 3 or 30-month-old rats was measured by fluorescence polarization and electron spin resonance. The viscosity of the hydrophobic core of the lipid bilayer was significantly increased in striatal but not in cortical membranes of old rats. In old rats the incorporation of methyl groups into phosphatidylcholine was significantly lower than in young rats. beta-Adrenergic specific binding sites were reduced in striata, cortex and pineal gland of old rats. In the striata of these animals, [3H] spiperone specific binding sites and dopamine-stimulated adenylate cyclase were also low. A chronic treatment of old rats with S-adenosyl-L-methionine, the cofactor in phospholipid methylation, lowered microviscosity to normal values in striata but not in the cortex. In the same old rats, the beta-receptor in striata and pineal gland returned towards juvenile values; in the cortex, the modification was in the same direction but did not reach significance. Neither putative dopaminergic receptors nor the adenylate cyclase age-dependent decrease was modified by treatment with S-adenosyl-L-methionine.

Aging↗

Duchenne muscular dystrophy: alterations in lymphocyte fluidity.

We have studied lymphocyte membrane from 7 patients with Duchenne Muscular Dystrophy (DMD) and 8 normal controls using fluorescence polarization of 1,6-diphenyl-hexatriene (DPH). The microviscosity of lymphocytes membrane is significatively higher in DMD patients than in normal controls. Our results support the hypothesis of a generalized membrane defect in Duchenne Muscular Dystrophy.

Chemical Phenomena↗

Temperature-dependent conformational changes in isolated oligomycin-sensitive ATPase.

Isolated oligomycin-sensitive ATPase undergoes a kinetic change at 20-25 degrees C with a higher activation energy and a lower Km for ATP below this temperature range. This observation has been correlated with temperature-dependent structural changes detected by circular dichroism in the UV region in the isolated enzyme. The negative ellipticities in the 208-225 nm region, which are proportional to the alpha-helix content, increase with rise in temperature to a maximum above 25 degrees C.

Adenosine Triphosphatases↗

Electron spin resonance studies of the effects of lipids on the environment of proteins in mitochondrial membranes.

The physical state of mitochondrial membranes has been investigated by means of stearic acid spin labels and of a maleimide spin label covalently bound to protein sulfhydryl groups. Stearic acid spin labels 5-NS and 16-NS show that n-butanol enhances the lipid fluidity of mitochondrial membranes in the whole temperature range between 4 and 37 degrees C; the effects in the hydrophobic membrane core, probed by 16-NS, are already apparent at 10 mM butanol. In liposomes formed of mitochondrial phospholipids, a fluidizing effect appears only at much higher concentration. Such results are compatible with the idea that butanol destabilizes lipid-protein interactions. On the other hand, the ratio between weakly and strongly immobilized SH groups probed by maleimide spin label is only slightly affected in the temperature range of 4-37 degrees C by addition of high concentrations of n-butanol, indicating that the environments probed are stable to agents inducing fluidity changes in the lipids. There are, however, indications that the environment probed by maleimide is affected by lipids, since the spin label, when bound to lipid-depleted mitochondria, becomes more immobilized, reconstitution of such lipid-depleted membranes with phospholipids restores the original spectra.

Adenosine Triphosphatases↗

The value of ultrasound as a screening procedure for urological disorders in renal failure.

Renal ultrasound was evaluated as a screening procedure for urological conditions (hydronephrosis or space-occupying lesions) in 175 patients (316 kidneys) with renal failure. The final diagnosis of renal disease was based on excretory urography, ascending or translumbar pyelography, open surgical findings, clinical and laboratory test results or renal biopsies. Ultrasonic images suggestive of renal disease requiring surgery were obtained in 83 kidneys and subsequently confirmed in 67. There were 16 falsely positive and 2 falsely negative results. Assuming a 22 per cent prevalence of renal disease requiring surgery in the azotemic population, Bayes' theorem predicts that an abnormal ultrasonic test will increase the probability of surgery in a patient with renal failure from 0.22 to 0.81, while a negative test will reduce the probability from 0.22 to 0.013. Renal ultrasound is a highly effective method to screen for patients with renal failure who require contrast studies. With this means the risks and cost of contrast investigations can be reduced markedly.

Adolescent↗

S-adenosylmethionine protects against erythrocyte membrane alterations induced in rabbits by cholesterol-rich diet.

In the erythrocyte membranes of rabbits fed a cholesterol rich diet for two months the cholesterol/phospholipid ratio was twice higher than in controls. The same diet induced a significant decrease in membrane fluidity and acetylcholinesterase activity. Two daily administrations of 25 mg X kg-1 b.w.-1 of S-adenosylmethionine (SAMe) have shown to protect against membrane alterations induced by hypercholesterolemic regimen. This observation would suggest the use of SAMe in those pathological conditions like hyperlipoproteinemias and liver diseases where membrane alterations are reported.

Acetylcholinesterase↗