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Biomedical subjects

A Di Giulio

Publications and source records attributed to A Di Giulio.

34 records · Page 2Linked to original sources

The ultraviolet derivative spectrophotometric determination of neutral liposome-entrapped beta-lactam antibiotics.

A simple, non-invasive spectrophotometric assay to measure the concentration of some beta-lactam antibiotics in turbid solutions containing liposomes was carried out. Since zero-order spectra gave strong interference because of sample turbidity, derivative spectrophotometry was used to enhance the spectral details. Derivative spectra showed bands in the ultraviolet region due to the presence of the cephalosporin and penicillin beta-lactams. A linear relationship between derivative amplitudes and antibiotic concentration was found when antibiotic-containing liposome solutions were measured. A saturative trend in the liposome-encapsulation was observed. The antibiotic entrapment was lowered by increasing the cholesterol-phospholipid ratio in the mixture used for liposome preparation. After treatment of antibiotic-loaded liposomes with beta-lactamase, a hydrolytic enzyme specific for beta-lactams, the remaining antibiotic concentration decreased significantly, showing that some of the antibiotic was retained on the outer surface of the vesicles.

Anti-Bacterial Agents↗

In vitro and in vivo effects of erythrocyte phototherapy on newborns.

The photodynamic action of the bilirubin is associated with severe consequences observed during 'in vitro' irradiation of the erythrocytes. This paper is designed to evaluate the bilirubin photodynamic effects which occur 'in vitro' and 'in vivo' on erythrocytes in healthy and jaundiced infants. The in vitro bilirubin sensitized photoreaction damages the erythrocytes mainly at the membrane level. In particular, a dramatic decrease of ATPase activity and an increased susceptibility to lipid peroxidation, expressed as malondialdehyde production, were observed. For in vivo studies, specific fluorescent probes have been used to verify probable changes on the functional architecture of the erythrocyte membrane in the phototherapy-treated infants. Our results showed that specific areas of the membrane are differently affected, mainly at lipid/protein interface. Although the role of the erythrocyte membrane is an important factor of the hemorheological behavior, the measurement of blood viscosity and erythrocyte aggregation and filtration did not show significant alterations during the overall time of phototherapy.

Adenosine Triphosphatases↗

PPIX induced photohemolysis of erythrocytes partially-depleted of cholesterol.

The protoporphyrin IX (PPIX)-sensitized hemolysis of erythrocytes depleted of cholesterol was investigated. From 20% to 30% of the total membrane cholesterol was removed from cells by incubation with old autologous plasma or by means of interaction with L-alpha-phosphatidylcholine dipalmitoyl (DPPC) liposomes. As expected, after this treatment, the cells show an overall increase in membrane fluidity revealed by means of specific fluorescent probes. The same cells are more susceptible to the photohemolysis induced by PPIX excited by visible light, but gave no lysis in the absence of the sensitizer. As a consequence, the primary oxidative damage which is produced during irradiation can be possibly assigned to the phospholipidic and/or proteic moiety instead of the steroidal moiety.

Anilino Naphthalenesulfonates↗

Controlled human RBC modifications affecting the binding of cationic liposomes.

Cationic liposomes were prepared either by sonication or by detergent dialysis and used to deliver the antioxidative enzyme glutathione peroxidase into human erythrocytes in vitro. The enrichment ability of these two preparations was similar, amounting to about 30% of the control cells. The lysis of enzyme-treated erythrocytes induced by photoirradiation in the presence of PPIX was compared with that of cells incubated with empty liposomes. Erythrocytes enriched with GPX appear to be more resistant toward photohemolysis. Pre-treatment of cells with neuraminidase or proteinase K suggests that: a) sialic acid seems to be essential for the cell-liposome fusion process, no enrichment being found with the neuraminidase-treated cells; b) hydrolysis of the outer membrane proteins leads to an increased fragility with respect to controls even in GPX-enriched cells. These results were confirmed by extrinsic fluorescence polarization experiments, using isolated erythrocyte membranes and specific fluorescent probes.

Endopeptidase K↗

Methionine-enkephalin, substance P, and homovanillic acid in the CSF of parkinsonian patients.

Methionine-enkephalin, substance P, and homovanillic acid concentrations were measured in the CSF of subjects not affected by neurologic disorders (group 1), and in parkinsonian patients who had a slight or moderate (group 2) or severe (group 3) disability. Homovanillic acid and substance P concentrations in the CSF of groups 2 and 3 were respectively lower and higher than in group 1. On the contrary, an increase in CSF methionine-enkephalin content was found only in group 2. Our results confirm in humans the close relation between the dopaminergic and peptidergic transmissions in the nigrostriatal system that has been observed in experimental animals.

Adult↗

Effect of hypophysectomy and growth hormone on tryptophan and serotonin metabolism.

The present study reports data on brain serotonin (5-HT) metabolism in two experimental conditions which have in common a lack of pituitary growth hormone (GH) and refers also the effect of a GH replacement therapy. In hypophysectomized female rats or genetically dwarf mice, which have a more selective deficiency of GH, brain tryptophan (TP) and 5-hydroxyindoleacetic acid (5-HIAA) concentrations were significantly higher than in age-matched controls. Brain levels of 5-HT were practically unchanged. A GH replacement therapy resulted in both hypophysectomized rats and dwarf mice in a significant reduction of brain TP and 5-HIAA concentrations. These results suggest that GH may exert an inhibitory action on 5-HT metabolism. Consonant with this view are the preliminary findings that in 7 individuals with elevated plasma GH levels for acromegaly, plasma TP levels were lower than in control subjects.

Acromegaly↗

Monitoring by cis-parinaric fluorescence of free radical induced lipid peroxidation in aqueous liposome suspensions.

Cis-parinaric acid is fluorescent when partioned into a lipid environment and its fluorescence is destroyed upon reaction with free radicals. In our study 1-palmitoyl-2-parinoyl-phosphatidylcholine (cis-PnA) has been used to monitor the time-course of liposomal lipid peroxidation, using reverse-phase evaporation vesicles (REV) of different composition exposed to oxidative stress in various conditions. This methodology allowed us to estimate the potential damage produced by two different oxidizing systems, namely hydrogen peroxide (H2O2), a water soluble oxidant, and t-butyl hydroperoxide (t-BHP), a hydrophobic hydroperoxide. Furthermore, we evaluated the protective effects of bilayer-associated antioxidants, namely alpha-tocopherol acetate (alpha-THA), vitamin K1 and beta-carotene, as well as of two antioxidants dissolved in the aqueous bulk solution, that is, biverdin and uric acid. Under our experimental conditions, the results suggest that (i) both oxidizing compounds were able to interact with liposomal PnA leading to decay either of the excitation and of emission spectra of the probe; (ii) hydrogen peroxide seemed to be of most effective among the two stressing agents, when employed at similar concentrations; (iii) the alpha-THA appeared to be a stronger antioxidant than vitamin K1 and beta-carotene, resulting in a decrease of the liposomal membrane stress caused by those two oxidizing agents; (iv) among the water soluble antioxidant compounds, biliverdin displayed a protective effect at least 10 x higher than uric acid; (v) the overall damage, as well as the protection mechanisms, seemed to be dependent either on the lipid composition of the vesicles and on the pH of the liposomal suspension. This relatively easy experimental approach suggests the validity of the use of the bilayer associated fluorescent probe PnA in the monitoring of spontaneous and/or chemically induced liposomal lipid damage.

Antioxidants↗

[Leukocyanidines and collagenases: in vitro enzyme inhibition activity].

OBJECTIVE: The matrix metalloproteinases (MMPs) play a key role in the extracellular matrix turnover. This protein family has been involved in some ocular pathologies such as glaucoma, diabetic retinopathy, macular degeneration, vitreous degeneration and corneal stroma ulceration cleaving all the matrix components. In the present study we evaluated the action of leucocyanidin from Vitis vinifera seeds as non toxic inhibitor of these proteinases. MATERIALS AND METHODS: To this purpose we used a fluorimetric method to evaluate the effect of this substance on the collagenase activity. We evaluated "in vitro" the inhibitory potency of the tested drug on type III collagenase activity, and the recover of the metalloprotease activity upon removal by dialysis of the inhibitor. RESULTS: The leucocyanidines extract (minimum procyanidines value of 95.0) resulted to be a good collagenase activity inhibitor showing an inhibition constant value, Ki, of 82 microM, evident index of affinity between the extract and the enzyme. Furthermore, the dialysis experiments demonstrated that the inhibitory effect persisted 24 h later, probably because the extract forms a stable complex with the enzyme. CONCLUSIONS: These results should be related to the pharmacokinetic profile of leucoanthocyanins, a family of natural polyphenols belonging to the class of bioflavonoids of grape seds extract (Vitis vinifera L.).

Collagenases↗

[Metalloproteinase inhibition: therapeutic application in rheumatic diseases].

PURPOSE: Matrix metalloproteinases (MMPs) play an important role in the degradation of articular cartilage in several diseases, including osteorthritis and rheumatoid arthritis. Aiming at developing new drugs with selective inhibiting action against enzyme damaging the extracellular matrix, research is mainly directed towards the: 1) development of new drugs with specific inhibitory effect on MMPs; 2) better understanding of the pharmacologic profile of drugs already used in the treatment of rheumatic diseases, in order to identify those having an inhibiting action on degradative enzymes. MATERIALS AND METHODS: The interaction between rifamycins and collagenase type XI were studied using a fluorogenic substrate MOCAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2. RESULTS: In our experimental conditions rifamycins showed a marked inhibition capacity with a IC50 ranging from 13 to 20.7 microM. This inhibition was reversible after extensive dialysis. CONCLUSIONS: Our results indicate that the effects of rifamycins in rheumatoid arthritis may correlate to the inhibitory activity of these molecules on collagenase activity.

Arthritis, Rheumatoid↗