Pasquale De Sole

Effect of smoking one cigarette on antioxidant metabolites in the saliva of healthy smokers

Concentrations of glutathione, uric acid and total antioxidant activity, expressed as Trolox (a water-soluble vitamin E analogue) equivalent, were measured in the saliva of healthy non-smokers and smokers before and just after smoking a single cigarette. There was no statistically significant difference between smokers and non-smokers in uric acid concentrations and total radical-trapping antioxidant capacity, but glutathione concentrations were significantly (p < 0.05) higher in smokers. Smoking of a single cigarette induced a significant reduction in glutathione concentration (p < 0.05). Salivary antioxidant power may affect individual sensitivity toward tobacco stress.

Antimicrobial and antioxidant activities of Feijoa sellowiana fruit

The present study was designed to evaluate the antibacterial and antioxidant activities of an aqueous extract from the tropical Feijoa sellowiana Berg. fruit which is widely used for human food. The extract was tested against gram-positive and gram-negative bacteria by a broth dilution test and on human whole blood leukocytes, as well as isolated neutrophils using a chemiluminescence (CL) assay. The extract inhibited bacterial growth; Pseudomonas aeruginosa, Enterobacter aerogenes and Enterobacter cloacae were the most sensitive. The fruit extract significantly decreased CL emission from human whole blood phagocytes and isolated polymorphonuclear leukocytes whether they were activated or not by soluble or phagocytic stimuli. F. sellowiana showed both antibacterial and antioxidant properties and therefore its extract might be used as a new multifaceted drug.

Possible relationship between Al/ferritin complex and Alzheimer's disease

OBJECTIVES Ferritin is the main iron-storage protein capable of containing thousands of iron atoms. However, ferritin can bind in vitro other atoms such as aluminum and it has been shown that also in vivo atoms other than iron, as aluminum and zinc, are present in large amounts in ferritin. Since aluminum appears to be involved in the development of Alzheimers disease, in the present study the specific content of aluminum in ferritin of Alzheimers patients was analyzed and compared with other control groups. DESIGN AND METHODS The content of Fe, Al and Zn of blood ferritin was measured by mass spectrometry in patients with Alzheimers disease and compared with other clinical and control groups. RESULTS The results obtained confirm the hypothesis of a functional role of ferritin as a regulatory protein of toxic metals and clearly indicate that ferritin from Alzheimers patients has a content of aluminum higher than that of controls. CONCLUSIONS The specific aluminum content of ferritin seems to be related to different disease stages of Alzheimers disease. This result confirms the hypothesis of aluminum as a possible factor inducing the Alzheimers disease and opens the ways to possible new diagnostic tests.

Is homocysteine a pro-oxidant?

High plasma homocysteine concentrations have been found to be associated with atherosclerosis and thrombosis of arteries and deep veins. The oxidative damage mediated by hydrogen peroxide production during the metal-catalyzed oxidation of homocysteine is to date considered to be one of the major pathophysiological mechanisms for this association. In this work, a very sensitive and accurate method was employed to measure the effective production of H2O2 during homocysteine oxidation. Furthermore, the interaction of homocysteine with powerful oxidizing species (hypochlorite, peroxynitrite, ferrylmyoglobin) was evaluated in order to ascertain the putative pro-oxidant role of homocysteine. Our findings indicate that homocysteine does not produce H2O2 in a significant amount (1/4000 mole/mole ratio of H2O2 to homocysteine). Moreover, homocysteine strongly inhibits the oxidation of luminol and dihydrorhodamine by hypochlorite or peroxynitrite and rapidly reduces back ferrylmyoglobin, the oxidizing species, to metmyoglobin. All these results should, in our opinion, lead to a rethinking of the commonly held view that homocysteine oxidation is one of the main causative mechanisms of cardiovascular damage.

A fast chemiluminescent method for H2O2 measurement in exhaled breath condensate.

BACKGROUND Breath condensate can give useful information on volatile compounds produced at alveolar level. Actual concentration of H(2)O(2) in breath condensate is dependent on its production at alveolar level and on the efficacy of the detoxifying systems, catalase, glutathione peroxidase, etc. METHODS In the present paper, a simple chemiluminescent method for the determination of the H(2)O(2) collected in exhaled breath is shown and data of both smokers and nonsmokers volunteers are presented. RESULTS The chemiluminescent response is linear up to 100 micromol/l H(2)O(2). The analytical sensitivity is about 0.01 micromol/l. Most of the nonsmokers have a H(2)O(2) content lower than 0.05 micromol/l, while smokers have a content ranging from 0.1 to 0.6 micromol/l.

Quenching of singlet oxygen by D-α-tocopherol in human granulocytes

The ability of D-α-tocopherol to act as a quencher of 1O2 (singlet oxygen) was tested with a biological source of 1O2, namely the phagocytosis activated myeloperoxidase contained in the homogenate of human circulating polymorphonuclear leukocytes. With this system, the 1O2 quenching efficiency of exogenously added D-α-tocopherol was estimated from its inhibitory effect on the luminol amplified chemiluminescence. This inhibitory effect was dose dependent. D-α-tocopherol was also efficient in quenching the chemiluminescence generated through the H2O2-horseradish system. In both systems the quenching effect may be almost entirely “physical”, since very little tocopherol was destroyed when compared to the relatively large amount of H2O2 consumed.

Serum Coenzyme Q10 in Uremic Patients on Chronic Hemodialysis

In a group of 48 chronic hemodialysis patients, serum levels of coenzyme Q10 (CoQ) have been measured and appeared abnormally low in 62% of cases. Figures were positively correlated to those of serum vitamin E (vit E), although the latter were within a normal range. The chronic hemodialysis (CHD) patients with normal serum values of CoQ exhibited higher blood triglycerides. Pathologically low levels of serum vit E were found only in uremic subjects on conservative regimen with dietary restrictions and low compliance to protein-caloric intake. The reduced CoQ levels may contribute to the defective serum antioxidant activity and the increased peroxidative damage in uremic patients on CHD.

Effect of aqueous extract of cigarette smoke on peripheral blood polymorphonuclear leukocytes chemiluminescence

Cigarette smoke induces a vast cohort of deleterious effects on biological structures. In the present paper, the effect of aqueous extract of cigarette smoke on the activity of polymorphonuclear leukocytes was studied. Although the aqueous extract of cigarette smoke inhibits the luminol oxidation catalysed by horseradish peroxidase, it strongly interacts with polymorphonuclear leukocytes and inhibits their phorbol-induced chemiluminescence in the presence of either luminol or lucigenin. The results indicate that at least some of the components of the aqueous extract of cigarette smoke may strongly interfere with polymorphonuclear cells, contributing to the deleterious effects of smoke products.

Effects of 1,4-butanediol dimethacrylate and urethane dimethacrylate on HL-60 cell metabolism.

The polymerization of methacrylic monomers present in dental composite resins never reaches completion and therefore the leakage of residual monomers into the oral cavity and into biological fluids can cause local and systemic adverse effects. This work was carried out to study the in vitro biochemical interactions of urethane dimethacrylate and 1,4-butanediol dimethacrylate monomers with HL-60 cells, a cell line assumed as an experimental model for simulating granulocyte behaviour. Our main finding was that both monomers induce cell differentiation at toxic concentrations and that cytotoxicity seems to be caused by alterations of glucose metabolism arising from mitochondrial dysfunction rather than from oxidative stress, which could not be altogether verified under our experimental conditions. Our study could be considered as a useful approach to investigate the biochemical mechanisms that contribute to the cytotoxicity of methacrylate compounds and it underlines the importance of assessing such parameters for testing biocompatibility in order to promote the development of better and safer dental materials.

Chemiluminescence activity in whole blood phagocytes of dogs naturally infected with Leishmania infantum.

Dogs are the domestic reservoir of Leishmania infantum, a vector-borne intracellular protozoan agent of human visceral leishmaniasis. The role of polymorphonuclear leukocytes (PMNs) in the immune defence against this parasite has been poorly studied. We have investigated the function of peripheral blood PMNs in naive beagle dogs that have been naturally exposed to phlebotomine vectors in an area highly endemic for canine leishmaniasis, and found infected by Leishmania at the end of the transmission season. Whole blood phagocyte oxidative metabolism was assessed by a rapid method that determines a luminol-amplified chemiluminescence (CL) emission. This was evaluated using either a soluble stimulant, phorbol mirystate acetate (PMA), or phagocytic stimuli, such as zymosan unopsonized (ZYM) or opsonized with autologous serum (OPZ). In blood samples taken 2 months after exposure to Leishmania transmission, data on CL emission revealed a significant decrease of reactive oxygen intermediates (ROI) production in the presence of both PMA and ZYM, compared with blood samples obtained from dogs before exposure. On the contrary, no variations in CL emission were detected in presence of OPZ. Our data indicate that immunological changes occur early in canine leishmaniasis and confirm that the role of PMNs and their products need to be clarified.