Maria Letizia Falcioni

Codrugs linking L-dopa and sulfur-containing antioxidants: new pharmacological tools against Parkinson's disease.

A series of multifunctional codrugs (1-6) were synthesized to overcome the pro-oxidant effect associated with L-dopa (LD) therapy. Target compounds release LD and dopamine (DA) in human plasma after enzymatic hydrolysis, displaying an antioxidant effect superior to that of N-acetylcysteine (NAC). After intracerebroventricular injection of codrug 4, the levels of DA in the striatum were higher than those in LD-treated groups, indicating that this compound has a longer half-life in brain than LD.

Effect of permethrin plus antioxidants on locomotor activity and striatum in adolescent rats

Pyrethroids are important insecticides used largely because of their high activity as an insecticide and their low mammalian toxicity. Some studies have demonstrated that these products show neurotoxic effects on the mammalian central nervous system. The aim of the present study was to investigate the propensity of permethrin to induce oxidative stress in adolescent rats and its possible attenuation by Vitamin E alone or+Coenzyme Q(10). Data indicated that adolescent rats exposed to permethrin exhibited alteration in the locomotor activity and plasma membrane fluidity of striatum. Vitamin E+Q(10) and Vitamin E alone supplementation reversed the negative effect on central nervous system. Permethrin alteration of striatum plasma membrane fluidity was restored by Vitamin E+Q(10). Data obtained from red blood cells showed that permethrin did not induce any modification of plasma membrane fluidity in adolescent rats, whereas antioxidants supplementation induced pro-oxidant effect. In summary some differences between antioxidant treatments were observed at striatum level: Coenzyme Q(10)+Vitamin E maintains plasma membrane fluidity, while Vitamin E is more effective to preserve GSH level.

Comparative toxicity of CuO nanoparticles and CuSO4 in rainbow trout.

This study compared the toxicity and accumulation of two different Cu compounds, CuO nanoparticles (NPs) and soluble CuSO4, in erythrocytes and different tissues in rainbow trout (Oncorhynchus mykiss). The crystal structure of CuO NP analysed by XRD indicates that the NP are Tenorite, a monoclinic CuO. The in vitro toxicity results indicate that both Cu compounds increase the haemolysis rate in a dose-dependent way, but the effect was reduced treating cells with CuO NP. Moreover, both Cu compounds induce DNA damage and the entity of the damage, similarly to haemolysis, was more marked in cells treated with CuSO4. In vivo results, obtained after intraperitoneal injection, showed that Cu concentrations were significantly higher in gills (p<0.0001), kidney (p=0.007) and liver (p<0.05) of exposed fish with a significant increase in plasma Cu concentration 15h after CuSO4 treatment. Cu concentrations were significantly higher in fish exposed to CuSO4 than CuO in kidney (p<0.05) and gills (p<0.0001). Significant DNA damage with respect to controls was detected only when Cu was injected as CuSO4. The present data could serve to evaluate environmental Cu toxicity in fish depending on Cu speciation.

The primary role of glutathione against nuclear DNA damage of striatum induced by permethrin in rats.

Pyrethroids are one of the most widely used class of insecticides and their toxicity is dominated by pharmacological actions upon the CNS. This study reports as the subchronic treatment (60 days) with permethrin (PERM) (1/10 of LD(50)) induced nuclear DNA damage in rat striatum cells. Comet assay outcomes showed that PERM produced single- and double-strand breaks in striatum cells, the DNA damage was not related to oxidation at pyrimidine and purine bases. Vitamin E (280 mg/kg body weight/day) and vitamin E+coenzyme Q(10) (10 mg/kg/3 ml) supplementation could protect PERM treated rats against nuclear DNA damage. With the aim to evaluate the cause of nuclear DNA damage observed in striatum of rat treated with PERM, in vitro studies on striatum submitochondrial particles (SMPs) and on striatum cells treated with 10 muM PERM alone or plus 16 or 32 nM GSH were performed. SMPs incubated with PERM showed a decrease in superoxide anion release from the electron transport chain by inhibition of mitochondrial complex I. The effect could be related to the decrease of membrane fluidity measured in the hydrophilic-hydrophobic region of the mitochondrial membrane. This result discarded the involvement of the mitochondrial reactive oxygen species in the nuclear DNA damage. On the contrary, GSH played a crucial role on striatum since it was able to protect the cells against nuclear DNA damage induced by PERM. In conclusion our outcomes suggested that nuclear DNA damage of striatum cells was directly related to GSH depletion due to PERM insecticide.

Interaction of tributyltin(IV) chloride and a related complex [Bu3Sn(LSM)] with rat leukocytes and erythrocytes : Effect on DNA and on plasma membrane

The discovery of the antitumor activity of cisplatin led several research groups to investigate the possible therapeutic applications of other metal-based compounds. Organotin(IV) complexes have been developed from organotin compounds that were employed in industry and agriculture as stabilizers and pesticides, respectively. A careful choice of the ligand coordinated to an organotin(IV) fragment can modulate the activity of the organotin(IV) complex and minimize its drawbacks. With this aim, the tributyltin(IV) complex [Bu(3)Sn(LSM)] (LSM=bis(1-methyl-1H-imidazol-2-ylthio)acetate) was synthesized and its in vitro effects on rat blood cells were compared with those of the analogous tributyltin(IV) compound without the anionic ligand. Comet-assay results show that both the tributyltin(IV) chloride (TBTC) and the complex [Bu(3)Sn(LSM)] can induce DNA damage in leukocytes, but a stronger effect was observed in the presence of the organotin(IV) complex. Moreover, lipid-hydroperoxide formation in leukocyte plasma membranes increases more in the presence of [Bu(3)Sn(LSM)] compared with TBTC, while TBTC can change the lipid order and packing of leukocytes and, partially, erythrocyte plasma membranes. The treatment of whole blood with these two compounds shows a preferential oxidative effect of TBTC on erythrocyte plasma membranes and erythrocyte oxidative processes, which influence the induction of DNA damage in leukocytes. The different hydrophobic characters and the different extents of steric hindrance of TBTC and [Bu(3)Sn(LSM)] influence the capacity of the two compounds to cross the plasma membrane and affect the pathways that lead to DNA damage.

Effect of permethrin insecticide on rat polymorphonuclear neutrophils.

Polymorphonuclear neutrophils are professional phagocytes whose efficacy depends on a multicomponent NADPH oxidase for generating superoxide anions and bacterial killing. They can be primed and activated by different agents that can impair oxidative burst and phagocytosis with opposite effects: reduced capability to destroy bacteria or hyperactivation that induces the generation of large quantities of toxic reactive oxygen species, which can damage surrounding tissue and participate in inflammation. The present study was designed to evaluate the effect of sub-chronic (60 days) permethrin treatment (1/10 DL(50)) on rat polymorphonuclear neutrophils respiratory burst. The results show that permethrin treatment increases superoxide anion production (33 times) and the activity of hydrogen peroxide-myeloperoxidase system (67 times). In vitro experiments suggest that this effect can be related to permethrin priming and to physico-chemical changes at the plasma membrane level of neutrophils. The antioxidant supplementation with Vitamin E and coenzyme Q(10) can protect against the abnormal respiratory burst in rat treated with permethrin. The in vitro studies show that neutrophil apoptosis begins soon after 1h of incubation with permethrin (0.725% of total cells) or its metabolites (3-phenoxybenzyl alcohol, 3-phenoxybenzaldehyde and 3-phenoxybenzoic acid 1.36, 2.26 and 1.3 of total cells, respectively) and that the level of apoptotic cells is very low. In conclusion, immunotoxicity of permethrin measured in rats could prompt future studies on the consequences of chronic insecticide exposure.

DNA damage and repair following In vitro exposure to two different forms of titanium dioxide nanoparticles on trout erythrocyte.

TiO2 has been widely used to promote organic compounds degradation on waste aqueous solution, however, data on TiO2 nanotoxicity to aquatic life are still limited. In this in vitro study, we compare the toxicity of two different families of TiO2 nanoparticles on erythrocytes from Oncorhynchus mykiss trout. The crystal structure of the two TiO2 nanoparticles was analyzed by XRD and the results indicated that one sample is composed of TiO2 in the anatase crystal phase, while the other sample contains a mixture of both the anatase and the rutile forms of TiO2 in a 2:8 ratio. Further characterization of the two families of TiO2 nanoparticles was determined by SEM high resolution images and BET technique. The toxicity results indicate that both TiO2 nanoparticles increase the hemolysis rate in a dose dependent way (1.6, 3.2, 4.8 μg mL−1) but they do not influence superoxide anion production due to NADH addition measured by chemiluminescence. Moreover, TiO2 nanoparticles (4.8 μg mL−1) induce DNA damage and the entity of the damage is independent from the type of TiO2 nanoparticles used. Modified comet assay (Endo III and Fpg) shows that TiO2 oxidizes not only purine but also pyrimidine bases. In our experimental conditions, the exposure to TiO2 nanoparticles does not affect the DNA repair system functionality. The data obtained contribute to better characterize the aqueous environmental risks linked to TiO2 nanoparticles exposure.

Permethrin induces lymphocyte DNA lesions at both Endo III and Fpg sites and changes in monocyte respiratory burst in rats.

Pyrethroids are widely used insecticides of low acute toxicity in mammals but the consequences of long‐term exposure are of concern. Their insecticidal action is related to neurotoxicity and, in addition, there are indications of mammalian immuno‐toxicity. In this work the effect of 60 days permethrin (150 mg kg−1 body weight/day) exposure on two types of leukocytes (monocytes and lymphocytes) in adolescent rats was investigated. In particular, the monocyte respiratory burst response was first investigated, followed by studies on the degree and type of lymphocyte DNA damage induced by permethrin at this stage of life. Permethrin treatment reduces the monocyte respiratory burst response to phorbol myristate acetate, thereby decreasing superoxide anion (65%) and hydrogen peroxide (37%) production. Moreover an increase [correction made here after initial online publication] in monocyte plasma membrane fluidity in the hydrophilic–hydrophobic interface of the lipid bilayer was measured. Data obtained from the comet assay show that permethrin induces lymphocyte DNA lesions at both formamido pyrimidine glycosylase (Fpg) and endonuclease III (Endo III) sites in adolescent rats. Our results indicate the key role of permethrin in oxidative stress whose consequences lead to biochemical and functional changes. The reduced phagocyte respiratory burst induced by permethrin treatment and the type of DNA damage measured could represent new relevant aspects of pyrethroid toxicity which should be considered for human health. Copyright

In vitro effect of AlCl3 on human erythrocytes: changes in membrane morphology and functionality.

Aluminum belongs to a group of potential toxic elements capable of penetrating the human body. In this paper, the effect of aluminum concentrations on red blood cell membranes using different fluorescent probes able to localize in various parts of the phospholipid bilayer (TMA-DPH, laurdan and pyrene) were studied. Our results confirm that human erythrocytes exposed to aluminum undergo physico-chemical modifications at the membrane level. A decrease in fluorescence anisotropy of TMA-DPH and in the polarity of the lipid bilayer with a concomitant shift toward a gel phase was observed, and the pyrene excimerization coefficient (kex) increased. Furthermore, the presence of aluminum induced lipid peroxidation and reduced the activity of erythrocyte antioxidant enzymes (SOD, CAT and GSHPx). Al-induced morphological changes on the erythrocyte membrane surface were monitored using atomic force microscopy. These results provide further information on the target of action of different aluminum amounts.

Erythrocyte antioxidants enzymes imbalance following subcutaneous pyrethroid treatments in rats of different sex .

Permethrin and deltamethrin are insecticides used all over the world in agriculture to control pests. The aim of this work is to investigate the effects of subcutaneous permethrin and/or deltamethrin treatments on erythrocyte antioxidant enzymes activity in rats of different sex. The results obtained show that permethrin decreases SOD and GPx activity in female rats, while it increases SOD activity in male rats. Deltamethrin treatment reduces GPx activity in male rats, while permethrin has a similar effect but only for short time treatments. A catalase activity decrease was observed in both sexes following pyrethroid treatment but when both the pyrethroids were administrated in male rats, the enzyme activity has increased. In conclusion, this study points out for the first time the importance of rat gender on erythrocyte antioxidant enzymes, which are influenced not only by the length and the type of insecticide treatment but importantly by sex.