Effects of glutathione S-transferase M1 and T1 deletions on Parkinson's disease risk among a North African population.

PURPOSE: In this study, the effects of glutathione S-transferase polymorphisms Mu1 (GSTM1) and glutathione S-transferase polymorphisms Theta1 (GSTT1) on Parkinson's disease (PD) risk factor were evaluated in a Tunisian population. METHODS: These polymorphisms were analyzed in 229 healthy Tunisian subjects and 64 Tunisian patients with PD, using a polymerase chain reaction (PCR). Statistical analysis was performed using SPSS 18.0. The relative associations between the GST genotypes and PD were assessed by calculating the odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: The study results demonstrated that the individuals with GSTM1 [OR=3.93, 95% CI: 1.98-7.92, P=10-6] and GSTT1 [OR=5.45, 95% CI: 2.90-10.30, p=10-6] were statistically associated with the risk of PD. A significant association was also found between the individuals with both GSTM1/T1 null genotypes and PD risk [OR=22.10, 95% CI: 6.99-73.75, P=10-6]. CONCLUSION: These genotyping findings suggest that the absence of both GSTM1 and GSTT1 activity could be a contributory factor for the development of PD.

Implications of metabolic parameters of carbamazepine in the therapeutic monitoring of Tunisian patients with epilepsy.

Carbamazepine (CBZ) is widely used in the control of simple and complex focal seizures and generalized tonic-clonic seizures in patients with epilepsy. The toxic effects of CBZ are not easily predicted, and this is due to the difficulty of delivering the optimal dose and/or plasma concentration of CBZ necessary to achieve beneficial effects, and especially to prevent the onset of toxicity associated with its use. Our study aimed to determine the relationship between the administered daily dose of CBZ and its pharmacokinetic parameters, including concentrations of CBZ and carbamazepine-10,11-epoxide (CBZ-E) plasma levels, and the metabolic ratio of CBZ-E to CBZ, in Tunisian patients with epilepsy. To accomplish this, a high-performance liquid chromatography method with ultraviolet detection was used for quantification in the simultaneous analysis of CBZ and one of its active metabolites, CBZ-E, in human plasma. A statistically significant positive correlation was found between the daily doses administered (mg/kg/day) and plasma concentrations of CBZ and CBZ-E, and the CBZ-E/CBZ ratio increased significantly as a function of the specific dose (in mg/kg/day). The increase in plasma concentrations of CBZ-E was non-linear in relation to plasma concentrations of CBZ, and there was no correlation between the CBZ-E/CBZ metabolic ratio and CBZ plasma concentrations. Our findings suggest that monitoring of CBZ as well as CBZ-E blood levels should be considered, as it may play a useful role in the therapeutic management of patients with epilepsy.

Polyphenol contents and antioxidant activities of extracts from flowers of two Crataegus azarolus L. varieties.

Phenolic contents of the ethyl acetate extracts prepared from floral buds and opened flowers harvested on Crataegus azarolus trees native in two localities were performed. The antioxidant activity was measured by DPPH' (2,2-diphenyl-picrylhydrazyl), ABTS+ (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radicals scavenging using spectrophotometric method. The C. azarolus var. aronia (Willd.) Batt., producing yellow fruits, was richer in total phenols (1638.7 +/- 89.9 mg acid gallic/100 g dry weight) according to C. azarolus var. eu-azarolus Maire (1415.5 +/- 23.8 mg acid gallic/100 g dry weight), producing red ones. Ethyl acetate extract from opened flowers has less content in total phenols, proanthocyanidins and flavonoids compared to this from floral buds. Floral buds from the two C. azarolus varieties occurring in Siliana-Djebel Serdj showed the highest radical scavenging activities (2431.8 +/- 32.7 and 2267.7 +/- 22.7 micromol Trolox/100 g dry weight). Hawthorn from Tunisia contains eight antioxidants of phenolic type (chlorogenic acid, hyperoside, rutin, spiraeoside, isoquercitrin, quercetin, (-)-epicatechin and the dimer procyanidin B2). These compounds identified specially in floral bud extracts presented a strong radical-scavenging activity.

The dihydrouracil/uracil ratio in plasma, clinical and genetic analysis for screening of dihydropyrimidine dehydrogenase deficiency in colorectal cancer patients treated with 5-fluorouracil.

A rapid and cost-effective reversed phase high performance liquid chromatography (HPLC) method for quantification of dihydrouracil to uracil ratio (UH2/U) in plasma has been developed and used to screen for dihydropyrimidine dehydrogenase (DPD) deficiency in nine patients treated with 5-fluorouracil (5-FU). This HPLC method is based on the use of a simultaneous UV detection at 205 and 268nm during the analysis run of the plasma extract and taking into account the particularity that UH2 shows no absorbance response at 268nm. The plasma UH2/U ratio values evaluated by the use of our HPLC assay were found to be highly correlated with the plasma 5-FU-half-life values and were significantly associated with the toxic side effects, whereas, data set provided from genetic analysis of the coding sequences of the DPD gene (DPYD) were found to be insufficient to explain all the cases of the 5-FU-related toxicity pattern. The proposed HPLC assay could be available for routine clinical use for DPD deficiency assessment in patients prior to 5-FU administration.

Marked differences between mutagenicity in Salmonella and tumour-initiating activities of dibenzo[a,e]fluoranthene proximate metabolites; initiation inhibiting activity of norharman.

Dibenzofluoranthene-12,13-dihydrodiol (DBF-12,13-DHD) is six times more mutagenic in Salmonella TA100 than dibenzofluoranthene-3,4-dihydrodiol (DBF-3,4-DHD). However, these two major dibenzo[a,e]fluoranthene (DBF) proximate metabolites, which are immediate precursors of the corresponding diolepoxides, showed on an equimolar basis nearly identical initiation activities on mouse skin; they induced three times more papillomas than the parent hydrocarbon. On the other hand the epithelioma initiation capacities, i.e. the number of papillomas progressing to malignant tumours, of DBF or the two dibenzofluoranthene dihydrodiols were equivalent. Norharman, a putative vicinal diolepoxidation inhibitor in DBF metabolism when administered topically together with the initiation dose (100 nmol), strongly inhibited the induction of tumours by DBF-3,4-DHD and DBF. The relationship between in vitro mutagenic activity in Salmonella and the carcinogenicity of DBF metabolites in mice appears to be qualitative rather than quantitative.

Determination of 23 polycyclic aromatic hydrocarbons in atmospheric particulate matter of the Paris area and photolysis by sunlight.

Twenty-three polycyclic aromatic hydrocarbons (PAH) were determined in atmospheric particulate matter in 4 places of the Paris area at several times of the year. Fractionation was performed by reversed-phase high-pressure liquid chromatography. Determination was done by recording emission or excitation fluorescence spectra via a stopped-flow technique. Triphenylene was also extemporaneously determined by its phosphorescence spectrum at low temperature. Among the PAH determined dibenz(e,ghi)perylene has not been detected before in atmospheric particulate matter. The 10 more abundant PAH ranged from 0.1 to 40 ng/m3 of filtered air. Concentrations in August are from 14 to 250 times less than in January depending on the PAH. The reasons for this difference of behaviour among the PAH were investigated with regard to their photochemical and non-photochemical reactivity.

Possible involvement of a vicinal, non-bay-region dihydrodiol-epoxide in the activation of dibenzo[a,e]fluoranthene into bacterial mutagens.

Dibenzo[a,e]fluoranthene, its 7-hydroxy, 3,4- and 12,13-dihydrodiol metabolic derivatives as well as three synthetic, structurally related hydrocarbons, were tested for mutagenicity towards Salmonella typhimurium TA100 strain in the presence of 3-methylcholanthrene-treated rat and mouse liver post-mitochondrial supernatants. Of these compounds, the 12,13-dihydrodiol showed the highest activity, being 6-10 times more mutagenic than the parent compound. Our data, in conjunction with those of previous studies on the liver microsomal metabolism and DNA binding of dibenzo[a,e]fluoranthene and its dihydrodiols, indicate that activation of dibenzo[a,e]fluoranthene to bacterial mutagens may occur predominantly through a vicinal, non-bay-region 12,13-dihydrodiol epoxide.

Multi-step metabolism of the carcinogen dibenzo[a,e]fluoranthene. I. Identification of the metabolites from rat microsomes.

The metabolism of the polycyclic hydrocarbon dibenzo[a,e]fluoranthene (DBF) has been investigated. Two new primary metabolites have been identified by proton n.m.r. as the trans diaxial dihydrodiols of the bay and of the pseudo bay regions of DBF. In addition, twelve new metabolites arising from secondary and tertiary metabolic transformations have been identified. The stereochemistry of eleven of these products has been established by proton n.m.r. spectroscopy. In contrast with other alternant carcinogenic polycyclic hydrocarbons, a vicinal diol epoxidation at the bay region of the trans diequatorial dihydrodiols is a minor reaction relative to attack at distant sites, which leads principally to phenolic derivatives of the dihydrodiols and, with a lower yield, to a bis-dihydrodiol (3,4,12,13-tetrahydro-3,4,12,13-tetrahydroxy-DBF). This offers a possibility of bifunctional metabolic activation in the carcinogenicity of DBF. A catechol of the benzanthracenic moiety of DBF has also be identified, dibenzo[a,e]fluoranthene 3,4-catechol. In summary, all three external rings of DBF can be attacked enzymatically, but apparently ring E does not undergo epoxidation while more usual trans diaxial and diequatorial dihydrodiols, as well as tetraols of rings A and D, were detected.

Multi-step metabolism of the carcinogen dibenzo[a,e]fluoranthene. II. Metabolic pathways.

The structural identification of nineteen metabolites of dibenzo[a,e]fluoranthene (DBF) obtained by incubation in rat and mouse liver microsomes, allows one to establish a qualitative and semi-quantitative metabolic chart, involving up to three distinct oxidative attacks. The primary steps lead to dihydrodiols on rings A and D and phenols on rings A and E. Secondary vicinal epoxidation of dihydrodiols is a minor route as compared to attack at a second peripheral ring. Even after a third oxidation, one of the peripheral rings A, D and E remains unsubstituted. A model for cytochrome P-450 enzymatic activity which takes into account most of the observations is proposed. It requires that the catalytic site for monooxygenation is 0.6 nm apart from the center of an hydrophobic protein site accommodating one of the unsubstituted peripheral benzenoid rings. both trans diequatorial dihydrodiols of ring A and D corresponding to the 'bay' and 'pseudo bay region'; of DBF appear in the activation pathways for the in vivo carcinogenesis. The ultimate metabolite reacting with DNA is thus, most probably, a vicinal dihydrodiol epoxide of ring A or D. The great complexity of the metabolic chart of DBF as compared to other carcinogenic polycyclic aromatic hydrocarbons leaves also the possibility of sequential reactions at these two distinct sites of the molecule.

Binding to DNA of bay region and pseudo bay region diol-epoxides of dibenzo[a,e]fluoranthene and comparison with adducts obtained with dibenzo[a,e]fluoranthene or its dihydrodiols in the presence of microsomes.

When dibenzo[a,e]fluoranthene (DBF) is incubated in vitro with mouse liver microsomes in the presence of calf thymus DNA, several metabolites bind covalently to DNA. The metabolite-nucleoside adducts were separated by h.p.l.c. after enzymatic hydrolysis. The elution profile of this chromatogram exhibits six main peaks, labeled from A to F in order of decreasing polarity. It was compared to those obtained by direct reaction of DNA with 3,4-dihydroxy-1,2-epoxy 1,2,3,4-tetrahydro DBF (the bay region diol-epoxide) or 12,13-dihydroxy 10,11-epoxy 10,11,12,13-tetrahydro DBF (the pseudo bay region diol-epoxide). In both cases the retention period of the peak of the adduct was identical to that of the main peak E. The fluorescence spectra of these two adducts were similar to those of the corresponding tetrols. When DNA is reacted in the presence of microsomes with 3,4-dihydrodihydroxy DBF, the elution profile of the adducts indicates that vicinal epoxidation of the dihydrodiol and direct reaction is dominant. The metabolic reaction with 12,13-dihydrodihydroxy DBF appears more complex as revealed by the observed number of adducts which correspond to vicinal epoxidation of dihydrodiol as well as further oxidation at other sites.