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Background The thiopurine S-methyltransferase (TPMT)/azathioprine (AZA) gene-drug pair is one of the most well-known pharmacogenetic markers. Despite this, few studies investigated the implementation of TPMT testing and the combined evaluation of genotype and phenotype in multidisciplinary clinical settings where patients are undergoing chronic therapy with AZA. Methods A total of 356 AZA-treated patients for chronic autoimmune diseases were enrolled. DNA was isolated from whole blood and the samples were analyzed for the c.460G>A and c.719A>G variants by the restriction fragment length polymorphism (RFLP) technique and sequenced for the c.238G>C variant. The TPMT enzyme activity was determined in erythrocytes by a high-performance liquid chromatography (HPLC) assay. Results All the patients enrolled were genotyped while the TPMT enzyme activity was assessed in 41 patients. Clinical information was available on 181 patients. We found no significant difference in the odds of having adverse drug reactions (ADRs) in wild-type patients and variant allele carriers, but the latter had an extra risk of experiencing hematologically adverse events. The enzyme activity was significantly associated to genotype. Conclusions TPMT variant allele carriers have an extra risk of experiencing hematologically adverse events compared to wild-type patients. Interestingly, only two out of 30 (6.6%) patients had discordant results between genotype, phenotype and onset of ADRs.

Onions (Allium cepa) are consumed worldwide and represent an important source of dietary phytochemicals with proven antioxidant properties, such as phenolic acids, flavonoids, thiosulfinates, and anthocyanins. Epidemiological and experimental data suggest that regular consumption of onions is associated with a reduced risk of degenerative disorders. Therefore, it is of interest to investigate the biological properties of different varieties of onions. Here, we characterized for the first time a variety of onion, called Ramata di Montoro (coppery onion from Montoro), grown in a niche area in southern Italy, and compared its phenolic profile and antioxidant properties to a commercial ecotype of red onion, Tropea, also present in southern Italy. An analytical method based on high-performance liquid chromatography coupled with UV detection and mass spectrometry was used to separate and characterize the phenolic fraction (anthocyanins and flavonols) extracted from both coppery and red types. The main compounds detected in the two ecotypes were quercetin and quercetin glucosides, isorhamnetin glucosides, kaempferol glucoside, and, among anthocyanins, cyanidin glucosides. Tropea ecotype onion showed a higher content of flavonols (632.82 mg/kg fresh weight) than Montoro type onion (252.91 mg/kg fresh weight). Accordingly, the antioxidant activity of the former was 2.8-fold higher compared to the latter. More pronounced were the differences existing between the four anthocyanins detected in the two ecotypes, with those in the Tropea ecotype onion present at concentrations 20-230-fold higher than in the Montoro type onion. Both extracts reduced LDL oxidation about 6-fold and protected human erythrocytes from oxidative damage induced by HClO by about 40%. In addition, as a consequence of HClO treatment, glutathione concentration in erythrocytes was reduced about 50% and pretreatment with onion extracts induced a recovery of glutathione level by about 15-22%. Qualitative differences highlighted in the chemical composition of the two phenolic extracts, especially the total content of anthocyanins, which was 30-fold higher in Montoro type onion compared to Tropea ecotype, can be associated with the protective effects measured against oxidative damage induced in human erythrocytes.

The chemical composition of the essential oils of Origanum vulgare ssp. hirtum, growing wild in three different localities in the Southern Apennines, was studied by GC-FID and GC/MS analyses. In total, 103 compounds were identified. The oils were mainly composed of phenolic compounds and all oils belonged to the chemotype carvacrol/thymol. The three essential oils were evaluated for their in vitro phytotoxic activity by determining their influence on the germination and initial radicle elongation of Sinapis arvensis L., Phalaris canariensis L., Lepidium sativum L., and Raphanus sativus L. The seed germination and radicle growth were affected in various degrees. Moreover, the antifungal activity of the three essential oils was assayed against three species causing pre- and postharvest fruit decay (Monilinia laxa, M. fructigena, and M. fructicola). At 1000 ppm, the three oils completely inhibited fungal growth. The hemolytic activity of the oils was assayed and showed no effect on the cell membranes of bovine erythrocytes., (Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.)

Selenium food fortification could be a cost-effective strategy to counteract the inadequacy of selenium intake among the Italian population. In this study, the effect of foliar fertilization with sodium selenate of an Italian rice cultivar and the increase of serum selenium and of erythrocyte glutathione peroxidase (GPx) activity after intake of fortified rice, have been evaluated. The effect of foliar fertilization with sodium selenate (50 g Se/ha) vs. water was studied. Moreover, in a randomized, double-blind study, 10 healthy women supplemented their usual diet with a daily dose of 80 g of Se-enriched-rice and 10 matched-women with 80 g of regular rice. Before, after 5 and 20 days of supplementation, serum Se and GPx-activity were evaluated. The mean selenium content in Se-enriched-rice was 1.64 ± 0.28 μg/g, while in regular rice it was 0.36 ± 0.15 μg/g (p < 0.001). A significant increase of serum Se and GPx-activity was observed only in the intervention group and only after 20 days. The results show that selenium fortification of rice can be achieved with foliar fertilization with sodium selenate and that the 20 days intake of this Se-enriched-rice increases the serum selenium levels and GPx-activity.

The oxidative action of copper on different trout Oncorhynchus mykiss erythrocyte components was studied. The results indicate that: --cupric ions differently influence the oxidative status of two trout hemoglobin components I and IV (HbI and HbIV) having different structural and functional properties; --reactive oxygen species (ROS) production associated with hemoglobins autoxidation is not influenced by the presence of copper sulfate; --the susceptibility to hemolysis increases in the presence of copper only when the erythrocyte suspension is incubated in air; the effect of copper is almost absent for carbon monoxide-saturated erythrocyte suspensions; --DNA damage due to copper was not observed in our experimental conditions. The data obtained are important for the analysis of the environmental risks produced by copper on fish., (Copyright 2009 Elsevier Ltd. All rights reserved.)

In this study we investigated some biological properties of flavonoids recovered in the aqueous (AqE) and ether (EtE) extracts from four Italian multifloral honeys. In particular, a cell-free assay was employed to detect direct reduction of ferricyanide, whereas an assay using intact human erythrocytes was used to measure the ability to donate electrons to a trans-plasma membrane oxidoreductase. It was found that the AqE displays greater "in vitro" ferricyanide-reducing activity than the EtE but, unlike the latter, is virtually ineffective in the cell-based assay. Uptake studies employing high-performance liquid chromatography/mass spectrometry (HPLC/MS) showed that the different results were explained by the inability of AqE components to cross the erythrocyte plasma membrane and by the excellent uptake of EtE flavonoids, which, once within the cell, donate electrons to the membrane oxidoreductase to efficiently reduce extracellular oxidants. The latter property appears to depend on the content of ether-soluble flavonoids in the starting honeys.

Erythrocyte micronucleus frequencies in wild fish from two riverine environments and in fish reproduced and reared under controlled conditions (control group) were compared, with the aim to evaluate the suitability of the MN test for the in situ detection of mutagens in freshwaters. Fish were caught in different months in two rivers of central Italy which have different pollution levels. As indicator species, the barbel (Barbus plebejus) was chosen because of its ecological significance. Blood samplings were performed on wild fish immediately after capture and repeated at different time intervals on the same individuals, which were maintained in controlled conditions after capture. A total of 10,000 erythrocytes per specimen were scored. No significant differences in micronucleus frequencies were observed between the control group and fish from the unpolluted river (Mignone). A significantly higher frequency of micronuclei was observed in fish caught in the polluted river (Tiber), in comparison to both the controls and the Mignone river fish. No significant seasonal differences were observed. Barbels examined 50 and 100 days after capture presented a remarkable decrease in micronucleus frequency in comparison with the frequency observed in barbels at capture. The micronucleus test in fish erythrocytes was shown to be a sensitive bioassay for detecting mutagenic pollution in fresh water environments.

The effect of short-term occupational exposure to lead on erythrocyte glucose-6-phosphate dehydrogenase (G6PD) activity and serum cholesterol was studied in 40 male workers of a lead and zinc foundry. All parameters were measured just before employment and after 172 +/- 21.3 days of work. Genetic deficiency of erythrocyte G6PD was observed in 5/40 subjects. Among G6PD normal subjects, increases in enzyme activity followed any change (increase or decrease) in blood lead. At the pre-employment test, serum cholesterol parameters did not show any correlation with GOD activity or blood lead, and they were not affected by exposure. Cholesterol values observed among all the GOD-deficient subjects were within the range of the rest of the study population.

Cholinesterase (ChE) levels (Ellman method) were monitored in 90 subjects (69 males and 21 females) exposed to carbamate and organophosphate pesticides (78 agricultural workers and 12 pesticide vendors). Pre-exposure baseline values of plasma and red blood cell cholinesterase activities were defined for each subject with two blood samples (23 workers) or three blood samples (59 workers) taken almost thirty days after the last exposure. After control of intra-individual variation, 8 subjects with only one pre-exposure value and 13 with a coefficient of variation above 30% were excluded. For the other 59 subjects, the intra-individual variation of erythrocyte ChE (16%) was similar to the inter-individual one (15%), whereas the inter-individual variation of plasma ChE (21%) was higher than the intra-individual one (14%). Laboratory variation for plasma ChE measurements was 8%. Baseline values were analyzed (ANOVA) for sex, age, task and hour and season of sampling. Both erythrocyte and plasma enzymes, corrected for hematocrit, were lower in females. Plasma cholinesterase activity was lower in "re-entry" agricultural workers and in pesticide vendors. Post-exposure cholinesterase activity was measured in 54 workers within a few (1-21) days after last handling. Average relative reduction was 15.2% (95% C.I. = 4.9%-25.5%) in erythrocyte cholinesterase activity and 29.1% (95% C.I. = 18.2%-40.1%) in plasma cholinesterase activity. The one-way variance analysis showed marked plasma ChE reduction in mixers, loaders and appliers (36%, 95% C.I. = 24%-48%) and in parathion handlers (35%, 95% C.I. = 21%-49%. No significant reduction in blood cell cholinesterase activity in relation to task and to pesticide handled was observed. We conclude that the intra-individual variations of the baseline values were higher for three repetitions (88% and 84% of the population were within a variability of less than 30%, for AChE and for ChE respectively) than for two repetitions (91% and 88% of the population were within 30% of variability for AChE and for ChE respectively). The figures show a greater sensitivity of plasma ChE activity in acute exposure, probably due to a poor reliability in detection of erythrocyte ChE by local laboratories. The maximum reduction (38%, 95% C.I. = 22%-53%) in plasma ChE activity was observed within six days of the last exposure in loaders and appliers.

Hb Altdorf alpha 2 beta 2 135 Ala leads to Pro is an unstable variant occurring near Lecce in Italy. The abnormal hemoglobin does not separate from Hb A in the electrophoresis. In vitro a marked Heinz body formation is produced with phenylhydrazin. In heterozygous individuals an almost compensated hemolysis and a slight splenomegaly are found. Hemolysis can be aggravated by exogenous factors. A rather severe hemolysis was induced by a viral infection in a 3 years old girl.