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ABSTRACT: Diphenylamine (DPA) is an antioxidant compound that occurs naturally in several vegetables. It is widely applied in agriculture for preservation of the quality of apples and pears, and used for controlling superficial scald, a disorder that renders fruits of a number of apple cultivars unfit for the market. Because of its anti-oxidative properties, DPA also has several industrial applications. The potential genotoxic effect of DPA on human lymphocytes has previously been investigated in only two studies, which focused on detection of chromosome aberrations and sister chromatid exchange, respectively. In the present analysis, we evaluated micronucleus (MN) formation in freshly isolated human peripheral lymphocytes exposed to different concentrations (0.625, 1.25, 2.50, 5.0 and 10.0μg/ml) of DPA. Peripheral venous blood was collected from ten healthy subjects, and a total of 10,000 bi-nucleated cells were analyzed. Results indicated that DPA significantly increased the micronucleus frequency at concentrations of 1.25μg/ml and higher. Significant differences in the MN frequency were also found between the lower dose (0.625μg/ml) and all other doses tested, with the exception of 1.25μg/ml. Our results indicate a potential cytogenetic effect of DPA on human cells in vitro and require further in vivo studies to clarify the actual genotoxicity of this compound and the consequent risks for human health.
Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 05/2012; 747(1):135-7. · 2.85 Impact Factor
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ABSTRACT: Endogenous nitric oxide (NO) is a molecule synthesized by endothelium nitric oxide synthase encoded by the ecNOS gene that plays an important role in regulating the systemic, cardiac and pulmonary circulation. Impairment in NO synthesis has been associated to many cardiovascular disorders, including coronary artery disease and hypertension. We investigated the frequency of the intron 4 VNTR ecNOS gene polymorphism in an Ivorian and an Italian population. The frequencies of the ecNOSb/b, ecNOSa/b and ecNOSa/a genotypes were 0.422, 0.476, and 0.102, respectively, for the Ivorian sample, and 0.712, 0.269, and 0.019, respectively, for the Italian population. The frequencies of ecNOS4b and ecNOS4a alleles were 0.660 and 0.340, respectively, for the Ivorian group, and 0.847 and 0.153, respectively, for the studied Italian population. Genotype frequencies were in Hardy-Weinberg equilibrium in both populations. The Ivorian population showed a significantly higher frequency of the ecNOS4a allele compared to other African and non-African populations, while the Italian sample confirmed the high genetic homogeneity of this polymorphism among Europeans. The maldistribution of endothelial ecNOS polymorphisms between populations could be the results of differential exposure to selection pressures in Africa and during the out-of-Africa expansion.
Molecular Biology Reports 03/2012; 39(6):6693-8. · 2.93 Impact Factor
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ABSTRACT: In vitro effects of different concentrations of Thiram were tested on human lymphocytes to determine, by means of the chromosome aberrations (CAs) assay, whether this fungicide could induce clastogenic damage. Evidences of the effect of Thiram on human lymphocytes were limited to sister chromatid exchange, micronuclei formation, and comet assays. We evaluated 0.01, 0.1, 1.2, and 12.0 μg/mL of Thiram, where 0.01 μg/mL represent the acceptable daily intake dose set by the World Health Organization and the Food and Agriculture Organization for fruit and vegetables, whereas 0.1, 1.2, and 12.0 μg/mL are its multiple values. Results indicated that human lymphocytes treated in vitro with Thiram at concentrations of 1.20 and 12.0 μg/mL significantly increased CAs frequency, compared with the negative control, whereas at lower concentrations (0.01 and 0.1 μg/mL), this effect was not observed. However, Thiram showed a clastogenic effect also at the concentration value of 1.2 μg/mL that represents a lower value with respect to the residue limits found in Italy for grapes, strawberries, potatoes, tobacco, and other fruits and vegetables. Finally, according to some evidence obtained from the study of other fungicides, Thiram produced a significant reduction in the mitotic index with increasing concentration.
Drug and Chemical Toxicology 01/2012; 35(3):347-51. · 1.08 Impact Factor
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ABSTRACT: The dipteran Chironomus riparius is found across the entire Palearctic region; its larvae are among the most abundant macroinvertebrates inhabiting inland waterbodies. Chironomid larvae have been extensively used in ecotoxicological and cytogenetic research, but relatively little is known on the population structure of this species. Transposable elements (TEs) are DNA sequences that are capable of autonomous replication; the number and genomic location of TE insertions varies across individuals; this variability is increasingly being used in population studies. Several TEs had been characterized in Chironomids; this enabled the analysis of insertional variability of four different TEs in six natural populations of C. riparius from Italy, Bulgaria and Russia using a PCR-based method, transposon insertion display (TID). The method allows to obtain dominant markers, similar to AFLP. In all populations, TE insertions showed high individual polymorphism, while median copy numbers of the same TEs did not vary between populations. Analysis of molecular variance (AMOVA) detected significant differentiation between populations for three of the TEs; although no correlation between genetic and geographic distances was found, the corresponding population structures were found to be significantly correlated and indicate a degree of isolation by distance. TEs belonging to different classes have different mechanisms of replication, resulting in different transposition rates of mobilization; the finding of mostly concordant population structuring for three of the TEs indicates that population dynamics contributed significantly in shaping the detected insertional polymorphism.
MGG Molecular & General Genetics 09/2011; 286(3-4):293-305. · 2.58 Impact Factor
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Clinical Chemistry and Laboratory Medicine 03/2011; 49(6):1065-7. · 2.15 Impact Factor
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ABSTRACT: The formaldehyde (FA) genotoxic potential in occupationally exposed individuals is conflicting. A relevant indoor-air FA pollution was found in hospitals and scientific institutions where FA is used as a bactericide and tissue preservative. In the present study, we evaluated the frequency of chromosomal aberrations (CAs) in peripheral blood lymphocytes from workers in pathology wards who have been exposed to FA, compared with a group of unexposed subjects. The subjects were also analyzed for the GSTM1 and GSTT1 metabolic gene polymorphisms. The exposed subjects showed a significant increase in the frequency of CA per cell and in the percentage of cells with aberrations compared to control subjects. The different GST genotypes did not affect the level of cytogenetic damage since CA frequencies were not statistically different between the GST "null" genotypes and the GST "positives". The generalized linear models showed that the number of CAs and cells with CAs increased with age, but, independent of age, it was significantly higher in the experimental rather than in the control group. Cubic-spline regression confirmed the linear relationship between CAs and age, but it provided evidence for a non-linear relationship between CAs and the number of years of FA exposure. Similar results were observed when the model included the number of cells with CAs as dependent variables. Our results demonstrate that air FA induces CAs even consequently to low levels of daily exposure, indicating an increased risk of genetic damage for workers exposed to this air pollutant.
Archive für Toxikologie 03/2011; 85(10):1295-302. · 4.67 Impact Factor
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ABSTRACT: Thiabendazole is a benzimidazole-derived compound widely employed in agriculture as anthelmintic and fungicide. It is also used as a post-harvest fungicide for imported citrus fruits during transport and storage, and thus, it was found at high concentration in fruits and vegetables. Several studies have analyzed the potential genotoxic effect of thiabendazole on different prokaryotic and eukaryotic systems, but in many cases, results were contradictory. In the present study, the genotoxic potential of thiabendazole have been evaluated, by micronucleus assay in freshly isolated human peripheral lymphocytes. The cells were incubated with 0.5, 5 and 50 μg/ml concentrations of the tested substance for 48 h at 37°C. Mitomycin C at final concentration of 0.01 μg/ml culture was used as a positive control. The results indicated that the thiabendazole significantly (P < 0.05) increased the micronucleus frequency compared with the negative control in all treatment concentrations, indicating a potential aneugenic hazard of thiabendazole in cultured human peripheral lymphocytes. The cytokinesis-block proliferation index value, however, was not decreased significantly compared with the negative control. Significant (P < 0.05) differences in the micronuclei frequency were also found between the lower dose (0.5 μg/ml) and the other two analyzed doses of thiabendazole. In contrast, no differences were found between 5 and 50 μg/ml of thiabendazole and between DMSO and negative control. Finally, control cultures treated with the known mutagen MMC showed a very consistent increase in MN with respect to the negative controls.
Archive für Toxikologie 10/2010; 85(6):689-93. · 4.67 Impact Factor
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ABSTRACT: In this study, the frequencies of CYP1A1, GSTM1, and GSTT1 gene polymorphisms were determined in 133 healthy individuals from Ouangolodougou, a small rural town situated in the north of the Ivory Coast. As appeared in several published studies, ethnic differences in these frequencies have been found to play an important role in the metabolism of a relevant number of human carcinogens. In the studied sample, the frequencies of Ile/Ile (wild type), Ile/Val (heterozygous variant), and Val/Val (homozygous variant) CYP1A1 genotypes were 0.271, 0.692, and 0.037, respectively. Frequencies of GSTM1 and GSTT1 null genotypes were 0.361 and 0.331, respectively. No significant differences were noted between men and women. In contrast to published data for Africans, CYP1A1 *Val Allele frequency (0.383) was significantly high (p < 0.001) in this specific population. For the GSTT1 null genotype, no differences were found between the studied and other African populations, the contrary to what occurred for the GSTM1 null genotype in relation to Gambia and Egypt.
Genetics and Molecular Biology 07/2010; 33(3):434-7. · 0.63 Impact Factor
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ABSTRACT: The present work attempts to determine the distribution of GSTM1 and GSTT1 genotype and allele frequencies in a sample of northern Italian population, and to examine the age-related association of these polymorphisms. The frequencies of the deleted GSTM1 and GSTT1 genotypes were 0.357 and 0.169, respectively. GSTT1 null-genotype frequency found in this work further confirms data obtained in previous studies of Italian populations, while for GSTM1 deletion our sample showed a significantly lower-frequency value with respect to other Italian and European populations, with exception of the Greek. No significant differences occurred between men and women in the frequency of each gene, which could suggest that, in the studied sample, there were no sex differences in susceptibility to diseases and in detoxifying enzymes such as GSTs. In order to analyze the relationship between GSTT1 and GSTM1 gene polymorphisms and age, the sample was subdivided into four age groups: 1-30 years (n = 101); 31-50 years (n = 160); 51-79 years (n = 144) and 80-100 years (n = 58). This age-related analysis showed a decreasing gradient of GSTs null genotypes between younger and older groups, with the 80-100 age group showing a significantly lower frequency of GSTT1 null, GSTM1 null and GSTT1/GSTM1 double null genotypes with respect to the younger group.
Archive für Toxikologie 06/2008; 82(12):903-7. · 4.67 Impact Factor
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ABSTRACT: Polymorphism frequencies of the dopamine transporter gene (DAT1) hypervariable region have been analyzed in a sample of Italian and Ivory Coast individuals. The 3' untranslated region (UTR) of DAT1 includes a variable number of tandem repeats (VNTR) of a 40-bp monomer, ranging from 3 to 13 repeats in Caucasian and African populations. In our sample we found alleles with 3 to 16 repeats, and the most common alleles were the 10-repeat (DAT1*10) and the 9-repeat (DAT1*9) alleles. We also found two rare alleles in the Italian population and four in the Ivory Coast population. For the first time the new allele DAT1*16 is described in the Ivorians. The Ivory Coast population was not in Hardy-Weinberg equilibrium for the DAT1 locus because of a deficit of heterozygote genotypes. The observed heterozygosity of the Ivorian population was half that of the Italians. The lower observed heterozygosity and deviation from Hardy-Weinberg equilibrium could be the result of microevolutionary trends, such as genetic drift and/or inbreeding, acting on the relatively small and isolated population sampled for this study, although some sort of selective pressures acting against the shorter alleles cannot be excluded. This evidence, in association with the reduced polymorphism shown by the DAT1 VNTR compared to other VNTRs, seems to indicate that the DAT1 locus may be under some selective pressure.
Human Biology 05/2008; 80(2):191-8. · 1.31 Impact Factor
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ABSTRACT: The present study investigated the morphological and genetic differentiation pattern between two sympatric dung beetle sister species, Onthophagus taurus and Onthophagus illyricus. The geometric morphometric approach coupled with the use of molecular markers allowed examination of the nature of interspecific relationships and an outline of the evolutionary and geographical processes that might have led to interspecific differentiation and the present-day partial sympatric and syntopic pattern of distribution. Geometric morphometrics failed to discrimininate the two species on the ground of external morphological traits, but revealed interspecific differences when the shape of the primary sexual traits was taken into account. The use of two different molecular markers (cytochrome oxidase subunit I gene and amplified fragment-length polymorphism) demonstrates that the two species are genetically well differentiated, forming two distinguishable lineages probably diverged during the Pliocene by allopatric speciation. No evidence of past or recent introgression and no support for hybridization were found, suggesting that sympatry was established subsequently, when speciation was accomplished. Both molecular markers and genital shape indicate that phenotypically intermediate individuals, despite their ambiguous appearance, are members of O. illyricus species rather than hybrids. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 89, 197–211.
Biological Journal of the Linnean Society 10/2006; 89(2):197 - 211. · 2.19 Impact Factor
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ABSTRACT: Aim To examine the phylogeography and population structure of three dung beetle species of the genus Trypocopris (Coleoptera, Geotrupidae). We wanted to test whether genetic differences and genealogies among populations were in accordance with morphologically described subspecies and we aimed to establish times of divergence among subspecies to depict the appropriate temporal framework of their phylogeographical differentiation. We also wished to investigate the historical demographic events and the relative influences of gene flow and drift on the distribution of genetic variability of the different populations.Location Europe (mostly Italy).Methods We collected adult males from dung pats from 15 Italian localities over the period 2000–2002. For sequence analysis, some dried specimens from Albania, Croatia, Slovakia and Spain were also used. We applied cytochrome oxidase I mitochondrial DNA sequencing and the amplified fragment length polymorphism (AFLP) technique to determine whether phylogeographical patterns within the three species support the proposed hypotheses of subspecies designations, and to detect further structure among populations that might mediate diversification.Results and main conclusions The results show a high concordance between the distribution of mtDNA variation and the main morphological groups recognized as subspecies, which thus may represent independent evolutionary units. The degree of mitochondrial divergence suggests that speciation events occurred during the Pliocene, while diversification of the main subspecific lineages took place in the Pleistocene, from c. 0.3 to 1.5 Ma. Mitochondrial and nuclear data also reveal that there is phylogeographical structuring among populations within each of the main groups and that both contemporary and historical processes determined this pattern of genetic structure. Geographical populations form monophyletic clades in both phylogenetic and network reconstructions. Despite the high levels of intrapopulational diversity, FST values indicate moderate but significant genetic differentiation among populations, and a Bayesian clustering analysis of the AFLP data clearly separates the geographical populations. Nucleotide and gene diversity estimates reveal interspecific differences in the degree of diversification among populations that may be related to the different ecological requirements of the three species.
Journal of Biogeography 06/2004; 31(7):1149 - 1162. · 4.54 Impact Factor
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ABSTRACT: Defensins are a recently described family of peptides that play an important role in innate immunity. Recent studies have shown that defensins exhibit a broad spectrum of antimicrobial activities against bacteria and fungi. Three families have been identified so far in mammals, alpha-defensins, beta-defensins and theta-defensins, presumably derived from a common ancestral defensin. A long-term study on the evolution of these multigene families among primates has been undertaken to investigate: (1) the degree of interspecific differentiation; (2) the genetic mechanisms responsible for the variability of these molecules; and (3) the possible role of different environmental factors in their evolution. Nucleotide sequences have been obtained from great and lesser apes, several African and Asian catarrhine monkeys and one New World monkey. A comparison of rates of synonymous and nonsynonymous (amino-acid changing) nucleotide substitution indicates that the primate beta-defensin 1 gene evolved under a pattern of random nucleotide substitution as predicted by the neutral theory of molecular evolution. These results are not consistent with the hypothesis that the primate beta-defensin 1 gene has diversified in response to changes in the microbial species to which a given host is exposed. Analyses of interpecific variability have yielded some insights about the pattern of molecular evolution of the gene among primates. Humans and great apes present high levels of sequence similarity, differing in only one amino acid residue in the mature peptide. Compared with these taxa, hylobatids and cercopithecids exhibit 3-4 amino acid substitutions, some of which increase the net charge of the active molecule.
Immunogenetics 03/2002; 53(10-11):907-13. · 2.93 Impact Factor
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ABSTRACT: The present work attempts to determine the distribution of ACE and LRPAP1 genotypes and allele frequencies in a sample of the population of north-western Italy and to examine the age-related association of these polymorphisms. ACE D allele frequency found in this work further confirms data obtained in previous studies of Northern Italian populations. Regarding the LRPAP1 gene, high frequencies of the deleted allele in European populations were also confirmed. In order to analyse the relationship between ACE and LRPAP1 gene polymorphisms and age, the sample was subdivided into four age groups: 1-30 (n= 99), 31-50 (n= 165), 51-79 (n= 146) and 80-100 years old (n= 57). For the ACE gene, significant difference in D/D genotype frequency was found only between the younger and the 51-79 age groups (p<0.05), the latter showing the lower frequency value. Significant differences were found, for both the I/D and D/D LRPAP1 genotypes, between the first and the second age group (p < 0.02) and between the first and the third age group (p < 0.01), with the 51-79 age group showing the higher D/D and the lower I/D genotype frequency values.
Annals of Human Biology 34(1):102-6. · 1.98 Impact Factor
