Valeria Maselli

Frogs, sentinels of DNA damage induced by pollution in Naples and the neighbouring provinces.

Many DNA mutation-based diseases recognised in Campania have recently been related to toxic substances in illegal dumping areas. We performed a comet assay on edible frog erythrocytes to evaluate DNA damage. Differences in genotoxic parameters were observed among populations. We show that severe DNA damage occurred in the north Campania where the emergence of environmental waste exploded recently. Although a similar magnitude of genotoxic damage was observed in some southern populations, it is attributable to a massive pesticide pollution related to intensive farming. The frog species analysed seems to be a good bioindicator for detecting genotoxic effects of chemical environmental hazards.

Poly(ADP-ribose) polymerase signaling of topoisomerase 1-dependent DNA damage in carcinoma cells

A molecular approach to enhance the antitumour activity of topoisomerase 1 (TOP1) inhibitors relies on the use of chemical inhibitors of poly(ADP-ribose)polymerases (PARP). Poly(ADP-ribosyl)ation is involved in the regulation of many cellular processes such as DNA repair, cell cycle progression and cell death. Recent findings showed that poly(ADP-ribosyl)ated PARP-1 and PARP-2 counteract camptothecin action facilitating resealing of DNA strand breaks. Moreover, repair of DNA strand breaks induced by poisoned TOP1 is slower in the presence of PARP inhibitors, leading to increased toxicity. In the present study we compared the effects of the camptothecin derivative topotecan (TPT), and the PARP inhibitor PJ34, in breast (MCF7) and cervix (HeLa) carcinoma cells either PARP-1 proficient or silenced, both BRCA1/2(+/+) and p53(+/+). HeLa and MCF7 cell lines gave similar results: (i) TPT-dependent cell growth inhibition and cell cycle perturbation were incremented by the presence of PJ34 and a 2 fold increase in toxicity was observed in PARP-1 stably silenced HeLa cells; (ii) higher levels of DNA strand breaks were found in cells subjected to TPT+PJ34 combined treatment; (iii) PARP-1 and -2 modification was evident in TPT-treated cells and was reduced by TPT+PJ34 combined treatment; (iv) concomitantly, a reduction of soluble/active TOP1 was observed. Furthermore, TPT-dependent induction of p53, p21 and apoptosis were found 24-72h after treatment and were increased by PJ34 both in PARP-1 proficient and silenced cells. The characterization of such signaling network can be relevant to a strategy aimed at overcoming acquired chemoresistance to TOP1 inhibitors.

An integrated study on antimicrobial activity and ecotoxicity of quantum dots and quantum dots coated with the antimicrobial peptide indolicidin

This study attempts to evaluate the antimicrobial activity and the ecotoxicity of quantum dots (QDs) alone and coated with indolicidin. To meet this objective, we tested the level of antimicrobial activity on Gram-positive and Gram-negative bacteria, and we designed an ecotoxicological battery of test systems and indicators able to detect different effects using a variety of end points. The antibacterial activity was analyzed against Staphylococcus aureus (ATCC 6538), Pseudomonas aeruginosa (ATCC 1025), Escherichia coli (ATCC 11229), and Klebsiella pneumoniae (ATCC 10031), and the results showed an improved germicidal action of QDs-Ind. Toxicity studies on Daphnia magna indicated a decrease in toxicity for QDs-Ind compared to QDs alone, lack of bioluminescence inhibition on Vibrio fisheri, and no mutations in Salmonella typhimurium TA 100. The comet assay and oxidative stress experiments performed on D. magna showed a genotoxic and an oxidative damage with a dose–response trend. Indolicidin retained its activity when bound to QDs. We observed an enhanced activity for QDs-Ind. The presence of indolicidin on the surface of QDs was able to decrease its QDs toxicity.

Landscape fragmentation and habitat suitability in endangered Italian hare (Lepus corsicanus) and European hare (Lepus europaeus) populations.

During the last years, the population of Italian hare decreased significantly in central and south Italy. This is imputable to harvest, poaching, habitat fragmentation, and the probable competition with congeneric European hare introduced in the last decades by man for hunt. The goal of our work is to define the ecological characteristics of the two aforementioned species in order to understand how landscape facilitates or impedes movement. Spatially explicit models are used to identify a species ecological niche and to build a landscape model of suitability. To validate ecological modeling of landscape, we performed a population genetic analysis. Results suggest that the Italian hare shows an ecological requirement close to average of available resources in the considered landscape. The genetic structure of this autochthonous species validates the habitat suitability model and highlights the differences with European hare. This work analyzes for the first time the ecological relationship between those two sympatric species.

Unexpected but welcome. Artificially selected traits may increase fitness in wild boar

Artificial selection affects phenotypes differently by natural selection. Domestic traits, which pass into the wild, are usually negatively selected. Yet, exceptionally, this axiom may fail to apply if genes, from the domestic animals, increase fertility in the wild. We studied a rare case of a wild boar population under the framework of Wrights interdemic selection model, which could explain gene flow between wild boar and pig, both considered as demes. We analysed the MC1R gene and microsatellite neutral loci in 62 pregnant wild boars as markers of hybridization, and we correlated nucleotide mutations on MC1R (which are common in domestic breeds) to litter size, as an evaluation of fitness in wild sow. Regardless of body size and phyletic effects, wild boar sows bearing nonsynonymous MC1R mutations produced larger litters. This directly suggests that artificially selected traits reaching wild populations, through interdemic gene flow, could bypass natural selection if and only if they increase the fitness in the wild.

Daphnia magna and Xenopus laevis as in vivo models to probe toxicity and uptake of quantum dots functionalized with gH625

The use of quantum dots (QDs) for nanomedicine is hampered by their potential toxicologic effects and difficulties with delivery into the cell interior. We accomplished an in vivo study exploiting Daphnia magna and Xenopus laevis to evaluate both toxicity and uptake of QDs coated with the membranotropic peptide gH625 derived from the glycoprotein H of herpes simplex virus and widely used for drug delivery studies. We evaluated and compared the effects of QDs and gH625-QDs on the survival, uptake, induction of several responsive pathways and genotoxicity in D. magna, and we found that QDs coating plays a key role. Moreover, studies on X. laevis embryos allowed to better understand their cell/tissue localization and delivery efficacy. X. laevis embryos raised in Frog Embryo Teratogenesis Assay-Xenopus containing QDs or gH625-QDs showed that both nanoparticles localized in the gills, lung and intestine, but they showed different distributions, indicating that the uptake of gH625-QDs was enhanced; the functionalized QDs had a significantly lower toxic effect on embryos’ survival and phenotypes. We observed that D. magna and X. laevis are useful in vivo models for toxicity and drug delivery studies.

Integrated analysis of the ecotoxicological and genotoxic effects of the antimicrobial peptide melittin on Daphnia magna and Pseudokirchneriella subcapitata.

Melittin is a major constituent of the bee venom of Apis mellifera with a broad spectrum of activities. Melittin therapeutical potential is subject to its toxicity and the assessment of ecotoxicity and genotoxicity is of particular interest for therapeutic use. Here we analyzed the biological effects of melittin on two aquatic species, which are representative of two different levels of the aquatic trophic chain: the invertebrate Daphnia magna and the unicellular microalgae Pseudokirchneriella subcapitata. The attention was focused on the determination of: i) ecotoxicity; ii) genotoxicity; iii) antigenotoxicity. Our main finding is that melittin is detrimental to D. magna reproduction and its sub-lethal concentrations create an accumulation dependent on exposition times and a negative effect on DNA. We also observed that melittin significantly delayed time to first eggs. Moreover, results showed that melittin exerted its toxic and genotoxic effects in both species, being a bit more aggressive towards P. subcapitata.

Wild boars’ social structure in the Mediterranean habitat

Abstract Wild boar Sus scrofa has become a pest throughout Europe, but its social system in this setting is unknown, making it difficult to implement an effective management strategy. We collected data on social grouping patterns by video-trapping the population living in an Italian Mediterranean National Park. We collected over 120 days of observations and the activities of 1431 wild boars. The seasonal variations in wild boar social organisation seem to be directly related to the biological cycle, with some exceptions, such as the unexpected presence of solitary or groups of piglets. The population was more active during the summer, and the mean group size (4.14 ± 0.21) was quite similar to other European boar group sizes. We confirmed that video traps are an appropriate method to monitor versatile wildlife, especially for assessing the population structure, sex and age ratios of wild boars.

Genotoxicity of gold nanoparticles functionalized with indolicidin towards Saccharomyces cerevisiae

The toxic effects of gold nanoparticles surface-functionalized with the antimicrobial peptide indolicidin (AuNPs-indolicidin) towards the yeast Saccharomyces cerevisiae, one of the major eukaryotic model organisms, have been evaluated. Growth and survival, genotoxicity, as measured by comet assay, and expression of the YCA1, an apoptosis indicating gene, following 72hr exposure of yeast to AuNPs-indolicidin, and to AuNPs and indolicidin alone have been examined. The gold nanoparticles exerted toxicity with DNA damage, accompanied by reactive oxygen species production (ROS), but they do not inhibit yeast growth and viability. Genotoxicity was less pronounced for surface-functionalized nanoparticles, showing that S. cerevisiae is quite resistant to the complex AuNPs-indolicidin. A progressive reduction of the genotoxic effect was observed along 72hr exposure, presumably due to the activation of DNA repair mechanisms. These findings suggest the occurrence of a physiological protective response of S. cerevisiae towards nanoparticles, thereby providing useful information to the assessment of the environmental impact of metal nanoparticles.

Efficiency of gold nanoparticles coated with the antimicrobial peptide indolicidin against biofilm formation and development of Candida spp. clinical isolates

Background This article examines the use of a novel nano-system, gold nanoparticles coated with indolicidin (AuNPs–indolicidin), against pathogenic Candida albicans biofilms. Candida species cause frequent infections owing to their ability to form biofilms, primarily on implant devices. Materials and methods We used an integrated approach, evaluating the effect of AuNPs-indolicidin on prevention and eradication of Candida biofilms formed in multi-well polystyrene plates, with relative gene expression assays. Four biofilm-associated genes (FG1, HWP1, ALS1 and ALS3, and CDR1 and CDR2) involved in efflux pump were analyzed using reverse transcription polymerase chain reaction. Results Treatment with the nano-complex significantly inhibits the capacity of C. albicans to form biofilms and impairs preformed mature biofilms. Treatment with AuNPs–indolicidin results in an increase in the kinetics of Rhodamine 6G efflux and a reduction in the expression of biofilm-related genes. Conclusion These data provide a chance to develop novel therapies against nosocomially acquired refractory C. albicans biofilms.