Genotoxicity Response of Fibroblast Cells and Human Epithelial Adenocarcinoma In Vitro Model Exposed to Bare and Ozone-Treated Silica Microparticles

Indoor air pollutants (IAP), which can pose a serious risk to human health, include biological pollutants, nitric oxide (NO), nitrogen dioxide (NO2 ), volatile organic compounds (VOC), sulfur dioxide (SO2 ), carbon monoxide (CO), carbon dioxide (CO2 ), silica, metals, radon, and particulate matter (PM). The aim of our work is to conduct a multidisciplinary study of fine silica particles (<2.5 µm) in the presence or absence of ozone (O3 ), and evaluate their potential cytotoxicity using MTS, micronucleus, and the comet test in two cell lines. We analyzed A549 (human basal alveolar epithelial cell adenocarcinoma) and Hs27 (human normal fibroblasts) exposed to dynamic conditions by an IRC simulator under ozone flow (120 ppb) and in the presence of silica particles (40 µg/h). The viability of A549 and Hs27 cells at 48 and 72 h of exposure to silica or silica/ozone decreases, except at 72 h in Hs27 treated with silica/ozone. The micronucleus and comet tests showed a significant increase in the number of micronuclei and the % of DNA in the queue, compared to the control, in both lines in all treatments, even if in different cell times/types. We found that silica alone or with more O3 causes more pronounced genotoxic effects in A549 tumor cells than in normal Hs27 fibroblasts

Tuber borchii mycelial protoplasts isolation, characterization and functional delivery of liposome content, a new step towards truffles biotechnology

The filametous ascomycete Tuber borchii is a plant-symbiotic ectomycorrhizal microrganism with an high value due to the production of hypogeous fruitbodies (truffles). The present work was undertaken to develop a procedure for the release of T. borchii viable protoplasts from Tuber mycelium, isolate ATTC 96540; several factors which affect the isolation, morphology and viability were examined and developed in order to improve applications of T. borchii protoplasts in morphological, biochemical and genetic investigations (protoplast fusion or transformation). Functional delivery of liposome content into T. borchii protoplasts has also been examined with a cytotoxic ribosome inactivator as saporin. T. borchii protoplasts incubation/fusion with saporin containing liposomes were made to demonstrate the absence of cell wall of 16 days cultured protoplasts. © 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved

Silica Nanoparticles and Ozone: an evaluation of in vitro cytotoxicity and genotoxicity in a model of indoor air exposition

Several studies point out on effects of indoor pollutants (silica particles, Pb, Ni, Zn, Si, ozone, carbon monoxide and nitrogen oxides). The work aim is a multidisciplinary study of silica ultrafine particles (&lt;2.5μ) in the presence or not of ozone O3 and their potential cyto-genotoxicity evaluated by MTS, Micronucleus and Comet test in two cell lines. We analyzed A549 (adenocarcinoma human alveolar basal epithelial cells) and HS 27(normal human fibroblasts) exposed under dynamic conditions by a simulator IRC under stream of ozone (120 ppb) and silica particles (40μg/h). A549 cells viability at 48 hours and 72 hours exposition to silica alone or silica plus ozone doesn’t differ in respect to the control. The viability trend in the Hs27 cells did not show any significant changes in treated samples compared to the control in all conditions. The micronucleus and the comet tests, showed in both the cell types exposed to silica alone or plus ozone, a significant increase in the number of micronuclei and in the tail DNA % in respect to the control even if at different times/cell type. We found that silica alone or plus O3 provokes genotoxic effects more evident in A549 cancer cells than in normal fibroblasts Hs27 ones. Final output will be to obtain a picture of the role of silica particles/ozone in the indoor air quality taking account of the potential simultaneous co-toxicant action

The use of hook plate in type III and V acromio-clavicular Rockwood dislocations: clinical and radiological midterm results and MRI evaluation in 42 patients

BACKGROUND: Acromio-clavicular (AC) joint dislocations are very common following falls on the shoulder or an overstretched hand. The best treatment for such lesions remains a matter of debate. Several studies have, however, lent support to the surgical role of the hook plate in Rockwood type III and V AC dislocations. The aim of this study was to evaluate the midterm clinical results and magnetic resonance imaging (MRI) features of coraco-clavicular ligaments 18 months after an AC dislocation treated with an AC Dreithaler hook plate. PATIENTS AND METHODS: The cohort was made up of a consecutive series of 42 patients, who underwent surgery between November 2002 and December 2006 for an AC dislocation. They were classified, according to the Rockwood classification, as 22 grade III and 20 grade V dislocations. Surgical treatment consisted of open reduction and stabilisation with an AC Dreithaler hook plate. A clinical and radiological follow-up examination was performed 1 and 3 months after surgery, that is, before removal of the plate, and 12 months following removal. Eighteen months after the trauma, an MRI and a clinical examination were performed and the Constant-Murley scores calculated. RESULTS: An acceptable joint alignment was achieved in all the patients after surgery; 1 year after plate removal, five cases (12%) of dislocation recurrence were reported. MRI showed the coraco-clavicular ligaments had healed in the remaining 37 cases (88%). CONCLUSION: An AC plate is a useful technique in acromio-clavicular dislocations because it is easy to implant, requires mini-invasive access and results in early resumption of normal activity. MRI can be used to evaluate healing of coraco-clavicular ligaments. A long-term follow-up study is, however, warranted to assess the likelihood of recurrence. LEVEL OF EVIDENCE: Level IV, therapeutic cases series

Analysis of bones ancient mtDNA from the medieval archeological site of Amiternum (L’Aquila), Italy

Introduction: The study of ancient DNA allows to analyze genetic relationships between individuals and populations of the past and the present. In this work we analyzed human bones remains datable between the 6th-9th century D.C. from burials of the archaeological site of Amiternum, L’Aquila. Materials and Methods: As a genetic marker, the hypervariable 1 region of mitochondrial DNA (HVR1) has been chosen. The HVR1 marker has been amplified by PCR, the amplicon have been cloned and sequenced. Sequences of the HVR1 region were compared with Anderson's sequence for the identification of polymorphisms. The data obtained were analyzed with different software and phylogenetic methods. For inter-populations comparisons, the known sequences in literature and found in ancient and modern databases have been used. Results and Conclusions: This work provides preliminary information on the correlation between the inhabitants of Amiternum and the Longobards populations of northern Italy and the Byzantines, migrant peoples transited and/or allocated in the territory of Amiternum. The study of the haplogroups, the analysis of genetic variability and the studies of phylogeny on the obtained sequences show a genetic proximity between individuals of Amiternum, the current population of north/central Italy and the Germanic tribe of Longobards, which dominated the Italian peninsula between 568 and 774 A. D. The match of ancient Byzantines sequences with one of the Amiternum samples highlights also a Byzantine genetic trait in the populations of Amiternum and L’Aquila

Extracorporeal shock-wave therapy compared with surgery for hypertrophic long-bone nonunions.

BACKGROUND: The authors of several studies have recommended extracorporeal shock-wave therapy as an alternative to surgical treatment for long-bone nonunions. This study was performed to compare the results of extracorporeal shock-wave therapy produced by two different devices with those of surgical treatment in the management of long-bone nonunions. METHODS: One hundred and twenty-six patients with a long-bone nonunion were randomly assigned to receive either extracorporeal shock-wave therapy (Groups 1 and 2) or surgical treatment (Group 3). The patients in the shock-wave groups received four treatments with 4000 impulses of shock waves with an energy flux density of 0.40 mJ/mm(2) (Group 1) or 0.70 mJ/mm(2) (Group 2). The patients in the three groups had similar demographic characteristics, durations of nonunion, and durations of follow-up. Radiographic results (the primary outcome) and clinical results (the secondary outcomes) were determined before and three, six, twelve, and twenty-four months after treatment. RESULTS: The radiographic findings did not differ among the three groups of patients. At six months, 70% of the nonunions in Group 1, 71% of the nonunions in Group 2, and 73% of the nonunions in Group 3 had healed. Three and six months after treatment, the clinical outcomes in the two shock-wave groups were significantly better than those in the surgical group (p < 0.001). However, at both twelve and twenty-four months after treatment, there were no differences among the three groups, with the exception of the DASH score, which differed significantly between Groups 1 and 3 (p = 0.038) and between Groups 2 and 3 (p = 0.021) at twelve months. CONCLUSIONS: Extracorporeal shock-wave therapy is as effective as surgery in stimulating union of long-bone hypertrophic nonunions and yields better short-term clinical outcomes

Influence of a low background environment on biochemical and biological responses in V79 cells.

We present the results of an experiment aimed at comparing the effects of different background radiation environments on metabolism and responses to gamma-rays and cycloheximide of cultured mammalian cells. Chinese hamster V79 cells were maintained in exponential growth in parallel for up to 9 months at the Istituto Superiore di Sanità (ISS) and at the INFN-Gran Sasso underground Laboratory (LNGS) where exposure due to gamma-rays and to radon was reduced by factors of about 70 and 25, respectively. After 9 months the cells grown at the LNGS (cumulative gamma dose about 30 microGy, average radon concentration around 5 Bq/m(3)), compared to the cells grown at the ISS (cumulative gamma-ray dose about 2 mGy, average radon concentration around 120 Bq/m(3)), exhibited i). a significant increase of the cell density at confluence, ii). a significantly higher capacity to scavenge organic and inorganic hydroperoxides but a reduced scavenging capacity towards superoxide anions and iii). an increase in both the basal hprt mutation frequency and sensitivity to the mutagenic effect of gamma-rays. The cells grown at the LNGS also showed a greater apoptotic sensitivity starting at the third month of culture, that was no longer detected after 9 months. Overall, these data suggest a role of background ionizing radiation in determining an adaptive response, although they cannot be considered conclusiv