TY - JOUR T1 - Ferruginous bodies resolved by synchrotron XRF in a dog with peritoneal malignant mesothelioma. JF - Environ Sci Pollut Res Int Y1 - 2018 A1 - Pascotto, Ernesto A1 - Gianoncelli, Alessandra A1 - Calligaro, Carla A1 - Marcuzzo, Thomas A1 - Melato, Mauro A1 - Rizzardi, Clara A1 - Pascolo, Lorella KW - Animals KW - Asbestos KW - Dogs KW - Environmental Exposure KW - Immunohistochemistry KW - Iron KW - Lung KW - Lung Neoplasms KW - Male KW - Mesothelioma KW - Peritoneal Neoplasms KW - Silicon KW - Spectrometry, X-Ray Emission KW - Synchrotrons AB -

Mesothelioma is a malignant tumor mainly correlated to occupational asbestos exposure. Rare reports describe its occurrence also in animals, mainly linked to asbestos in the environment. Asbestos exposure is demonstrated by the appearance of characteristic histological hallmarks: asbestos containing ferruginous bodies that are iron-based structures forming around fibers and also other dust particles. Here we present a clinical case of a suspect of mesothelioma in the peritoneum of a dog with parallel histological observation of ferruginous bodies. To possibly correlate the dog tumor to environmental exposure, we performed X-ray fluorescence (XRF) analyses at two different synchrotrons to resolve the ferruginous bodies' composition. While the histological examination diagnoses a tubulo-papillary mesothelioma, the XRF analyses show that ferruginous bodies contain Si particles, resembling formations of exogenous origin; however, the morphology is unlikely that of asbestos fibers. We speculate that the peritoneal mesothelioma of this dog could be related to environmental exposure to non-asbestos material.

VL - 25 IS - 35 U1 - http://www.ncbi.nlm.nih.gov/pubmed/30357666?dopt=Abstract ER - TY - JOUR T1 - Iron-related toxicity of single-walled carbon nanotubes and crocidolite fibres in human mesothelial cells investigated by Synchrotron XRF microscopy. JF - Sci Rep Y1 - 2018 A1 - Cammisuli, Francesca A1 - Giordani, Silvia A1 - Gianoncelli, Alessandra A1 - Rizzardi, Clara A1 - Radillo, Lucia A1 - Zweyer, Marina A1 - Da Ros, Tatiana A1 - Salomè, Murielle A1 - Melato, Mauro A1 - Pascolo, Lorella KW - Asbestos, Crocidolite KW - Cell Line KW - Epithelial Cells KW - Humans KW - Iron KW - Microscopy, Fluorescence KW - Nanotubes, Carbon AB -

Carbon nanotubes (CNTs) are promising products in industry and medicine, but there are several human health concerns since their fibrous structure resembles asbestos. The presence of transition metals, mainly iron, in the fibres seems also implicated in the pathogenetic mechanisms. To unravel the role of iron at mesothelial level, we compared the chemical changes induced in MeT-5A cells by the exposure to asbestos (crocidolite) or CNTs at different content of iron impurities (raw-SWCNTs, purified- and highly purified-SWCNTs). We applied synchrotron-based X-Ray Fluorescence (XRF) microscopy and soft X-ray imaging (absorption and phase contrast images) to monitor chemical and morphological changes of the exposed cells. In parallel, we performed a ferritin assay. X-ray microscopy imaging and XRF well localize the crocidolite fibres interacting with cells, as well as the damage-related morphological changes. Differently, CNTs presence could be only partially evinced by low energy XRF through carbon distribution and sometimes iron co-localisation. Compared to controls, the cells treated with raw-SWCNTs and crocidolite fibres showed a severe alteration of iron distribution and content, with concomitant stimulation of ferritin production. Interestingly, highly purified nanotubes did not altered iron metabolism. The data provide new insights for possible CNTs effects at mesothelial/pleural level in humans.

VL - 8 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/29335462?dopt=Abstract ER - TY - JOUR T1 - Combined use of AFM and soft X-ray microscopy to reveal fibres' internalization in mesothelial cells. JF - Analyst Y1 - 2017 A1 - Gianoncelli, Alessandra A1 - Kourousias, George A1 - Cammisuli, Francesca A1 - Cassese, Damiano A1 - Rizzardi, Clara A1 - Radillo, Oriano A1 - Lazzarino, Marco A1 - Pascolo, Lorella KW - Asbestos KW - Cell Line KW - Epithelial Cells KW - Epithelium KW - Humans KW - Microscopy, Atomic Force KW - X-Rays AB -

Nanotoxicology and nanomedicine investigations often require the probing of nano-objects such as fibres and particles in biological samples and cells, whilst internalization and intracellular destiny are the main issues for in vitro cellular studies. Various high resolution microscopy techniques are well suited for providing this highly sought-after information. However, sample preparation, nanomaterial composition and sectioning challenges make it often difficult to establish whether the fibres or particles have been internalized or they are simply overlaying or underlying the biological matter. In this paper we suggest a novel suitable combination of two different microscopic techniques to reveal in intact cells the uptake of asbestos fibres by mesothelial cells. After exposure to asbestos fibres and fixation, cells were first analysed under the AFM instrument and then imaged under the TwinMic soft X-ray microscope at Elettra Sincrotrone. The suggested approach combines standard soft X-ray microscopy imaging and AFM microscopy, with a common non-invasive sample preparation protocol which drastically reduces the experimental uncertainty and provides a quick and definitive answer to the nanoparticle cellular and tissue uptake.

VL - 142 IS - 11 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28509933?dopt=Abstract ER - TY - JOUR T1 - Puzzling Results from Germline Mutations Analysis in a Group of Asbestos-Exposed Patients in a High-risk Area of Northeast Italy. JF - Anticancer Res Y1 - 2017 A1 - Rizzardi, Clara A1 - Athanasakis, Emmanouil A1 - Cammisuli, Francesca A1 - Monego, Simeone Dal A1 - DE Spelorzi, Yeraldin Chiquinquira Castillo A1 - Costantinides, Fulvio A1 - Giudici, Fabiola A1 - Pinamonti, Maurizio A1 - Canzonieri, Vincenzo A1 - Melato, Mauro A1 - Pascolo, Lorella KW - Aged KW - Aged, 80 and over KW - Asbestos KW - Environmental Exposure KW - Female KW - Germ-Line Mutation KW - Humans KW - Italy KW - Lung Neoplasms KW - Male KW - Mesothelioma KW - Middle Aged KW - Risk KW - Tumor Suppressor Proteins KW - Ubiquitin Thiolesterase AB -

BACKGROUND: Germline mutations of the oncosuppressor gene breast cancer 1-associated protein 1 (BAP1) were recently related to an autosomal-dominant tumor predisposition syndrome (BAP1-TPDS), characterized by uveal melanoma, malignant mesothelioma (MM), cutaneous melanoma, and other malignancies. The demonstration that BAP1 mutations are strongly associated with MM has provided a real breakthrough in the study of genetic predisposition in MM, that may explain why only a fraction of asbestos-exposed individuals go on to develop MM.

MATERIALS AND METHODS: To evaluate the possible role of BAP1 mutations in the epidemiology of sporadic MM, and their relationship with asbestos exposure, we determined the prevalence of germline BAP1 mutations by the Sanger method in a group of 29 asbestos-exposed patients, 21 of which were diagnosed with MM. They were residents of Trieste, a ship-building town in Northeast Italy with a very high incidence of mesothelioma.

RESULTS: We identified non-obviously pathogenetic germline sequence variants of BAP1 in 3/29 patients and in 2/21 MM cases (10%).

CONCLUSION: Non obviously pathogenic germline sequence variants of BAP1 were found. Nevertheless, limitations of predictive web tools allowed us to comment on some interesting peculiarities of our findings.

VL - 37 IS - 6 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28551647?dopt=Abstract ER - TY - JOUR T1 - Differential protein folding and chemical changes in lung tissues exposed to asbestos or particulates. JF - Sci Rep Y1 - 2015 A1 - Pascolo, Lorella A1 - Borelli, Violetta A1 - Canzonieri, Vincenzo A1 - Gianoncelli, Alessandra A1 - Birarda, Giovanni A1 - Bedolla, Diana E A1 - Salomè, Murielle A1 - Vaccari, Lisa A1 - Calligaro, Carla A1 - Cotte, Marine A1 - Hesse, Bernhard A1 - Luisi, Fernando A1 - Zabucchi, Giuliano A1 - Melato, Mauro A1 - Rizzardi, Clara AB -

Environmental and occupational inhalants may induce a large number of pulmonary diseases, with asbestos exposure being the most risky. The mechanisms are clearly related to chemical composition and physical and surface properties of materials. A combination of X-ray fluorescence (μXRF) and Fourier Transform InfraRed (μFTIR) microscopy was used to chemically characterize and compare asbestos bodies versus environmental particulates (anthracosis) in lung tissues from asbestos exposed and control patients. μXRF analyses revealed heterogeneously aggregated particles in the anthracotic structures, containing mainly Si, K, Al and Fe. Both asbestos and particulates alter lung iron homeostasis, with a more marked effect in asbestos exposure. μFTIR analyses revealed abundant proteins on asbestos bodies but not on anthracotic particles. Most importantly, the analyses demonstrated that the asbestos coating proteins contain high levels of β-sheet structures. The occurrence of conformational changes in the proteic component of the asbestos coating provides new insights into long-term asbestos effects.

VL - 5 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26159651?dopt=Abstract ER - TY - JOUR T1 - Peroxidase-like activity of ferruginous bodies isolated by exploiting their magnetic property. JF - J Toxicol Environ Health A Y1 - 2012 A1 - Borelli, Violetta A1 - Trevisan, Elisa A1 - Vita, Francesca A1 - Bottin, Cristina A1 - Melato, Mauro A1 - Rizzardi, Clara A1 - Zabucchi, Giuliano KW - Air Pollutants, Occupational KW - Asbestos KW - Asbestosis KW - Benzidines KW - Catalysis KW - Cell Line KW - Chromogenic Compounds KW - Cytotoxins KW - Ferric Compounds KW - Ferritins KW - Ferrosoferric Oxide KW - Humans KW - Hydrogen-Ion Concentration KW - Lung KW - Magnetic Phenomena KW - Mesothelioma KW - Mineral Fibers KW - Oxidation-Reduction KW - Peroxidases KW - Respiratory Mucosa AB -

Ferruginous bodies (FB) are polymorphic structures whose formation is macrophage dependent, and are composed of a core, which may consist of an asbestos fiber coated with proteins, among which ferritin is the main component. Within ferritin, the ferric and ferrous ions are coordinated as ferrihydrite, which is the main iron (Fe) storage compound. However, when ferritin accumulates in some tissues following Fe overload it also contains magnetite along with ferrihydrite, which endows it with magnetic properties. Recently studies showed that magnetite exerts peroxidase-like activity, and since ferruginous bodies display magnetic properties, it was postulated that these particular structures may also contain magnetite within the ferritin coating, and thus may also exert peroxidase-like activity. Histochemical analysis for peroxidase of isolated FB smears demonstrated positive staining. Samples isolated from 4 different autopsy lung fragments were also able to oxidize 3,3',5,5'-tetramethyl-benzidine to a blue colored compound that absorbs at 655 nm. This activity was (1) azide and heat insensitive with optimal pH from 5 to 6, and (2) highly variable, changing more than 25-fold from one sample to another. These findings, together with evidence that the peroxidase-like activity of ferruginous bodies has a hydrogen peroxide and substrate requirement different from that of human myeloperoxidase, can exclude that this enzyme gives a significant contribution to the formation of FB. Standard Fe-rich asbestos fibers also express a peroxidase-like activity, but this appears negligible compared to that of ferruginous bodies. Strong acidification of standard Fe-containing asbestos fibers or magnetically isolated ferruginous bodies liberates a high amount of peroxidase-like activity, which is probably accounted for by the release of Fe ions. Further, FB also damage mesothelial cells in vitro. Data suggest that FB exert peroxidase-like activity and cytotoxic activity against mesothelial cells, and hence may be an important factor in pathogenesis of asbestos-related diseases.

VL - 75 IS - 11 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22712847?dopt=Abstract ER -