@article {10442, title = {Light element distribution in fresh and frozen-thawed human ovarian tissues: a preliminary study.}, journal = {Reprod Biomed Online}, volume = {37}, year = {2018}, month = {2018 08}, pages = {153-162}, abstract = {

RESEARCH QUESTION: Does synchrotron X-ray fluorescence (XRF) provide novel chemical information for the evaluation of human ovarian tissue cryopreservation protocols?

DESIGN: Tissues from five patients undergoing laparoscopic surgery for benign gynaecological conditions were fixed for microscopic analysis either immediately or after cryopreservation. After fixation, fresh and slowly frozen samples were selected by light microscopy and transmission electron microscopy, and subsequently analysed with synchrotron XRF microscopy at different incident energies.

RESULTS: The distributions of elements detected at 7.3 keV (S, P, K, Cl, Fe, and Os) and 1.5 keV (Na and Mg) were related to the changes revealed by light microscopy and transmission electron microscopy analyses. The light elements showed highly informative findings. The S distribution was found to be an indicator of extracellular component changes in the stromal tissues of the freeze-stored samples, further revealed by the transmission electron microscopy analyses. Low-quality follicles, frequent in the freeze-thawed tissues, showed a high Na level in the ooplasm. On the contrary, good-quality follicles were detected by a homogeneous Cl distribution. The occurrence of vacuolated follicles increased after cryopreservation, and the XRF analyses showed that the vacuolar structures contained mainly Cl and Na.

CONCLUSIONS: The study demonstrates that elemental imaging techniques, particularly revealing the distribution of light elements, could be useful in establishing new cryopreservation protocols.

}, keywords = {Cryopreservation, Female, Humans, Microscopy, Electron, Transmission, Organ Preservation, Ovarian Follicle, Ovary}, issn = {1472-6491}, doi = {10.1016/j.rbmo.2018.04.051}, author = {Pascolo, Lorella and Venturin, Irene and Gianoncelli, Alessandra and Bortul, Roberta and Zito, Gabriella and Giolo, Elena and Salom{\`e}, Murielle and Bedolla, Diana E and Altissimo, Matteo and Zweyer, Marina and Ricci, Giuseppe} } @article {8317, title = {Pitfalls and promises in FTIR spectromicroscopy analyses to monitor iron-mediated DNA damage in sperm.}, journal = {Reprod Toxicol}, volume = {61}, year = {2016}, month = {2016 Jun}, pages = {39-46}, abstract = {

Many drugs, chemicals, and environmental factors can impair sperm functionality by inducing DNA damage, one of the important causes of reduced fertility potential. The use of vibrational spectromicroscopy represents a promising approach for monitoring DNA integrity in sperm, although some limitations exist, depending from the experimental conditions. Here, we report that when using FTIR spectromicroscopy to reveal oxidative stress mediated by Fenton{\textquoteright}s reaction on hydrated sperm samples, DNA damage interpretation is partially compromised by unexpected cell surface precipitates. The precipitates give a broad band in the 1150-1000cm(-1) infrared region, which partially covers one of the signatures of DNA (phosphate stretching bands), and are detected as iron and oxygen containing material when using XRF spectroscopy. On the other hand, the analyses further support the potential of FTIR spectromicroscopy to reveal cellular oxidative damage events such as lipid peroxidation, protein misfolding and aggregations, as well as DNA strain breaks.

}, issn = {1873-1708}, doi = {10.1016/j.reprotox.2016.02.011}, author = {Pascolo, Lorella and Bedolla, Diana E and Vaccari, Lisa and Venturin, Irene and Cammisuli, Francesca and Gianoncelli, Alessandra and Mitri, Elisa and Giolo, Elena and Luppi, Stefania and Martinelli, Monica and Zweyer, Marina and Ricci, Giuseppe} } @article {7711, title = {Differential protein folding and chemical changes in lung tissues exposed to asbestos or particulates.}, journal = {Sci Rep}, volume = {5}, year = {2015}, month = {2015}, pages = {12129}, abstract = {

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.

}, issn = {2045-2322}, doi = {10.1038/srep12129}, author = {Pascolo, Lorella and Borelli, Violetta and Canzonieri, Vincenzo and Gianoncelli, Alessandra and Birarda, Giovanni and Bedolla, Diana E and Salom{\`e}, Murielle and Vaccari, Lisa and Calligaro, Carla and Cotte, Marine and Hesse, Bernhard and Luisi, Fernando and Zabucchi, Giuliano and Melato, Mauro and Rizzardi, Clara} }