title: Pitfalls and promises in FTIR spectromicroscopy analyses to monitor iron-mediated DNA damage in sperm.
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'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.
authors.affiliation: Department of Medical, Surgical, and Health Sciences, University of Trieste, Strada di Fiume 447, 34149 Trieste, Italy. Electronic address: lorella.pascolo@gmail.com.
authors.forename: Lorella
authors.initials: L
authors.lastname: Pascolo
authors.affiliation: Elettra - Sincrotrone Trieste, SS.14 Km 163.5, 34149 Basovizza, Trieste, Italy; Physics Department, University of Trieste, Via Valerio 2, 34127 Trieste, Italy. Electronic address: dianabedollaelettra@gmail.com.
authors.forename: Diana E
authors.initials: DE
authors.lastname: Bedolla
authors.affiliation: Elettra - Sincrotrone Trieste, SS.14 Km 163.5, 34149 Basovizza, Trieste, Italy. Electronic address: lisa.vaccari@elettra.eu.
authors.forename: Lisa
authors.initials: L
authors.lastname: Vaccari
authors.affiliation: Department of Medical, Surgical, and Health Sciences, University of Trieste, Strada di Fiume 447, 34149 Trieste, Italy. Electronic address: ireneventurin@hotmail.it.
authors.forename: Irene
authors.initials: I
authors.lastname: Venturin
authors.affiliation: Department of Medical, Surgical, and Health Sciences, University of Trieste, Strada di Fiume 447, 34149 Trieste, Italy; Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell'Istria 65, 34137 Trieste, Italy. Electronic address: francesca.cammisuli@gmail.com.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cammisuli
authors.affiliation: Elettra - Sincrotrone Trieste, SS.14 Km 163.5, 34149 Basovizza, Trieste, Italy. Electronic address: alessandra.gianoncelli@elettra.eu.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Gianoncelli
authors.affiliation: Elettra - Sincrotrone Trieste, SS.14 Km 163.5, 34149 Basovizza, Trieste, Italy. Electronic address: elisa.mitri@elettra.eu.
authors.forename: Elisa
authors.initials: E
authors.lastname: Mitri
authors.affiliation: Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell'Istria 65, 34137 Trieste, Italy. Electronic address: elena.giolo@burlo.trieste.it.
authors.forename: Elena
authors.initials: E
authors.lastname: Giolo
authors.affiliation: Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell'Istria 65, 34137 Trieste, Italy. Electronic address: stefania.luppi@burlo.trieste.it.
authors.forename: Stefania
authors.initials: S
authors.lastname: Luppi
authors.affiliation: Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell'Istria 65, 34137 Trieste, Italy. Electronic address: monica.martinelli@burlo.trieste.it.
authors.forename: Monica
authors.initials: M
authors.lastname: Martinelli
authors.affiliation: Department of Medical, Surgical, and Health Sciences, University of Trieste, Strada di Fiume 447, 34149 Trieste, Italy. Electronic address: zweyer@units.it.
authors.forename: Marina
authors.initials: M
authors.lastname: Zweyer
authors.affiliation: Department of Medical, Surgical, and Health Sciences, University of Trieste, Strada di Fiume 447, 34149 Trieste, Italy; Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell'Istria 65, 34137 Trieste, Italy. Electronic address: giuseppe.ricci@burlo.trieste.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Ricci
mesh_terms: D004249:DNA Damage; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007501:Iron / Q000633:toxicity*; D008297:Male; D008853:Microscopy; D018384:Oxidative Stress; D013052:Spectrometry, X-Ray Emission; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013094:Spermatozoa / Q000187:drug effects* / Q000378:metabolism
chemical_list: C045076:Fenton's reagent; D006861:Hydrogen Peroxide; D007501:Iron
keywords: DNA damage; Fenton’s reaction; Fourier transform infraRed (FTIR); Oxidative stress; Phosphate stretching bands; Sperm integrity; Vibrational spectromicroscopy; X-ray fluorescence