Iron content (PIXE) in competent and incompetent veins is related to the vein wall morphology and tissue antioxidant enzymes ☆ Wirginia Krzyściak a, b, ⁎, Joanna Kowalska c , Mariusz Kózka d, e , Monika A. Papież f , Wojciech M. Kwiatek c a Department of Medical Diagnostics, Jagiellonian University, Medical College, Pharmacy Faculty, Krakow, Poland b Department of Radioligand, Chair of Pharmacobiology, Jagiellonian University, Medical College, Pharmacy Faculty, Krakow, Poland c Department of Applied Spectroscopy, The Henryk Niewodniczanski Institute of Nuclear Physics, Polish Academy of Sciences, Krakow, Poland d II Department of General Surgery, Jagiellonian University, Medical College, Krakow, Poland e Department of General and Vascular Surgery at 5th Army Hospital, Krakow, Poland f Department of Cytobiology, Jagiellonian University Medical College, Pharmacy Faculty, Krakow, Poland abstract article info Article history: Received 6 July 2011 Received in revised form 18 December 2011 Accepted 30 December 2011 Available online xxxx Keywords: Proton Induced X-ray Emission (PIXE) Reactive oxygen species (ROS) Chronic venous insufficiency (CVI) Tissue iron Blood oxidized DNA (boxDNA) Impaired venous drainage of the lower extremities determines a cascade of pathologic events leading to chronic venous disease (CVD). It is believed that the one cause of CVD is red blood cell extravasation and local iron overload that could generate free radicals and iron-dependent inflammation. The aim of this study was to investigate the relationship between: the intracellular iron deposits in varicose veins and tissue oxidative state measured by: the Proton Induced X-ray Emission Spectroscopy (Fe PIXE ), (tSOD), (tGPx), (tTBARs) and (boxDNA). Patients with diagnosed CVD were qualified for surgical procedure. Entire trunk of the great saphenous vein (GSV) was extracted. Part located near medial ankle was considered competent (C) in duplex ultrasonography (USG) examination. The incompetent (I) part was extracted from GSV where USG showed incompetent valves and massive venous reflux. The difference between local tFe PIXE , tTBARS, boxDNA, tGPx, tSOD in incompetent and competent part of vein tissue was statistically significant. Intima/ media ratio directly correlated with Fe PIXE C/I concentration. Iron deposition in competent vs incompetent part of vein was also related to the oxidative stress parameters (boxDNA). The findings from this pilot study suggest that Fe PIXE measurement may be useful for explaining the progression of chronic venous disease. © 2012 Elsevier B.V. All rights reserved. 1. Introduction Chronic venous diseases (CVDs) represent one of the oldest known chronic human illnesses. Earliest data describing varicose veins, one of the main syndromes of chronic venous insufficiency (CVI) and its treatment are dated to antiquity [1]. The biggest multicentric study including 40,095 people in Poland [2,3] showed that teleangiectasias and varicose veins are included in the most frequently occurring syndromes in women and men, 65% and 50%, respectively. Varices are placed second in relation to the prevalence of CVI syndromes concerning women and men, 25–33% and 10–20%, respectively [4,5]. The size of this phenomenon provides for proofs that this disease has a social character [6]. Lack of effective therapy not only results in development of serious complications, such as thrombosis but also leads to significant lowering of patient lifestyle. Currently, CVD treatment makes 2% of the total budget of the healthcare system of Western Europe Countries [7] and the USA. There are reports, which predict that there will be 400 million people suffering from CVD globally in 2020 [8–10]. This places the dynamics of CVD on the first place within social diseases. This fact justifies the intensive studies on the pathogenesis, diagnostics, and treatment methods of CVD. Bioelectrochemistry xxx (2012) xxx–xxx Abbreviations: 8-oxo-Gua, 8-oxoguanine; API, active pharmaceutical ingredient; BCS, bathocuproine disulfonate disodium salt; BER, base-excision repair; BHT, Butylated hydroxytoluene; BSA, bovine serum albumin; CEAP, clinical state, etiology, anatomy and pathophysiology; CO, carbon monoxide; CVDs, Chronic venous diseases; CVI, chronic venous insufficiency; DETAPAC, diethylenetriaminepentaacetic acid; DMSO, Dimethyl sulfoxide; EtBr, ethidium bromide; FBS, Fetal bovine serum; FDA, fluorescein diacetate; Fpg, formamidopyrimidine DNA glycosylase; GE, General Electric; GST, glutathione S-transferase; GSV, great saphenous vein; H 2 O 2 , hydroxy peroxide; HEPES, [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid]; HmUra, 5- hydroxymethyluracil; HO 2 • , hydroperoxide radical; K 2 EDTA, ethylenediaminetetraacetic acid dipotassium salt; KCl, potassium chloride; LMPA, low melting point agarose; MAPK, mitogen activated protein kinases; MMPs, matrix metalloproteinases; NBT, nitroblue tet- razolium; NBT-BCS, nitroblue tetrazolium-bathocuproine disulfonate disodium salt; NMPA, normal melting point agarose; O 2 - • , superoxide anion; O 2 • , oxygen singlet; OH • , hy- droxyl radical; PBS, phosphate buffered saline; RNS, reactive nitrogen species; ROS, reac- tive oxygen species; SOD, superoxide dismutase activity; TBA, thiobarbituric acid; TBARs, thiobarbituric acid reactive substances; Tris THAM, tris(hydroxymethyl)amino- methane; USG, ultrasonography. ☆ Statement of authorship. WK conceived of the study, and participated in its design, carried out samples, analyzed the samples, made all experiments during the study, performed the statistical analysis and data analyses and wrote the manuscript. JK participated in study design, made the sample preparation and performed the PIXE experiment with the data analysis. MK participated in study design and collect clinical material. WMK participated in study design, conduct and subject enrollment. MAP participated in study design and made the histological sample preparation. All authors read and approved the final manuscript. ⁎ Corresponding author at: Department of Medical Diagnostics, Pharmacy Faculty, UJCM, 9 Medyczna Str. 30-688 Krakow, Poland. Tel.: +48 12 6205760; fax: +48 12 6205400. E-mail address: wirginiakrzysciak@cm-uj.krakow.pl (W. Krzyściak). BIOJEC-06579; No of Pages 10 1567-5394/$ – see front matter © 2012 Elsevier B.V. All rights reserved. doi:10.1016/j.bioelechem.2011.12.011 Contents lists available at SciVerse ScienceDirect Bioelectrochemistry journal homepage: www.elsevier.com/locate/bioelechem Please cite this article as: W. Krzyściak, et al., Iron content (PIXE) in competent and incompetent veins is related to the vein wall morphology and tissue antioxidant enzymes , Bioelectrochemistry (2012), doi:10.1016/j.bioelechem.2011.12.011