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A Comparison of the Salivary Aldehyde Dehydrogenase (ALDH3A1) Activity in Patients with Radicular Cysts, Keratocystic Odontogenic Tumours and Healthy Subjects

Author(s): Joanna Giebułtowicz | Piotr Wroczyński | Danuta Samolczyk-Wanyura

Journal: International Journal of Collaborative Research on Internal Medicine & Public Health
ISSN 1840-4529

Volume: 3;
Issue: 3;
Start page: 249;
Date: 2011;
Original page

Keywords: Aldehyde dehydrogenese | saliva | keratocystic odontogenic tumor | odontogenic cyst

Background: The salivary ALDH3A1 (E.C. is a homodimeric enzyme oxidizing mainly longandmedium-chain aliphatic and aromatic aldehydes. The protective role of ALDH3A1 in the oxidativestress is considered because of its relatively high affinity (Km = 45 μM) for 4-hydroxynonenal (4-HNE),the concentration of which increases as a result of the oxidative stress and the lipid peroxidation.Therefore, the total ALDH3A1 activity increases as a result of oxidative stress and inflammation causedby such factors as e.g. cigarette smoking, alcohol or a proinflammatory diet.The up-regulation of ALDH3A1 transcription could be the result of the activation of xenobioticresponsive elements (XRE) or electrophile responsive element (EpRE) which are present in the 5'-upstream regions of this gene. Moreover this up-regulation might also be related to such transcriptionfactors as NF-κB, Oct1 and Pax6.The inflammation is also related to certain pathologies of the oral cavity. The radicular cyst (RC)develops in response to an inflammatory stimulus. Moreover the inflammation mediators (e.g.prostaglandins, interleukins) also contribute to the cyst enlargement. Interestingly the cytokine expressionpattern of cells originating from RC is similar to that derived from the keratocystic odontogenic tumuor(KCOT). Even though the KCOT was classified as neoplasms (by WHO) in 2005, it is still considered tobe a pathology with features of both odontogenic cysts and neoplasms.Aim & Objectives: The aim of the present study was to compare the activity of ALDH3A1 in the salivaof healthy subjects and patients with RC and KCOT.Methods: Saliva samples were collected from healthy volunteers (N=130) and patients with RC (N=74)and KCOT (N=11) in the morning and immediately transferred to test tubes with 50mM pyrophosphatebuffer solution of 1mM EDTA, 1mM GSH, pH=8.1.Salivary ALDH3A1 activity was measured fluorometrically. Fluorimetric assays were run in the 50 mMpyrophosphate buffer, pH 8.1, at 25°C, in the presence of 1 mM EDTA and 0.5 mM DTT. The assaysutilize a highly fluorogenic naphthaldehyde substrate, 6-methoxy-2-naphthaldehyde (5 μM), reacting withNAD+ (100 μM) as co-substrate. 6-methoxy-2-naphthoic acid (1.5 μM), was added as internal standard.The saliva samples were diluted 50-fold with buffer and an increase in the fluorescence of the naphthoatewas recorded at 360 nm, with excitation at 315 nm for 6 – 10 minutes.One enzyme unit is defined as the amount oxidizing 1 micromole of 6-methoxy-2-naphthaldehyde perminute. This unit is approximately twice as large as the commonly used benzaldehyde unit.Specific activities were calculated as the ratio of the reaction rate to protein concentration, the latterdetermined by the Bradford method.Normal distribution and homogeneity of variance of the data were assessed with the Shapiro-Wilk testand Levene’s test respectively.Results: Because of the lack of normal distribution and/or homogeneity of variance of the data (p
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