DNA damage levels in systemic lupus erythematosus patients with low disease activity: An evaluation by comet assay

Montalvão, Thaís Muniz; Miranda-Vilela, Ana Luisa; Roll, Mariana Matos; Grisólia, César Koppe; Santos-Neto, Leopoldo Luiz dos

doi:10.4236/abb.2012.327121

Cited by 7 publications

(11 citation statements)

References 31 publications

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“…This is the first study to use a buccal micronucleus cytome assay to identify potential biomarkers associated with SLE. Interestingly, this study showed a statistically significant and clinically relevant increase in levels of biomarkers of DNA damage (micronuclei) in patients as compared to controls, which supports results of several studies that used other methods to assess DNA damage, including a comet assay (), a colony survival assay, and kinetics of phosphorylation of structural maintenance of chromosomes protein 1 (). Micronuclei are formed mainly from chromatids or chromosome fragments, which remain excluded from the main cell nucleus following mitosis ().…”

Section: Discussionsupporting

confidence: 86%

Brief Report: A Novel Application of Buccal Micronucleus Cytome Assay in Systemic Lupus Erythematosus: A Case–Control Study

Al-Rawi

Gorial

Tawfiq³

et al. 2014

Arthritis & Rheumatology

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Objective. Systemic lupus erythematous (SLE) is a systemic autoimmune disease, and both genetic and environmental factors are implicated in its pathogenesis. The aim of this study was to assess cellular and nuclear morphology of SLE using a buccal micronucleus cytome assay.Methods. This study included 58 SLE patients and 58 healthy age-and sex-matched controls. Patients' baseline characteristics were recorded, and a buccal micronucleus cytome assay was used to measure cellular and nuclear abnormalities in both groups.Results. Significantly higher frequencies of micronuclei, nuclear buds, binucleated cells, basal cells, condensed chromatin cells, karyorrhectic cells, pyknotic cells, and karyolytic cells were seen in SLE patients as compared to controls. A buccal micronucleus cytome assay score of >4 had the highest accuracy (93.1%), almost perfect positive predictive value (98.1% at 50% pretest probability and 99.8% at 90% pretest probability), 87.9% sensitivity, and 98.3% specificity for the diagnosis of SLE.Conclusion. Biomarkers of DNA damage, proliferative potential, and cell death were significantly increased in SLE patients. The buccal micronucleus cytome assay was a valid and easy way for clinicians to assess individuals at high risk of developing SLE.Systemic lupus erythematosus (SLE) is a multisystem autoimmune disorder with a variety of proposed pathogeneses. Apoptosis is one of several important mechanisms involved in the pathogenesis of SLE. A previous study has demonstrated that lymphocyte apoptosis is increased and removal of apoptotic cells is impaired in SLE. Furthermore, apoptosis was found to be higher in active SLE as compared to inactive SLE (1).Several lines of evidence have demonstrated the close relationship between apoptosis and autoantibodies. Delayed clearance of apoptotic residue, such as histone H3, may result in the activation of antigenpresenting cells and the formation of autoantibodies; further, the antigenicity of autoantibodies is mostly against apoptotic cells (2). Studies have linked lymphocyte and leukocyte apoptosis with SLE and titers of autoantibodies (3).The buccal micronucleus cytome assay is a noninvasive method used to study DNA damage, chromosomal instability, cell death, and the regenerative potential of human buccal mucosal tissue (4). To our knowledge, no studies have as yet assessed buccal cell micronuclei in patients with connective tissue diseases or inflammatory arthritis. This study was designed to assess cellular and nuclear morphology in SLE using a buccal micronucleus cytome assay. PATIENTS AND METHODSStudy design. This case-control study was conducted at the Baghdad Teaching Hospital Rheumatology Unit from September 2012 to May 2013. We screened SLE patients who were eligible for inclusion in the study and then compared patients to matched healthy controls; buccal micronucleus cytome biomarkers were measured in both groups. Informed consent was obtained from all participants, and this study was approved by the ethics committee of Baghdad University, College...

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Section: Discussionsupporting

confidence: 86%

Brief Report: A Novel Application of Buccal Micronucleus Cytome Assay in Systemic Lupus Erythematosus: A Case–Control Study

Al-Rawi

Gorial

Tawfiq³

et al. 2014

Arthritis & Rheumatology

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“…In fact elevated MDA levels and reduced TAS levels also point towards a state of oxidative stress. Shah et al [9] have recently reported inflammatory response in SLE which aggravates oxidative stress with potential to provoke damage to lipids, proteins and DNA [12,13]. The observations of the present study on significantly increased genomic damage also find reflections in the results documented by Montalvao et al [13] who reported 1.3x significantly increased (p=0.000) DNA damage index in 25 SLE patients compared to the value in equal number of healthy controls.…”

Section: Discussionsupporting

confidence: 82%

“…The inflammatory response in SLE exacerbates oxidative stress [9] which tends to promote cellular macromolecular damage, lipid peroxidation and protein oxidation [10,11] and damage to DNA [12,13]. Health can further be compromised as any unrepaired DNA damage can initiate neoplasia [14].…”

Section: Introductionmentioning

confidence: 99%

DNA damage in a patient with Systemic Lupus Erythematosus and Nephropathy- A Case Report

Tung

Gandhi

2015

Int Jour of Biomed Res

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Objectives: Systemic Lupus Erythematosus (SLE) is a complex, multisystem autoimmune inflammatory disease. The inflammatory response in SLE exacerbates oxidative stress which tends to promote lipid peroxidation, protein oxidation and damage to DNA. Documentation of manifestation of DNA damage in SLE patients is scarce and related studies have not come to attention from this part of the world, therefore the present study showcases a case report on SLE. Methods: Assessment of DNA damage in peripheral blood leukocytes and oxidative stress biomarkers in blood sera of a 12y-old male presenting with SLE and nephropathy (on dialysis therapy) was made. Results: Basal DNA damage in PBL scored as percent DNA in tail was higher (51.48%) compared to levels (40.38%) in an age-and sex-matched healthy control. Quantitative measures of DNA damage revealed DF of 96 vs. 94 and DI of 317 vs. 208 in the case and the control, respectively. Serum lipid peroxidation as estimated from MDA level was 3x higher (1.759µmol/l) compared to its level in a healthy control (0.571µmol/l). OSI was slightly higher in the patient (0.134 arbitrary units) compared to that in the control (0.132 arbitrary units). The atherogenic indices were higher in the present case compared to ratios in the control. Conclusion:The results from the case report on increased oxidative stress, dyslipidemia, genetic damage and atherogenic indices support observations from earlier studies and imply onset of other complications in patients with SLE. The assessment of various blood-based biomarkers (as carried out in the present case) in SLE patients may assist in disease diagnosis, prognosis and optimal disease management.

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“…Determinations of DNA damage and DNA damage responses is rapidly expanding in research related to disease states [ 38 – 40 ] and in the investigation of the specific mechanisms of action of ionizing radiation, viral infections, and chemotherapeutic drugs [ 41 – 45 ] but is currently less understood in cryobiology. There is also an ever-increasing understanding of the mechanisms by which endogenous DNA damage signals can activate innate immune responses, especially effects mediated by increased expression of interferons [ 46 – 51 ].…”

Section: Discussionmentioning

confidence: 99%

The effects of storage temperature on PBMC gene expression

et al. 2016

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BackgroundCryopreservation of peripheral blood mononuclear cells (PBMCs) is a common and essential practice in conducting research. There are different reports in the literature as to whether cryopreserved PBMCs need to only be stored ≤ −150 °C or can be stored for a specified time at −80 °C. Therefore, we performed gene expression analysis on cryopreserved PBMCs stored at both temperatures for 14 months and PBMCs that underwent temperature cycling 104 times between these 2 storage temperatures. Real-time RT-PCR was performed to confirm the involvement of specific genes associated with identified cellular pathways. All cryopreserved/stored samples were compared to freshly isolated PBMCs and between storage conditions.ResultsWe identified a total of 1,367 genes whose expression after 14 months of storage was affected >3 fold in PBMCs following isolation, cryopreservation and thawing as compared to freshly isolated PBMC aliquots that did not undergo cryopreservation. Sixty-six of these genes were shared among two or more major stress-related cellular pathways (stress responses, immune activation and cell death). Thirteen genes involved in these pathways were tested by real-time RT-PCR and the results agreed with the corresponding microarray data. There was no significant change on the gene expression if the PBMCs experienced brief but repetitive temperature cycling as compared to those that were constantly kept ≤ −150 °C. However, there were 18 genes identified to be different when PBMCs were stored at −80 °C but did not change when stored < −150 °C. A correlation was also found between the expressions of 2′–5′- oligoadenylate synthetase (OAS2), a known interferon stimulated gene (IFSG), and poor PBMC recovery post-thaw. PBMC recovery and viability were better when the cells were stored ≤ −150 °C as compared to −80 °C.ConclusionsNot only is the viability and recovery of PBMCs affected during cryopreservation but also their gene expression pattern, as compared to freshly isolated PBMCs. Different storage temperature of PBMCs can activate or suppress different genes, but the cycling between −80 °C and −150 °C did not produce significant alterations in gene expression when compared to PBMCs stored ≤ −150 °C. Further analysis by gene expression of various PBMC processing and cryopreservation procedures is currently underway, as is identifying possible molecular mechanisms.Electronic supplementary materialThe online version of this article (doi:10.1186/s12865-016-0144-1) contains supplementary material, which is available to authorized users.

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DNA damage levels in systemic lupus erythematosus patients with low disease activity: An evaluation by comet assay

Cited by 7 publications

References 31 publications

Brief Report: A Novel Application of Buccal Micronucleus Cytome Assay in Systemic Lupus Erythematosus: A Case–Control Study

Brief Report: A Novel Application of Buccal Micronucleus Cytome Assay in Systemic Lupus Erythematosus: A Case–Control Study

DNA damage in a patient with Systemic Lupus Erythematosus and Nephropathy- A Case Report

The effects of storage temperature on PBMC gene expression

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