scholarly journals Effect of cryopreservation on DNA damage and DNA repair activity in human blood samples in the comet assay

2021 ◽  
Author(s):  
Ezgi Eyluel Bankoglu ◽  
Franzisca Stipp ◽  
Johanna Gerber ◽  
Florian Seyfried ◽  
August Heidland ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Dna Damage ◽  
Comet Assay ◽  
Human Blood ◽  
Ex Vivo ◽  
Human Biomonitoring ◽  
Dna Damage And Repair ◽  
Blood Samples ◽  
Repair Activity ◽  
Human Blood Samples

AbstractThe comet assay is a commonly used method to determine DNA damage and repair activity in many types of samples. In recent years, the use of the comet assay in human biomonitoring became highly attractive due to its various modified versions, which may be useful to determine individual susceptibility in blood samples. However, in human biomonitoring studies, working with large sample numbers that are acquired over an extended time period requires some additional considerations. One of the most important issues is the storage of samples and its effect on the outcome of the comet assay. Another important question is the suitability of different blood preparations. In this study, we analysed the effect of cryopreservation on DNA damage and repair activity in human blood samples. In addition, we investigated the suitability of different blood preparations. The alkaline and FPG as well as two different types of repair comet assay and an in vitro hydrogen peroxide challenge were applied. Our results confirmed that cryopreserved blood preparations are suitable for investigating DNA damage in the alkaline and FPG comet assay in whole blood, buffy coat and PBMCs. Ex vivo hydrogen peroxide challenge yielded its optimal effect in isolated PBMCs. The utilised repair comet assay with either UVC or hydrogen peroxide-induced lesions and an aphidicolin block worked well in fresh PBMCs. Cryopreserved PBMCs could not be used immediately after thawing. However, a 16-h recovery with or without mitotic stimulation enabled the application of the repair comet assay, albeit only in a surviving cell fraction.

scholarly journals Characterization of Five Zoonotic Streptococcus suis Strains from Germany, Including One Isolate from a Recent Fatal Case of Streptococcal Toxic Shock-Like Syndrome in a Hunter

2015 ◽  
Vol 53 (12) ◽  
pp. 3912-3915 ◽  
Author(s):  
Tobias Eisenberg ◽  
Christoph Hudemann ◽  
Hamid M. Hossain ◽  
Angela Hewer ◽  
Khodr Tello ◽  
...  
Keyword(s):  
Human Blood ◽  
Ex Vivo ◽  
Fatal Case ◽  
Streptococcus Suis ◽  
Toxic Shock ◽  
Blood Samples ◽  
Content Type ◽  
Human Blood Samples ◽  
Low Levels

AStreptococcus suisisolate from a German hunter with streptococcal toxic shock-like syndrome (STSLS) and four additional zoonotic isolates were genotyped asmrp+epf* (variant 1890)sly+cps2+. All five zoonotic German strains were characterized by high multiplication in human blood samplesex vivo, but induction of only low levels of proinflammatory cytokines compared to a Chinese STSLS strain.

scholarly journals New Application of the Comet Assay

2011 ◽  
Vol 59 (7) ◽  
pp. 655-660 ◽  
Author(s):  
Elva I. Cortés-Gutiérrez ◽  
Martha I. Dávila-Rodríguez ◽  
José Luís Fernández ◽  
Carmen López-Fernández ◽  
Altea Gosálbez ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Single Cells ◽  
White Blood Cells ◽  
Human Biomonitoring ◽  
Cell Types ◽  
Dna Damage And Repair ◽  
Dna Migration ◽  
Sensitive Tool ◽  
Chromosome Isolation

The comet assay is a well-established, simple, versatile, visual, rapid, and sensitive tool used extensively to assess DNA damage and DNA repair quantitatively and qualitatively in single cells. The comet assay is most frequently used to analyze white blood cells or lymphocytes in human biomonitoring studies, although other cell types have been examined, including buccal, nasal, epithelial, and placental cells and even spermatozoa. This study was conducted to design a protocol that can be used to generate comets in subnuclear units, such as chromosomes. The new technique is based on the chromosome isolation protocols currently used for whole chromosome mounting in electron microscopy, coupled to the alkaline variant of the comet assay, to detect DNA damage. The results show that migrant DNA fragments can be visualized in whole nuclei and isolated chromosomes and that they exhibit patterns of DNA migration that depend on the level of DNA damage produced. This protocol has great potential for the highly reproducible study of DNA damage and repair in specific chromosomal domains.

Enhancement of genotoxic effects in the comet assay with human blood samples by aphidicolin

2004 ◽  
Vol 153 (3) ◽  
pp. 303-310 ◽  
Author(s):  
Günter Speit ◽  
Petra Schütz ◽  
Heike Hoffmann
Keyword(s):  
Comet Assay ◽  
Human Blood ◽  
Genotoxic Effects ◽  
Blood Samples ◽  
Human Blood Samples

Ex vivo induction of cytokine mRNA expression in human blood samples

2001 ◽  
Vol 249 (1-2) ◽  
pp. 63-71 ◽  
Author(s):  
Christoph Härtel ◽  
Gregor Bein ◽  
Michael Müller-Steinhardt ◽  
Harald Klüter
Keyword(s):  
Mrna Expression ◽  
Human Blood ◽  
Ex Vivo ◽  
Cytokine Mrna ◽  
Blood Samples ◽  
Cytokine Mrna Expression ◽  
Human Blood Samples

A study of DNA protective effect of orange juice supplementation

2013 ◽  
Vol 38 (5) ◽  
pp. 533-536 ◽  
Author(s):  
Yim Tong Szeto ◽  
Tai Lun To ◽  
Sok Cheon Pak ◽  
Wouter Kalle
Keyword(s):  
Hydrogen Peroxide ◽  
Dna Damage ◽  
Vitamin C ◽  
Comet Assay ◽  
Protective Effect ◽  
Orange Juice ◽  
Healthy Subjects ◽  
Venous Blood ◽  
Blood Samples ◽  
And Control

The potential acute genoprotective effect of orange juice supplementation was investigated. Six healthy subjects (aged 33 to 60 years; 3 women and 3 men) were asked to drink 400 mL of commercial orange juice, which contained 100 mg vitamin C and 40.8 g sugar. Venous blood (2 mL) was taken before and 2 h after ingestion (test trial). A week later, the subjects were asked to repeat the trial by drinking 400 mL water with 100 mg vitamin C and 40.8 g glucose (control trial). Lymphocytes isolated from blood samples underwent comet assay on the day of collection. Pre- and postingestion DNA damage scores were measured in both the test and control trials. Results showed that there was a significant decrease in DNA damage induced by hydrogen peroxide after 2 h of supplementation with orange juice, and no change in baseline DNA damage. There was no significant decrease in the DNA damage in lymphocytes in the control trial.

Application of the comet assay for the evaluation of DNA damage from frozen human whole blood samples: Implications for human biomonitoring

2020 ◽  
Vol 319 ◽  
pp. 58-65 ◽  
Author(s):  
Goran Gajski ◽  
Marko Gerić ◽  
Tanja Živković Semren ◽  
Blanka Tariba Lovaković ◽  
Višnja Oreščanin ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Whole Blood ◽  
Human Biomonitoring ◽  
Blood Samples ◽  
Human Whole Blood ◽  
Whole Blood Samples

scholarly journals Collection and storage of human white blood cells for analysis of DNA damage and repair activity using the comet assay in molecular epidemiology studies

Mutagenesis ◽  
2021 ◽  
Author(s):  
Peter Møller ◽  
Ezgi Eylül Bankoglu ◽  
Helga Stopper ◽  
Lisa Giovannelli ◽  
Carina Ladeira ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Molecular Epidemiology ◽  
Blood Cells ◽  
Mononuclear Cells ◽  
White Blood Cells ◽  
Minor Effect ◽  
Dna Damage And Repair ◽  
Repair Activity ◽  
Epidemiology Studies

Abstract DNA damage and repair activity are often assessed in blood s#38les from humans in different types of molecular epidemiology studies. However, it is not always feasible to analyse the s#38les on the day of collection without any type of storage. For instance, certain studies use repeated s#38ling of cells from the same subject or s#38les from different subjects collected at different time-points, and it is desirable to analyse all these s#38les in the same comet assay experiment. In addition, flawless comet assay analyses on frozen s#38les opens up for the possibility of using this technique on biobank material. In this article we discuss the use of cryopreserved peripheral blood mononuclear cells (PBMCs), buffy coat (BC) and whole blood (WB) for analysis of DNA damage and repair using the comet assay. The published literature and the authors’ experiences indicate that various types of blood s#38les can be cryopreserved with only minor effect on the basal level of DNA damage. There is evidence to suggest that WB and PBMCs can be cryopreserved for several years without much effect on the level of DNA damage. However, care should be taken when cryopreserving WB and BCs. It is possible to use either fresh or frozen s#38les of blood cells, but results from fresh and frozen cells should not be used in the same dataset. The article outlines detailed protocols for the cryopreservation of PBMCs, BCs and WB s#38les.

scholarly journals A frogspawn-like Ag@C core–shell structure for an ultrasensitive label-free electrochemical immunosensing of carcinoembryonic antigen in blood plasma

RSC Advances ◽  
2021 ◽  
Vol 11 (27) ◽  
pp. 16339-16350
Author(s):  
Mengkui Ding ◽  
Ling Zha ◽  
Hui Wang ◽  
Jinyao Liu ◽  
Peiwu Chen ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Carcinoembryonic Antigen ◽  
Human Blood ◽  
Shell Structure ◽  
Core Shell ◽  
Label Free ◽  
Blood Samples ◽  
Human Blood Samples ◽  
Core Shell Structure ◽  
Electrochemical Immunosensing

Novel frogspawn-like Ag@C nanoparticles were successfully used to fabricate an ultrasensitive electrochemical immunosensing platform toward CEA in human blood samples.

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