Migration Groups: A Poorly Explored Point of View for Genetic Damage Assessment Using Comet Assay in Human Lymphocytes

A new point of view for genetic damage assessment using the comet assay is proposed based on the number of migration groups, the number of comets in each group, and the groups with the highest number of comets. Human lymphocytes were exposed to different concentrations of Methyl Methane Sulfonate (MMS), Maleic Hydrazide (MH), 2,4-Dichlorophenoxyacetic (2,4-D), and N-nitroso diethylamine (NDEA). Using comet assay, the migration means of the comets were determined and later grouped arbitrarily in migration groups with no higher differences than 1 µc. The number of migration groups, the number of comets in each group, and the groups with the highest number of comets (modes) were determined. All four of the genotoxic agents studied showed a significant increase (p < 0.05) in the tail length and the number of migration groups compared to the negative control. The number of migration groups did not show a significant variation between the four-genotoxic agents nor within their different concentrations. However, the comparison of the modes did show differences between the genotoxic agents, but not within the concentrations of a same genotoxic agent, which indicated a determined chemical interaction on the DNA. These parameters can improve the detection of genetic damage associated with certain genotoxic agents.

Oxidal® Ameliorates the Ty1 Retrotransposition Induced by Methyl Methanesulfonate in Saccharomyces cerevisiae

Aim: The aim of the study was to evaluate the potential of Oxidal® to decrease the Ty1 retrotransposition rate in a model system Saccharomyces cerevisiae. Study Design: Saccharomyces cerevisiae cell suspensions were pre-treated with different concentrations Oxidal® and subsequently treated with 16mM methyl methanesulfonate. (MMS) Methodology: The potential of various concentrations Oxidal® was evaluated based on “spot” test and Ty1 retro-transposition test. Results: Data revealed that only 5% Oxidal® possesses some cytotoxic properties. Lack of Ty1 retro-transposition was observed after single treatment with 1, 2.5 and 5% Oxidal® concentrations. On the other hand, all the tested concentrations showed promising results against the standard carcinogen methyl methane sulfonate. The most pronounced anti-carcinogenic and cytoprotective effects were observed after pre-treatment with 2.5% Oxidal®, which could be attributed to the antioxidant properties of the combination of ingredients; methylene blue, salicylic acid and caffeine. Further studies could reveal the exact mechanism of action of the supplement and the role of the antioxidant potential. Conclusion: New data is provided concerning the potential of Oxidal® at low concentrations to protect Saccharomyces cerevisiae cells from MMS-induced Ty1 retro-transposition. The cytoprotective properties of the supplement were also obtained. These results could be considered as a basis for further studies revealing the exact mechanisms of cell protection of the Oxidal®. Additionally, our data could also serve as an important step of the in-depth research of a potential antiviral activity.

Regulation of Structure-Specific Endonucleases in Replication Stress

Replication stress results in various forms of aberrant replication intermediates that need to be resolved for faithful chromosome segregation. Structure-specific endonucleases (SSEs) recognize DNA secondary structures rather than primary sequences and play key roles during DNA repair and replication stress. Holliday junction resolvase MUS81 (methyl methane sulfonate (MMS), and UV-sensitive protein 81) and XPF (xeroderma pigmentosum group F-complementing protein) are a subset of SSEs that resolve aberrant replication structures. To ensure genome stability and prevent unnecessary DNA breakage, these SSEs are tightly regulated by the cell cycle and replication checkpoints. We discuss the regulatory network that control activities of MUS81 and XPF and briefly mention other SSEs involved in the resolution of replication intermediates.

Quantitative proteome-level analysis of paulownia witches’ broom disease with methyl methane sulfonate assistance reveals diverse metabolic changes during the infection and recovery processes

Paulownia witches’ broom (PaWB) disease caused by phytoplasma is a fatal disease that leads to considerable economic losses. Although there are a few reports describing studies of PaWB pathogenesis, the molecular mechanisms underlying phytoplasma pathogenicity in Paulownia trees remain uncharacterized. In this study, after building a transcriptome database containing 67,177 sequences, we used isobaric tags for relative and absolute quantification (iTRAQ) to quantify and analyze the proteome-level changes among healthy P. fortunei (PF), PaWB-infected P. fortunei (PFI), and PaWB-infected P. fortunei treated with 20 mg L−1 or 60 mg L−1 methyl methane sulfonate (MMS) (PFI-20 and PFI-60, respectively). A total of 2,358 proteins were identified. We investigated the proteins profiles in PF vs. PFI (infected process) and PFI-20 vs. PFI-60 (recovered process), and further found that many of the MMS-response proteins mapped to “photosynthesis” and “ribosome” pathways. Based on our comparison scheme, 36 PaWB-related proteins were revealed. Among them, 32 proteins were classified into three functional groups: (1) carbohydrate and energy metabolism, (2) protein synthesis and degradation, and (3) stress resistance. We then investigated the PaWB-related proteins involved in the infected and recovered processes, and discovered that carbohydrate and energy metabolism was inhibited, and protein synthesis and degradation decreased, as the plant responded to PaWB. Our observations may be useful for characterizing the proteome-level changes that occur at different stages of PaWB disease. The data generated in this study may serve as a valuable resource for elucidating the pathogenesis of PaWB disease during phytoplasma infection and recovery stages.