scholarly journals UV rays induced DNA damage: Protection by polyphenols enriched extract of mint leaves

2020 ◽  
Vol 8 (2) ◽  
pp. 152-157
Author(s):  
Rajesh Kowti ◽  
Vedamurthy Joshi ◽  
Rupesh Kumar Mani ◽  
B. Ramesh ◽  
Santhosh Kumar Nune ◽  
...  
Keyword(s):  
Dna Damage ◽  
Gel Electrophoresis ◽  
Positive Control ◽  
Live Cells ◽  
Ultraviolet Rays ◽  
Cytotoxicity Studies ◽  
Calf Thymus ◽  
Thymine Dimers ◽  
Damage Protection ◽  
Uv Rays

DNA of live cells is damaged when exposed to harmful Ultraviolet rays. If the cells are exposed to UV rays exposure time more duration of time, more thymine dimers are formed in the DNA and the greater the risk of an incorrect repair or a missed dimer. To analyze the effect of UV rays Calf thymus DNA and prevention by polyphenol enriched extract of Mint leaves and also its non toxic nature. The above study was done by using submarine gel electrophoresis where UV rays are used to damage the DNA and BHA (400µM) used as positive control. The polyphenol enriched extract was taken at 15μg concentration prevent UV rays induced DNA fragmentation in submarine agarose gel electrophoresis which provides same protection when compared to standard antioxidant BHA (400μM). The cytotoxicity studies showed that, the extract and BHA provides a protection of 54% whereas, the extract of Mint leaves extract showed 72%. In conclusion, the Mint leaves extract showed a promising DNA protectant activity against UV rays induced DNA damage.

Do We have a Satisfactory Cell Viability Assay? Review of the Currently Commercially-Available Assays

2020 ◽  
Vol 17 (1) ◽  
pp. 2-22 ◽  
Author(s):  
Abdel-Baset Halim
Keyword(s):  
Cell Viability ◽  
Data Interpretation ◽  
Positive Control ◽  
Live Cells ◽  
Proof Of Concept ◽  
Cell Viability Assay ◽  
Viability Assay ◽  
Current Cell ◽  
Dying Cells ◽  
Differential Permeability

:Cell-based assays are an important part of the drug discovery process and clinical research. One of the main hurdles is to design sufficiently robust assays with adequate signal to noise parameters while maintaining the inherent physiology of the cells and not interfering with the pharmacology of target being investigated.:A plethora of assays that assess cell viability (or cell heath in general) are commercially available and can be classified under different categories according to their concepts and principle of reactions. The assays are valuable tools, however, suffer from a large number of limitations. Some of these limitations can be procedural or operational, but others can be critical as those related to a poor concept or the lack of proof of concept of an assay, e.g. those relying on differential permeability of dyes in-and-out of viable versus compromised cell membranes. While the assays can differentiate between dead and live cells, most, if not all, of them can just assess the relative performance of cells rather than providing a clear distinction between healthy and dying cells. The possible impact of relatively high molecular weight dyes, used in most of the assay, on cell viability has not been addressed. More innovative assays are needed, and until better alternatives are developed, setup of current cell-based studies and data interpretation should be made with the limitations in mind. Negative and positive control should be considered whenever feasible. Also, researchers should use more than one orthogonal method for better assessment of cell health.

scholarly journals Monitoring S. pombe genome stress by visualizing end-binding protein Ku

2020 ◽  
Author(s):  
Chance Jones ◽  
Susan L Forsburg
Keyword(s):  
Dna Damage ◽  
Fission Yeast ◽  
Protein Complex ◽  
Genome Stability ◽  
Binding Protein ◽  
Dna Lesions ◽  
Live Cells ◽  
Dna Damage Checkpoint ◽  
Unique Pattern ◽  
Ku Protein

AbstractStudies of genome stability have exploited visualization of fluorescently tagged proteins in live cells to characterize DNA damage, checkpoint, and repair responses. In this report, we describe a new tool for fission yeast, a tagged version of the end-binding protein Pku70 which is part of the KU protein complex. We compare Pku70 localization to other markers upon treatment to various genotoxins, and identify a unique pattern of distribution. Pku70 provides a new tool to define and characterize DNA lesions and the repair response.

scholarly journals Streamlined histone-based fluorescence lifetime imaging microscopy reveals ATM regulation of chromatin compaction

2017 ◽  
Author(s):  
Alice Sherrard ◽  
Paul Bishop ◽  
Melanie Panagi ◽  
Maria Beatriz Villagomez ◽  
Dominic Alibhai ◽  
...  
Keyword(s):  
Dna Damage ◽  
Fluorescence Lifetime ◽  
Genotoxic Stress ◽  
Live Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Chromatin Compaction ◽  
Lifetime Imaging ◽  
Detailed Method ◽  
Chromatin Organisation

AbstractChanges in chromatin compaction are crucial during genomic responses. Thus, methods that enable such measurements are instrumental for investigating genome function. Here, we address this challenge by developing, validating, and streamlining histone-based fluorescence lifetime imaging microscopy (FLIM) that robustly detects chromatin compaction states in fixed and live cells; in 2D and 3D. We present quality-controlled and detailed method that is simpler and faster than previous approches, and uses FLIMfit open-source software. We demonstrate the versatility of our method through its combination with immunofluorescence and its implementation in immortalised cells and primary neurons. Owing to these developments, we applied this method to elucidate the function of the DNA damage response kinase, ATM, in regulating chromatin organisation after genotoxic-stress. We unravelled a role for ATM in regulating chromatin compaction independently of DNA damage. Collectively, we present an adaptable chromatin FLIM method for examining chromatin structure in cells, and establish its broader utility.

Silver Nanoparticles and Carbon Nanotubes Induced DNA Damage in Mice Evaluated by Single Cell Gel Electrophoresis

2015 ◽  
Vol 357 (1) ◽  
pp. 210-217 ◽  
Author(s):  
Kumud Kant Awasthi ◽  
Anjali Awasthi ◽  
Rajbala Verma ◽  
Inderpal Soni ◽  
Kamlendra Awasthi ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Dna Damage ◽  
Silver Nanoparticles ◽  
Single Cell ◽  
Gel Electrophoresis ◽  
Single Cell Gel Electrophoresis
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