We present a single molecule visualization approach for the quantitative analysis of reactive oxygen species (ROS) induced DNA damage, such as base oxidation and single stranded breaks in large DNA molecules. We utilized the Fenton reaction to generate DNA damage with subsequent enzymatic treatment using a mixture of three types of glycosylases to remove oxidized bases, and then fluorescent labeling on damaged lesions via nick translation. This single molecule analytical platform provided the capability to count one or two damaged sites per λ DNA molecule (48.5 kb), which were reliably dependent on the concentrations of hydrogen peroxide and ferrous ion at the micromolar level. More importantly, the labeled damaged sites that were visualized under a microscope provided positional information, which offered the capability of comparing DNA damaged sites with the in silico genomic map to reveal sequence specificity that GTGR is more sensitive to oxidative damage. Consequently, single DNA molecule analysis provides a sensitive analytical platform for ROS-induced DNA damage and suggests an interesting biochemical insight that the genome primarily active during the lysogenic cycle may have less probability for oxidative DNA damage.
Ultraviolet radiation induced DNA damage such as double stranded breaks and single stranded breaks is visualized at the level of single DNA molecules. Molecular observations provide a map of DNA radiation-mediated breakages, revealing the sequence dependency of DNA damage rather than random occurrences.
Based on the advances made by artificial intelligence (AI) technologies in drug discovery, including target identification, hit molecule identification, and lead optimization, this study investigated natural compounds that could act as transient receptor potential vanilloid 1 (TRPV1) channel protein antagonists. Using a molecular transformer drug–target interaction (MT-DTI) model, troxerutin was predicted to be a TRPV1 antagonist at IC50 582.73 nM. In a TRPV1-overexpressing HEK293T cell line, we found that troxerutin antagonized the calcium influx induced by the TRPV1 agonist capsaicin in vitro. A structural modeling and docking experiment of troxerutin and human TRPV1 confirmed that troxerutin could be a TRPV1 antagonist. A small-scale clinical trial consisting of 29 participants was performed to examine the efficacy of troxerutin in humans. Compared to a vehicle lotion, both 1% and 10% w/v troxerutin lotions reduced skin irritation, as measured by skin redness induced by capsaicin, suggesting that troxerutin could ameliorate skin sensitivity in clinical practice. We concluded that troxerutin is a potential TRPV1 antagonist based on the deep learning MT-DTI model prediction. The present study provides a useful reference for target-based drug discovery using AI technology and may provide useful information for the integrated research field of AI technology and biology.
Consumption of alcohol injures DNA, and such damage is considered to be a primary cause for the development of cancer and many other diseases essentially due to reactive oxygen species generated from alcohol. To sensitively detect alcohol-induced DNA lesions in a biological system, we introduced a novel analytical platform for visualization of single genomic DNA molecules using E. coli. By fluorescently labelling the DNA lesions, our approach demonstrated, with the highest sensitivity, that we could count the number of DNA lesions induced by alcohol metabolism in a single bacterial cell. Moreover, our results showed a linear relationship between ethanol concentration and the number of DNA lesions: 0.88 lesions per 1% ethanol. Using this approach, we quantitatively analysed the DNA damage induced by exposure to alcoholic beverages such as beer (5% ethanol), rice wine (13%), soju (20%), and whisky (40%).
Background: The accumulation of advanced glycation end products has been proposed as a causative agent of skin aging, but there are no conventional devices for quantifying advanced glycation end‐product accumulation in facial skin.Aims: This study aimed to develop a convenient and accurate in situ advanced glycation end‐product measurement system for the human face.Methods: We developed a facial glycation imaging system, which consisted of illumination (white light‐emitting diode, ultraviolet light‐emitting diode) and image acquisition modules to capture face images. Advanced glycation end product–related autofluorescence and total skin reflectance were calculated to obtain the skin glycation index using an image analysis algorithm. Correlations between the skin glycation index and facial skin elasticity and age were examined in 36 healthy Korean women.Results: The facial glycation imaging system was validated against a volar forearm skin autofluorescence measurement device, that is, the AGE Reader mu, with forearm skin glycation index (R = 0.64, P < .01). Cheek elasticity was negatively correlated with cheek skin glycation index (R = −0.56, R = −0.57, and R = −0.61, P < .01 for R2, R5, and R7, respectively). Age was significantly correlated with forearm skin glycation index (R = 0.44, P < .01) and cheek skin glycation index (R = 0.48, P < .01).Conclusion: We successfully developed a novel in situ facial skin glycation index measurement device. Our convenient and accurate system enables in situ skin glycation index monitoring for skin aging studies such as those on anti‐glycation cosmetics.