Generation of 5-(2′-deoxycytidyl)methyl radical and the formation of intrastrand cross-link lesions in oligodeoxyribonucleotides

Hydroxyl radical is one of the major reactive oxygen species (ROS) formed from γ-radiolysis of water or Fenton reaction, and it can abstract one hydrogen atom from the methyl carbon atom of thymine and 5-methylcytosine to give the 5-methyl radical of the pyrimidine bases. The latter radical can also be induced from Type-I photo-oxidation process. Here, we examined the reactivity of the independently generated 5-(2′-deoxycytidyl)methyl radical (I) in single- and double-stranded oligodeoxyribonucleotides (ODNs). It was found that an intrastrand cross-link lesion, in which the methyl carbon atom of 5-methylcytosine and the C8 carbon atom of guanine are covalently bonded, could be formed from the independently generated radical at both GmC and mCG sites, with the yield being much higher at the former site. We also showed by LC-MS/MS that the same cross-link lesions were formed in mC-containing duplex ODNs upon γ irradiation under both aerobic and anaerobic conditions, and the yield was ∼10-fold higher under the latter conditions. The independently generated radical allows for the availability of pure, sufficient and well-characterized intrastrand cross-link lesion-bearing ODN substrates for future biochemical and biophysical characterizations. This was also the first demonstration that the coupling of radical I with its 5′ neighboring guanine can occur in the presence of molecular oxygen, suggesting that the formation of this and other types of intrastrand cross-link lesions might have important implications in the cytotoxic effects of ROS

Tandem mass spectrometryfor the determination of the sitesof DNA interstrand cross-link

Formation of DNA interstrand cross-link is implicated in the mechanism of anticancer activity of some drugs. Here we examined the fragmentation of deprotonated ions of double-stranded oligodeoxynucleotides (ODNs) that are covalently held together with either a mitomycin C or a 4,5′,8-trimethylpsoralen. Our results showed that, upon collisional activation, the covalently-bound duplex ODNs cleaved to give a series of wn and [an − base] ions; the sites of interstrand cross-linking could be determined from the mass shifts of some product ions. In addition, compared with the product-ion spectra acquired on an ion trap, those obtained from sustained off-resonance irradiation-collisionally activated dissociation (SORI-CAD) on a Fourier transform mass spectrometer offered high mass-resolving power, which facilitated unambiguous assignment of product ions and made it an effective method for locating the cross-linking sites

Arsenite binds to the RING finger domains of RNF20-RNF40 histone E3 ubiquitin ligase and inhibits DNA double-strand break repair.

Arsenic is a widespread environmental contaminant. However, the exact molecular mechanisms underlying the carcinogenic effects of arsenic remain incompletely understood. Core histones can be ubiquitinated by RING finger E3 ubiquitin ligases, among which the RNF20-RNF40 heterodimer catalyzes the ubiquitination of histone H2B at lysine 120. This ubiquitination event is important for the formation of open and biochemically accessible chromatin fiber that is conducive for DNA repair. Herein, we found that arsenite could bind directly to the RING finger domains of RNF20 and RNF40 in vitro and in cells, and treatment with arsenite resulted in substantially impaired H2B ubiquitination in multiple cell lines. Exposure to arsenite also diminished the recruitment of BRCA1 and RAD51 to laser-induced DNA double-strand break (DSB) sites, compromised DNA DSB repair in human cells, and rendered cells sensitive toward a radiomimetic agent, neocarzinostatin. Together, the results from the present study revealed, for the first time, that arsenite may exert its carcinogenic effect by targeting cysteine residues in the RING finger domains of histone E3 ubiquitin ligase, thereby altering histone epigenetic mark and compromising DNA DSB repair. Our results also suggest arsenite as a general inhibitor for RING finger E3 ubiquitin ligases

BMAD: Benchmarks for Medical Anomaly Detection

Anomaly detection (AD) is a fundamental research problem in machine learning and computer vision, with practical applications in industrial inspection, video surveillance, and medical diagnosis. In medical imaging, AD is especially vital for detecting and diagnosing anomalies that may indicate rare diseases or conditions. However, there is a lack of a universal and fair benchmark for evaluating AD methods on medical images, which hinders the development of more generalized and robust AD methods in this specific domain. To bridge this gap, we introduce a comprehensive evaluation benchmark for assessing anomaly detection methods on medical images. This benchmark encompasses six reorganized datasets from five medical domains (i.e. brain MRI, liver CT, retinal OCT, chest X-ray, and digital histopathology) and three key evaluation metrics, and includes a total of fourteen state-of-the-art AD algorithms. This standardized and well-curated medical benchmark with the well-structured codebase enables comprehensive comparisons among recently proposed anomaly detection methods. It will facilitate the community to conduct a fair comparison and advance the field of AD on medical imaging. More information on BMAD is available in our GitHub repository: https://github.com/DorisBao/BMA

Structural basis for DNMT3A-mediated de novo DNA methylation.

DNA methylation by de novo DNA methyltransferases 3A (DNMT3A) and 3B (DNMT3B) at cytosines is essential for genome regulation and development. Dysregulation of this process is implicated in various diseases, notably cancer. However, the mechanisms underlying DNMT3 substrate recognition and enzymatic specificity remain elusive. Here we report a 2.65-ångström crystal structure of the DNMT3A-DNMT3L-DNA complex in which two DNMT3A monomers simultaneously attack two cytosine-phosphate-guanine (CpG) dinucleotides, with the target sites separated by 14 base pairs within the same DNA duplex. The DNMT3A-DNA interaction involves a target recognition domain, a catalytic loop, and DNMT3A homodimeric interface. Arg836 of the target recognition domain makes crucial contacts with CpG, ensuring DNMT3A enzymatic preference towards CpG sites in cells. Haematological cancer-associated somatic mutations of the substrate-binding residues decrease DNMT3A activity, induce CpG hypomethylation, and promote transformation of haematopoietic cells. Together, our study reveals the mechanistic basis for DNMT3A-mediated DNA methylation and establishes its aetiological link to human disease

Third-order nonlinearity in Ge–Sb–Se glasses at mid-infrared wavelengths

International audienceThe optical properties of Ge–Sb–Se glasses have been extensively studied at telecom wavelengths in recent years. However, the understanding of nonlinearity in Ge–Sb–Se glasses at mid-infrared wavelengths still remains limited. In this work, a series of Ge20SbxSe80−x (x = 0, 5, 10) glasses were prepared by conventional melt–quenching method. The absorption spectra and the refractive index of glasses were recorded. The third order nonlinearity, n2, and nonlinear absorption coefficient were measured for Ge–Sb–Se glass samples at the wavelengths of 1550, 2000 and 2500 nm by Z-scan technique, respectively. With the increasing of Sb contents, the linear refractive index of glass increased. Among the three operating wavelengths, all the three glass samples have a highest n2 at 2000 nm. By using the figure of merit (FOM) to evaluate the studied three glasses, the Ge20Sb10Se70 glass shows the greatest potential for mid-IR all optical switching device