Dehydration, deamination and enzymatic repair of cytosine glycols from oxidized poly(dG-dC) and poly(dI-dC)

Cytosine glycols (5,6-dihydroxy-5,6-dihydrocytosine) are initial products of cytosine oxidation. Because these products are not stable, virtually all biological studies have focused on the stable oxidation products of cytosine, including 5-hydroxycytosine, uracil glycols and 5-hydroxyuracil. Previously, we reported that the lifetime of cytosine glycols was greatly enhanced in double-stranded DNA, thus implicating these products in DNA repair and mutagenesis. In the present work, cytosine and uracil glycols were generated in double-stranded alternating co-polymers by oxidation with KMnO4. The half-life of cytosine glycols in poly(dG-dC) was 6.5 h giving a ratio of dehydration to deamination of 5:1. At high substrate concentrations, the excision of cytosine glycols from poly(dG-dC) by purified endonuclease III was comparable to that of uracil glycols, whereas the excision of these substrates was 5-fold greater than that of 5-hydroxycytosine. Kinetic studies revealed that the Vmax was several fold higher for the excision of cytosine glycols compared to 5-hydroxycytosine. In contrast to cytosine glycols, uracil glycols did not undergo detectable dehydration to 5-hydroxyuracil. Replacing poly(dG-dC) for poly(dI-dC) gave similar results with respect to the lifetime and excision of cytosine glycols. This work demonstrates the formation of cytosine glycols in DNA and their removal by base excision repair

Drop Formation in Non-Newtonian Fluids

International audienceWe study the pinch-off dynamics of droplets of yield stress and shear thinning fluids. To separate the two non-Newtonian effects, we use a yield stress material for which the yield stress can be tuned without changing the shear thinning behavior, and a shear thinning system (without a yield stress) for which the shear thinning can be controlled over a large range, without introducing too much elasticity into the system. We find that the pinch-off remains very similar to that of constant viscosity Newtonian liquids, and consequently thinning in shear flow does not imply a thinning in elongational flow

Low-frequency chatter genesis during inclined surface copy-milling with ball-end mill: experimental study

In this study low-frequency chatter during machining of inclined surfaces with ball-end mills is experimentally investigated. An explanation of genesis of low-frequency vibrations have been proposed for various conditions: cutting direction, lead angle values, spindle speed, depth of cut. As a result, it has been proven that low-frequency chatter has more significant effect on machined surface than usual chatter. Low-frequency chatter occurs during downward milling, rather than upward milling, especially when lead angle increases. Furthermore, low-frequency chatter takes place in the beginning of cutting process, thereafter develops into steady state of usual chatter, which has no such significant effect on machined surface, as it has been shown. The results are in line with the supposition that low frequency vibrations are caused by sudden and irregular nature of shearing process, when magnitude is small

Stability of ball-end milling on warped surface: semi-analytical and experimental analysis

This paper presents a study of warped surface machining with ball-end milling. A specific model based on the classical stability lobe theory was used and improved with important aspects. The non-linear effect of radial allowance on the contact angle was integrated by an original averaging method. The cutting coefficients are updated in order to follow effective radius and cutting velocity. An original experimental procedure was developed in order to compute the cutting coefficient for various inclined surfaces. More complete experimental analysis was conducted in order to study the effect of machining parameters on the stability of inclined surface milling. The comparison between experiment and simulation shows good correlation for the prediction of stable cutting condition

In situ targeted activation of an anticancer agent using ultrasound-triggered release of composite droplets

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Retour d'expériences national sur les bassins d'orage en réseau unitaire

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Short- and Long-term Evolutionary Dynamics of Bacterial Insertion Sequences: Insights from Wolbachia Endosymbionts

Transposable elements (TE) are one of the major driving forces of genome evolution, raising the question of the long-term dynamics underlying their evolutionary success. Long-term TE evolution can readily be reconstructed in eukaryotes, thanks to many degraded copies constituting genomic fossil records of past TE proliferations. By contrast, bacterial genomes usually experience high sequence turnover and short TE retention times, thereby obscuring ancient TE evolutionary patterns. We found that Wolbachia bacterial genomes contain 52–171 insertion sequence (IS) TEs. IS account for 11% of Wolbachia wRi, which is one of the highest IS genomic coverage reported in prokaryotes to date. We show that many IS groups are currently expanding in various Wolbachia genomes and that IS horizontal transfers are frequent among strains, which can explain the apparent synchronicity of these IS proliferations. Remarkably, >70% of Wolbachia IS are nonfunctional. They constitute an unusual bacterial IS genomic fossil record providing direct empirical evidence for a long-term IS evolutionary dynamics following successive periods of intense transpositional activity. Our results show that comprehensive IS annotations have the potential to provide new insights into prokaryote TE evolution and, more generally, prokaryote genome evolution. Indeed, the identification of an important IS genomic fossil record in Wolbachia demonstrates that IS elements are not always of recent origin, contrary to the conventional view of TE evolution in prokaryote genomes. Our results also raise the question whether the abundance of IS fossils is specific to Wolbachia or it may be a general, albeit overlooked, feature of prokaryote genomes

Speech Synthesis Evaluation—State-of-the-Art Assessment and Suggestion for a Novel Research Program

Wagner P, Beskow J, Betz S, et al. Speech Synthesis Evaluation—State-of-the-Art Assessment and Suggestion for a Novel Research Program. In: Proceedings of the 10th Speech Synthesis Workshop (SSW10). 2019.Speech synthesis applications have become an ubiquity, in navigation systems, digital assistants or as screen or audio book readers. Despite their impact on the acceptability of the systems in which they are embedded, and despite the fact that different applications probably need different types of TTS voices, TTS evaluation is still largely treated as an isolated problem. Even though there is strong agreement among researchers that the mainstream approaches to Text-to-Speech (TTS) evaluation are often insufficient and may even be misleading, there exist few clear-cut suggestions as to (1) how TTS evaluations may be realistically improved on a large scale, and (2) how such improvements may lead to an informed feedback for system developers and, ultimately, better systems relying on TTS. This paper reviews the current state-of-the-art in TTS evaluation, and suggests a novel user-centered research program for this area

Dynamic proteomic analysis of Aedes aegypti Aag-2 cells infected with Mayaro virus

Background Mayaro virus (MAYV) is responsible for a mosquito-borne tropical disease with clinical symptoms similar to dengue or chikungunya virus fevers. In addition to the recent territorial expansion of MAYV, this virus may be responsible for an increasing number of outbreaks. Currently, no vaccine is available. Aedes aegypti is promiscuous in its viral transmission and thus an interesting model to understand MAYV-vector interactions. While the life-cycle of MAYV is known, the mechanisms by which this arbovirus affects mosquito host cells are not clearly understood. Methods After defining the best conditions for cell culture harvesting using the highest virus titer, Ae. aegypti Aag-2 cells were infected with a Brazilian MAYV isolate at a MOI of 1 in order to perform a comparative proteomic analysis of MAYV-infected Aag-2 cells by using a label-free semi-quantitative bottom-up proteomic analysis. Time-course analyses were performed at 12 and 48 h post-infection (hpi). After spectrum alignment between the triplicates of each time point and changes of the relative abundance level calculation, the identified proteins were annotated and using Gene Ontology database and protein pathways were annotated using the Kyoto Encyclopedia of Genes and Genomes. Results After three reproducible biological replicates, the total proteome analysis allowed for the identification of 5330 peptides and the mapping of 459, 376 and 251 protein groups, at time 0, 12 hpi and 48 hpi, respectively. A total of 161 mosquito proteins were found to be differentially abundant during the time-course, mostly related to host cell processes, including redox metabolism, translation, energy metabolism, and host cell defense. MAYV infection also increased host protein expression implicated in viral replication. Conclusions To our knowledge, this first proteomic time-course analysis of MAYV-infected mosquito cells sheds light on the molecular basis of the viral infection process and host cell response during the first 48 hpi. Our data highlight several mosquito proteins modulated by the virus, revealing that MAYV manipulates mosquito cell metabolism for its propagation