m6A RNA methylation of major satellite repeat transcripts facilitates chromatin association and RNA:DNA hybrid formation in mouse heterochromatin

Heterochromatin has essential functions in maintaining chromosome structure, in protecting genome integrity and in stabilizing gene expression programs. Heterochromatin is often nucleated by underlying DNA repeat sequences, such as major satellite repeats (MSR) and long interspersed nuclear elements (LINE). In order to establish heterochromatin, MSR and LINE elements need to be transcriptionally competent and generate non-coding repeat RNA that remain chromatin associated. We explored whether these heterochromatic RNA, similar to DNA and histones, may be methylated, particularly for 5-methylcytosine (5mC) or methyl-6-adenosine (m6A). Our analysis in mouse ES cells identifies only background level of 5mC but significant enrichment for m6A on heterochromatic RNA. Moreover, MSR transcripts are a novel target for m6A RNA modification, and their m6A RNA enrichment is decreased in ES cells that are mutant for Mettl3 or Mettl14, which encode components of a central RNA methyltransferase complex. Importantly, MSR transcripts that are partially deficient in m6A RNA methylation display impaired chromatin association and have a reduced potential to form RNA:DNA hybrids. We propose that m6A modification of MSR RNA will enhance the functions of MSR repeat transcripts to stabilize mouse heterochromatin

β-Microseminoprotein Endows Post Coital Seminal Plasma with Potent Candidacidal Activity by a Calcium- and pH-Dependent Mechanism

The innate immune factors controlling Candida albicans are mostly unknown. Vulvovaginal candidiasis is common in women and affects approximately 70–75% of all women at least once. Despite the propensity of Candida to colonize the vagina, transmission of Candida albicans following sexual intercourse is very rare. This prompted us to investigate whether the post coital vaginal milieu contained factors active against C. albicans. By CFU assays, we found prominent candidacidal activity of post coital seminal plasma at both neutral and the acid vaginal pH. In contrast, normal seminal plasma did not display candidacidal activity prior to acidification. By antifungal gel overlay assay, one clearing zone corresponding to a protein band was found in both post coital and normal seminal plasma, which was subsequently identified as β-microseminoprotein. At neutral pH, the fungicidal activity of β-microseminoprotein and seminal plasma was inhibited by calcium. By NMR spectroscopy, amino acid residue E71 was shown to be critical for the calcium coordination. The acidic vaginal milieu unleashed the fungicidal activity by decreasing the inhibitory effect of calcium. The candidacidal activity of β-microseminoprotein was mapped to a fragment of the C-terminal domain with no structural similarity to other known proteins. A homologous fragment from porcine β-microseminoprotein demonstrated calcium-dependent fungicidal activity in a CFU assay, suggesting this may be a common feature for members of the β-microseminoprotein family. By electron microscopy, β-microseminoprotein was found to cause lysis of Candida. Liposome experiments demonstrated that β-microseminoprotein was active towards ergosterol-containing liposomes that mimic fungal membranes, offering an explanation for the selectivity against fungi. These data identify β-microseminoprotein as an important innate immune factor active against C. albicans and may help explain the low sexual transmission rate of Candida

Epigenetic regulation of prostate cancer

Prostate cancer is a commonly diagnosed cancer in men and a leading cause of cancer deaths. Whilst the underlying mechanisms leading to prostate cancer are still to be determined, it is evident that both genetic and epigenetic changes contribute to the development and progression of this disease. Epigenetic changes involving DNA hypo- and hypermethylation, altered histone modifications and more recently changes in microRNA expression have been detected at a range of genes associated with prostate cancer. Furthermore, there is evidence that particular epigenetic changes are associated with different stages of the disease. Whilst early detection can lead to effective treatment, and androgen deprivation therapy has a high response rate, many tumours develop towards hormone-refractory prostate cancer, for which there is no successful treatment. Reliable markers for early detection and more effective treatment strategies are, therefore, needed. Consequently, there is a considerable interest in the potential of epigenetic changes as markers or targets for therapy in prostate cancer. Epigenetic modifiers that demethylate DNA and inhibit histone deacetylases have recently been explored to reactivate silenced gene expression in cancer. However, further understanding of the mechanisms and the effects of chromatin modulation in prostate cancer are required. In this review, we examine the current literature on epigenetic changes associated with prostate cancer and discuss the potential use of epigenetic modifiers for treatment of this disease

Molecular mechanism(s) of prostate cancer progression : potential of therapeutic modalities

Prostate cancer remains one of the most commonly diagnosed cancers in men and is a leading cause of cancer death. While great success has been achieved at curing early stage prostate cancer, limited success has been obtained when treating late-stage hormone independent prostate cancer. This is due to the increased propensity for skeletal and non-skeletal metastases. Thus development of novel effective therapeutic modalities against late stage prostate cancer is of critical importance.Towards these objectives, I have focused my attention on the role of prostate secretory protein (PSP-94) which is expressed in normal individuals and in patients with early stage prostate cancer. Using our well established in vivo models of prostate cancer, I have evaluated the ability of PSP-94 and its amino acids 31-45 required (PCK3145) to decrease tumor growth and skeletal metastases in vivo and evaluated the potential mechanism(s) associated with PCK3145 anti-cancer actions.Prostatic cancer can also develop as a result of epigenetic activation of tumor promoting genes. To evaluate the role of methylation in prostate cancer, late stage prostate cancer cells were treated with the universal methylating agent S-adenosylmethionine (SAM) and an anti-sense oligonucleotide directed against MBD2 (AS). Scrambled oligonucleotide was included as a control (S). Both SAM and MBD2-AS resulted in inhibition in uPA, MMP-2 and VEGF production leading to decreased tumor cell invasive capacity. However, SAM and MBD2-AS were not able to either further repress partially methylated genes (GSTP1) or reactivate already methylated genes (AR). Furthermore, SAM and MBD2-AS treatment resulted in significant reduction in tumor growth in vivo . Immunohistochemical and RT-PCR analyses carried out on SAM and MBD2-AS tumors revealed decreased protein and mRNA expression of uPA and MMP-2 which was partially due to increased methylation of the respective promoters even after 10 weeks post in vitro treatment as analyzed by bisulfate sequencing. In addition decreased levels of angiogenesis and tumor survival markers were observed.Collectively, these studies are aimed at the development of novel reliable approached to diagnose and treat advanced, hormone refractory prostate cancer to reduce tumor associated morbidity and mortality

Étude par simulations de dynamique des dislocations des effets d'irradiation sur la plasticité de la ferrite : proposition d'une loi de plasticité cristalline

This work aims to reproduce the individual interactions between screw dislocations and radiation-induced loops using dislocation dynamics in good agreement with molecular dynamics simulations. Such agreement is characterized by reproducing the dynamics of the reaction and obtaining the critical resolved stress to overcome the obstacles. This approach provides the mean to calibrate our dislocation dynamics code with parameters from the molecular dynamics simulations. Consequently, it permits to perform massive simulations at the mesoscopic scale. In this scope, this work consists of two parts, an identification of the energetic model and identification of elementary mechanisms. In the first part we propose a procedure to calibrate the line tension based on Orowan's mechanism using a sensibility study. In the second part, we have identified the cross-slip and twining/anti-twinning mechanisms to be essential to reproduce the individual dislocation-loop interactions. The dislocation dynamics simulations are done using a 3D nodal code called NUMODIS, where the recent developments in this code are presented. The uniqueness of this code is its ability to manage and control collisions and core reactions between dislocation segments. This is done through a set of generic algorithms with the minimum amount of local rules.Ce travail vise à reproduire les interactions individuelles entre les dislocations vis et les boucles induites par l'irradiation en utilisant les simulations de dynamique des dislocations en accord avec les simulations de dynamique moléculaire. Un tel accord se caractérise par la reproduction de la réaction et avoir un valeur des contraintes critiques résolues pour franchir les obstacles. Cette approche fournit le moyen de calibrer notre code de dynamique des dislocations avec les paramètres des simulations de dynamique moléculaire. Par conséquent, il permet d'effectuer des simulations massives à l'échelle mésoscopique. Dans ce cadre, ce travail se compose de deux parties, une identification du modèle énergétique et une identification des mécanismes élémentaires. Dans la première partie, nous proposons une procédure de calibrage de la tension ligne basée sur le mécanisme d'Orowan en utilisant une étude de sensibilité. Dans la deuxième partie, nous avons identifié les mécanismes de glissement dévié et le maclage/antimaclage comme étant essentiels pour reproduire les interactions individuelles de dislocation-boucle. Les simulations de la dynamique des dislocations sont réalisées à l'aide d'un code nodal 3D appelé NUMODIS, où les développements récents dans ce code sont présentés. Un des caractéristiques de ce code est sa capacité à gérer et contrôler les collisions entre les segments des dislocations. Cela se fait au moyen en utilisant un ensemble d'algorithmes génériques avec un minimum de règles locales

Evaluation of prostate secretory protein (PSP-94) as a novel therapeutic agent for blocking prostate cancer progression and hypercalcemia of malignancy

Human prostate cancer is one of the most common malignancies affecting men. It is associated with a high degree of mortality and morbidity due to the development of non-skeletal and skeletal metastases. A common complication in patients suffering from prostate cancer is the development of hypercalcemia of malignancy. While determination of PSA and PSP-94 levels can serve as diagnostic/prognostic markers, PSP-94 can also serve as a therapeutic agent. The efficacy of PSP-94 to block tumor progression and hypercalcemia of malignancy was tested in our syngeneic in vivo rat model of prostate cancer. Rat prostate cancer Mat Ly Lu cells were transfected with full length cDNA encoding PTHrP [Mat Ly Lu-PTHrP] which is known to be the main factor responsible for hypercalcemia of malignancy. Mat Ly Lu-PTHrP cells were inoculated subcutaneously into the right flank or via intracardiac injection into the left ventricle of male Copenhagen rats. Animals were treated with different doses of PSP-94. Tumor volume, time of hind-limb paralysis, plasma calcium and plasma and tumoral PTHrP levels were determined. PSP-94 caused a significant delay in the development of hind-limb paralysis, reduction in tumor volume, plasma calcium levels, plasma and tumoral PTHrP levels as compared to control animals receiving vehicle alone. These effects were due to the induction of tumor cell apoptosis. In conclusion, our studies illustrate the ability of PSP-94 to block prostate cancer growth and skeletal metastases

Simulation of Interactions Between Screw Dislocations and Radiation-induced Defects in et945;-Iron Using Dislocation Dynamics

International audienceLow-alloy ferritic steel (16MND5) is used for pressure vessels in French nuclear reactors, where radiationinduced defects are continuously nucleated during their operational lifetime. More specifically, such defects have a major influence on the collective behavior of dislocations, leading to a significant hardening and embrittlement. A multiscale modeling approach based on a combination of molecular dynamics and dislocation dynamics simulations is therefore adopted. Interactions of edge dislocations with radiation-induced defects have been widely studied compared to those of screw dislocations. The later are nevertheless of great interest as their specific mobility has a significant impact on the mechanical properties of steels at low and moderate temperatures. Furthermore, prior molecular dynamics(MD) studies[1, 2] have shown that their interactions with irradiation loopslead to significant hardening. In this study, a systematic investigation is made of the interactions between screw dislocations with [100] and [111] loops using a three-dimensional nodal dislocation dynamics (DD) code NUMODIS. The same configurations that were previously simulated with MD are successfully reproduced. This allows for a direct comparison between atomic scale and DD simulations to validate our multiscale strategy. Ultimately, the modeling of the collective behavior of dislocations properties controlling irradiation strengthening will be undertaken in order to identify and incorporate the relevant parameters in a crystal plasticity simulation

Simulation of Orowan's mechanism using dislocation dynamics based on the non-singular elastic theory

The motion of dislocations, as computed by dislocation dynamics simulations, depends on theunderlying energetic model casted within a continuum approach. This model is nevertheless stilldebated due to the diculty in capturing the behavior of the atoms in the core of dislocations.Here, we investigate the influence of the corresponding material core parameters on the outcomeof dislocation dynamics simulations of the Orowan bypassing mechanism. A parametric studyfirst reveals a large dispersion of the critical Orowan stress. Within a semi empirical approach,a new predictive equation is then motivated to encompass the core parameters, and extend theoriginal formula proposed by Bacon, Kocks and Scattergood. Emphasizing the need to carefullyselecting these parameters, we finally advocate the use of the Orowan mechanism to calibratedislocation dynamics simulations