Arp2p, a potential substrate for NatB acetyltransferase in fission yeast, is important in actin patch nucleation and motility in arp2-1 and arm1 ts mutants

The actin cytoskeleton is a dynamic and complex structure in fission yeast that plays a major function in many cell processes including cellular growth, septa formation, endocytosis and cellular division. Computational studies have shown that Arp2p, which forms part of the Arp2/3 complex, is a potential substrate of NatB acetyltransferase which has specificity for proteins possessing an N-terminal Met-Asp or Met-Glu sequence motif. In arm1- mutants the loss of function of Arm1p, an auxillary subunit required for NatB activity, results in a temperature sensitive phenotype characterized by multiple septa, failure of endocytosis, and the inability to form actin cables. A temperature sensitive mutant of Schizosaccharomyces pombe arp2 gene exhibits a similar phenotype as seen by the formation of improper septa, slow growth, and the delocalization of actin patches. Four expression vectors encoding the open reading frames of arp2 and cdc8 (tropomyosin) were constructed with a modification changing the second residue to a Histidine, believed to mimic the charge distribution of natural acetylation by NatB. Constructs tested in normal yeast strains remained viable and grew normally in the presence of Met-His Arp2p and tropomyosin. Analysis of their ability to suppress the mutant phenotypes of arp2-1 and arm1- mutants is an area of research to be explored in future studies.

Two-hybrid analysis of the interaction between the fission yeast proteins Sal3 and Cdc25

Cdc25 is a mitosis triggering phosphatase in Schizosaccharomyces pombe, and is transported in to the nucleus during G2 phase by the importin-β protein Sal3. Cdc25 triggers mitosis and cell division by dephosphorylating tyrosine 15 of Cdc2. In sal3 mutants, Cdc25 is not transported into the nucleus and the cells halt in G2. The purpose of this study is to use a two-hybrid system to determine the nature of the relationship between Sal3 and Cdc25. Previous research has failed to detect any interaction between the two proteins, but specific modifications were made to the two-hybrid system in this study including the separation of Sal3 into its two binding domains, the addition of fluorescent tags to the fusion protein, and the reversal of plasmids in the fusion proteins. Unique PCR primers were successfully designed, based on a multiple alignment of Sal3 and its homologues, to separate Sal3 into its two domains.

Development of the nervous system in Dugesia tigrina during regeneration after fission and decapitation

In the present study of Dugesia tigrina the development of the nervous system is followed and compared during regeneration after fission and after decapitation. Immunocytochemistry was used, with antisera raised against the biogenic amine, 5-hydroxytryptamine (5-HT) and the two neuropeptides, neuropeptide F (NPF), and FMRF amide. The results indicate that two processes are involved in the formation of the new cerebral ganglion. First, new processes sprouting from the original main longitudinal nerve cords bend transversely, indicating the position of the developing horseshoe-shaped anterior cerebral commissure. Then new nerve cells in front of the commissure differentiate from neoblasts and their growth cones fasciculate with the fibres from the old main longitudinal nerve cords. In the cerebral ganglion, 5-HT-IR cells appear before NPF-IR cells, in contrast to the pharynx where NPF-IR cells differentiate before the 5-HT-IR cells. In the peripheral nervous system, NPF-IR fibres and cells appear at a very early stage and dominate the whole regeneration process. A role for the PNS in early pattern formation is suggested.

Temporal Narrowing of Neutrons Produced by High-Intensity Short-Pulse Lasers

The production of neutron beams having short temporal duration is studied using ultraintense laser pulses. Laser-accelerated protons are spectrally filtered using a laser-triggered microlens to produce a short duration neutron pulse via nuclear reactions induced in a converter material (LiF). This produces a similar to 3 ns duration neutron pulse with 10(4) n/MeV/sr/shot at 0.56 m from the laser-irradiated proton source. The large spatial separation between the neutron production and the proton source allows for shielding from the copious and undesirable radiation resulting from the laser-plasma interaction. This neutron pulse compares favorably to the duration of conventional accelerator sources and should scale up with, present and future, higher energy laser facilities to produce brighter and shorter neutron beams for ultrafast probing of dense materials.

Novel role of androgens in mitochondrial fission and apoptosis

Androgen and androgen receptors (AR) play critical roles in the proliferation of prostate cancer through transcriptional regulation of target genes. Here, we found that androgens upregulated the expression of dynamin-related protein 1 (Drp1), which is involved in the induction of mitochondrial fission, a common event in mitosis and apoptosis. Clinical tissue samples and various prostate cancer cell lines revealed a positive correlation between Drp1 and AR levels. Treatment of androgen-sensitive cells with an AR agonist, R1881, and antagonist, bicalutamide, showed that Drp1 is transcriptionally regulated by androgens, as confirmed by an AR ChIP-seq assay. Live imaging experiments using pAcGFP1-Mito stably transfected LNCaP (mito-green) cells revealed that androgen did not induce significant mitochondrial fission by itself, although Drp1 was upregulated. However, when treated with CGP37157 (CGP), an inhibitor of mitochondrial Ca²⁺ efflux, these cells exhibited mitochondrial fission, which was further enhanced by pretreatment with R1881, suggesting that androgen-induced Drp1 expression facilitated CGP-induced mitochondrial fission. This enhanced mitochondrial fission was correlated with increased apoptosis. Transfection with dominant-negative (DN-Drp1, K38A) rescued cells from increased apoptosis, confirming the role of androgen-induced Drp1 in the observed apoptosis with combination treatment. Furthermore, we found that CGP reduced the expression of Mfn1, a protein that promotes mitochondrial fusion, a process which opposes fission. We suggest that androgen-increased Drp1 enhanced mitochondrial fission leading to apoptosis. The present study shows a novel role for androgens in the regulation of mitochondrial morphology that could potentially be utilized in prostate cancer therapy.

Membrane fission is promoted by insertion of amphipathic helices and is restricted by crescent BAR domains

Shallow hydrophobic insertions and crescent-shaped BAR scaffolds promote membrane curvature. Here, we investigate membrane fission by shallow hydrophobic insertions quantitatively and mechanistically. We provide evidence that membrane insertion of the ENTH domain of epsin leads to liposome vesiculation, and that epsin is required for clathrin-coated vesicle budding in cells. We also show that BAR-domain scaffolds from endophilin, amphiphysin, GRAF, and β2-centaurin limit membrane fission driven by hydrophobic insertions. A quantitative assay for vesiculation reveals an antagonistic relationship between amphipathic helices and scaffolds of N-BAR domains in fission. The extent of vesiculation by these proteins and vesicle size depend on the number and length of amphipathic helices per BAR domain, in accord with theoretical considerations. This fission mechanism gives a new framework for understanding membrane scission in the absence of mechanoenzymes such as dynamin and suggests how Arf and Sar proteins work in vesicle scission.

Cement As a Waste Form for Nuclear Fission Products: The Case of 90Sr and Its Daughters

One of the main challenges faced by the nuclear industry is the long-term confinement of nuclear waste. Because it is inexpensive and easy to manufacture, cement is the material of choice to store large volumes of radioactive materials, in particular the low-level medium-lived fission products. It is therefore of utmost importance to assess the chemical and structural stability of cement containing radioactive species. Here, we use ab initio calculations based on density functional theory (DFT) to study the effects of ⁹⁰Sr insertion and decay in C-S-H (calcium-silicate-hydrate) in order to test the ability of cement to trap and hold this radioactive fission product and to investigate the consequences of its β-decay on the cement paste structure. We show that ⁹⁰Sr is stable when it substitutes the Ca²⁺ ions in C-S-H, and so is its daughter nucleus ⁹⁰Y after β-decay. Interestingly, ⁹⁰Zr, daughter of ⁹⁰Y and final product in the decay sequence, is found to be unstable compared to the bulk phase of the element at zero K but stable when compared to the solvated ion in water. Therefore, cement appears as a suitable waste form for ⁹⁰Sr storage.

Laser Driven Nuclear Physics at ELI–NP

High power lasers have proven being capable to produce high energy γ-rays, charged particles and neutrons, and to induce all kinds of nuclear reactions. At ELI, the studies with high power lasers will enter for the first time into new domains of power and intensities: 10 PW and 10^23 W/cm^2. While the development of laser based radiation sources is the main focus at the ELI-Beamlines pillar of ELI, at ELI-NP the studies that will benefit from High Power Laser System pulses will focus on Laser Driven Nuclear Physics (this TDR, acronym LDNP, associated to the E1 experimental area), High Field Physics and QED (associated to the E6 area) and fundamental research opened by the unique combination of the two 10 PW laser pulses with a gamma beam provided by the Gamma Beam System (associated to E7 area). The scientific case of the LDNP TDR encompasses studies of laser induced nuclear reactions, aiming for a better understanding of nuclear properties, of nuclear reaction rates in laser-plasmas, as well as on the development of radiation source characterization methods based on nuclear techniques. As an example of proposed studies: the promise of achieving solid-state density bunches of (very) heavy ions accelerated to about 10 MeV/nucleon through the RPA mechanism will be exploited to produce highly astrophysical relevant neutron rich nuclei around the N~126 waiting point, using the sequential fission-fusion scheme, complementary to any other existing or planned method of producing radioactive nuclei.

The studies will be implemented predominantly in the E1 area of ELI-NP. However, many of them can be, in a first stage, performed in the E5 and/or E4 areas, where higher repetition laser pulses are available, while the harsh X-ray and electromagnetic pulse (EMP) environments are less damaging compared to E1.

A number of options are discussed through the document, having an important impact on the budget and needed resources. Depending on the TDR review and subsequent project decisions, they may be taken into account for space reservation, while their detailed design and implementation will be postponed.

The present TDR is the result of contributions from several institutions engaged in nuclear physics and high power laser research. A significant part of the proposed equipment can be designed, and afterwards can be built, only in close collaboration with (or subcontracting to) some of these institutions. A Memorandum of Understanding (MOU) is currently under preparation with each of these key partners as well as with others that are interested to participate in the design or in the future experimental program.

Functional differentiation of tbf1 orthologues in fission and budding yeasts.

In Saccharomyces cerevisiae, TBF1, an essential gene, influences telomere function but also has other roles in the global regulation of transcription. We have identified a new member of the tbf1 gene family in the mammalian pathogen Pneumocystis carinii. We demonstrate by transspecies complementation that its ectopic expression can provide the essential functions of Schizosaccharomyces pombe tbf1 but that there is no rescue between fission and budding yeast orthologues. Our findings indicate that an essential function of this family of proteins has diverged in the budding and fission yeasts and suggest that effects on telomere length or structure are not the primary cause of inviability in S. pombe tbf1 null strains.

Cold fission: splitting the pombe cell at room temperature.

The mechanisms responsible for cytokinesis and its coordination with other events of the cell cycle are poorly understood. Genetic studies of cytokinesis in fission yeast are one useful approach to this problem. A number of conditional mutants of fission yeast that show defects in the formation of the septum of cytokinesis have been identified. Cloning of the genes affected in these mutants has begun to shed light upon the elements required to direct the construction of the division septum and also upon how the initiation of septum formation may be coordinated with mitosis.

Étude de la réponse du détecteur ATLAS-MPX aux neutrons rapides

Les détecteurs ATLAS-MPX sont des détecteurs Medipix2-USB recouverts de convertisseurs de fluorure de lithium et de polyéthylène pour augmenter l’efficacité de détection des neutrons lents et des neutrons rapides respectivement. Un réseau de quinze détecteurs ATLAS-MPX a été mis en opération dans le détecteur ATLAS au LHC du CERN. Deux détecteurs ATLAS-MPX de référence ont été exposés à des sources de neutrons rapides 252 Cf et 241 AmBe ainsi qu’aux neutrons rapides produits par la réaction 7Li(p, xn) pour l’étude de la réponse du détecteur à ces neutrons. Les neutrons rapides sont principalement détectés à partir des protons de recul des collisions élastiques entre les neutrons et l’hydrogène dans le polyéthylène. Des réactions nucléaires entre les neutrons et le silicium produisent des particules-α. Une étude de l’efficacité de reconnaissance des traces des protons et des particules-α dans le détecteur Medipix2-USB a été faite en fonction de l’énergie cinétique incidente et de l’angle d’incidence. L’efficacité de détection des neutrons rapides a été évaluée à deux seuils d’énergie (8 keV et 230 keV) dans les détecteurs ATLAS-MPX. L’efficacité de détection des neutrons rapides dans la région du détecteur couverte avec le polyéthylène augmente en fonction de l’énergie des neutrons : (0.0346 ± 0.0004) %, (0.0862 ± 0.0018) % et (0.1044 ± 0.0026) % pour des neutrons rapides de 2.13 MeV, 4.08 MeV et 27 MeV respectivement. L’étude pour déterminer l’énergie des neutrons permet donc d’estimer le flux des neutrons quand le détecteur ATLAS-MPX est dans un champ de radiation inconnu comme c’est le cas dans le détecteur ATLAS au LHC.

Détection des événements de "Minimum Bias" et neutrons avec les détecteurs ATLAS-MPX par simulations

Un réseau de seize détecteurs ATLAS-MPX a été mis en opération dans le détecteur ATLAS au LHC du CERN. Les détecteurs ATLAS-MPX sont sensibles au champ mixte de radiation de photons et d’électrons dans la caverne d’ATLAS et sont recouverts de convertisseurs de fluorure de lithium et de polyéthylène pour augmenter l’efficacité de détection des neutrons thermiques et des neutrons rapides respectivement. Les collisions à haute énergie sont dominées par des interactions partoniques avec petit moment transverse pT , associés à des événements de “minimum bias”. Dans notre cas la collision proton-proton se produit avec une énergie de 7 TeV dans le centre de masse avec une luminosité de 10³⁴cm⁻²s⁻¹ telle que fixée dans les simulations. On utilise la simulation des événements de "minimum bias" générés par PYTHIA en utilisant le cadre Athena qui fait une simulation GEANT4 complète du détecteur ATLAS pour mesurer le nombre de photons, d’électrons, des muons qui peuvent atteindre les détecteurs ATLASMPX dont les positions de chaque détecteur sont incluses dans les algorithmes d’Athena. Nous mesurons les flux de neutrons thermiques et rapides, générés par GCALOR, dans les régions de fluorure de lithium et de polyéthylène respectivement. Les résultats des événements de “minimum bias” et les flux de neutrons thermiques et rapides obtenus des simulations sont comparés aux mesures réelles des détecteurs ATLAS-MPX.

Caractérisation de la réponse de détecteurs aux neutrons de très basses énergies dans le cadre du projet PICASSO

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal