Interpretation of the centromere epigenetic mark to maintain genome stability

Le centromère est la région chromosomique où le kinétochore s'assemble en mitose. Contrairement à certaines caractéristiques géniques, la séquence centromérique n'est ni conservée entre les espèces ni suffisante à la fonction centromérique. Il est donc bien accepté dans la littérature que le centromère est régulé épigénétiquement par une variante de l'histone H3, CENP-A. KNL-2, aussi connu sous le nom de M18BP1, ainsi que ces partenaires Mis18α et Mis18β sont des protéines essentielles pour l'incorporation de CENP-A nouvellement synthétisé aux centromères. Des évidences expérimentales démontrent que KNL-2, ayant un domaine de liaison à l'ADN nommé Myb, est la protéine la plus en amont pour l'incorporation de CENP-A aux centromères en phase G1. Par contre, sa fonction dans le processus d'incorporation de CENP-A aux centromères n'est pas bien comprise et ces partenaires de liaison ne sont pas tous connus. De nouveaux partenaires de liaison de KNL-2 ont été identifiés par des expériences d'immunoprécipitation suivies d'une analyse en spectrométrie de masse. Un rôle dans l'incorporation de CENP-A nouvellement synthétisé aux centromères a été attribué à MgcRacGAP, une des 60 protéines identifiées par l'essai. MgcRacGAP ainsi que les protéines ECT-2 (GEF) et la petite GTPase Cdc42 ont été démontrées comme étant requises pour la stabilité de CENP-A incorporé aux centromères. Ces différentes observations ont mené à l'identification d'une troisième étape au niveau moléculaire pour l'incorporation de CENP-A nouvellement synthétisé en phase G1, celle de la stabilité de CENP-A nouvellement incorporé aux centromères. Cette étape est importante pour le maintien de l'identité centromérique à chaque division cellulaire. Pour caractériser la fonction de KNL-2 lors de l'incorporation de CENP-A nouvellement synthétisé aux centromères, une technique de microscopie à haute résolution couplée à une quantification d'image a été utilisée. Les résultats générés démontrent que le recrutement de KNL-2 au centromère est rapide, environ 5 minutes après la sortie de la mitose. De plus, la structure du domaine Myb de KNL-2 provenant du nématode C. elegans a été résolue par RMN et celle-ci démontre un motif hélice-tour-hélice, une structure connue pour les domaines de liaison à l'ADN de la famille Myb. De plus, les domaines humain (HsMyb) et C. elegans (CeMyb) Myb lient l'ADN in vitro, mais aucune séquence n'est reconnue spécifiquement par ces domaines. Cependant, il a été possible de démontrer que ces deux domaines lient préférentiellement la chromatine CENP-A-YFP comparativement à la chromatine H2B-GFP par un essai modifié de SIMPull sous le microscope TIRF. Donc, le domaine Myb de KNL-2 est suffisant pour reconnaître de façon spécifique la chromatine centromérique. Finalement, l'élément reconnu par les domaines Myb in vitro a potentiellement été identifié. En effet, il a été démontré que les domaines HsMyb et CeMyb lient l'ADN simple brin in vitro. De plus, les domaines HsMyb et CeMyb ne colocalisent pas avec CENP-A lorsqu'exprimés dans les cellules HeLa, mais plutôt avec les corps nucléaires PML, des structures nucléaires composées d'ARN. Donc, en liant potentiellement les transcrits centromériques, les domaines Myb de KNL-2 pourraient spécifier l'incorporation de CENP-A nouvellement synthétisé uniquement aux régions centromériques.

Rest-activity cycle disturbances in the acute phase of moderate to severe traumatic brain injury

Background. Sleep-wake disturbances are among the most persistent sequelae after traumatic brain injury (TBI) and probably arise during the hospital stay following TBI. These disturbances are characterized by difficulties sleeping at night and staying awake during the day. Objective. The aim of the present study was to document rest-activity cycle consolidation in acute moderate/severe TBI using actigraphy and to assess its association with injury severity and outcome. Methods. In all, 16 hospitalized patients (27.1 ± 11.3 years) with moderate/severe TBI wore actigraphs for 10 days, starting in the intensive care unit (ICU) when continuous sedation was discontinued and patients had reached medical stability. Activity counts were summed for daytime (7:00-21:59 hours) and nighttime periods (22:00-6:59 hours). The ratio of daytime period activity to total 24-hour activity was used to quantify rest-activity cycle consolidation. An analysis of variance was carried out to characterize the evolution of the daytime activity ratio over the recording period. Results. Rest-activity cycle was consolidated only 46.6% of all days; however, a significant linear trend of improvement was observed over time. Greater TBI severity and longer ICU and hospital lengths of stay were associated with poorer rest-activity cycle consolidation and evolution. Patients with more rapid return to consolidated rest-activity cycle were more likely to have cleared posttraumatic amnesia and have lower disability at hospital discharge. Conclusions. Patients with acute moderate/ severe TBI had an altered rest-activity cycle, probably reflecting severe fragmentation of sleep and wake episodes, which globally improved over time. A faster return to rest-activity cycle consolidation may predict enhanced brain recovery.

Effets des recuits ultra-rapides (10^5 K/s) sur la formation des siliciures métalliques en phase solide

La synthèse de siliciures métalliques sous la forme de films ultra-minces demeure un enjeu majeur en technologie CMOS. Le contrôle du budget thermique, afin de limiter la diffusion des dopants, est essentiel. Des techniques de recuit ultra-rapide sont alors couramment utilisées. Dans ce contexte, la technique de nanocalorimétrie est employée afin d'étudier, in situ, la formation en phase solide des siliciures de Ni à des taux de chauffage aussi élevés que 10^5 K/s. Des films de Ni, compris entre 9.3 et 0.3 nm sont déposés sur des calorimètres avec un substrat de a-Si ou de Si(100). Des mesures de diffraction de rayons X, balayées en température à 3 K/s, permettent de comparer les séquences de phase obtenues à bas taux de chauffage sur des échantillons de contrôle et à ultra-haut taux de chauffage sur les calorimètres. En premier lieu, il est apparu que l'emploi de calorimètres de type c-NC, munis d'une couche de 340 nm de Si(100), présente un défi majeur : un signal endothermique anormal vient fausser la mesure à haute température. Des micro-défauts au sein de la membrane de SiNx créent des courts-circuits entre la bande chauffante de Pt du calorimètre et l'échantillon métallique. Ce phénomène diminue avec l'épaisseur de l'échantillon et n'a pas d'effet en dessous de 400 °C tant que les porteurs de charge intrinsèques au Si ne sont pas activés. Il est possible de corriger la mesure de taux de chaleur en fonction de la température avec une incertitude de 12 °C. En ce qui a trait à la formation des siliciures de Ni à ultra-haut taux de chauffage, l'étude montre que la séquence de phase est modifiée. Les phases riches en m étal, Ni2Si et théta, ne sont pas détectées sur Si(100) et la cinétique de formation favorise une amorphisation en phase solide en début de réaction. Les enthalpies de formation pour les couches de Ni inférieures à 10 nm sont globalement plus élevées que dans le cas volumique, jusqu' à 66 %. De plus, les mesures calorimétriques montrent clairement un signal endothermique à haute température, témoignant de la compétition que se livrent la réaction de phase et l'agglomération de la couche. Pour les échantillons recuits a 3 K/s sur Si(100), une épaisseur critique telle que décrite par Zhang et Luo, et proche de 4 nm de Ni, est supposée. Un modèle est proposé, basé sur la difficulté de diffusion des composants entre des grains de plus en plus petits, afin d'expliquer la stabilité accrue des couches de plus en plus fines. Cette stabilité est également observée par nanocalorimétrie à travers le signal endothermique. Ce dernier se décale vers les hautes températures quand l'épaisseur du film diminue. En outre, une 2e épaisseur critique, d'environ 1 nm de Ni, est remarquée. En dessous, une seule phase semble se former au-dessus de 400 °C, supposément du NiSi2.

Phase synchronization in an array of driven Josephson junctions

We consider an array of N Josephson junctions connected in parallel and explore the condition for chaotic synchronization. It is found that the outer junctions can be synchronized while they remain uncorrelated to the inner ones when an external biasing is applied. The stability of the solution is found out for the outer junctions in the synchronization manifold. Symmetry considerations lead to a situation wherein the inner junctions can synchronize for certain values of the parameter. In the presence of a phase difference between the applied fields, all the junctions exhibit phase synchronization. It is also found that chaotic motion changes to periodic in the presence of phase differences.

Temperature and moist-stability effects on midlatitude orographic precipitation

Idealized, convection-resolving simulations of moist orographic flows are conducted to investigate the influence of temperature and moist stability on the drying ratio (DR), defined as the fraction of the impinging water mass removed as orographic precipitation. In flow past a long ridge, where most of the air rises over the barrier rather than detouring around it, DR decreases as the surface temperature (Ts) increases, even as the orographic cap cloud becomes statically unstable at higher Ts and develops embedded convection. This behaviour is explained by a few physical principles: (1) the Clausius–Clapeyron equation dictates that the normalized condensation rate decreases as the flow gets warmer, (2) the replacement of ice-phase precipitation growth with warm-rain processes decreases the efficiency by which condensate is converted to precipitation, thereby lowering precipitation efficiency, and (3) embedded convection acts more to vertically redistribute moisture than to enhance precipitation. Over an isolated mountain, the effects of (1) and (2) are counteracted by moisture deflection around the barrier, which is stronger in the colder, more stable flows.

Effects of chlorogenic acid and bovine serum albumin on the oxidative stability of low density lipoproteins in vitro

The ability of chlorogenic acid to inhibit oxidation of human low-density lipoprotein (LDL) was studied by in vitro copper-induced LDL oxidation. The effect of chlorogenic acid on the lag time before LDL oxidation increased in a dose dependent manner by up to 176% of the control value when added at concentrations of 0.25 -1.0 μM. Dose dependent increases in lag time of LDL oxidation were also observed, but at much higher concentrations, when chlorogenic acid was incubated with LDL (up to 29.7% increase in lag phase for 10 μM chlorogenic acid) or plasma (up to 16.6% increase in lag phase for 200 μM chlorogenic acid) prior to isolation of LDL, and this indicated that chlorogenic acid was able to bind, at least weakly, to LDL. Bovine serum albumin (BSA) increased the oxidative stability of LDL in the presence of chlorogenic acid. Fluorescence spectroscopy showed that chlorogenic acid binds to BSA with a binding constant of 3.88 x 104 M-1. BSA increased the antioxidant effect of chlorogenic acid, and this was attributed to copper ions binding to BSA, thereby reducing the amount of copper available for inducing lipid peroxidation.

Bode's maximum available feedback and phase margin

'Maximum Available Feedback' is Bode's term for the highest possible loop gain over a given bandwidth, with specified stability margins, in a single loop feedback system. His work using asymptotic analysis allowed Bode to develop a methodology for achieving this. However, the actual system performance differs from that specified, due to the use of asymptotic approximations, and the author[2] has described how, for instance, the actual phase margin is often much lower than required when the bandwidth is high, and proposed novel modifications to the asymptotes to address the issue. This paper gives some new analysis of such systems, showing that the method also contravenes Bode's definition of phase margin, and shows how the author's modifications can be used for different amounts of bandwidth.

Monte Carlo phase diagram for a polydisperse diblock copolymer melt

The phase diagram for an AB diblock copolymer melt with polydisperse A blocks and monodisperse B blocks is evaluated using lattice-based Monte Carlo simulations. Experiments on this system have shown that the A-block polydispersity shifts the order-order transitions (OOTs) towards higher A-monomer content, while the order-disorder transition (ODT) moves towards higher temperatures when the A blocks form the minority domains and lower temperatures when the A blocks form the matrix. Although self-consistent field theory (SCFT) correctly accounts for the change in the OOTs, it incorrectly predicts the ODT to shift towards higher temperatures at all diblock copolymer compositions. In contrast, our simulations predict the correct shifts for both the OOTs and the ODT. This implies that polydispersity amplifies the fluctuation-induced correction to the mean-field ODT, which we attribute to a reduction in packing frustration. Consistent with this explanation, polydispersity is found to enhance the stability of the perforated-lamellar phase.

Stability of dobutamine 500 mg in 50 ml syringes prepared using a Central Intravenous Additive Service

Objectives A pharmacy Central Intravenous Additives Service (CIVAS) provides ready to use injectable medicines. However, manipulation of a licensed injectable medicine may significantly alter the stability of drug(s) in the final product. The aim of this study was to develop a stability indicating assay for CIVAS produced dobutamine 500 mg in 50 ml dextrose 1% (w/v) prefilled syringes, and to allocate a suitable shelf life. Methods A stability indicating high performance liquid chromatography (HPLC) assay was established for dobutamine. The stability of dobutamine prefilled syringes was evaluated under storage conditions of 4°C (protected from light), room temperature (protected from light), room temperature (exposed to light) and 40°C (protected from light) at various time points (up to 42 days). Results An HPLC method employing a Hypersil column, mobile phase (pH=4.0) consisting of 82:12:6 (v/v/v) 0.05 M KH2PO4:acetonitrile:methanol plus 0.3% (v/v) triethylamine with UV detection at λ=280 nm was specific for dobutamine. Under different storage conditions only samples stored at 40°C showed greater than 5% degradation (5.08%) at 42 days and had the shortest T95% based on this criterion (44.6 days compared with 111.4 days for 4°C). Exposure to light also reduced dobutamine stability. Discolouration on storage was the limiting factor in shelf life allocation, even when dobutamine remained within 5% of the initial concentration. Conclusions A stability indicating HPLC assay for dobutamine was developed. The shelf life recommended for the CIVAS product was 42 days at 4°C and 35 days at room temperature when protected from light.

Sea surface temperature datasets for climate applications from Phase 1 of the European Space Agency Climate Change Initiative (SST CCI)

Sea surface temperature (SST) datasets have been generated from satellite observations for the period 1991–2010, intended for use in climate science applications. Attributes of the datasets specifically relevant to climate applications are: first, independence from in situ observations; second, effort to ensure homogeneity and stability through the time-series; third, context-specific uncertainty estimates attached to each SST value; and, fourth, provision of estimates of both skin SST (the fundamental measure- ment, relevant to air-sea fluxes) and SST at standard depth and local time (partly model mediated, enabling comparison with his- torical in situ datasets). These attributes in part reflect requirements solicited from climate data users prior to and during the project. Datasets consisting of SSTs on satellite swaths are derived from the Along-Track Scanning Radiometers (ATSRs) and Advanced Very High Resolution Radiometers (AVHRRs). These are then used as sole SST inputs to a daily, spatially complete, analysis SST product, with a latitude-longitude resolution of 0.05°C and good discrimination of ocean surface thermal features. A product user guide is available, linking to reports describing the datasets’ algorithmic basis, validation results, format, uncer- tainty information and experimental use in trial climate applications. Future versions of the datasets will span at least 1982–2015, better addressing the need in many climate applications for stable records of global SST that are at least 30 years in length.

Biomineralisation by earthworms: an investigation into the stability and distribution of amorphous calcium carbonate

Background Many biominerals form from amorphous calcium carbonate (ACC), but this phase is highly unstable when synthesised in its pure form inorganically. Several species of earthworm secrete calcium carbonate granules which contain highly stable ACC. We analysed the milky fluid from which granules form and solid granules for amino acid (by liquid chromatography) and functional group (by Fourier transform infrared (FTIR) spectroscopy) compositions. Granule elemental composition was determined using inductively coupled plasma-optical emission spectroscopy (ICP-OES) and electron microprobe analysis (EMPA). Mass of ACC present in solid granules was quantified using FTIR and compared to granule elemental and amino acid compositions. Bulk analysis of granules was of powdered bulk material. Spatially resolved analysis was of thin sections of granules using synchrotron-based μ-FTIR and EMPA electron microprobe analysis. Results The milky fluid from which granules form is amino acid-rich (≤ 136 ± 3 nmol mg−1 (n = 3; ± std dev) per individual amino acid); the CaCO3 phase present is ACC. Even four years after production, granules contain ACC. No correlation exists between mass of ACC present and granule elemental composition. Granule amino acid concentrations correlate well with ACC content (r ≥ 0.7, p ≤ 0.05) consistent with a role for amino acids (or the proteins they make up) in ACC stabilisation. Intra-granule variation in ACC (RSD = 16%) and amino acid concentration (RSD = 22–35%) was high for granules produced by the same earthworm. Maps of ACC distribution produced using synchrotron-based μ-FTIR mapping of granule thin sections and the relative intensity of the ν2: ν4 peak ratio, cluster analysis and component regression using ACC and calcite standards showed similar spatial distributions of likely ACC-rich and calcite-rich areas. We could not identify organic peaks in the μ-FTIR spectra and thus could not determine whether ACC-rich domains also had relatively high amino acid concentrations. No correlation exists between ACC distribution and elemental concentrations determined by EMPA. Conclusions ACC present in earthworm CaCO3 granules is highly stable. Our results suggest a role for amino acids (or proteins) in this stability. We see no evidence for stabilisation of ACC by incorporation of inorganic components.

New insights into the compressibility and high-pressure stability of Ni(CN)2: a combined study of neutron diffraction, Raman spectroscopy, and inelastic neutron scattering

Nickel cyanide is a layered material showing markedly anisotropic behaviour. High-pressure neutron diffraction measurements show that at pressures up to 20.1 kbar, compressibility is much higher in the direction perpendicular to the layers, c, than in the plane of the strongly chemically bonded metal-cyanide sheets. Detailed examination of the behaviour of the tetragonal lattice parameters, a and c, as a function of pressure reveal regions in which large changes in slope occur, for example, in c(P) at 1 kbar. The experimental pressure dependence of the volume data is fitted to a bulk modulus, B0, of 1050 (20) kbar over the pressure range 0–1 kbar, and to 124 (2) kbar over the range 1–20.1 kbar. Raman spectroscopy measurements yield additional information on how the structure and bonding in the Ni(CN)2 layers change with pressure and show that a phase change occurs at about 1 kbar. The new high-pressure phase, (Phase PII), has ordered cyanide groups with sheets of D4h symmetry containing Ni(CN)4 and Ni(NC)4 groups. The Raman spectrum of phase PII closely resembles that of the related layered compound, Cu1/2Ni1/2(CN)2, which has previously been shown to contain ordered C≡N groups. The phase change, PI to PII, is also observed in inelastic neutron scattering studies which show significant changes occurring in the phonon spectra as the pressure is raised from 0.3 to 1.5 kbar. These changes reflect the large reduction in the interlayer spacing which occurs as Phase PI transforms to Phase PII and the consequent increase in difficulty for out-of-plane atomic motions. Unlike other cyanide materials e.g. Zn(CN)2 and Ag3Co(CN)6, which show an amorphization and/or a decomposition at much lower pressures (~100 kbar), Ni(CN)2 can be recovered after pressurising to 200 kbar, albeit in a more ordered form.

The effect of homogenisation on the stability of pineapple pulp

P>Pineapple pulp was homogenised at different pressures, and its stability investigated by way of flow curves, particle size distribution, morphology, cloudiness and sedimentation. The particle size of the homogenised pulp ranged from 400 to 100 mu m for homogenisation pressures of between 0 and 700 bar. The pineapple pulp showed shear thinning behaviour with increasing flow index (n) after processing at higher pressures. In addition, the pulps with smaller particles showed less serum cloudiness, even though the sedimentation tests showed the highest stability for pulp homogenised between 200 and 300 bar. Above 400 bar, the pulp showed phase separation and higher sedimentation indexes, similar to that observed for the untreated samples, which was attributed to the formation of aggregates because of interparticle attraction.

Phase transitions of frozen camu-camu (Myrciaria dubia (HBK) McVaugh) pulp: Effect of cryostabilizer addition

Camu-camu is a tropical fruit with very high vitamin C content and commercialized as frozen pulp. Enthalpies of freezing, temperatures of the onset of ice melting, and glass transition temperatures of the maximally freeze-concentrated phase (T`(g)) of camu-camu pulp and of samples containing maltodextrin (DE20) and sucrose were measured by differential scanning calorimetry. Maltodextrin exhibited the largest freeze stabilization potential, increasing T`(g) from -58.2 degrees C (natural pulp) to -39.6 degrees C when 30% (w/w) maltodextrin DE 20 was added. Sucrose showed negligible effect on T`(g) but enhanced considerably the freezing point depression and less amount of ice was formed.

Ab initio calculation of the BCC Fe-Al-Mo (Iron-Aluminum-Molybdenum) phase diagram: Implications for the nature of the tau(2) phase

The metastable phase diagram of the BCC-based ordering equilibria in the Fe-Al-Mo system has been calculated via a truncated cluster expansion, through the combination of Full-Potential-Linear augmented Plane Wave (FP-LAPW) electronic structure calculations and of Cluster Variation Method (CVM) thermodynamic calculations in the irregular tetrahedron approximation. Four isothermal sections at 1750 K, 2000 K, 2250 K and 2500 K are calculated and correlated with recently published experimental data on the system. The results confirm that the critical temperature for the order-disorder equilibrium between Fe(3)Al-D0(3) and FeAl-B2 is increased by Mo additions, while the critical temperature for the FeAl-B2/A2 equilibrium is kept approximately invariant with increasing Mo contents. The stabilization of the Al-rich A2 phase in equilibrium with overstoichiometric B2-(Fe,Mo)Al is also consistent with the attribution of the A2 structure to the tau(2) phase, stable at high temperatures in overstoichiometric B2-FeAl. (C) 2009 Elsevier Ltd. All rights reserved.