Phase separation of p53 precedes aggregation and is affected by oncogenic mutations and ligands

Mutant p53 tends to form aggregates with amyloid properties, especially amyloid oligomers inside the nucleus, which are believed to cause oncogenic gain-of-function (GoF). The mechanism of the formation of the aggregates in the nucleus remains uncertain. The present study demonstrated that the DNA-binding domain of p53 (p53C) underwent phase separation (PS) on the pathway to aggregation under various conditions. p53C phase separated in the presence of the crowding agent polyethylene glycol (PEG). Similarly, mutant p53C (M237I and R249S) underwent PS; however, the process evolved to a solid-like phase transition faster than that in the case of wild-type p53C. The data obtained by microscopy of live cells indicated that transfection of mutant full-length p53 into the cells tended to result in PS and phase transition (PT) in the nuclear compartments, which are likely the cause of the GoF effects. Fluorescence recovery after photobleaching (FRAP) experiments revealed liquid characteristics of the condensates in the nucleus. Mutant p53 tended to undergo gel- and solid-like phase transitions in the nucleus and in nuclear bodies demonstrated by slow and incomplete recovery of fluorescence after photobleaching. Polyanions, such as heparin and RNA, were able to modulate PS and PT in vitro. Heparin apparently stabilized the condensates in a gel-like state, and RNA apparently induced a solid-like state of the protein even in the absence of PEG. Conditions that destabilize p53C into a molten globule conformation also produced liquid droplets in the absence of crowding. The disordered transactivation domain (TAD) modulated both phase separation and amyloid aggregation. In summary, our data provide mechanistic insight into the formation of p53 condensates and conditions that may result in the formation of aggregated structures, such as mutant amyloid oligomers, in cancer. The pathway of mutant p53 from liquid droplets to gel-like and solid-like (amyloid) species may be a suitable target for anticancer therapy.

Mutant p53 tends to form aggregates with amyloid properties, especially amyloid oligomers inside the nucleus, which are believed to cause oncogenic gain-of-function (GoF).     

The LHCf experiment at LHC

The LHCf experiment will be installed in 2007 on the LHC collider in the forward direction at ±140m from the ATLAS interaction point. The purpose of LHCf is to precisely measure the pion production cross section near zero degrees through the measurement of the photons produced in neutral pion decay. This measurement is crucial for the simulation of the showers induced in the atmosphere by very high energy cosmic rays; the 14 TeV energy available in the center of mass frame corresponds in fact to an equivalent energy of 10¹⁷ eV in the laboratory system. The paper focus on the proposed experiment and on the physics results that we expect from it.     

Solar modulation of cosmic deuteron fluxes in the PAMELA experiment

The preliminary results from measurements of deuteron fluxes in galactic cosmic rays (GCR) in the vicinity of the Earth in 2006–2009 are presented. The results are obtained by analyzing data from the PAMELA experiment aboard the Resurs DK-1 satellite. High-precision detection instruments provided an opportunity to identify GCR deuterons and measure their spectrum in the energy interval of 90–650MeV/nucleon. Spectra averaged over six-month intervals from the summer of 2006 to the summer of 2009 (the solar activity minimum) are presented. The influence of solar modulation on the observed spectrum is clearly seen in the results.     

Trapped Positrons and Electrons in the Inner Radiation Belt According to Data of the PAMELA Experiment

Measurements of secondary-electron and secondary-positron fluxes below the geomagnetic cutoff in near-Earth space were performed by means of the PAMELA magnetic spectrometer installed on board the Resurs-DK1 satellite launched on June 15, 2006, in an elliptical orbit of inclination 70° and altitude 350 to 600 km. This spectrometer permits measuring the fluxes of electrons and positrons over a wide energy range, as well as determining their spatial distributions to a precision of about 2°. A calculation of particle trajectories in the geomagnetic field makes it possible to separate electrons and positrons originating from cosmic-ray interactions in the Earth’s magnetosphere. The spatial distributions of quasitrapped, trapped, and short-lived albedo positrons and electrons of energy above 70 MeV in the radiation belt were analyzed. The ratio of the electron-to-positron fluxes and the energy spectra of the electrons and positrons in question are indicative of different productionmechanisms for stably trapped and quasitrapped secondary particles.     

Combined Effect of Temperature and Ternary Mobile Phase Composition on the Retention in Ternary Isocratic and Gradient Elution RP-LC under Isothermal Conditions. Application to the Retention Prediction of Four Macrolide Antibiotics

Retention models considering simultaneously ternary mobile phase organic contents and column temperature (T) were developed by a direct combination of equations expressing separately the dependence of the retention upon each of these factors. Thus, a combination of a linear dependence of the logarithm of the solute retention factor, ln k (T), against 1/T, i.e. ln k (T) = a + b/T, and of either a linear or a quadratic dependence of ln k upon the compositions of a binary mobile phase, gave two different expressions for the logarithm of the solute retention factor in terms of both temperature and organic contents in the ternary mobile phase. The effectiveness of the above models was tested in the prediction of isothermal retention of a mixture of four macrolide antibiotics under ternary isocratic and gradient elution conditions using mobile phases modified by methanol and acetonitrile. The limiting case of using ternary eluents with constant ratio of the concentrations of the two organic modifiers was also tested.     

Direct RP‐HPLC determination of underivatized amino acids with online dual UV absorbance,fluorescence, and multiple electrochemical detection

The combined use of a dual‐UV detector, a fluorimetric one and of a multiple electrochemical (EC) detector equipped with a dual electrode, consisting of a conventional size 3 mm diameter glassy carbon electrode (GCE) and of a pair of 30 μm thick carbon microfibers, is proposed for the determination of 15 amino acids, two dipeptides and creatinine. This online coupling of the above detection modes could partially replace amino acid analysis by derivatization methods, since it solves problems concerning the direct detection of selected underivatized amino acids. Additionally, it was proved that the use of multiple‐detection allows positive peak identification in a single chromatographic run, yields more information for free amino acids and solves in some cases the problem of chromatographic resolution. In order to optimize the detection conditions of the underivatized amino acids and related compounds by different detectors, their detection characteristics were determined by adequate preliminary experiments. The electro‐oxidation characteristics of the underivatized compounds of interest were determined by hydrodynamic voltammetry using a flow cell with a macrodisc GCE and by ex‐situ voltammetry using both a GCE of conventional size and a carbon fiber disk microelectrode. Important practical advantages of microfiber and microdisk electrodes with respect to macroelectrodes were demonstrated.     

Delay aversion but preference for large and rare rewards in two choice tasks: implications for the measurement of self-control parameters

Background  

Impulsivity is defined as intolerance/aversion to waiting for reward. In intolerance-to-delay (ID) protocols, animals must choose between small/soon (SS) versus large/late (LL) rewards. In the probabilistic discount (PD) protocols, animals are faced with choice between small/sure (SS) versus large/luck-linked (LLL) rewards. It has been suggested that PD protocols also measure impulsivity, however, a clear dissociation has been reported between delay and probability discounting.