Development and Validation of a Fast and Homogeneous Cell-Based Fluorescence Screening Assay for Divalent Metal Transporter 1 (DMT1/SLC11A2) Using the FLIPR Tetra.

Divalent metal ion transporter 1 (DMT1) is a proton-coupled Fe(2+) transporter that is essential for iron uptake in enterocytes and for transferrin-associated endosomal iron transport in many other cell types. DMT1 dysfunction is associated with several diseases such as iron overload disorders and neurodegenerative diseases. The main objective of the present work is to develop and validate a fluorescence-based screening assay for DMT1 modulators. We found that Fe(2+) or Cd(2+) influx could be reliably monitored in calcium 5-loaded DMT1-expressing HEK293 cells using the FLIPR Tetra fluorescence microplate reader. DMT1-mediated metal transport shows saturation kinetics depending on the extracellular substrate concentration, with a K0.5 value of 1.4 µM and 3.5 µM for Fe(2+) and Cd(2+), respectively. In addition, Cd(2+) was used as a substrate for DMT1, and we find a Ki value of 2.1 µM for a compound (2-(3-carbamimidoylsulfanylmethyl-benzyl)-isothiourea) belonging to the benzylisothioureas family, which has been identified as a DMT1 inhibitor. The optimized screening method using this compound as a reference demonstrated a Z' factor of 0.51. In summary, we developed and validated a sensitive and reproducible cell-based fluorescence assay suitable for the identification of compounds that specifically modulate DMT1 transport activity.

Genome Sequence of Mycoplasma feriruminatoris sp. nov., a Fast-Growing Mycoplasma Species

Members of the “Mycoplasma mycoides cluster” represent important livestock pathogens worldwide. We report the genome sequence of Mycoplasma feriruminatoris sp. nov., the closest relative to the “Mycoplasma mycoides cluster” and the fastest-growing Mycoplasma species described to date.

Mycoplasma feriruminatoris sp. nov., a fast growing Mycoplasma species isolated from wild Caprinae

Five Mycoplasma strains from wild Caprinae were analyzed: four from Alpine ibex (Capra ibex) which died at the Berlin Zoo between 1993 and 1994, one from a Rocky Mountain goat collected in the USA prior to 1987. These five strains represented a population different from the populations belonging to the 'Mycoplasma mycoides cluster' as tested using multi locus sequence typing, Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and DNA-DNA hybridization. Analysis of the 16S rRNA gene (rrs), genomic sequence based in silico as well as laboratory DNA-DNA hybridization, and the analysis of phenotypic traits in particular their exceptionally rapid growth all confirmed that they do not belong to any Mycoplasma species described to date. We therefore suggest these strains represent a novel species, for which we propose the name Mycoplasma feriruminatoris sp. nov. The type strain is G5847(T) (=DSM 26019(T)=NCTC 1362(T)).

THE SOLAR WIND AS A POSSIBLE SOURCE OF FAST TEMPORAL VARIATIONS OF THE HELIOSPHERIC RIBBON

We present a possible source of pickup ions (PUIs) the ribbon observed by the Interstellar Boundary EXplorer (IBEX). We suggest that a gyrating solar wind and PUIs in the ramp and in the near downstream region of the termination shock (TS) could provide a significant source of energetic neutral atoms (ENAs) in the ribbon. A fraction of the solar wind and PUIs are reflected and energized during the first contact with the TS. Some of the solar wind may be reflected propagating toward the Sun but most of the solar wind ions form a gyrating beam-like distribution that persists until it is fully thermalized further downstream. Depending on the strength of the shock, these gyrating distributions can exist for many gyration periods until they are scattered/thermalized due to wave-particle interactions at the TS and downstream in the heliosheath. During this time, ENAs can be produced by charge exchange of interstellar neutral atoms with the gyrating ions. In order to determine the flux of energetic ions, we estimate the solar wind flux at the TS using pressure estimates inferred from in situ measurements. Assuming an average path length in the radial direction of the order of a few AU before the distribution of gyrating ions is thermalized, one can explain a significant fraction of the intensity of ENAs in the ribbon observed by IBEX. With a localized source and such a short integration path, this model would also allow fast time variations of the ENA flux.

Theoretical models of planetary system formation: mass vs. semi-major axis

Context. Planet formation models have been developed during the past years to try to reproduce what has been observed of both the solar system and the extrasolar planets. Some of these models have partially succeeded, but they focus on massive planets and, for the sake of simplicity, exclude planets belonging to planetary systems. However, more and more planets are now found in planetary systems. This tendency, which is a result of radial velocity, transit, and direct imaging surveys, seems to be even more pronounced for low-mass planets. These new observations require improving planet formation models, including new physics, and considering the formation of systems. Aims: In a recent series of papers, we have presented some improvements in the physics of our models, focussing in particular on the internal structure of forming planets, and on the computation of the excitation state of planetesimals and their resulting accretion rate. In this paper, we focus on the concurrent effect of the formation of more than one planet in the same protoplanetary disc and show the effect, in terms of architecture and composition of this multiplicity. Methods: We used an N-body calculation including collision detection to compute the orbital evolution of a planetary system. Moreover, we describe the effect of competition for accretion of gas and solids, as well as the effect of gravitational interactions between planets. Results: We show that the masses and semi-major axes of planets are modified by both the effect of competition and gravitational interactions. We also present the effect of the assumed number of forming planets in the same system (a free parameter of the model), as well as the effect of the inclination and eccentricity damping. We find that the fraction of ejected planets increases from nearly 0 to 8% as we change the number of embryos we seed the system with from 2 to 20 planetary embryos. Moreover, our calculations show that, when considering planets more massive than ~5 M⊕, simulations with 10 or 20 planetary embryos statistically give the same results in terms of mass function and period distribution.

Measurement of Neutrino Oscillation Parameters from Muon Neutrino Disappearance with an Off-Axis Beam

The T2K collaboration reports a precision measurement of muon neutrino disappearance with an off-axis neutrino beam with a peak energy of 0.6 GeV. Near detector measurements are used to constrain the neutrino flux and cross section parameters. The Super-Kamiokande far detector, which is 295 km downstream of the neutrino production target, collected data corresponding to 3.01×1020 protons on target. In the absence of neutrino oscillations, 205±17 (syst.) events are expected to be detected and only 58 muon neutrino event candidates are observed. A fit to the neutrino rate and energy spectrum assuming three neutrino flavors, normal mass hierarchy and θ23≤π/4 yields a best-fit mixing angle sin2(2θ23)=1.000 and mass splitting |Δm232|=2.44×10−3 eV2/c4. If θ23≥π/4 is assumed, the best-fit mixing angle changes to sin2(2θ23)=0.999 and the mass splitting remains unchanged.

A convolution model for energy transport in a therapeutic fast neutron beam

A three-dimensional model has been proposed that uses Monte Carlo and fast Fourier transform convolution techniques to calculate the dose distribution from a fast neutron beam. This method transports scattered neutrons and photons in the forward, lateral, and backward directions and protons, electrons, and positrons in the forward and lateral directions by convolving energy spread kernels with initial interaction available energy distributions. The primary neutron and photon spectrums have been derived from narrow beam attenuation measurements. The positions and strengths of the effective primary neutron, scattered neutron, and photon sources have been derived from dual ion chamber measurements. The size of the effective primary neutron source has been measured using a copper activation technique. Heterogeneous tissue calculations require a weighted sum of two convolutions for each component since the kernels must be invariant for FFT convolution. Comparisons between calculations and measurements were performed for several water and heterogeneous phantom geometries. ^

Feasibility of post mortem cardiac proton density weighted fast field echo imaging in two cases of sudden death

OBJECTIVE The aim of this work is to investigate and compare cardiac proton density (PD) weighted fast field echo (FFE) post-mortem magnetic resonance (PMMR) imaging with standard cardiac PMMR imaging (T1-weighted and T2-weighted turbo spin-echo (TSE)), postmortem CT (PMCT) as well as autopsy. MATERIALS AND METHODS Two human cadavers sequentially underwent cardiac PMCT and PMMR imaging (PD-weighted FFE, T1-weighted and T2-weighted TSE) and autopsy. The cardiac PMMR images were compared to each other as well as to PMCT and autopsy findings. RESULTS For the first case, cardiac PMMR exhibited a focal region of low signal in PD-weighted FFE and T2-weighted TSE images, surrounded by a signal intense rim in the T2-weighted images. T1-weighted TSE and PMCT did not appear to identify any focal abnormality. Macroscopic inspection identified a blood clot; histology confirmed this to be a thrombus with an adhering myocardial infarction. In the second case, a myocardial rupture with heart tamponade was identified in all PMMR images, located at the anterior wall of the left ventricle; PMCT excluded additional ruptures. In PD-weighted FFE and T2-weighted TSE images, it occurred hypo-intense, while resulting in small clustered hyper-intense spots in T1-weighted TSE. Autopsy confirmed the PMMR and PMCT findings. CONCLUSIONS Presented initial results have shown PD-weighted FFE to be a valuable imaging sequence in addition to traditional T2-weighted TSE imaging for blood clots and myocardial haemorrhage with clearer contrast between affected and healthy myocardium.

Growth and Persistence of Diverse Intertidal Crusts: Survival of the Slow in a Fast-Paced World

Encrusting algae are conspicuous components of hard-substratum benthic communities in the photic zone despite being poor competitors and slow growers. Little is known about their growth rates or about mechanisms controlling key processes such as wound healing and surviving overgrowth. We manipulated 12 crustose species (including red and brown algae and a lichen) from the intertidal zone of Washington, USA, studying their varying responses to identical experimental conditions. Three of 8 crust species tested showed increased growth rates with size. Species healed from standardized wounds at different rates and using different mechanisms (e.g. lateral vs vertical regeneration) as seen in cross-sections. Three species showed altered growth rates at unwounded margins of wounded crusts, suggesting intrathallus communication. Year-long experiments involving simulated overgrowth showed that some species can maintain healthy tissue in a covered area, and one (the coralline Lithothamnion phymatodeum) even grew new tissue there. Other species gradually lost color, thickness, and area in covered areas. Hildenbrandia occidentalis survived remarkably well when covered, possibly due to its very slow growth and low metabolic demand. One suggested mechanism underlying the high variation in responses among crusts is the degree to which their thalli may be anatomically integrated by features such as cell fusions; physiological work testing translocation via these features is needed. Other mechanisms allowing persistence include rapid wound healing and frequent recruitment.

Microwave radiometer to retrieve temperature profiles from the surface to the stratopause

TEMPERA (TEMPERature RAdiometer) is a new ground-based radiometer which measures in a frequency range from 51–57 GHz radiation emitted by the atmosphere. With this instrument it is possible to measure temperature profiles from ground to about 50 km. This is the first ground-based instrument with the capability to retrieve temperature profiles simultaneously for the troposphere and stratosphere. The measurement is done with a filterbank in combination with a digital fast Fourier transform spectrometer. A hot load and a noise diode are used as stable calibration sources. The optics consist of an off-axis parabolic mirror to collect the sky radiation. Due to the Zeeman effect on the emission lines used, the maximum height for the temperature retrieval is about 50 km. The effect is apparent in the measured spectra. The performance of TEMPERA is validated by comparison with nearby radiosonde and satellite data from the Microwave Limb Sounder on the Aura satellite. In this paper we present the design and measurement method of the instrument followed by a description of the retrieval method, together with a validation of TEMPERA data over its first year, 2012.