Structure of the mammalian ribosome as it decodes the selenocysteine UGA codon

The elongation of eukaryotic selenoproteins relies on a poorly understood process of interpreting in-frame UGA stop codons as selenocysteine (Sec). We used cryo-electron microscopy to visualize Sec UGA recoding in mammals. A complex between the noncoding Sec-insertion sequence (SECIS), SECIS-binding protein 2 (SBP2), and 40S ribosomal subunit enables Sec-specific elongation factor eEFSec to deliver Sec. eEFSec and SBP2 do not interact directly but rather deploy their carboxyl-terminal domains to engage with the opposite ends of the SECIS. By using its Lys-rich and carboxyl-terminal segments, the ribosomal protein eS31 simultaneously interacts with Sec-specific transfer RNA (tRNASec) and SBP2, which further stabilizes the assembly. eEFSec is indiscriminate toward l-serine and facilitates its misincorporation at Sec UGA codons. Our results support a fundamentally distinct mechanism of Sec UGA recoding in eukaryotes from that in bacteria

Space- and time-resolved observation of extreme laser frequency upshifting during ultrafast-ionization

A 65-fs, 800-nm, 2-TW laser pulse propagating through a nitrogen gas jet has been experimentally studied by 90 Thomson scattering. Time-integrated spectra of scattered light show unprecedented broadening towards the blue which exceeds 300 nm. Images of the scattering region provide for the first time a space- and time-resolved description of the process leading quite regularly to such a large upshift. The mean shifting rate was as high as dk/dt3A ̊/fs, never observed before. Interferometry shows that it occurs after partial laser defocusing. Numerical simulations prove that such an upshift is consistent with a laser-gas late interaction, when laser intensity has decreased well below relativistic values (a0 1) and ionization process involves most of the laser pulse. This kind of interaction makes spectral tuning of ultrashort intense laser pulses possible in a large spectral range. VC 2013 AIP Publishing LLC. [http://dx.doi.org/10.1063/1.4818602

Anti-PF4 immunothrombosis without proximate heparin or adenovirus vector vaccine exposure.

Platelet-activating anti-platelet factor 4 (PF4)/heparin antibodies and anti-PF4 antibodies cause heparin-induced thrombocytopenia (HIT) and vaccine-induced immune thrombocytopenia and thrombosis (VITT), respectively. Diagnostic and treatment considerations differ somewhat between HIT and VITT. We identified patients with thrombocytopenia and thrombosis without proximate heparin exposure or adenovirus-based vaccination who tested strongly positive by PF4/polyanion enzyme-immunoassays and negative/weakly positive by heparin-induced platelet activation (HIPA) test but strongly positive by PF4-induced platelet activation (PIPA) test (ie, VITT-like profile). We tested these patients by a standard chemiluminescence assay that detects anti-PF4/heparin antibodies found in HIT (HemosIL AcuStar HIT-IgG(PF4-H)) as well as a novel chemiluminescence assay for anti-PF4 antibodies found in VITT. Representative control sera included an exploratory anti-PF4 antibody-positive but HIPA-negative/weak cohort obtained before 2020 (n = 188). We identified 9 patients with a clinical-pathological profile of a VITT-like disorder in the absence of proximate heparin or vaccination, with a high frequency of stroke (arterial, n = 3; cerebral venous sinus thrombosis, n = 4), thrombocytopenia (median platelet count nadir, 49 × 109/L), and hypercoagulability (greatly elevated D-dimer levels). VITT-like serological features included strong reactivity by PIPA (aggregation <10 minutes in 9/9 sera) and positive testing in the novel anti-PF4 chemiluminescence assay (3/9 also tested positive in the anti-PF4/heparin chemiluminescence assay). Our exploratory cohort identified 13 additional patient sera obtained before 2020 with VITT-like anti-PF4 antibodies. Platelet-activating VITT-like anti-PF4 antibodies should be considered in patients with thrombocytopenia, thrombosis, and very high D-dimer levels, even without a proximate exposure to heparin or adenovirus vector vaccines

Plasma density profile reconstruction of a gas cell for Ionization Induced Laser Wakefield Acceleration

Laser-driven plasma wakefields can provide hundreds of MeV electron beam in mm-range distances potentially shrinking the dimension of the actual particle accelerators. The plasma density plays a fundamental role in the control and stability of the acceleration process, which is a key development for the future electron injector proposed by EuPRAXIA. A gas cell was designed by LPGP and LIDYL teams, with variable length and backing pressure, to confine the gas and tailor the gas density profile before the arrival of the laser. This cell was used during an experimental campaign with the multi TW-class laser at the Lund Laser Centre. Ionization assisted injection in a tailored density profile is used to tune the electron beam properties. During the experiment, we filled the gas cell with hydrogen mixed with different concentration of nitrogen. We also varied the backing pressure of the gas and the geometrical length of the gas cell. We used a transverse probe to acquire shadowgraphic images of the plasma and to measure the plasma electron density. Methods and results of the analysis with comparisons between shadowgraphic and interferometric images will be discussed

Non-adiabatic cluster expansion after ultrashort laser interaction

AbstractWe used X-ray spectroscopy as a diagnostic tool for investigating the properties of laser-cluster interactions at the stage in which non-adiabatic cluster expansion takes place and a quasi-homogeneous plasma is produced. The experiment was carried out with a 10 TW, 65 fs Ti:Sa laser focused on CO2 cluster jets. The effect of different laser-pulse contrast ratios and cluster concentrations was investigated. The X-ray emission associated to the Rydberg transitions allowed us to retrieve, through the density and temperature of the emitting plasma, the time after the beginning of the interaction at which the emission occurred. The comparison of this value with the estimated time for the "homogeneous" plasma formation shows that the degree of adiabaticity depends on both the cluster concentration and the pulse contrast. Interferometric measurements support the X-ray data concerning the plasma electron density

Constraints on Charon's Orbital Elements from the Double Stellar Occultation of 2008 June 22

The original publication is available at http://iopscience.iop.org/1538-3881/International audiencePluto and its main satellite, Charon, occulted the same star on 2008 June 22. This event was observed from Australia and La Réunion Island, providing the east and north Charon Plutocentric offset in the sky plane (J2000): X= + 12,070.5 ± 4 km (+ 546.2 ± 0.2 mas), Y= + 4,576.3 ± 24 km (+ 207.1 ± 1.1 mas) at 19:20:33.82 UT on Earth, corresponding to JD 2454640.129964 at Pluto. This yields Charon's true longitude L= 153.483 ± 0fdg071 in the satellite orbital plane (counted from the ascending node on J2000 mean equator) and orbital radius r= 19,564 ± 14 km at that time. We compare this position to that predicted by (1) the orbital solution of Tholen & Buie (the "TB97" solution), (2) the PLU017 Charon ephemeris, and (3) the solution of Tholen et al. (the "T08" solution). We conclude that (1) our result rules out solution TB97, (2) our position agrees with PLU017, with differences of ΔL= + 0.073 ± 0fdg071 in longitude, and Δr= + 0.6 ± 14 km in radius, and (3) while the difference with the T08 ephemeris amounts to only ΔL= 0.033 ± 0fdg071 in longitude, it exhibits a significant radial discrepancy of Δr= 61.3 ± 14 km. We discuss this difference in terms of a possible image scale relative error of 3.35 × 10-3in the 2002-2003 Hubble Space Telescope images upon which the T08 solution is mostly based. Rescaling the T08 Charon semi-major axis, a = 19, 570.45 km, to the TB97 value, a = 19636 km, all other orbital elements remaining the same ("T08/TB97" solution), we reconcile our position with the re-scaled solution by better than 12 km (or 0.55 mas) for Charon's position in its orbital plane, thus making T08/TB97 our preferred solution

Human Umbilical Cord Blood Treatment in a Mouse Model of ALS: Optimization of Cell Dose

Amyotrophic Lateral Sclerosis (ALS) is a multicausal disease characterized by motor neuron degeneration in the spinal cord and brain. Cell therapy may be a promising new treatment for this devastating disorder. We recently showed that a single low dose (10(6) cells) of mononuclear human umbilical cord blood (MNC hUCB) cells administered intravenously to G93A mice delayed symptom progression and modestly prolonged lifespan. The aim of this pre-clinical translation study is to optimize the dose of MNC hUCB cells to retard disease progression in G93A mice. Three different doses of MNC hUCB cells, 10x10(6), 25x10(6) and 50x10(6), were administered intravenously into pre-symptomatic G93A mice. Motor function tests and various assays to determine cell effects were performed on these mice.Our results showed that a cell dose of 25x10(6) cells significantly increased lifespan of mice by 20-25% and delayed disease progression by 15%. The most beneficial effect on decreasing pro-inflammatory cytokines in the brain and spinal cord was found in this group of mice. Human Th2 cytokines were found in plasma of mice receiving 25x10(6) cells, although prevalent human Th1 cytokines were indicated in mice with 50x10(6) cells. High response of splenic cells to mitogen (PHA) was indicated in mice receiving 25x10(6) (mainly) and 10x10(6) cells. Significantly increased lymphocytes and decreased neutrophils in the peripheral blood were found only in animals receiving 25x10(6) cells. Stable reduction in microglia density in both cervical and lumbar spinal cords was also noted in mice administered with 25x10(6) cells.These results demonstrate that treatment for ALS with an appropriate dose of MNC hUCB cells may provide a neuroprotective effect for motor neurons through active involvement of these cells in modulating the host immune inflammatory system response

Identification and characterization of novel rapidly mutating Y-chromosomal short tandem repeat markers

Short tandem repeat polymorphisms on the male‐specific part of the human Y‐chromosome (Y‐STRs) are valuable tools in many areas of human genetics. Although their paternal inheritance and moderate mutation rate (~10−3 mutations per marker per meiosis) allow detecting paternal relationships, they typically fail to separate male relatives. Previously, we identified 13 Y‐STR markers with untypically high mutation rates (>10−2 ), termed rapidly mutating (RM) Y‐STRs, and showed that they improved male relative differentiation over standard Y‐STRs. By applying a newly developed in silico search approach to the Y‐chromosome reference sequence, we identified 27 novel RM Y‐STR candidates. Genotyping them in 1,616 DNA‐confirmed father–son pairs for mutation rate estimation empirically highlighted 12 novel RM Y‐STRs. Their capacity to differentiate males related by 1, 2, and 3 meioses was 27%, 47%, and 61%, respectively, while for all 25 currently known RM Y‐STRs, it was 44%, 69%, and 83%. Of the 647 Y‐STR mutations o

Functional features of gene expression profiles differentiating gastrointestinal stromal tumours according to KIT mutations and expression

<p>Abstract</p> <p>Background</p> <p>Gastrointestinal stromal tumours (GISTs) represent a heterogeneous group of tumours of mesenchymal origin characterized by gain-of-function mutations in <it>KIT </it>or <it>PDGFRA </it>of the type III receptor tyrosine kinase family. Although mutations in either receptor are thought to drive an early oncogenic event through similar pathways, two previous studies reported the mutation-specific gene expression profiles. However, their further conclusions were rather discordant. To clarify the molecular characteristics of differentially expressed genes according to GIST receptor mutations, we combined microarray-based analysis with detailed functional annotations.</p> <p>Methods</p> <p>Total RNA was isolated from 29 frozen gastric GISTs and processed for hybridization on GENECHIP^®HG-U133 Plus 2.0 microarrays (Affymetrix). <it>KIT </it>and <it>PDGFRA </it>were analyzed by sequencing, while related mRNA levels were analyzed by quantitative RT-PCR.</p> <p>Results</p> <p>Fifteen and eleven tumours possessed mutations in <it>KIT </it>and <it>PDGFRA</it>, respectively; no mutation was found in three tumours. Gene expression analysis identified no discriminative profiles associated with clinical or pathological parameters, even though expression of hundreds of genes differentiated tumour receptor mutation and expression status. Functional features of genes differentially expressed between the two groups of GISTs suggested alterations in angiogenesis and G-protein-related and calcium signalling.</p> <p>Conclusion</p> <p>Our study has identified novel molecular elements likely to be involved in receptor-dependent GIST development and allowed confirmation of previously published results. These elements may be potential therapeutic targets and novel markers of <it>KIT </it>mutation status.</p

Toward Male Individualization with Rapidly Mutating Y-Chromosomal Short Tandem Repeats

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