Blue laser cooling transitions in Tm I

We have studied possible candidates for laser cooling transitions in $^{169}$Tm in the spectral region 410 -- 420 nm. By means of saturation absorption spectroscopy we have measured the hyperfine structure and rates of two nearly closed cycling transitions from the ground state $4\textrm{f}^{13}6\textrm{s}^2(^2\textrm{F}_0)(J_g=7/2)$ to upper states $4\textrm{f}^{12}(^3\textrm{H}_5)5\textrm{d}_{3/2}6\textrm{s}^2(J_e=9/2)$ at 410.6 nm and $4\textrm{f}^{12}(^3\textrm{F}_4)5\textrm{d}_{5/2}6\textrm{s}^2(J_e=9/2)$ at 420.4 nm and evaluated the life times of the excited levels as 15.9(8) ns and 48(6) ns respectively. Decay rates from these levels to neighboring opposite-parity levels are evaluated by means of Hartree-Fock calculations. We conclude, that the strong transition at 410.6 nm has an optical leak rate of less then $2\cdot10^{-5}$ and can be used for efficient laser cooling of $^{169}$Tm from a thermal atomic beam. The hyperfine structure of two other even-parity levels which can be excited from the ground state at 409.5 nm and 418.9 nm is also measured by the same technique. In addition we give a calculated value of $7(2)$ s$^{-1}$ for the rate of magnetic-dipole transition at 1.14 $\mu$m between the fine structure levels $(J_g=7/2)\leftrightarrow(J'_g=5/2)$ of the ground state which can be considered as a candidate for applications in atomic clocks.Comment: 8 pages, 5 figure

Tracing the evolution of NGC6397 through the chemical composition of its stellar populations

With the aim to constrain multiple populations in the metal-poor globular cluster NGC6397, we analyse and discuss the chemical compositions of a large number of elements in 21 red giant branch stars. High-resolution spectra were obtained with the FLAMES/UVES spectrograph on VLT. We have determined non-LTE abundances of Na and LTE abundances for the remaining 21 elements, including O, Mg, Al, alpha, iron-peak, and neutron-capture elements, many of which have not previously been analysed for this cluster. We have also considered the influence of possible He enrichment in the analysis of stellar spectra. We find that the Na abundances of evolved, as well as unevolved, stars show a distinct bimodality, which suggests the presence of two stellar populations; one primordial stellar generation with composition similar to field stars, and a second generation that is enriched in material processed through hydrogen-burning (enriched in Na and Al and depleted in O and Mg). The cluster is dominated (75%) by the second generation. The red giant branch show a similar bimodal distribution in the Stroemgren colour index c_y=c_1-(b-y), implying a large difference also in N abundance. The two populations have the same composition of all analysed elements heavier than Al, within the measurement uncertainty of the analysis, with the possible exception of [Y/Fe]. Using two stars with close to identical stellar parameters, one from each generation, we estimate the difference in He content, Delta Y=0.01+-0.06, given the assumption that the mass fraction of iron is the same for the stars. Finally, we show that winds from fast rotating massive stars of the first generation can be held responsible for the abundance patterns observed in NGC6397 second generation long-lived stars and estimate that the initial mass of the cluster were at least ten times higher than its present-day value.Comment: 13 pages + appendix with two tables. Accepted for publication in A&A. v2: minor language corrections and Table A.2. correcte

Elevated mitochondrial genome variation after 50 generations of radiation exposure in a wild rodent

Currently, the effects of chronic, continuous low dose environmental irradiation on the mitochondrial genome of resident small mammals are unknown. Using the bank vole (Myodes glareolus) as a model system, we tested the hypothesis that approximately 50 generations of exposure to the Chernobyl environment has significantly altered genetic diversity of the mitochondrial genome. Using deep sequencing, we compared mitochondrial genomes from 131 individuals from reference sites with radioactive contamination comparable to that present in northern Ukraine before the 26 April 1986 meltdown, to populations where substantial fallout was deposited following the nuclear accident. Population genetic variables revealed significant differences among populations from contaminated and uncontaminated localities. Therefore, we rejected the null hypothesis of no significant genetic effect from 50 generations of exposure to the environment created by the Chernobyl meltdown. Samples from contaminated localities exhibited significantly higher numbers of haplotypes and polymorphic loci, elevated genetic diversity, and a significantly higher average number of substitutions per site across mitochondrial gene regions. Observed genetic variation was dominated by synonymous mutations, which may indicate a history of purify selection against nonsynonymous or insertion/deletion mutations. These significant differences were not attributable to sample size artifacts. The observed increase in mitochondrial genomic diversity in voles from radioactive sites is consistent with the possibility that chronic, continuous irradiation resulting from the Chernobyl disaster has produced an accelerated mutation rate in this species over the last 25 years. Our results, being the first to demonstrate this phenomenon in a wild mammalian species, are important for understanding genetic consequences of exposure to low-dose radiation sources. © 2017 John Wiley & Sons Ltd

Molecular basis of Lys11-polyubiquitin specificity in the deubiquitinase Cezanne

The post-translational modification of proteins with polyubiquitin regulates virtually all aspects of cell biology. Eight distinct chain linkage types in polyubiquitin co-exist and are independently regulated in cells. This ‘ubiquitin code’ determines the fate of the modified protein1. Deubiquitinating enzymes of the Ovarian Tumour (OTU) family regulate cellular signalling by targeting distinct linkage types within polyubiquitin2, and understanding their mechanisms of linkage specificity gives fundamental insights into the ubiquitin system. We here reveal how the deubiquitinase Cezanne/OTUD7B specifically targets Lys11-linked polyubiquitin. Crystal structures of Cezanne alone and in complex with mono- and Lys11-linked diubiquitin, in combination with hydrogen-deuterium exchange mass spectrometry, enable reconstruction of the enzymatic cycle in exquisite detail. An intricate mechanism of ubiquitin-assisted conformational changes activate the enzyme, and while all chain types interact with the enzymatic S1 site, only Lys11-linked chains can bind productively across the active site and stimulate catalytic turnover. Our work highlights the fascinating plasticity of deubiquitinases, and indicates that new conformational states can occur when a true substrate, such as diubiquitin, is bound at the active site

Perturbing the Ubiquitin Pathway Reveals How Mitosis Is Hijacked to Denucleate and Regulate Cell Proliferation and Differentiation In Vivo

The eye lens presents a unique opportunity to explore roles for specific molecules in cell proliferation, differentiation and development because cells remain in place throughout life and, like red blood cells and keratinocytes, they go through the most extreme differentiation, including removal of nuclei and cessation of protein synthesis. Ubiquitination controls many critical cellular processes, most of which require specific lysines on ubiquitin (Ub). Of the 7 lysines (K) least is known about effects of modification of K6.We replaced K6 with tryptophan (W) because K6 is the most readily modified K and W is the most structurally similar residue to biotin. The backbone of K6W-Ub is indistinguishable from that of Wt-Ub. K6W-Ub is effectively conjugated and deconjugated but the conjugates are not degraded via the ubiquitin proteasome pathways (UPP). Expression of K6W-ubiquitin in the lens and lens cells results in accumulation of intracellular aggregates and also slows cell proliferation and the differentiation program, including expression of lens specific proteins, differentiation of epithelial cells into fibers, achieving proper fiber cell morphology, and removal of nuclei. The latter is critical for transparency, but the mechanism by which cell nuclei are removed has remained an age old enigma. This was also solved by expressing K6W-Ub. p27(kip), a UPP substrate accumulates in lenses which express K6W-Ub. This precludes phosphorylation of nuclear lamin by the mitotic kinase, a prerequisite for disassembly of the nuclear membrane. Thus the nucleus remains intact and DNAseIIβ neither gains entry to the nucleus nor degrades the DNA. These results could not be obtained using chemical proteasome inhibitors that cannot be directed to specific tissues.K6W-Ub provides a novel, genetic means to study functions of the UPP because it can be targeted to specific cells and tissues. A fully functional UPP is required to execute most stages of lens differentiation, specifically removal of cell nuclei. In the absence of a functional UPP, small aggregate prone, cataractous lenses are formed

Inflammasome sensor NLRP1 controls rat macrophage susceptibility to Toxoplasma gondii

Toxoplasma gondii is an intracellular parasite that infects a wide range of warm-blooded species. Rats vary in their susceptibility to this parasite. The Toxo1 locus conferring Toxoplasma resistance in rats was previously mapped to a region of chromosome 10 containing Nlrp1. This gene encodes an inflammasome sensor controlling macrophage sensitivity to anthrax lethal toxin (LT) induced rapid cell death (pyroptosis). We show here that rat strain differences in Toxoplasma infected macrophage sensitivity to pyroptosis, IL-1β/IL-18 processing, and inhibition of parasite proliferation are perfectly correlated with NLRP1 sequence, while inversely correlated with sensitivity to anthrax LT-induced cell death. Using recombinant inbred rats, SNP analyses and whole transcriptome gene expression studies, we narrowed the candidate genes for control of Toxoplasma-mediated rat macrophage pyroptosis to four genes, one of which was Nlrp1. Knockdown of Nlrp1 in pyroptosis-sensitive macrophages resulted in higher parasite replication and protection from cell death. Reciprocally, overexpression of the NLRP1 variant from Toxoplasma-sensitive macrophages in pyroptosis-resistant cells led to sensitization of these resistant macrophages. Our findings reveal Toxoplasma as a novel activator of the NLRP1 inflammasome in rat macrophages

TLR2/MyD88/NF-κB Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

Human respiratory syncytial virus (RSV) constitute highly pathogenic virus that cause severe respiratory diseases in newborn, children, elderly and immuno-compromised individuals. Airway inflammation is a critical regulator of disease outcome in RSV infected hosts. Although “controlled” inflammation is required for virus clearance, aberrant and exaggerated inflammation during RSV infection results in development of inflammatory diseases like pneumonia and bronchiolitis. Interleukin-1β (IL-1β) plays an important role in inflammation by orchestrating the pro-inflammatory response. IL-1β is synthesized as an immature pro-IL-1β form. It is cleaved by activated caspase-1 to yield mature IL-1β that is secreted extracellularly. Activation of caspase-1 is mediated by a multi-protein complex known as the inflammasome. Although RSV infection results in IL-1β release, the mechanism is unknown. Here in, we have characterized the mechanism of IL-1β secretion following RSV infection. Our study revealed that NLRP3/ASC inflammasome activation is crucial for IL-1β production during RSV infection. Further studies illustrated that prior to inflammasome formation; the “first signal” constitutes activation of toll-like receptor-2 (TLR2)/MyD88/NF-κB pathway. TLR2/MyD88/NF-κB signaling is required for pro-IL-1β and NLRP3 gene expression during RSV infection. Following expression of these genes, two “second signals” are essential for triggering inflammasome activation. Intracellular reactive oxygen species (ROS) and potassium (K+) efflux due to stimulation of ATP-sensitive ion channel promote inflammasome activation following RSV infection. Thus, our studies have underscored the requirement of TLR2/MyD88/NF-κB pathway (first signal) and ROS/potassium efflux (second signal) for NLRP3/ASC inflammasome formation, leading to caspase-1 activation and subsequent IL-1β release during RSV infection

The Hamburg/ESO R-process Enhanced Star survey (HERES) III. HE 0338-3945 and the formation of the r+s stars

We have derived abundances of 33 elements and upper limits for 6 additional elements for the metal-poor ([Fe/H] = -2.42) turn-off star HE 0338-3945 from high-quality VLT-UVES spectra. The star is heavily enriched, by about a factor of 100 relative to iron and the Sun, in the heavy s-elements (Ba, La, ..). It is also heavily enriched in Eu, which is generally considered an r-element, and in other similar elements. It is less enriched, by about a factor of 10, in the lighter s-elements (Sr, Y and Zr). C is also strongly enhanced and, to a somewhat lesser degree, N and O. These abundance estimates are subject to severe uncertainties due to NLTE and thermal inhomogeneities which are not taken into detailed consideration. However, an interesting result, which is most probably robust in spite of these uncertainties, emerges: the abundances derived for this star are very similar to those of other stars with an overall enhancement of all elements beyond the iron peak. We have defined criteria for this class of stars, r+s stars, and discuss nine different scenarios to explain their origin. None of these explanations is found to be entirely convincing. The most plausible hypotheses involve a binary system in which the primary component goes through its giant branch and asymptotic giant branch phases and produces CNO and s-elements which are dumped onto the observed star. Whether the r-element Eu is produced by supernovae before the star was formed (perhaps triggering the formation of a low-mass binary), by a companion as it explodes as a supernova (possibly triggered by mass transfer), or whether it is possibly produced in a high-neutron-density version of the s-process is still unclear. Several suggestions are made on how to clarify this situation.Comment: Accepted for A&A; 22 pages, 9 figures, 2 tables. Table 2 is in electronic form and available at http://www.astro.uu.se/~karin/table2.dat with description at http://www.astro.uu.se/~karin/jonsellReadMe

Structural insights into the catalysis and regulation of E3 ubiquitin ligases

Covalent attachment (conjugation) of one or more ubiquitin molecules to protein substrates governs numerous eukaryotic cellular processes, including apoptosis, cell division and immune responses. Ubiquitylation was originally associated with protein degradation, but it is now clear that ubiquitylation also mediates processes such as protein–protein interactions and cell signalling depending on the type of ubiquitin conjugation. Ubiquitin ligases (E3s) catalyse the final step of ubiquitin conjugation by transferring ubiquitin from ubiquitin-conjugating enzymes (E2s) to substrates. In humans, more than 600 E3s contribute to determining the fates of thousands of substrates; hence, E3s need to be tightly regulated to ensure accurate substrate ubiquitylation. Recent findings illustrate how E3s function on a structural level and how they coordinate with E2s and substrates to meticulously conjugate ubiquitin. Insights regarding the mechanisms of E3 regulation, including structural aspects of their autoinhibition and activation are also emerging

Specificity and disease in the ubiquitin system

Post-translational modification (PTM) of proteins by ubiquitination is an essential cellular regulatory process. Such regulation drives the cell cycle and cell division, signalling and secretory pathways, DNA replication and repair processes and protein quality control and degradation pathways. A huge range of ubiquitin signals can be generated depending on the specificity and catalytic activity of the enzymes required for attachment of ubiquitin to a given target. As a consequence of its importance to eukaryotic life, dysfunction in the ubiquitin system leads to many disease states, including cancers and neurodegeneration. This review takes a retrospective look at our progress in understanding the molecular mechanisms that govern the specificity of ubiquitin conjugation