Watching gene expression in color.

A combination of two fluorescent proteins with different half-lives allows gene expression to be followed with improved time resolution

ARPES studies of cuprate Fermiology: superconductivity, pseudogap, and quasiparticle dynamics

We present angle-resolved photoemission spectroscopy (ARPES) studies of the cuprate high-temperature superconductors which elucidate the relation between superconductivity and the pseudogap and highlight low-energy quasiparticle dynamics in the superconducting state. Our experiments suggest that the pseudogap and superconducting gap represent distinct states, which coexist below T$_c$. Studies on Bi-2212 demonstrate that the near-nodal and near-antinodal regions behave differently as a function of temperature and doping, implying that different orders dominate in different momentum-space regions. However, the ubiquity of sharp quasiparticles all around the Fermi surface in Bi-2212 indicates that superconductivity extends into the momentum-space region dominated by the pseudogap, revealing subtlety in this dichotomy. In Bi-2201, the temperature dependence of antinodal spectra reveals particle-hole asymmetry and anomalous spectral broadening, which may constrain the explanation for the pseudogap. Recognizing that electron-boson coupling is an important aspect of cuprate physics, we close with a discussion of the multiple 'kinks' in the nodal dispersion. Understanding these may be important to establishing which excitations are important to superconductivity.Comment: To appear in a focus issue on 'Fermiology of Cuprates' in New Journal of Physic

Small extracellular vesicles secreted from human amniotic fluid mesenchymal stromal cells possess cardioprotective and promigratory potential

Mesenchymal stromal cells (MSCs) exhibit antiapoptotic and proangiogenic functions in models of myocardial infarction which may be mediated by secreted small extracellular vesicles (sEVs). However, MSCs have frequently been harvested from aged or diseased patients, while the isolated sEVs often contain high levels of impurities. Here, we studied the cardioprotective and proangiogenic activities of size-exclusion chromatography-purified sEVs secreted from human foetal amniotic fluid stem cells (SS-hAFSCs), possessing superior functional potential to that of adult MSCs. We demonstrated for the first time that highly pure (up to 1.7 × 1010 particles/µg protein) and thoroughly characterised SS-hAFSC sEVs protect rat hearts from ischaemia–reperfusion injury in vivo when administered intravenously prior to reperfusion (38 ± 9% infarct size reduction, p < 0.05). SS-hAFSC sEVs did not protect isolated primary cardiomyocytes in models of simulated ischaemia–reperfusion injury in vitro, indicative of indirect cardioprotective effects. SS-hAFSC sEVs were not proangiogenic in vitro, although they markedly stimulated endothelial cell migration. Additionally, sEVs were entirely responsible for the promigratory effects of the medium conditioned by SS-hAFSC. Mechanistically, sEV-induced chemotaxis involved phosphatidylinositol 3-kinase (PI3K) signalling, as its pharmacological inhibition in treated endothelial cells reduced migration by 54 ± 7% (p < 0.001). Together, these data indicate that SS-hAFSC sEVs have multifactorial beneficial effects in a myocardial infarction setting

Robust inference of neutron-star parameters from thermonuclear burst observations

Thermonuclear (type-I) bursts arise from unstable ignition of accumulated fuel on the surface of neutron stars in low-mass X-ray binaries. Measurements of burst properties in principle enable observers to infer the properties of the host neutron star and mass donors, but a number of confounding astrophysical effects contribute to systematic uncertainties. Here we describe some commonly-used approaches for determining system parameters, including composition of the burst fuel, and introduce a new suite of software tools, concord, intended to fully account for astrophysical uncertainties. Comparison of observed burst properties with the predictions of numerical models is a complementary method of constraining host properties, and the tools presented here are intended to make comprehensive model-observation comparisons straightforward. When combined with the extensive samples of burst observations accumulated by X-ray observatories, these software tools will provide a step-change in the amount of information that can be inferred about typical burst sources.Comment: 22 pages, 11 figures, 1 table & 1 machine-readable table as supplementary data; submitted to ApJS. Accompanying software package available at https://github.com/outs1der/concor

Cryo-electron microscopy structure of the di-domain core of <em>Mycobacterium tuberculosis</em> polyketide synthase 13, essential for mycobacterial mycolic acid synthesis

Mycobacteria are known for their complex cell wall, which comprises layers of peptidoglycan, polysaccharides and unusual fatty acids known as mycolic acids that form their unique outer membrane. Polyketide synthase 13 (Pks13) of Mycobacterium tuberculosis, the bacterial organism causing tuberculosis, catalyses the last step of mycolic acid synthesis prior to export to and assembly in the cell wall. Due to its essentiality, Pks13 is a target for several novel anti-tubercular inhibitors, but its 3D structure and catalytic reaction mechanism remain to be fully elucidated. Here, we report the molecular structure of the catalytic core domains of M. tuberculosis Pks13 (Mt-Pks13), determined by transmission cryo-electron microscopy (cryoEM) to a resolution of 3.4 \uc5. We observed a homodimeric assembly comprising the ketoacyl synthase (KS) domain at the centre, mediating dimerization, and the acyltransferase (AT) domains protruding in opposite directions from the central KS domain dimer. In addition to the KS–AT di-domains, the cryoEM map includes features not covered by the di-domain structural model that we predicted to contain a dimeric domain similar to dehydratases, yet likely lacking catalytic function. Analytical ultracentrifugation data indicate a pH-dependent equilibrium between monomeric and dimeric assembly states, while comparison with the previously determined structures of M. smegmatis Pks13 indicates architectural flexibility. Combining the experimentally determined structure with modelling in AlphaFold2 suggests a structural scaffold with a relatively stable dimeric core, which combines with considerable conformational flexibility to facilitate the successive steps of the Claisen-type condensation reaction catalysed by Pks13

Integrable models and quantum spin ladders: comparison between theory and experiment for the strong coupling ladder compounds

(abbreviated) This article considers recent advances in the investigation of the thermal and magnetic properties of integrable spin ladder models and their applicability to the physics of real compounds. The ground state properties of the integrable two-leg spin-1/2 and the mixed spin-(1/2,1) ladder models at zero temperature are analyzed by means of the Thermodynamic Bethe Ansatz. Solving the TBA equations yields exact results for the critical fields and critical behaviour. The thermal and magnetic properties of the models are investigated in terms of the recently introduced High Temperature Expansion method, which is discussed in detail. It is shown that in the strong coupling limit the integrable spin-1/2 ladder model exhibits three quantum phases: (i) a gapped phase in the regime $H<H_{c1}$, (ii) a fully polarised phase for $H>H_{c2}$, and (iii) a Luttinger liquid magnetic phase in the regime $H_{c1}<H<H_{c2}$. The critical behaviour in the vicinity of the critical points is of the Pokrovsky-Talapov type. The temperature-dependent thermal and magnetic properties are directly evaluated from the exact free energy expression and compared to known experimental results for a range of strong coupling ladder compounds. Similar analysis of the mixed spin-(1/2,1) ladder model reveals a rich phase diagram, with a 1/3 and a full saturation magnetisation plateau within the strong antiferromagnetic rung coupling regime. For weak rung coupling, the fractional magnetisation plateau is diminished and a new quantum phase transition occurs. The phase diagram can be directly deduced from the magnetisation curve obtained from the exact result derived from the HTE. The thermodynamics of the spin-orbital model with different single-ion anisotropies is also investigated.Comment: 90 pages, 33 figures, extensive revisio

Cellular uptake of ribonuclease A-functionalised core-shell silica microspheres

Analysis of protein function in a cellular context ideally requires physiologically representative levels of that protein. Thus conventional nucleic acid-based transfection methods are far from ideal owing to the over expression that generally results. Likewise fusions with protein transduction domains can be problematic whilst delivery via liposomes/nanoparticles typically results in endosomal localisation. Recently polymer microspheres have been reported to be highly effective at delivering proteins into cells and thus provide a viable new alternative for protein delivery (protein transduction). Herein we describe the successful delivery of active ribonuclease A into HeLa cells via novel polymer core-silica shell microspheres. Specifically, poly(styrene-co-vinylbenzylisothiouronium chloride) core particles, generated by dispersion polymerisation, were coated with a poly(styrene-co-trimethoxysilylpropyl methacrylate) shell. The resultant core-shell morphology was characterised by transmission electron, scanning electron and fluorescence confocal microscopies, whilst size and surface charge was assessed by dynamic light scattering and zeta-potential measurements, respectively. Subsequently ribonuclease A was coupled to the microspheres using simple carbodiimide chemistry. Gel electrophoresis confirmed and quantified the activity of the immobilised enzyme against purified HeLa RNA. Finally, the polymer-protein particles were evaluated as protein-transduction vectors in vitro to deliver active ribonuclease A to HeLa cells. Cellular uptake of the microspheres was successful and resulted in reduced levels of both intracellular RNA and cell viability

Evidence for reduced immune gene diversity and activity during the evolution of termites

The evolution of biological complexity is associated with the emergence of bespoke immune systems that maintain and protect organism integrity. Unlike the well-studied immune systems of cells and individuals, little is known about the origins of immunity during the transition to eusociality, a major evolutionary transition comparable to the evolution of multicellular organisms from single-celled ancestors. We aimed to tackle this by characterizing the immune gene repertoire of 18 cockroach and termite species, spanning the spectrum of solitary, subsocial and eusocial lifestyles. We find that key transitions in termite sociality are correlated with immune gene family contractions. In cross-species comparisons of immune gene expression, we find evidence for a caste-specific social defence system in termites, which appears to operate at the expense of individual immune protection. Our study indicates that a major transition in organismal complexity may have entailed a fundamental reshaping of the immune system optimized for group over individual defence

IgH gene rearrangements as plasma biomarkers in Non-Hodgkin's Lymphoma patients

New biomarkers with improved accuracy could be helpful for monitoring disease in patients with Non-Hodgkin's lymphomas (NHL). Towards this end, we have explored the feasibility of identifying the sequence of rearranged IgH genes using next-generation sequencing, then using PCR to detect specific rearranged DNA fragments in patients' plasma. By capturing and sequencing the IgH genomic regions (IgCap), we were able to detect and precisely determine the sequence of rearranged IgH loci in the tumors of three NHL patients. Moreover, circulating rearranged DNA fragments could be identified in the plasma of all three patients. Even in cases wherein tumor biopsies were unavailable, we were able to use the IgH capture approach to identify rearranged DNA loci in the plasma of 8 of 14 patients. IgCap may enable a more informed management of selected patients with NHL and other B-cell malignancies in the future

A School-Based Environmental Intervention to Reduce Smoking among High School Students: The Acadiana Coalition of Teens against Tobacco (ACTT)

A school-based environmental program to reduce adolescent smoking was conducted in 20 schools (10 intervention; 10 control) in south central Louisiana. The 9th grade cohort (n = 4,763; mean age = 15.4 yrs; 51% female; 61% Caucasian; 30-day smoking prevalence at baseline = 25%) was followed over four years for 30-day smoking prevalence with the school as the unit of analysis. Although prevalence decreased in intervention schools and increased in control schools in Year 2 the significant difference between the two groups at baseline was not overcome by the intervention and increases in prevalence were observed in both groups in Years 3 and 4. The higher the percentage of white students in a school the higher the prevalence rates regardless of intervention/control status. Boys’ and girls’ smoking rates were similar. These outcome data, student feedback and process evaluation provide a basis for continuing to create more innovative adolescent tobacco control programs