Chaperone-mediated native folding of a β-scorpion toxin in the periplasm of E.coli

Background: Animal neurotoxin peptides are valuable probes for investigating ion channel structure/function relationships and represent lead compounds for novel therapeutics and insecticides. However, misfolding and aggregation are common outcomes when toxins containing multiple disulfides are expressed in bacteria. Methods: The ß-scorpion peptide toxin Bj-xtrIT from Hottentotta judaica and four chaperone enzymes (DsbA, DsbC, SurA and FkpA) were co-secreted into the oxidizing environment of the E.coli periplasm. Expressed Bj-xtrIT was purified and analyzed by HPLC and FPLC chromatography. Its thermostability was assessed using synchrotron radiation circular dichroism spectroscopy and its crystal structure was determined. Results: Western blot analysis showed that robust expression was only achieved when cells co-expressed the chaperones. The purified samples were homogenous and monodisperse and the protein was thermostable. The crystal structure of the recombinant toxin confirmed that it adopts the native disulfide connectivity and fold. Conclusions: The chaperones enabled correct folding of the four-disulfide-bridged Bj-xtrIT toxin. There was no apparent sub-population of misfolded Bj-xtrIT, which attests to the effectiveness of this expression method. General Significance: We report the first example of a disulfide-linked scorpion toxin natively folded during bacterial expression. This method eliminates downstream processing steps such as oxidative refolding or cleavage of a fusion-carrier and therefore enables efficient production of insecticidal Bj-xtrIT. Periplasmic chaperone activity may produce native folding of other extensively disulfide-reticulated proteins including animal neurotoxins. This work is therefore relevant to venomics and studies of a wide range of channels and receptors

Folding factors and partners for the intrinsically disordered protein Micro-Exon Gene 14 (MEG-14)

The micro-exon genes (MEG) of Schistosoma mansoni, a parasite responsible for the second most widely spread tropical disease, code for small secreted proteins with sequences unique to the Schistosoma genera. Bioinformatics analyses suggest the soluble domain of the MEG-14 protein will be largely disordered, and using synchrotron radiation circular dichroism spectroscopy, its secondary structure was shown to be essentially completely unfolded in aqueous solution. It does, however, show a strong propensity to fold into more ordered structures under a wide range of conditions. Partial folding was produced by increasing temperature (in a reversible process), contrary to the behavior of most soluble proteins. Furthermore, significant folding was observed in the presence of negatively charged lipids and detergents, but not in zwitterionic or neutral lipids or detergents. Absorption onto a surface followed by dehydration stimulated it to fold into a helical structure, as it did when the aqueous solution was replaced by nonaqueous solvents. Hydration of the dehydrated folded protein was accompanied by complete unfolding. These results support the identification of MEG-14 as a classic intrinsically disordered protein, and open the possibility of its interaction/folding with different partners and factors being related to multifunctional roles and states within the host

Developing mHealth Apps with researchers: multi-stakeholder design considerations

The authors have been involved with developing a number of mHealth smartphone Apps for use in health or wellness research in collaboration with researchers, clinicians and patient groups for clinical areas including Sickle Cell Disease, Attention Deficit Hyperactivity Disorder, asthma and infertility treatment. In these types of applications, end-users self-report their symptoms and quality of life or conduct psychometric tests. Physiological data may also be captured using sensors that are internal or external to the device. Following a discussion of the multiple stakeholders that are typically involved in small scale research projects involving end-user data collection, four Apps are used as case studies to explore the issue of non-functional requirements

Radiation monitoring with CVD Diamonds and PIN Diodes at BaBar

The BaBar experiment at the Stanford Linear Accelerator Center has been using two polycrystalline chemical vapor deposition (pCVD) diamonds and 12 silicon PIN diodes for radiation monitoring and protection of the Silicon Vertex Tracker (SVT). We have used the pCVD diamonds for more than 3 years, and the PIN diodes for 7 years. We will describe the SVT and SVT radiation monitoring system as well as the operational difficulties and radiation damage effects on the PIN diodes and pCVD diamonds in a high-energy physics environment

SO₂ emission rates and incorporation into the air pollution dispersion forecast during the 2021 eruption of Fagradalsfjall, Iceland

During the low-effusion rate Fagradalsfjall eruption (19 March – 18 September 2021), the emission of sulfur dioxide (SO₂) was frequently measured using ground-based UV spectrometers. The total SO₂ emitted during the entire eruption was 970 ± 540 kt, which is only about 6% of the SO₂ emitted during the similar length Holuhraun eruption (2014–2015). The eruption was divided into five phases based on visual observations, including the number of active vents and the occurrence of lava fountaining. The SO₂ emission rate ranged from 44 ± 19 kg/s in Phase 2 to 85 ± 29 kg/s in Phase 5, with an average of 64 ± 34 kg/s for the entire eruption. There was notable variability in SO₂ on short timescales, with measurements on 11 August 2021 ranging from 17 to 78 kg/s. SO₂ flux measurements were made using scanning DOAS instruments located at different distances from and orientations relative to the eruption site augmented by traverses. Four hundred and forty-four scan and traverse measurements met quality criteria and were used, along with plume height and meteorological data, to calculate SO₂ fluxes while accounting for wind-related uncertainties. A tendency for stronger SO₂ flux concurrent with higher amplitude seismic tremor and the occurrence of lava fountaining was observed during Phases 4 and 5 which were characterized by intermittent crater activity including observable effusion of lava and gas release interspersed with long repose times. This tendency was used to refine the calculation of the amount of SO₂ emitted during variably vigorous activity. The continuous seismic tremor time series was used to quantify how long during these eruption phases strong/weak activity was exhibited to improve the calculated SO₂ flux during these Phases. The total SO₂ emissions derived from field measurements align closely with results obtained by combining melt inclusion and groundmass glass analyses with lava effusion rate measurements (910 ± 230 kt SO₂). Specifically, utilizing the maximum S content found in evolved melt inclusions and the least remaining S content in accompanying quenched groundmasses provides an identical result between field measurements and the petrological calculations. This suggests that the maximum SO₂ release calculated from petrological estimates should be preferentially used to initialize gas dispersion models for basaltic eruptions when other measurements are lacking. During the eruption, the CALPUFF dispersion model was used to forecast ground-level exposure to SO₂. The SO₂ emission rates measured by DOAS were used as input for the dispersion model, with updates made when a significant change was measured. A detailed analysis of one mid-distance station over the entire eruption shows that the model performed very well at predicting the presence of volcanic SO₂ when it was measured. However, it frequently predicted the presence of SO₂ that was not measured and the concentrations forecasted had no correlation with the concentrations measured. Various approaches to improve the model forecast were tested, including updating plume height and SO₂ flux source terms based on measurements. These approaches did not unambiguously improve the model performance but suggest that improvements might be achieved in more-polluted conditions

Prostate cancer genetic risk and associated aggressive disease in men of African ancestry

African ancestry is a significant risk factor for prostate cancer and advanced disease. Yet, genetic studies have largely been conducted outside the context of Sub-Saharan Africa, identifying 278 common risk variants contributing to a multiethnic polygenic risk score, with rare variants focused on a panel of roughly 20 pathogenic genes. Based on this knowledge, we are unable to determine polygenic risk or differentiate prostate cancer status interrogating whole genome data for 113 Black South African men. To further assess for potentially functional common and rare variant associations, here we interrogate 247,780 exomic variants for 798 Black South African men using a case versus control or aggressive versus non-aggressive study design. Notable genes of interest include HCP5, RFX6 and H3C1 for risk, and MKI67 and KLF5 for aggressive disease. Our study highlights the need for further inclusion across the African diaspora to establish African-relevant risk models aimed at reducing prostate cancer health disparities.Co-founding SAPCS members the late Professor Philip A. Venter and retired urologist Dr. Richard Monare from the University of Limpopo in South Africa, as well as the Medical Research Council (MRC) of South Africa for SAPCS seed-funding; supported by a Cancer Association of South Africa (CANSA), the National Health and Medical Research Council (NHMRC) of Australia, Ideas grants, the U.S.A. Congressionally Directed Medical Research Programs (CDMRP) Prostate Cancer Research Program (PCRP) through an Idea Development Award, HEROIC Consortium Award, and the Petre Foundation via the University of Sydney Foundation, Australia.https://www.nature.com/ncomms/am2024School of Health Systems and Public Health (SHSPH)SDG-03:Good heatlh and well-bein

Observation of Scaling Violations in Scaled Momentum Distributions at HERA

Charged particle production has been measured in deep inelastic scattering (DIS) events over a large range of $x$ and $Q^2$ using the ZEUS detector. The evolution of the scaled momentum, $x_p$, with $Q^2,$ in the range 10 to 1280 $GeV^2$, has been investigated in the current fragmentation region of the Breit frame. The results show clear evidence, in a single experiment, for scaling violations in scaled momenta as a function of $Q^2$.Comment: 21 pages including 4 figures, to be published in Physics Letters B. Two references adde

Search for direct production of charginos and neutralinos in events with three leptons and missing transverse momentum in √s = 7 TeV pp collisions with the ATLAS detector

A search for the direct production of charginos and neutralinos in final states with three electrons or muons and missing transverse momentum is presented. The analysis is based on 4.7 fb−1 of proton–proton collision data delivered by the Large Hadron Collider and recorded with the ATLAS detector. Observations are consistent with Standard Model expectations in three signal regions that are either depleted or enriched in Z-boson decays. Upper limits at 95% confidence level are set in R-parity conserving phenomenological minimal supersymmetric models and in simplified models, significantly extending previous results

Jet size dependence of single jet suppression in lead-lead collisions at sqrt(s(NN)) = 2.76 TeV with the ATLAS detector at the LHC

Measurements of inclusive jet suppression in heavy ion collisions at the LHC provide direct sensitivity to the physics of jet quenching. In a sample of lead-lead collisions at sqrt(s) = 2.76 TeV corresponding to an integrated luminosity of approximately 7 inverse microbarns, ATLAS has measured jets with a calorimeter over the pseudorapidity interval |eta| < 2.1 and over the transverse momentum range 38 < pT < 210 GeV. Jets were reconstructed using the anti-kt algorithm with values for the distance parameter that determines the nominal jet radius of R = 0.2, 0.3, 0.4 and 0.5. The centrality dependence of the jet yield is characterized by the jet "central-to-peripheral ratio," Rcp. Jet production is found to be suppressed by approximately a factor of two in the 10% most central collisions relative to peripheral collisions. Rcp varies smoothly with centrality as characterized by the number of participating nucleons. The observed suppression is only weakly dependent on jet radius and transverse momentum. These results provide the first direct measurement of inclusive jet suppression in heavy ion collisions and complement previous measurements of dijet transverse energy imbalance at the LHC.Comment: 15 pages plus author list (30 pages total), 8 figures, 2 tables, submitted to Physics Letters B. All figures including auxiliary figures are available at http://atlas.web.cern.ch/Atlas/GROUPS/PHYSICS/PAPERS/HION-2011-02

Observation of a new chi_b state in radiative transitions to Upsilon(1S) and Upsilon(2S) at ATLAS

The chi_b(nP) quarkonium states are produced in proton-proton collisions at the Large Hadron Collider (LHC) at sqrt(s) = 7 TeV and recorded by the ATLAS detector. Using a data sample corresponding to an integrated luminosity of 4.4 fb^-1, these states are reconstructed through their radiative decays to Upsilon(1S,2S) with Upsilon->mu+mu-. In addition to the mass peaks corresponding to the decay modes chi_b(1P,2P)->Upsilon(1S)gamma, a new structure centered at a mass of 10.530+/-0.005 (stat.)+/-0.009 (syst.) GeV is also observed, in both the Upsilon(1S)gamma and Upsilon(2S)gamma decay modes. This is interpreted as the chi_b(3P) system.Comment: 5 pages plus author list (18 pages total), 2 figures, 1 table, corrected author list, matches final version in Physical Review Letter