p21-activated kinase (PAK) regulates cytoskeletal reorganization and directional migration in human neutrophils

Neutrophils serve as a first line of defense in innate immunity owing in part to their ability to rapidly migrate towards chemotactic factors derived from invading pathogens. As a migratory function, neutrophil chemotaxis is regulated by the Rho family of small GTPases. However, the mechanisms by which Rho GTPases orchestrate cytoskeletal dynamics in migrating neutrophils remain ill-defined. In this study, we characterized the role of p21-activated kinase (PAK) downstream of Rho GTPases in cytoskeletal remodeling and chemotactic processes of human neutrophils. We found that PAK activation occurred upon stimulation of neutrophils with f-Met-Leu-Phe (fMLP), and PAK accumulated at the actin-rich leading edge of stimulated neutrophils, suggesting a role for PAK in Rac-dependent actin remodeling. Treatment with the pharmacological PAK inhibitor, PF3758309, abrogated the integrity of RhoA-mediated actomyosin contractility and surface adhesion. Moreover, inhibition of PAK activity impaired neutrophil morphological polarization and directional migration under a gradient of fMLP, and was associated with dysregulated Ca2+ signaling. These results suggest that PAK serves as an important effector of Rho-family GTPases in neutrophil cytoskeletal reorganization, and plays a key role in driving efficient directional migration of human neutrophils

Tan's contact and the phase distribution of repulsive Fermi gases: Insights from quantum chromodynamics noise analyses

Path-integral analyses originally pioneered in the study of the complex-phase problem afflicting lattice calculations of finite-density quantum chromodynamics are generalized to nonrelativistic Fermi gases with repulsive interactions. Using arguments similar to those previously applied to relativistic theories, we show that the analogous problem in nonrelativistic systems manifests itself naturally in Tan's contact as a nontrivial cancellation between terms with varied dependence on extensive thermodynamic quantities. We analyze that case under the assumption of a Gaussian phase distribution, which is supported by our Monte Carlo calculations and perturbative considerations. We further generalize these results to observables other than the contact, as well as to polarized systems and systems with fixed particle number. Our results are quite general in that they apply to repulsive multicomponent fermions, they are independent of dimensionality or trapping potential, and they hold in the ground state as well as at finite temperature

Convexity of the entanglement entropy of Su(2n)-symmetric fermions with attractive interactions

The positivity of the probability measure of attractively interacting systems of 2N-component fermions enables the derivation of an exact convexity property for the ground-state energy of such systems. Using analogous arguments, applied to path-integral expressions for the entanglement entropy derived recently, we prove nonperturbative analytic relations for the Rényi entropies of those systems. These relations are valid for all subsystem sizes, particle numbers, and dimensions, and in arbitrary external trapping potentials

Dynamics of entanglement entropy of interacting fermions in a 1D driven harmonic trap

Following up on a recent analysis of two cold atoms in a time-dependent harmonic trap in one dimension, we explore the entanglement entropy of two and three fermions in the same situation when driven through a parametric resonance. We find that the presence of such a resonance in the two-particle system leaves a clear imprint on the entanglement entropy. We show how the signal is modified by attractive and repulsive contact interactions, and how it remains present for the three-particle system. Additionaly, we extend the work of recent experiments to demonstrate how restricting observation to a limited subsystem gives rise to locally thermal behavior

A comparison of precipitation and filtration-based SARS-CoV-2 recovery methods and the influence of temperature, turbidity, and surfactant load in urban wastewater

Wastewater-based epidemiology (WBE) has become a complimentary surveillance tool during the SARS-CoV-2 pandemic. Viral concentration methods from wastewater are still being optimised and compared, whilst viral recovery under different wastewater characteristics and storage temperatures remains poorly understood. Using urban wastewater samples, we tested three viral concentration methods; polyethylene glycol precipitation (PEG), ammonium sulphate precipitation (AS), and CP select™ InnovaPrep® (IP) ultrafiltration. We found no major difference in SARS-CoV-2 and faecal indicator virus (crAssphage) recovery from wastewater samples (n = 46) using these methods, PEG slightly (albeit non-significantly), outperformed AS and IP for SARS-CoV-2 detection, as a higher genome copies per litre (gc/l) was recorded for a larger proportion of samples. Next generation sequencing of 8 paired samples revealed non-significant differences in the quality of data between AS and IP, though IP data quality was slightly better and less variable. A controlled experiment assessed the impact of wastewater suspended solids (turbidity; 0–400 NTU), surfactant load (0–200 mg/l), and storage temperature (5–20 °C) on viral recovery using the AS and IP methods. SARS-CoV-2 recoveries were >20% with AS and  0.05), whilst surfactant and storage temperature combined were significant negative correlates (p < 0.001 and p < 0.05, respectively). In conclusion, our results show that choice of methodology had small effect on viral recovery of SARS-CoV-2 and crAssphage in wastewater samples within this study. In contrast, sample turbidity, storage temperature, and surfactant load did affect viral recovery, highlighting the need for careful consideration of the viral concentration methodology used when working with wastewater samples

Development of a high-throughput assay for identifying inhibitors of TBK1 and IKKε

IKKε and TBK1 are noncanonical IKK family members which regulate inflammatory signaling pathways and also play important roles in oncogenesis. However, few inhibitors of these kinases have been identified. While the substrate specificity of IKKε has recently been described, the substrate specificity of TBK1 is unknown, hindering the development of high-throughput screening technologies for inhibitor identification. Here, we describe the optimal substrate phosphorylation motif for TBK1, and show that it is identical to the phosphorylation motif previously described for IKKε. This information enabled the design of an optimal TBK1/IKKε substrate peptide amenable to high-throughput screening and we assayed a 6,006 compound library that included 4,727 kinase-focused compounds to discover in vitro inhibitors of TBK1 and IKKε. 227 compounds in this library inhibited TBK1 at a concentration of 10 μM, while 57 compounds inhibited IKKε. Together, these data describe a new high-throughput screening assay which will facilitate the discovery of small molecule TBK1/IKKε inhibitors possessing therapeutic potential for both inflammatory diseases and cancer

Assessing the accuracy of current near infra-red reflectance spectroscopy analysis for fresh grass-clover mixture silages and development of new equations for this purpose

The purpose of this study was to ascertain whether Near Infra-Red Reflectance Spectroscopy (NIRS) prediction equations calibrated on grass silage samples, could accurately predict the chemical composition of mixed grass-clover silage samples, and furthermore, to develop and calibrate new grass-clover equations should the grass-based equations be insufficiently accurate for these silages. A set of 94 silage samples from mixed grass-clover swards (clover concentration (CC) ranging from 4 to 1000 g/kg as fed; determined manually) were analysed for chemical composition using reference laboratory techniques, in vivo digestible organic matter in the dry matter (DOMD, in sheep), and in situ degradability of dry matter and crude protein (in cows). The same samples were scanned fresh (undried and unmilled, as is standard practice for silage analysis within UK laboratories) using NIRS (at AFBI, Northern Ireland) and grass-based prediction equations applied. Predicted and observed results were compared. Of 15 chemical components that were tested for prediction accuracy, only volatile-corrected dry matter and nitrogen were well predicted (RPD values of 4.9 and 2.4 respectively, with low root mean square errors of prediction (RMSEP)). Neutral detergent fibre and DOMD showed low RPD values, however the predicted and observed datasets had no significant bias between them and were therefore also considered as fit for purpose. Variables with significant bias between predicted and observed datasets that were not considered suitably accurate included crude protein, acid detergent fibre, microbial dry matter yield and the effective degradability of protein. For many components, bias could be attributed at least in part to CC and changes in the fractionation of nutrients present. For some variables such as crude protein, grass-based equations were sufficiently accurate at low CCs but became inaccurate as CC increased, as expected. In response to inadequate prediction accuracy of certain nutrients, new grass-clover equations were calibrated using the obtained spectra. These were validated and results indicated that the grass-clover-based equations outperformed their grass-based counterparts. The adoption of new grass-clover equations, or alternatively, with further development, the use of a CC correction factor to the existing grass-based equations, is recommended for commercial laboratories offering undried and unmilled silage analysis on samples containing clover

Plasma sterols and vitamin D are correlates and predictors of ozone-induced inflammation in the lung: A pilot study

BACKGROUND: Ozone (O3) exposure causes respiratory effects including lung function decrements, increased lung permeability, and airway inflammation. Additionally, baseline metabolic state can predispose individuals to adverse health effects from O3. For this reason, we conducted an exploratory study to examine the effect of O3 exposure on derivatives of cholesterol biosynthesis: sterols, oxysterols, and secosteroid (25-hydroxyvitamin D) not only in the lung, but also in circulation. METHODS: We obtained plasma and induced sputum samples from non-asthmatic (n = 12) and asthmatic (n = 12) adult volunteers 6 hours following exposure to 0.4ppm O3 for 2 hours. We quantified the concentrations of 24 cholesterol precursors and derivatives by UPLC-MS and 30 cytokines by ELISA. We use computational analyses including machine learning to determine whether baseline plasma sterols are predictive of O3 responsiveness. RESULTS: We observed an overall decrease in the concentration of cholesterol precursors and derivatives (e.g. 27-hydroxycholesterol) and an increase in concentration of autooxidation products (e.g. secosterol-B) in sputum samples. In plasma, we saw a significant increase in the concentration of secosterol-B after O3 exposure. Machine learning algorithms showed that plasma cholesterol was a top predictor of O3 responder status based on decrease in FEV1 (>5%). Further, 25-hydroxyvitamin D was positively associated with lung function in non-asthmatic subjects and with sputum uteroglobin, whereas it was inversely associated with sputum myeloperoxidase and neutrophil counts. CONCLUSION: This study highlights alterations in sterol metabolites in the airway and circulation as potential contributors to systemic health outcomes and predictors of pulmonary and inflammatory responsiveness following O3 exposure

Azimuthal anisotropy and correlations in p+p, d+Au and Au+Au collisions at 200 GeV

We present the first measurement of directed flow ($v_1$) at RHIC. $v_1$ is found to be consistent with zero at pseudorapidities $\eta$ from -1.2 to 1.2, then rises to the level of a couple of percent over the range $2.4 < |\eta| < 4$. The latter observation is similar to data from NA49 if the SPS rapidities are shifted by the difference in beam rapidity between RHIC and SPS. Back-to-back jets emitted out-of-plane are found to be suppressed more if compared to those emitted in-plane, which is consistent with {\it jet quenching}. Using the scalar product method, we systematically compared azimuthal correlations from p+p, d+Au and Au+Au collisions. Flow and non-flow from these three different collision systems are discussed.Comment: Quark Matter 2004 proceeding, 4 pages, 3 figure

Azimuthal anisotropy: the higher harmonics

We report the first observations of the fourth harmonic (v_4) in the azimuthal distribution of particles at RHIC. The measurement was done taking advantage of the large elliptic flow generated at RHIC. The integrated v_4 is about a factor of 10 smaller than v_2. For the sixth (v_6) and eighth (v_8) harmonics upper limits on the magnitudes are reported.Comment: 4 pages, 6 figures, contribution to the Quark Matter 2004 proceeding