Peptide nanofiber hydrogel adjuvanted live virus vaccine enhances cross-protective immunity to porcine reproductive and respiratory syndrome virus

Porcine reproductive and respiratory syndrome virus (PRRSV) is prevalent in swine farms worldwide and is a major source of economic loss and animal suffering. Rapid genetic variation of PRRSV makes it difficult for current vaccines to confer protection against newly emerging strains. We recently demonstrated that a novel peptide nanofiber hydrogel (H9e) could act as a potent adjuvant for killed H1N1 vaccines. Therefore, the objective of this study was to evaluate H9e as an adjuvant for PRRSV modified live virus (MLV) vaccines. Pigs were vaccinated with Ingelvac PRRSV MLV with or without H9e adjuvant before being challenged with the VR-2332 (parental vaccine strain) or MN184A (genetically diverse strain) PRRSV. Pigs vaccinated with MLV+H9e had higher levels of circulating vaccine virus. More importantly, pigs vaccinated with MLV+H9e had improved protection against challenge by both PRRSV strains, as demonstrated by reduced challenge-induced viremia compared with pigs vaccinated with MLV alone. Pigs vaccinated with MLV+H9e had lower frequency of T-regulatory cells and IL-10 production but higher frequency of Th/memory cells and IFN-γ secretion than that in pigs vaccinated with MLV alone. Taken together, our studies suggest that the peptide nanofiber hydrogel H9e, when combined with the PRRSV MLV vaccine, can enhance vaccine efficacy against two different PRRSV strains by modulating both host humoral and cellular immune responses

EvryFlare. II. Rotation Periods of the Cool Flare Stars in TESS across Half the Southern Sky

We measure rotation periods and sinusoidal amplitudes in Evryscope light curves for 122 two-minute K5-M4 TESS targets selected for strong flaring. The Evryscope array of telescopes has observed all bright nearby stars in the south, producing 2-minute cadence light curves since 2016. Long-term, high-cadence observations of rotating flare stars probe the complex relationship between stellar rotation, starspots, and superflares. We detect periods from 0.3487 to 104 days and observe amplitudes from 0.008 to 0.216 g′ mag. We find that the Evryscope amplitudes are larger than those in TESS with the effect correlated to stellar mass (p-value = 0.01). We compute the Rossby number (R o ) and find that our sample selected for flaring has twice as many intermediate rotators (0.04 0.44) rotators; this may be astrophysical or a result of period detection sensitivity. We discover 30 fast, 59 intermediate, and 33 slow rotators. We measure a median starspot coverage of 13% of the stellar hemisphere and constrain the minimum magnetic field strength consistent with our flare energies and spot coverage to be 500 G, with later-type stars exhibiting lower values than earlier-type stars. We observe a possible change in superflare rates at intermediate periods. However, we do not conclusively confirm the increased activity of intermediate rotators seen in previous studies. We split all rotators at R o ∼ 0.2 into bins of P Rot 10 days to confirm that short-period rotators exhibit higher superflare rates, larger flare energies, and higher starspot coverage than do long-period rotators, at p-values of 3.2 × 10-5, 1.0 × 10-5, and 0.01, respectively

EvryFlare. III. Temperature Evolution and Habitability Impacts of Dozens of Superflares Observed Simultaneously by Evryscope and TESS

Superflares may provide the dominant source of biologically relevant UV radiation to rocky habitable-zone M-dwarf planets (M-Earths), altering planetary atmospheres and conditions for surface life. The combined line and continuum flare emission has usually been approximated by a 9000 K blackbody. If superflares are hotter, then the UV emission may be 10 times higher than predicted from the optical. However, it is unknown for how long M-dwarf superflares reach temperatures above 9000 K. Only a handful of M-dwarf superflares have been recorded with multiwavelength high-cadence observations. We double the total number of events in the literature using simultaneous Evryscope and Transiting Exoplanet Survey Satellite observations to provide the first systematic exploration of the temperature evolution of M-dwarf superflares. We also increase the number of superflaring M dwarfs with published time-resolved blackbody evolution by ∼10×. We measure temperatures at 2 minutes cadence for 42 superflares from 27 K5-M5 dwarfs. We find superflare peak temperatures (defined as the mean of temperatures corresponding to flare FWHM) increase with flare energy and impulse. We find the amount of time flares emit at temperatures above 14,000 K depends on energy. We discover that 43% of the flares emit above 14,000 K, 23% emit above 20,000 K and 5% emit above 30,000 K. The largest and hottest flare briefly reached 42,000 K. Some do not reach 14,000 K. During superflares, we estimate M-Earths orbiting <200 Myr stars typically receive a top-of-atmosphere UV-C flux of ∼120 W m-2 and up to 103 W m-2, 100-1000 times the time-averaged X-ray and UV flux from Proxima Cen

Identification of Kinases Regulating Prostate Cancer Cell Growth Using an RNAi Phenotypic Screen

As prostate cancer progresses to castration-resistant disease, there is an increase in signal transduction activity. Most castration-resistant prostate tumors continue to express the androgen receptor (AR) as well as androgen-responsive genes, despite the near absence of circulating androgen in these patients. The AR is regulated not only by its cognate steroid hormone, but also by interactions with a constellation of co-regulatory and signaling molecules. Thus, the elevated signaling activity that occurs during progression to castration resistance can affect prostate cancer cell growth either through the AR or independent of the AR. In order to identify signaling pathways that regulate prostate cancer cell growth, we screened a panel of shRNAs targeting 673 human kinases against LNCaP prostate cancer cells grown in the presence and absence of hormone. The screen identified multiple shRNA clones against known and novel gene targets that regulate prostate cancer cell growth. Based on the magnitude of effect on growth, we selected six kinases for further study: MAP3K11, DGKD, ICK, CIT, GALK2, and PSKH1. Knockdown of these kinases decreased cell growth in both androgen-dependent and castration-resistant prostate cancer cells. However, these kinases had different effects on basal or androgen-induced transcriptional activity of AR target genes. MAP3K11 knockdown most consistently altered transcription of AR target genes, suggesting that MAP3K11 affected its growth inhibitory effect by modulating the AR transcriptional program. Consistent with MAP3K11 acting on the AR, knockdown of MAP3K11 inhibited AR Ser 650 phosphorylation, further supporting stress kinase regulation of AR phosphorylation. This study demonstrates the applicability of lentiviral-based shRNA for conducting phenotypic screens and identifies MAP3K11, DGKD, ICK, CIT, GALK2, and PSKH1 as regulators of prostate cancer cell growth. The thorough evaluation of these kinase targets will pave the way for developing more effective treatments for castration-resistant prostate cancer

Pro-asthmatic cytokines regulate unliganded and ligand-dependent glucocorticoid receptor signaling in airway smooth muscle

To elucidate the regulation of glucocorticoid receptor (GR) signaling under pro-asthmatic conditions, cultured human airway smooth muscle (HASM) cells were treated with proinflammatory cytokines or GR ligands alone and in combination, and then examined for induced changes in ligand-dependent and -independent GR activation and downstream signaling events. Ligand stimulation with either cortisone or dexamethsone (DEX) acutely elicited GR translocation to the nucleus and, comparably, ligand-independent stimulation either with the Th2 cytokine, IL-13, or the pleiotropic cytokine combination, IL-1β/TNFα, also acutely evoked GR translocation. The latter response was potentiated by combined exposure of cells to GR ligand and cytokine. Similarly, treatment with either DEX or IL-13 alone induced GR phosphorylation at its serine-211 residue (GRSer211), denoting its activated state, and combined treatment with DEX+IL-13 elicited heightened and sustained GRSer211phosphorylation. Interestingly, the above ligand-independent GR responses to IL-13 alone were not associated with downstream GR binding to its consensus DNA sequence or GR transactivation, whereas both DEX-induced GR:DNA binding and transcriptional activity were significantly heightened in the presence of IL-13, coupled to increased recruitment of the transcriptional co-factor, MED14. The stimulated GR signaling responses to DEX were prevented in IL-13-exposed cells wherein GRSer211 phosphorylation was suppressed either by transfection with specific serine phosphorylation-deficient mutant GRs or treatment with inhibitors of the MAPKs, ERK1/2 and JNK. Collectively, these novel data highlight a heretofore-unidentified homeostatic mechanism in HASM cells that involves pro-asthmatic cytokine-driven, MAPK-mediated, non-ligand-dependent GR activation that confers heightened glucocorticoid ligand-stimulated GR signaling. These findings raise the consideration that perturbations in this homeostatic cytokine-driven GR signaling mechanism may be responsible, at least in part, for the insensirtivity to glucocorticoid therapy that is commonly seen in individuals with severe asthma

Analysis and characterization of heparin impurities

This review discusses recent developments in analytical methods available for the sensitive separation, detection and structural characterization of heparin contaminants. The adulteration of raw heparin with oversulfated chondroitin sulfate (OSCS) in 2007–2008 spawned a global crisis resulting in extensive revisions to the pharmacopeia monographs on heparin and prompting the FDA to recommend the development of additional physicochemical methods for the analysis of heparin purity. The analytical chemistry community quickly responded to this challenge, developing a wide variety of innovative approaches, several of which are reported in this special issue. This review provides an overview of methods of heparin isolation and digestion, discusses known heparin contaminants, including OSCS, and summarizes recent publications on heparin impurity analysis using sensors, near-IR, Raman, and NMR spectroscopy, as well as electrophoretic and chromatographic separations

Revisiting the HD 21749 planetary system with stellar activity modelling

HD 21749 is a bright (V = 8.1 mag) K dwarf at 16 pc known to host an inner terrestrial planet HD 21749c as well as an outer sub-Neptune HD 21749b, both delivered by Transiting Exoplanet Survey Satellite (TESS). Follow-up spectroscopic observations measured the mass of HD 21749b to be 22.7 ± 2.2 M with a density of 7.0^{+1.6}_{-1.3} g cm-3, making it one of the densest sub-Neptunes. However, the mass measurement was suspected to be influenced by stellar rotation. Here, we present new high-cadence PFS RV data to disentangle the stellar activity signal from the planetary signal. We find that HD 21749 has a similar rotational time-scale as the planet's orbital period, and the amplitude of the planetary orbital RV signal is estimated to be similar to that of the stellar activity signal. We perform Gaussian process regression on the photometry and RVs from HARPS and PFS to model the stellar activity signal. Our new models reveal that HD 21749b has a radius of 2.86 ± 0.20 R, an orbital period of 35.6133 ± 0.0005 d with a mass of Mb = 20.0 ± 2.7 M and a density of 4.8^{+2.0}_{-1.4} g cm-3 on an eccentric orbit with e = 0.16 ± 0.06, which is consistent with the most recent values published for this system. HD 21749c has an orbital period of 7.7902 ± 0.0006 d, a radius of 1.13 ± 0.10 R, and a 3σ mass upper limit of 3.5 M. Our Monte Carlo simulations confirm that without properly taking stellar activity signals into account, the mass measurement of HD 21749b is likely to arrive at a significantly underestimated error bar

Antidepressants increase human hippocampal neurogenesis by activating the glucocorticoid receptor

Antidepressants increase adult hippocampal neurogenesis in animal models, but the underlying molecular mechanisms are unknown. In this study, we used human hippocampal progenitor cells to investigate the molecular pathways involved in the antidepressant-induced modulation of neurogenesis. Because our previous studies have shown that antidepressants regulate glucocorticoid receptor (GR) function, we specifically tested whether the GR may be involved in the effects of these drugs on neurogenesis. We found that treatment (for 3–10 days) with the antidepressant, sertraline, increased neuronal differentiation via a GR-dependent mechanism. Specifically, sertraline increased both immature, doublecortin (Dcx)-positive neuroblasts (+16%) and mature, microtubulin-associated protein-2 (MAP2)-positive neurons (+26%). This effect was abolished by the GR-antagonist, RU486. Interestingly, progenitor cell proliferation, as investigated by 5′-bromodeoxyuridine (BrdU) incorporation, was only increased when cells were co-treated with sertraline and the GR-agonist, dexamethasone, (+14%) an effect which was also abolished by RU486. Furthermore, the phosphodiesterase type 4 (PDE4)-inhibitor, rolipram, enhanced the effects of sertraline, whereas the protein kinase A (PKA)-inhibitor, H89, suppressed the effects of sertraline. Indeed, sertraline increased GR transactivation, modified GR phosphorylation and increased expression of the GR-regulated cyclin-dependent kinase-2 (CDK2) inhibitors, p27Kip1 and p57Kip2. In conclusion, our data suggest that the antidepressant, sertraline, increases human hippocampal neurogenesis via a GR-dependent mechanism that requires PKA signaling, GR phosphorylation and activation of a specific set of genes. Our data point toward an important role for the GR in the antidepressant-induced modulation of neurogenesis in humans