Combined Pharmacological Inhibition of Cyclophilins, FK506-Binding Proteins, Hsp90, and Hsp70 Protects Cells From Clostridium botulinum C2 Toxin

The Clostridium botulinum C2 toxin is an exotoxin causing severe enterotoxic symptoms. The C2 toxin consists of the binding/translocation component C2II, and the enzymatic active component C2I. After proteolytic activation, C2IIa forms heptamers that bind C2I. The C2IIa/C2I complex is taken up into mammalian target cells via receptor-mediated endocytosis. Acidification of endosomes leads to conformational changes in both components. C2IIa heptamers form a pore into the endosomal membrane, and C2I becomes unfolded and translocates through the narrow C2IIa pores into the cytosol of the cell. Here, C2I covalently transfers an ADP-ribose moiety from its co-substrate NAD+ onto G-actin, which leads to depolymerization of F-actin resulting in rounding up of adherent cells. Translocation of C2I into the cytosol depends on the activity of the chaperones Hsp90 and Hsp70 and peptidyl-prolyl cis/trans isomerases of the cyclophilin (Cyp) and FK506-binding protein (FKBP) families. Here, we demonstrated that C2I is detected in close proximity with Hsp90, Cyp40, and FKBP51 in cells, indicating their interaction. This interaction was dependent on the concentration of C2 toxin and detected in mammalian Vero and human HeLa cells. Moreover, the present study reveals that combination of radicicol, VER-155008, cyclosporine A, and FK506, which are specific pharmacological inhibitors of Hsp90, Hsp70, Cyps, and FKBPs, respectively, resulted in a stronger inhibition of intoxication of cells with C2 toxin compared to application of the single inhibitors. Thus, the combination of inhibitors showed enhanced protection of cells against the cytotoxic effects of C2 toxin. Cell viability was not significantly impaired by application of the inhibitor combination. Moreover, we confirmed that the combination of radicicol, VER-155008, CsA, and FK506 in particular inhibit the membrane translocation step of C2I into the cytosol whereas receptor binding and enzyme activity of the toxin were not affected. Our findings further characterize the mode of action of Hsp90, Hsp70, Cyps, and FKBPs during membrane translocation of bacterial toxins and furthermore supply starting points for developing of novel therapeutic strategies against diseases caused by bacterial toxins that depend on Hsp90, Hsp70, Cyps, and FKBPs

Human Peptides α-Defensin-1 and -5 Inhibit Pertussis Toxin

Bordetella pertussis causes the severe childhood disease whooping cough, by releasing several toxins, including pertussis toxin (PT) as a major virulence factor. PT is an AB(5)-type toxin, and consists of the enzymatic A-subunit PTS1 and five B-subunits, which facilitate binding to cells and transport of PTS1 into the cytosol. PTS1 ADP-ribosylates alpha-subunits of inhibitory G-proteins (G alpha i) in the cytosol, which leads to disturbed cAMP signaling. Since PT is crucial for causing severe courses of disease, our aim is to identify new inhibitors against PT, to provide starting points for novel therapeutic approaches. Here, we investigated the effect of human antimicrobial peptides of the defensin family on PT. We demonstrated that PTS1 enzyme activity in vitro was inhibited by alpha-defensin-1 and -5, but not beta-defensin-1. The amount of ADP-ribosylated G alpha i was significantly reduced in PT-treated cells, in the presence of alpha-defensin-1 and -5. Moreover, both alpha-defensins decreased PT-mediated effects on cAMP signaling in the living cell-based interference in the G alpha i-mediated signal transduction (iGIST) assay. Taken together, we identified the human peptides alpha-defensin-1 and -5 as inhibitors of PT activity, suggesting that these human peptides bear potential for developing novel therapeutic strategies against whooping cough

Robotic reverberation mapping of the broad-line radio galaxy 3C 120

We carried out photometric and spectroscopic observations of the well-studied broad-line radio galaxy 3C 120 with the Las Cumbres Observatory (LCO) global robotic telescope network from 2016 December to 2018 April as part of the LCO AGN Key Project on Reverberation Mapping of Accretion Flows. Here, we present both spectroscopic and photometric reverberation mapping results. We used the interpolated cross-correlation function (ICCF) to perform multiple-line lag measurements in 3C 120. We find the H$\gamma$, He II $\lambda 4686$, H$\beta$ and He I $\lambda 5876$ lags of $\tau_{\text{cen}} = 18.8_{-1.0}^{+1.3}$, $2.7_{-0.8}^{+0.7}$, $21.2_{-1.0}^{+1.6}$, and $16.9_{-1.1}^{+0.9}$ days respectively, relative to the V-band continuum. Using the measured lag and rms velocity width of the H$\beta$ emission line, we determine the mass of the black hole for 3C 120 to be $M=\left(6.3^{+0.5}_{-0.3}\right)\times10^7\,(f/5.5)$ M$_\odot$. Our black hole mass measurement is consistent with similar previous studies on 3C 120, but with small uncertainties. In addition, velocity-resolved lags in 3C 120 show a symmetric pattern across the H$\beta$ line, 25 days at line centre decreasing to 17 days in the line wings at $\pm4000$ km s$^{-1}$. We also investigate the inter-band continuum lags in 3C 120 and find that they are generally consistent with $\tau\propto\lambda^{4/3}$ as predicted from a geometrically-thin, optically-thick accretion disc. From the continuum lags, we measure the best fit value $\tau_{\rm 0} = 3.5\pm 0.2$ days at $\lambda_{\rm 0} = 5477$A. It implies a disc size a factor of $1.6$ times larger than prediction from the standard disc model with $L/L_{\rm Edd} = 0.4$. This is consistent with previous studies in which larger than expected disc sizes were measured

The first spectroscopic IR reverberation programme on Mrk 509

Near IR spectroscopic reverberation of Active Galactic Nuclei (AGN) potentially allows the IR broad line region (BLR) to be reverberated alongside the disc and dust continua, while the spectra can also reveal details of dust astro-chemistry. Here we describe results of a short pilot study (17 near-IR spectra over a 183 day period) for Mrk 509. The spectra give a luminosity-weighted dust radius of 〈Rd, lum〉 = 186 ± 4 light-days for blackbody (large grain dust), consistent with previous (photometric) reverberation campaigns, whereas carbon and silicate dust give much larger radii. We develop a method of calibrating spectral data in objects where the narrow lines are extended beyond the slit width. We demonstrate this by showing our resultant photometric band lightcurves are consistent with previous results, with a hot dust lag at >40 days in the K-band, clearly different from the accretion disc response at <20 days in the z-band. We place this limit of 40 days by demonstrating clearly that the modest variability that we do detect in the H and K-band does not reverberate on timescales of less than 40 days. We also extract the Paβ line lightcurve, and find a lag which is consistent with the optical BLR Hβ line of ∼70 − 90 days. This is important as direct imaging of the near-IR BLR is now possible in a few objects, so we need to understand its relation to the better studied optical BLR

More data, more problems: Strategically addressing data ethics and policy issues in LIS curricula and courses

Library and information science (LIS) schools are revising undergraduate and graduate curricula and individual courses to prepare students for data-centric careers, as well as to participate in a data-driven society. To meet these new challenges, programs are developing courses on, among other things, data curation, analytics, visualization, algorithm design, and artificial intelligence. While such changes reflect new workforce and society needs, it remains to be seen whether or not such efforts adequately address the very real and serious ethics and policy issues associated with related data practices (e.g., privacy, bias, fairness, and justice). The Information Ethics SIG and the Information Policy SIG have merged to present a panel on data ethics and policy issues in LIS education. In this session, two recent books on information ethics and information policy will be discussed to bring context to the panel, three papers will be presented, and the audience will have an opportunity to participate in a structured discussion. The papers will address three topics that explore the implications and concerns of living in a data-driven society: collaborative strategies for contributing to the data ethics education landscape, young adult information privacy concerns when using mobile devices, and artificial intelligence and social responsibility. The structured discussion will invite participation on issues raised by the papers, as well as implications for practice in LIS education

Pharmacological targeting of host chaperones protects from pertussis toxin in vitro and in vivo

Whooping cough is caused by Bordetella pertussis that releases pertussis toxin (PT) which comprises enzyme A-subunit PTS1 and binding/transport B-subunit. After receptor-mediated endocytosis, PT reaches the endoplasmic reticulum from where unfolded PTS1 is transported to the cytosol. PTS1 ADP-ribosylates G-protein alpha -subunits resulting in increased cAMP signaling. Here, a role of target cell chaperones Hsp90, Hsp70, cyclophilins and FK506-binding proteins for cytosolic PTS1-uptake is demonstrated. PTS1 specifically and directly interacts with chaperones in vitro and in cells. Specific pharmacological chaperone inhibition protects CHO-K1, human primary airway basal cells and a fully differentiated airway epithelium from PT-intoxication by reducing intracellular PTS1-amounts without affecting cell binding or enzyme activity. PT is internalized by human airway epithelium secretory but not ciliated cells and leads to increase of apical surface liquid. Cyclophilin-inhibitors reduced leukocytosis in infant mouse model of pertussis, indicating their promising potential for developing novel therapeutic strategies against whooping cough

Bimodal antagonism of PKA signalling by ARHGAP36

Protein kinase A is a key mediator of cAMP signalling downstream of G-protein-coupled receptors, a signalling pathway conserved in all eukaryotes. cAMP binding to the regulatory subunits (PKAR) relieves their inhibition of the catalytic subunits (PKAC). Here we report that ARHGAP36 combines two distinct inhibitory mechanisms to antagonise PKA signalling. First, it blocks PKAC activity via a pseudosubstrate motif, akin to the mechanism employed by the protein kinase inhibitor proteins. Second, it targets PKAC for rapid ubiquitin-mediated lysosomal degradation, a pathway usually reserved for transmembrane receptors. ARHGAP36 thus dampens the sensitivity of cells to cAMP. We show that PKA inhibition by ARHGAP36 promotes derepression of the Hedgehog signalling pathway, thereby providing a simple rationale for the upregulation of ARHGAP36 in medulloblastoma. Our work reveals a new layer of PKA regulation that may play an important role in development and disease

Grundwasser - Altlasten - Boden aktuell

Neun Fachbeiträge dokumentieren Ergebnisse der aktuellen Projekt- und Forschungsarbeit des Landesamtes in den Themenbereichen Grundwasser, Altlasten und Boden. Die Inhalte reichen vom Interesse aus Namibia für diese Themenbereiche über verschiedene Auswertungen von Hintergrundwerten und Hochwasser 2013 bis hin zu Planungen im Bereich Bodendauerbeobachtung in Sachsen. Einzelerkenntnisse zu Wülknitz bzw. zum Tagebau Witznitz werden in dem Heft fortgeführt

Gender Gap in Parental Leave Intentions: Evidence from 37 Countries

Despite global commitments and efforts, a gender-based division of paid and unpaid work persists. To identify how psychological factors, national policies, and the broader sociocultural context contribute to this inequality, we assessed parental-leave intentions in young adults (18–30 years old) planning to have children (N = 13,942; 8,880 identified as women; 5,062 identified as men) across 37 countries that varied in parental-leave policies and societal gender equality. In all countries, women intended to take longer leave than men. National parental-leave policies and women’s political representation partially explained cross-national variations in the gender gap. Gender gaps in leave intentions were paradoxically larger in countries with more gender-egalitarian parental-leave policies (i.e., longer leave available to both fathers and mothers). Interestingly, this cross-national variation in the gender gap was driven by cross-national variations in women’s (rather than men’s) leave intentions. Financially generous leave and gender-egalitarian policies (linked to men’s higher uptake in prior research) were not associated with leave intentions in men. Rather, men’s leave intentions were related to their individual gender attitudes. Leave intentions were inversely related to career ambitions. The potential for existing policies to foster gender equality in paid and unpaid work is discussed.Gender Gap in Parental Leave Intentions: Evidence from 37 CountriespublishedVersio

Gender Gap in Parental Leave Intentions: Evidence from 37 Countries

Despite global commitments and efforts, a gender-based division of paid and unpaid work persists. To identify how psychological factors, national policies, and the broader sociocultural context contribute to this inequality, we assessed parental-leave intentions in young adults (18–30 years old) planning to have children (N = 13,942; 8,880 identified as women; 5,062 identified as men) across 37 countries that varied in parental-leave policies and societal gender equality. In all countries, women intended to take longer leave than men. National parental-leave policies and women’s political representation partially explained cross-national variations in the gender gap. Gender gaps in leave intentions were paradoxically larger in countries with more gender-egalitarian parental-leave policies (i.e., longer leave available to both fathers and mothers). Interestingly, this cross-national variation in the gender gap was driven by cross-national variations in women’s (rather than men’s) leave intentions. Financially generous leave and gender-egalitarian policies (linked to men’s higher uptake in prior research) were not associated with leave intentions in men. Rather, men’s leave intentions were related to their individual gender attitudes. Leave intentions were inversely related to career ambitions. The potential for existing policies to foster gender equality in paid and unpaid work is discussed