Viral Capsid Is a Pathogen-Associated Molecular Pattern in Adenovirus Keratitis

Human adenovirus (HAdV) infection of the human eye, in particular serotypes 8, 19 and 37, induces the formation of corneal subepithelial leukocytic infiltrates. Using a unique mouse model of adenovirus keratitis, we studied the role of various virus-associated molecular patterns in subsequent innate immune responses of resident corneal cells to HAdV-37 infection. We found that neither viral DNA, viral gene expression, or viral replication was necessary for the development of keratitis. In contrast, empty viral capsid induced keratitis and a chemokine profile similar to intact virus. Transfected viral DNA did not induce leukocyte infiltration despite CCL2 expression similar to levels in virus infected corneas. Mice without toll-like receptor 9 (Tlr9) signaling developed clinical keratitis upon HAdV-37 infection similar to wild type mice, although the absolute numbers of activated monocytes in the cornea were less in Tlr9−/− mice. Virus induced leukocytic infiltrates and chemokine expression in mouse cornea could be blocked by treatment with a peptide containing arginine glycine aspartic acid (RGD). These results demonstrate that adenovirus infection of the cornea induces chemokine expression and subsequent infiltration by leukocytes principally through RGD contact between viral capsid and the host cell, possibly through direct interaction between the viral capsid penton base and host cell integrins

Cosmic kidney disease: an integrated pan-omic, physiological and morphological study into spaceflight-induced renal dysfunction.

Missions into Deep Space are planned this decade. Yet the health consequences of exposure to microgravity and galactic cosmic radiation (GCR) over years-long missions on indispensable visceral organs such as the kidney are largely unexplored. We performed biomolecular (epigenomic, transcriptomic, proteomic, epiproteomic, metabolomic, metagenomic), clinical chemistry (electrolytes, endocrinology, biochemistry) and morphometry (histology, 3D imaging, miRNA-ISH, tissue weights) analyses using samples and datasets available from 11 spaceflight-exposed mouse and 5 human, 1 simulated microgravity rat and 4 simulated GCR-exposed mouse missions. We found that spaceflight induces: 1) renal transporter dephosphorylation which may indicate astronauts' increased risk of nephrolithiasis is in part a primary renal phenomenon rather than solely a secondary consequence of bone loss; 2) remodelling of the nephron that results in expansion of distal convoluted tubule size but loss of overall tubule density; 3) renal damage and dysfunction when exposed to a Mars roundtrip dose-equivalent of simulated GCR

Assessment of a panel of interleukin-8 reporter lung epithelial cell lines to monitor the pro-inflammatory response following zinc oxide nanoparticle exposure under different cell culture conditions

Stably transfected lung epithelial reporter cell lines pose an advantageous alternative to replace complex experimental techniques to monitor the pro-inflammatory response following nanoparticle (NP) exposure. Previously, reporter cell lines have been used under submerged culture conditions, however, their potential usefulness in combination with air-liquid interface (ALI) exposures is currently unknown. Therefore, the aim of the present study was to compare a panel of interleukin-8 promoter (pIL8)-reporter cell lines (i.e. green or red fluorescent protein (GFP, RFP), and luciferase (Luc)), originating from A549 lung epithelial type II-like cells cells, following NPs exposure under both submerged and ALI conditions. All cell lines were exposed to zinc oxide (ZnO) NPs at 0.6 and 6.2 μg/cm 2 for 3 and 16 hours under both submerged and ALI conditions. Following physicochemical characterization, the cytotoxic profile of the ZnO-NPs was determined for each exposure scenario. Expression of IL-8 from all cell types was analyzed at the promoter level and compared to the mRNA (qRT-PCR) and protein level (ELISA). In summary, each reporter cell line detected acute pro-inflammatory effects following ZnO exposure under each condition tested. The pIL8-Luc cell line was the most sensitive in terms of reporter signal strength and onset velocity following TNF-α treatment. Both pIL8-GFP and pIL8-RFP also showed a marked signal induction in response to TNF-α, although only after 16 hrs. In terms of ZnO-NP-induced cytotoxicity pIL8-RFP cells were the most affected, whilst the pIL8-Luc were found the least responsive. In conclusion, the use of fluorescence-based reporter cell lines can provide a useful tool in screening the pro-inflammatory response following NP exposure in both submerged and ALI cell cultures. The online version of this article (doi:10.1186/s12989-015-0104-6) contains supplementary material, which is available to authorized users

Making the Anscombe-Aumann approach to ambiguity suitable for descriptive applications

The Anscombe-Aumann (AA) model, originally introduced to give a normative basis to expected utility, is nowadays mostly used for another purpose: to analyze deviations from expected utility due to ambiguity (unknown probabilities). The AA model makes two ancillary assumptions that do not refer to ambiguity: expected utility for risk and backward induction. These assumptions, even if normatively appropriate, fail descriptively. This paper relaxes these ancillary assumptions to avoid the descriptive violations, while maintaining AA\xe2\x80\x99s convenient mixture operation. Thus, it becomes possible to test and apply all AA-based ambiguity theories descriptively while avoiding confounds due to violated ancillary assumptions. The resulting tests use only simple stimuli, avoiding noise due to complexity. We demonstrate the latter in a simple experiment where we find that three assumptions about ambiguity, commonly made in AA theories, are violated: reference independence, universal ambiguity aversion, and weak certainty independence. The second, theoretical, part of the paper accommodates the violations found for the first ambiguity theory in the AA model\xe2\x80\x94Schmeidler\xe2\x80\x99s CEU theory\xe2\x80\x94by introducing and axiomatizing a reference dependent generalization. That is, we extend the AA ambiguity model to prospect theory

Abstract P6-15-17: Synergy of Ixabepilone and Dasatinib in Triple Negative Breast Cancer Cell Lines

Abstract Background: Treatment options are very limited for breast cancers that do not express HER2 or estrogen and progesterone receptors (Triple Negative Breast Cancer; TNBC). Dasatinib is an orally active small molecule inhibitor of the src kinase. It has been proposed that dasatinib inhibits growth of breast cancer cells by modulating epidermal growth factor receptor (EGFR) signaling. Approximately 70% of breast cancers over express EGFR family members as well as c-Src. Src is co-expressed with at least one of the EGFRs. Both src and EGFR1 expression have been associated with TNBC. Dasatinib has previously shown activity as a single agent in TNBC cell lines. However, dasatinib has limited single agent activity in a cohort of chemotherapy refractory TNBC patients according to a phase I/II study. We believe that dasatinib may sensitize breast cancer cells to anti-mitotic agents. Ixabepilone is a potent anti-mitotic agent that has shown significant response in TNBC patients, both in the neoadjuvant setting and in the metastatic setting, including those cancers that are taxane refractory. Therefore, combining these two drugs may have utility in this setting and may lead to inhibition of tumor cell proliferation, invasion, metastasis, angiogenesis, and tumor cell survival. Objectives: To study the activity of dasatinib combined with Ixabepilone in breast cancer cell lines. Methods: A panel of 8 breast cancer cell lines was used in these experiments. These included MCF-7(ER+), T-47D (ER +), SKBR3 (HER2+), and five triple negative cell lines: BT-549, HS578T, MDA-MB-157, MDA-MB-231, MDA-MB-436. First, these cell lines were evaluated for src expression by Western blot as well as sensitivity to dasatinib alone, ixabepilone alone and then the combination of these two drugs using a high-throughput, 384- well CellTiter-Blue cell viability assay. Percent viability was calculated from four replicate cell wells after 72-hour drug incubation and IC50s were determined. Experiments were repeated a minimum of three times. Synergistic, additive, or antagonistic effects were determined using the combination index (CI) method developed by Chou and Talalay. Results: TNBC cell lines expressed higher levels of src protein than cell lines with positive expression of HER2, ER and/or PR by Western blot analysis. Viability assays suggested a correlation between src expression and dasatinib IC50. Combinations of dasatinib and ixabepilone were determined to have combinational indexes &amp;lt;1 in all cell lines tested, suggestive of synergistic activity. Conclusion: This study demonstrates that in a range of breast cell lines with varying levels of src expression, dasatinib combined with ixabepilone is a synergistic combination and may prove to be beneficial to patients with very limited treatment options. Figure available in online version. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-15-17.</jats:p

Use of neutron reflectivity to measure the dynamics of solvation and structural changes in polyvinylferrocene films during electrochemically controlled redox cycling

Time-resolved specular neutron reflectivity measurements are presented and interpreted for electroactive polyvinylferrocene (PVF) films subject to potentiodynamic electrochemical control. New data acquisition methodology allows an effective measurement time scale on the order of seconds, which is an improvement over conventional methodology by 2 to 3 orders of magnitude. Reflectivity profiles were obtained for PVF films exposed to aqueous 0.1 M NaClO4 in which PVF films are thermodynamically permselective, with contrast variation via H2O and D2O. Irrespective of any model, the raw profiles show chemically reversible film "breathing" due to redox-driven solvent entry and exit during polymer oxidation and reduction, respectively. Modeling reveals three compositionally distinct regions within the polymer film: interfacial regions at the electrode and solution interfaces and a "bulk" interior. The new methodology, supported by simultaneous in situ visible transmission spectroscopy, reveals an unprecedented level of insight into the temporal and spatial mechanistic details of film solvation changes, including a two-stage (de)solvation mechanism for redox switching, differences in interior (in)homogeneity for reduced and oxidized films, and permselectivity failure under dynamic electrochemical conditions for the reduced (but not oxidized) state, in contrast to static conditions that allow permselectivity for both states