Low pH gel intranasal sprays inactivate influenza viruses in vitro and protect ferrets against influenza infection

<p>Abstract</p> <p>Background</p> <p>Developing strategies for controlling the severity of pandemic influenza is a global public health priority. In the event of a pandemic there may be a place for inexpensive, readily available, effective adjunctive therapies to support containment strategies such as prescription antivirals, vaccines, quarantine and restrictions on travel. Inactivation of virus in the intranasal environment is one possible approach. The work described here investigated the sensitivity of influenza viruses to low pH, and the activity of low pH nasal sprays on the course of an influenza infection in the ferret model.</p> <p>Methods</p> <p>Inactivation of influenza A and avian reassortment influenza was determined using <it>in vitro </it>solutions tests. Low pH nasal sprays were tested using the ferret model with an influenza A Sydney/5/97 challenge. Clinical measures were shed virus, weight loss and body temperature.</p> <p>Results</p> <p>The virus inactivation studies showed that influenza viruses are rapidly inactivated by contact with acid buffered solutions at pH 3.5. The titre of influenza A Sydney/5/97 [H3N2] was reduced by at least 3 log cycles with one minute contact with buffers based on simple acid mixtures such as L-pyroglutamic acid, succinic acid, citric acid and ascorbic acid. A pH 3.5 nasal gel composition containing pyroglutamic acid, succinic acid and zinc acetate reduced titres of influenza A Hong Kong/8/68 [H3N2] by 6 log cycles, and avian reassortment influenza A/Washington/897/80 X A Mallard/New York/6750/78 [H3N2] by 5 log cycles, with 1 min contact.</p> <p>Two ferret challenge studies, with influenza A Sydney/5/97, demonstrated a reduction in the severity of the disease with early application of low pH nasal sprays versus a saline control. In the first study there was decreased weight loss in the treatment groups. In the second study there were reductions in virus shedding and weight loss, most notably when a gelling agent was added to the low pH formulation.</p> <p>Conclusion</p> <p>These findings indicate the potential of a low pH nasal spray as an adjunct to current influenza therapies, and warrant further investigation in humans.</p

Activation of Toll-like receptor 5 in microglia modulates their function and triggers neuronal injury

Microglia are the primary immune-competent cells of the central nervous system (CNS) and sense both pathogen- and host-derived factors through several receptor systems including the Toll-like receptor (TLR) family. Although TLR5 has previously been implicated in different CNS disorders including neurodegenerative diseases, its mode of action in the brain remained largely unexplored. We sought to determine the expression and functional consequences of TLR5 activation in the CNS. Quantitative real-time PCR and immunocytochemical analysis revealed that microglia is the major CNS cell type that constitutively expresses TLR5. Using Tlr5(-/-) mice and inhibitory TLR5 antibody we found that activation of TLR5 in microglial cells by its agonist flagellin, a principal protein component of bacterial flagella, triggers their release of distinct inflammatory molecules, regulates chemotaxis, and increases their phagocytic activity. Furthermore, while TLR5 activation does not affect tumor growth in an ex vivo GL261 glioma mouse model, it triggers microglial accumulation and neuronal apoptosis in the cerebral cortex in vivo. TLR5-mediated microglial function involves the PI3K/Akt/mammalian target of rapamycin complex 1 (mTORC1) pathway, as specific inhibitors of this signaling pathway abolish microglial activation. Taken together, our findings establish TLR5 as a modulator of microglial function and indicate its contribution to inflammatory and injurious processes in the CNS

Neurofibromatosis 1 (NF1) mutation results in impaired function of human induced pluripotent stem cell-derived microglia

Neurofibromatosis type 1 (NF1) is an autosomal dominant condition caused by germline mutations in the NF1 gene. Children with NF1 are prone to the development of multiple nervous system abnormalities, including autism and brain tumors, which could reflect the effect of NF1 mutation on microglia function. Using heterozygous Nf1-mutant mice, we previously demonstrated that impaired purinergic signaling underlies deficits in microglia process extension and phagocytosis in situ. To determine whether these abnormalities are also observed in human microglia in the setting of NF1, we leveraged an engineered isogenic series of human induced pluripotent stem cells to generate human microglia-like (hiMGL) cells heterozygous for three different NF1 patient-derived NF1 gene mutations. While all NF1-mutant and isogenic control hiMGL cells expressed classical microglia markers and exhibited similar transcriptomes and cytokine/chemokine release profiles, only NF1-mutant hiMGL cells had defects in P2X receptor activation, phagocytosis and motility. Taken together, heterozygous NF1 mutation impairs a subset of human microglia functional properties, which could contribute to the neurological abnormalities seen in children with NF1

Microglia/macrophage derived human CCL18 promotes glioma progression via CCR8-ACP5 axis analyzed in humanized slice model

Factors released from glioma-associated microglia/macrophages (GAMs) play a crucial role in glioblastoma multiforme (GBM) progression. Here, we study the importance of CCL18, a cytokine expressed in human but not in rodent GAMs, as a modulator of glioma growth. Since CCL18 signaling could not be studied in classical mouse glioma models, we developed an approach by transplanting induced pluripotent stem cell-derived human microglia and human glioma cells into mouse brain slices depleted of their intrinsic microglia. We observe that CCL18 promotes glioma cell growth and invasion. Chemokine (C-C motif) receptor 8 (CCR8) is identified as a functional receptor for CCL18 on glioma cells, and ACP5 (acid phosphatase 5) is revealed as an important part of the downstream signaling cascade for mediating glioma growth. We conclude, based on the results from an in vitro, ex vivo humanized glioma model and an in vivo GBM model that microglia/macrophage-derived CCL18 promotes glioma growth

Characterization of OSIRIS NAC filters for the interpretation of multispectral data of comet 67P/Churyumov-Gerasimenko

We interpret multicolor data from OSIRISNAC for the remote-sensing exploration of comet 67P/Churyumov-Gerasimenko. We determine the most meaningful definition of color maps for the characterization of surface variegation with filters available on OSIRIS NAC. Methods. We analyzed laboratory spectra of selected minerals and olivine-pyroxene mixtures seen through OSIRIS NAC filters, with spectral methods existing in the literature: reflectance ratios, minimum band wavelength, spectral slopes, band tilt, band curvature, and visible tilt. Results. We emphasize the importance of reflectance ratios and particularly the relation of visible tilt vs. band tilt. This technique provides a reliable diagnostic of the presence of silicates. Color maps constructed by red-green-blue colors defined with the green, orange, red, IR, and Fe2O3 filters let us define regions that may significantly differ in composition

Regional unit definition for the nucleus of comet 67P/Churyumov-Gerasimenko on the SHAP7 model

The previously defined regions on the nucleus of comet 67P/Churyumov-Gerasimenko have been mapped back onto the 3D SHAP7 model of the nucleus (Preusker et al., 2017). The resulting regional definition is therefore self-consistent with boundaries that are well defined in 3 dimensions. The facets belonging to each region are provided as supplementary material. The shape model has then been used to assess inhomogeneity of nucleus surface morphology within individual regions. Several regions show diverse morphology. We propose sub-division of these regions into clearly identifiable units (sub-regions) and a comprehensive table is provided. The surface areas of each sub-region have been computed and statistics based on grouping of unit types are provided. The roughness of each region is also provided in a quantitative manner using a technique derived from computer graphics applications. The quantitative method supports the sub-region definition by showing that differences between sub-regions can be numerically justified

Shape model, reference system definition, and cartographic mapping standards for comet 67P/Churyumov-Gerasimenko - Stereo-photogrammetric analysis of Rosetta/OSIRIS image data

We analyzed more than 200 OSIRIS NAC images with a pixel scale of 0.9-2.4 m/pixel of comet 67P/Churyumov-Gerasimenko (67P) that have been acquired from onboard the Rosetta spacecraft in August and September 2014 using stereo-photogrammetric methods (SPG). We derived improved spacecraft position and pointing data for the OSIRIS images and a high-resolution shape model that consists of about 16 million facets (2 m horizontal sampling) and a typical vertical accuracy at the decimeter scale. From this model, we derive a volume for the northern hemisphere of 9.35 km3 ± 0.1 km3. With the assumption of a homogeneous density distribution and taking into account the current uncertainty of the position of the comet's center-of-mass, we extrapolated this value to an overall volume of 18.7 km3± 1.2 km3, and, with a current best estimate of 1.0 × 1013 kg for the mass, we derive a bulk density of 535 kg/m3± 35 kg/m3. Furthermore, we used SPG methods to analyze the rotational elements of 67P. The rotational period for August and September 2014 was determined to be 12.4041 ± 0.0004 h. For the orientation of the rotational axis (z-axis of the body-fixed reference frame) we derived a precession model with a half-cone angle of 0.14°, a cone center position at 69.54°/64.11° (RA/Dec J2000 equatorial coordinates), and a precession period of 10.7 days. For the definition of zero longitude (x-axis orientation), we finally selected the boulder-like Cheops feature on the big lobe of 67P and fixed its spherical coordinates to 142.35° right-hand-rule eastern longitude and -0.28° latitude. This completes the definition of the new Cheops reference frame for 67P. Finally, we defined cartographic mapping standards for common use and combined analyses of scientific results that have been obtained not only within the OSIRIS team, but also within other groups of the Rosetta mission

Search for satellites near comet 67P/Churyumov-Gerasimenko using Rosetta/OSIRIS images

The European Space Agency Rosetta mission reached and started escorting its main target, the Jupiter-family comet 67P/Churyumov-Gerasimenko, at the beginning of August 2014. Within the context of solar system small bodies, satellite searches from approaching spacecraft were extensively used in the past to study the nature of the visited bodies and their collisional environment. During the approaching phase to the comet in July 2014, the OSIRIS instrument onboard Rosetta performed a campaign aimed at detecting objects in the vicinity of the comet nucleus and at measuring these objects' possible bound orbits. In addition to the scientific purpose, the search also focused on spacecraft security to avoid hazardous material in the comet's environment. Methods. Images in the red spectral domain were acquired with the OSIRIS Narrow Angle Camera, when the spacecraft was at a distance between 5785 km and 5463 km to the comet, following an observational strategy tailored to maximize the scientific outcome. From the acquired images, sources were extracted and displayed to search for plausible displacements of all sources from image to image. After stars were identified, the remaining sources were thoroughly analyzed. To place constraints on the expected displacements of a potential satellite, we performed Monte Carlo simulations on the apparent motion of potential satellites within the Hill sphere. Results. We found no unambiguous detections of objects larger than ~6 m within ~20 km and larger than ~1 m between ~20 km and ~110 km from the nucleus, using images with an exposure time of 0.14 s and 1.36 s, respectively. Our conclusions are consistent with independent works on dust grains in the comet coma and on boulders counting on the nucleus surface. Moreover, our analysis shows that the comet outburst detected at the end of April 2014 was not strong enough to eject large objects and to place them into a stable orbit around the nucleus. Our findings underline that it is highly unlikely that large objects survive for a long time around cometary nuclei

Insolation, erosion, and morphology of comet 67P/Churyumov-Gerasimenko

The complex shape of comet 67P and its oblique rotation axis cause pronounced seasonal effects. Irradiation and hence activity vary strongly. Aims. We investigate the insolation of the cometary surface in order to predict the sublimation of water ice. The strongly varying erosion levels are correlated with the topography and morphology of the present cometary surface and its evolution. Methods. The insolation as a function of heliocentric distance and diurnal (spin dependent) variation is calculated using &gt;105 facets of a detailed digital terrain model. Shading, but also illumination and thermal radiation by facets in the field of view of a specific facet are iteratively taken into account. We use a two-layer model of a thin porous dust cover above an icy surface to calculate the water sublimation, presuming steady state and a uniform surface. Our second model, which includes the history of warming and cooling due to thermal inertia, is restricted to a much simpler shape model but allows us to test various distributions of active areas. Results. Sublimation from a dirty ice surface yields maximum erosion. A thin dust cover of 50 μm yields similar rates at perihelion. Only about 6% of the surface needs to be active to match the observed water production rates at perihelion. A dust layer of 1 mm thickness suppresses the activity by a factor of 4 to 5. Erosion on the south side can reach more than 10 m per orbit at active spots. The energy input to the concave neck area (Hapi) during northern summer is enhanced by about 50% owing to self-illumination. Here surface temperatures reach maximum values along the foot of the Hathor wall. Integrated over the whole orbit this area receives the least energy input. Based on the detailed shape model, the simulations identify "hot spots" in depressions and larger pits in good correlation with observed dust activity. Three-quarters of the total sublimation is produced while the sub-solar latitude is south, resulting in a distinct dichotomy in activity and morphology. Conclusions. The northern areas display a much rougher morphology than what is seen on Imhotep, an area at the equator that will be fully illuminated when 67P is closer to the Sun. Self-illumination in concave regions enhance the energy input and hence erosion. This explains the early activity observed at Hapi. Cliffs are more prone to erosion than horizontal, often dust covered, areas, which leads to surface planation. Local activity can only persist if the forming cliff walls are eroding. Comet 67P has two lobes and also two distinct sides. Transport of material from the south to the north is probable. The morphology of the Imhotep plain should be typical for the terrains of the yet unseen southern hemisphere