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Deer are the most abundant large herbivores in temperate and boreal forests across the Northern Hemisphere. They are ecosystem engineers known to alter understory vegetation and future tree species composition by selective browsing. Also, deer have strong impacts on faunistic groups, often mediated by vegetation. The ongoing loss of faunal biodiversity in forests worldwide can be exacerbated by high deer population densities. Adapted deer management for the purpose of forest biodiversity conservation requires a holistic understanding of deer–fauna relationships. In this systematic map, we examine the existing literature assessing the effects of deer on faunal communities in boreal and temperate forests. Our aim is to synthesize currently described trends and identify research gaps for our understanding of deer as biotic drivers of forest communities. We reviewed 64 studies on how the abundance, species richness, or diversity of faunal taxa responded to different levels of deer abundance or density in forest ecosystems across the Northern Hemisphere. In total, we found almost 400 individual reported effects of nine native deer species on forest‐dwelling faunal communities. However, our systematic map reveals that comprehensive synthesis of the current literature remains a challenge. Published studies often do not report contextual data essential for comparison and meta‐analysis, for example, deer density, forest management, and predation pressure. Moreover, the methodological approaches of the included studies often only account for potential linear effects of deer on fauna, likely oversimplifying the complexity of direct and indirect effects that deer can have on their ecosystem. We recommend that multi‐level enclosure experiments be applied to assess the impact on faunal taxa. This approach combines robust causal inference with the potential straightforward comparison and replication between deer species, forest types, and system productivity, which will facilitate the utilization of the results in future research and management. [ABSTRACT FROM AUTHOR]

Purpose: Adaptive load-bearing structures pursue the approach of saving mass within a load-bearing structure by adding external energy, thus saving materials and resources. This paper provides an overview of current research developments and shows some examples of existing prototypes. Design/methodology/approach: First, basic terms and definitions from the research field of adaptive structures are introduced. After a brief historical insight, the numerical methods and prototypes used are presented as examples. The paper concludes with a summary of the state-of-the-art and open questions. Findings: The current state of the art shows that the idea of adaptive structures offers great potential for more sustainability and resource efficiency in the construction industry. However, it also shows that research is still at the basic stage and that there are still some gaps in research. Originality/value: The implementation of adaptive load-bearing structures is just one of many different approaches to greater sustainability in the construction sector. The issue of adaptive structures is a highly interdisciplinary field of research. The following paper is a literature review intended to summarize and critically evaluate the state-of-the-art research in this field. In the final section, some open questions are addressed, indicating that this research topic is still evolving. [ABSTRACT FROM AUTHOR]

Biodiversity loss due to intensive timber production is a ubiquitous conservation issue across temperate and boreal forest ecosystems. Retention forestry, the retention of deadwood and old-growth features within production forest, is one management strategy that has been implemented in various countries around the world to conserve a wide range of taxa within managed forests. The success and ecological implications of retention forestry are currently subject to intensive investigation and while some taxa like birds and insects have already been studied frequently, larger mammals have received less attention. Pine martens are one of the few larger mammals in central Europe preferring older forest and potentially profiting directly from deadwood retention as a consequence of implemented retention forestry. The goal of our study was to assess the response of European marten species to deadwood retention in montane mixed forests. Using marten detection rates from camera traps on 135 research plots we assessed the response of martens to deadwood at three different spatial scales using generalized linear mixed models. We found no effect of lying deadwood on marten detections at the plot scale (1 ha) or in a 10 m radius around the camera traps. However, we found a significant increase of marten detections if logs (> 10 cm in diameter) were directly in front and in view of the camera trap. Our results show that deadwood retention as a measure of retention forestry does affect microhabitat use of martens, but not stand selection during the growing season. Logs directly in view of the camera trap increase marten detection rates as martens choose to move and forage along fallen trees when they are available. When using camera trapping to collect data on martens, trap positioning in front of logs can heavily bias trapping results when unaccounted for. [ABSTRACT FROM AUTHOR]

Carrion is a valuable resource in forests, providing sustenance for vertebrate and invertebrate scavenger communities and contributing to ecosystem functions, such as nutrient cycling. Intensive ungulate hunting, and thereby extraction of carcasses, removes large quantities of potential carrion from the system, denying a valuable resource from scavenger fauna. It may be possible to reduce the loss and negative consequences to forest biodiversity by retaining evisceration residues from hunted deer, where full carcasses cannot be retained. However, what roll evisceration residues play as a resource for scavengers in temperate forests is not well understood. In this study, we exposed 47 carrion samples from hunted roe deer, in front of triple sets of camera traps, to examine how hunting remains are removed and fed upon by vertebrate scavengers. Overall, 70% of the samples were completely removed from experimental sites by vertebrates. We detected twelve vertebrate taxa feeding on evisceration residues, including martens (Martes spp.), red kites Milvus milvus and garden dormice Eliomys quercinus. Common buzzards Buteo buteo and Eurasian jays Garrulus glandarius were the most frequent feeders on carrion samples, while red foxes Vulpes vulpes displaced the largest proportion of samples. Finally, we found a range of insectivorous bird and mammal species using hunting remains as a source for invertebrate prey, while not scavenging on the remains directly. We demonstrate that evisceration residues can be a valuable resource for a wide range of taxa and suggest that viscera retention from hunted game may contribute to resource provisioning for scavengers in forest ecosystems. [ABSTRACT FROM AUTHOR]

Selective browsing by deer on young trees may impede the management goal of increasing forest resilience against climate change and other disturbances. Deer population density is often considered the main driver of browsing impacts on young trees, however, a range of other variables such as food availability also affect this relationship. In this study, we use browsing survey data from 135 research plots to explore patterns of roe deer (Capreolus capreolus) browsing pressure on woody plants in mountainous forests in central Europe. We fitted species‐specific generalised linear mixed models for eight woody taxa, assessing the potential effects of understory characteristics, roe deer abundance and lying deadwood on browsing intensity. Our study reveals conspecific and associational effects for woody taxa that are intermediately browsed by roe deer. Selective browsing pressure was mediated by preferences of plants, in that, browsing of strongly preferred woody taxa as for example mountain ash (Sorbus aucuparia) and of least preferred woody taxa, for example Norway spruce (Picea abies) was not affected by the surrounding understory vegetation, while browsing pressure on intermediately browsed species like for example silver fir (Abies alba) was affected by understory characteristics. Contrary to our expectations, roe deer abundance was only positively associated with browsing pressure on silver fir and bilberry (Vaccinium myrtillus), while all other plants were unaffected by deer abundance. Finally, we did not find an influence of lying deadwood volume on the browsing pressure on any woody‐plant species. Overall, our results indicate that patterns in browsing preference and intensity are species‐specific processes and are partly affected by the surrounding understory vegetation. Current management strategies that aim to reduce browsing pressure through culling may be inefficient as they do not address other drivers of browsing pressure. However, managers also need to consider the characteristics of the local understory vegetation in addition to deer abundance and design species‐specific plans to reduce browsing on woody plant taxa. [ABSTRACT FROM AUTHOR]

The focus of this study is on the geography of robotics Research and Development (R&D) activities. The objectives are, first, to identify hotspots in robotics R&D worldwide, and second, to characterise structures and dynamics of global robotics R&D collaboration networks through detailed geographical lenses of global urban areas. We use patents as marker for R&D activities, and accordingly focus on technologically oriented R&D, drawing on information from patents applied for between 2002 and 2016. We employ an appropriate search strategy to identify relevant robotics patents based on detailed levels of the Cooperative Patent Classification (CPC) and assign patents to more than 900 global urban areas based on the inventor addresses. The co-patent networks are examined from a Social Network Analysis (SNA) perspective by means of robotics co-patents, contributing to a global network where urban areas are the nodes inter-linked by joint inventive activities recorded in robotics patents. Global SNA measures illustrate structures and dynamics of the network as a whole, while local measures indicate the specific positioning and roles of urban areas in the network. The results are original in characterising the global spatial emergence of this generic new industry, highlighting prominent urban hotspots in terms of specialisation in robotics R&D, pointing to a global shift reflected by the increasing role of emerging economies, in particular China. The global robotics R&D has grown significantly both in total patenting and also in terms of R&D collaboration activities between urban areas. Also, for the networks, growth is not equally distributed, but is rather characterised by significant spatial shifts, both in terms of cities declining or climbing up the specialisation ranking, but even more in terms of the spatial network structure. [ABSTRACT FROM AUTHOR]

Carrion of wild ungulates is an important resource for microbes, invertebrates and vertebrates, and affects local plant communities and soils. Most wild ungulates in central Europe, however, are intensely hunted and removed from the ecosystem, thus not available as carrion. This study explores the use of evisceration residues as a resource by necrophagous insects in a temperate mountainous forest in Germany. We experimentally compared the relative abundance of necrophagous insects between sites of caged roe deer carcasses and evisceration residues of roe deer using pitfall-traps. While evisceration residues were used by necrophagous insects, significantly more individuals were sampled at entire deer carcasses; when corrected for biomass, however, no signifi- cant difference in abundance for Coleoptera groups was found. Overall, evisceration residues were used earlier and for a shorter period than entire deer carcasses. Leaving evisceration residues on site where game animals are obtained might help maintain specialized necrophagous insect communities, although their ecological role cannot completely replace that of whole carcasses of wild ruminants. [ABSTRACT FROM AUTHOR]

Carrion of wild ungulates is an important resource for microbes, invertebrates and vertebrates, and affects local plant communities and soils. Most wild ungulates in central Europe, however, are intensely hunted and removed from the ecosystem, thus not available as carrion. This study explores the use of evisceration residues as a resource by necrophagous insects in a temperate mountainous forest in Germany. We experimentally compared the relative abundance of necrophagous insects between sites of caged roe deer carcasses and evisceration residues of roe deer using pitfall‐traps. While evisceration residues were used by necrophagous insects, significantly more individuals were sampled at entire deer carcasses; when corrected for biomass, however, no significant difference in abundance for Coleoptera groups was found. Overall, evisceration residues were used earlier and for a shorter period than entire deer carcasses. Leaving evisceration residues on site where game animals are obtained might help maintain specialized necrophagous insect communities, although their ecological role cannot completely replace that of whole carcasses of wild ruminants. [ABSTRACT FROM AUTHOR]

Investigates the stability and reactivity of the surface and mixed structures via various nitrogen oxide-induced reconstructions, in titration experiments as well as under stationary reaction conditions using work function, rate measurements and thermal desorption spectroscopy; Drawback of the oscillation mechanism.

Knowledge about Antarctic sea-ice volume and its changes over the past decades has been sparse due to the lack of systematic sea-ice thickness measurements in this remote area. Recently, first attempts have been made to develop a sea-ice thickness product over the Southern Ocean from space-borne radar altimetry and results look promising. Today, more than 20 years of radar altimeter data are potentially available for such products. However, the characteristics of individual radar types differ for the available altimeter missions. Hence, it is important and our goal to study the consistency between single sensors in order to develop long and consistent time series. Here, the consistency between freeboard measurements of the Radar Altimeter 2 on board Envisat and freeboard measurements from the Synthetic- Aperture Interferometric Radar Altimeter on board CryoSat- 2 is tested for their overlap period in 2011. Results indicate that mean and modal values are in reasonable agreement over the sea-ice growth season (May-October) and partly also beyond. In general, Envisat data show higher freeboards in the first-year ice zone while CryoSat-2 freeboards are higher in the multiyear ice zone and near the coasts. This has consequences for the agreement in individual sectors of the Southern Ocean, where one or the other ice class may dominate. Nevertheless, over the growth season, mean freeboard for the entire (regionally separated) Southern Ocean differs generally by not more than 3 cm (8 cm, with few exceptions) between Envisat and CryoSat-2, and the differences between modal freeboards lie generally within ± 10 cm and often even below. [ABSTRACT FROM AUTHOR]

The spike protein S of transmissible gastroenteritis virus, an Alphacoronavirus , contains a tyrosine-based sorting signal that is responsible for ERGIC retention and may be important for a correct viral assembly process. To find out whether the S protein interacts with cellular proteins via this sorting signal, a pulldown assay with GST fusion proteins was performed. Filamin A has been identified as a putative interaction candidate. Immunofluorescence assays confirmed a co-localization between the TGEV S protein and filamin A. Further experiments have to be performed to prove a significant impact of filamin A on TGEV infection. Different approaches of several researchers for the identification of cellular interaction candidates relevant for coronavirus replication are summarized. These results may help in the future to identify the role of cellular proteins during coronavirus assembly at the ER-Golgi intermediate compartment. [ABSTRACT FROM AUTHOR]

Many flying insects, such as flies, wasps and bees, pursue a saccadic flight and gaze strategy. This behavioral strategy is thought to separate the translational and rotational components of self-motion and, thereby, to reduce the computational efforts to extract information about the environment from the retinal image flow. Because of the distinguishing dynamic features of this active flight and gaze strategy of insects, the present study analyzes systematically the spatiotemporal statistics of image sequences generated during saccades and intersaccadic intervals in cluttered natural environments. We show that, in general, rotational movements with saccade-like dynamics elicit fluctuations and overall changes in brightness, contrast and spatial frequency of up to two orders of magnitude larger than translational movements at velocities that are characteristic of insects. Distinct changes in image parameters during translations are only caused by nearby objects. Image analysis based on larger patches in the visual field reveals smaller fluctuations in brightness and spatial frequency composition compared to small patches. The temporal structure and extent of these changes in image parameters define the temporal constraints imposed on signal processing performed by the insect visual system under behavioral conditions in natural environments. [ABSTRACT FROM AUTHOR]

Der Offshore-Windpark Amrumbank West liegt 35 km nördlich der Insel Helgoland und 35 km westlich der Insel Amrum in der Ausschließlichen Wirtschaftzone Deutschlands in der Nordsee und befindet sich derzeit in der Realisierungsphase. Nach mehr als dreijähriger Planungs- und Genehmigungsphase berichtet dieser Beitrag über die Planung und Genehmigung des Offshore-Windparks. Neben den für die 80 Monopile-Tragstrukturen in knapp 25 m Wassertiefe maßgebenden und bereits in der statischen Vorplanungsphase berücksichtigten Beanspruchungen werden Besonderheiten in der Genehmigungsphase beschrieben und die erforderlichen Stahltonnagen für die Projektumsetzung vorgestellt. Abschließend werden erste Meilensteine in der Projektrealisierung erläutert und ein Überblick über die ausgeführten Sonderlösungen gegeben. Planning, certification and installation of Amrumbank West offshore wind farm. The offshore wind farm Amrumbank West is located 35 km in the north of Helgoland and 35 km kilometers in the west of the island of Amrum in the exclusive economic zone of Germany. The installation of Amrumbank West is currently ongoing. After more than three years of development and certification, this paper deals with the main topics of planning and certification of the offshore wind farm. Besides the design driving parameters which have been already considered during the conceptual design phase of the 80 monopile substructures in nearly 25 m water depths, specific features of the certification process will be discussed. Furthermore, first milestones of the installation and special technical solutions are presented. [ABSTRACT FROM AUTHOR]

Future generations should not be burdened with debt, but their security needs must also be met. [ABSTRACT FROM AUTHOR]

Abstract: A photocatalytic disinfection with P25 TiO2 was conducted at three pH-values (4, 7 and 10) in NaCl solution (c = 10 mmol L−1). Neither the osmotic stress nor solely the pH-value had a significant impact on the survival of the Escherichia coli cells. The cells were more damaged at pH 4 than at pH 7 and 10 in irradiation experiments with TiO2. The smallest disinfection rates occurred at pH 10. SEM images revealed that the surface of the cells was covered with TiO2 to a greater extent at pH 4 compared to pH 10. At pH 10, the disinfection rates increased when increasing the ionic strength and changing the NaCl solution to CaCl2. The results indicate that the disinfection rates depend on the electrostatic interaction between cells and TiO2. The differences could be qualitatively better explained by the XDLVO theory in comparison to the DLVO theory. [Copyright &y& Elsevier]

Environmental protection is close related with environmental education. Environmental education is as necessary in the educational system as outside it. The present study, starting from the perception of the representatives of the local rural public administration and the citizens of the rural communities on environmental protection issues: (i) highlights the need for information and education in the field of environmental protection, especially with regard to environmental legislation and waste management; (ii) demonstrates the usefulness of environmental educational projects and the use of open educational resources, as resources that can be easily adapted to the real needs of education and information. [ABSTRACT FROM AUTHOR]

Abstract: The influence of the zeta potential on the sorption between microorganisms (Saccharomyces cerevisiae and Escherichia coli) and iron oxide nanoparticles is demonstrated in a model salt solution at two different pH-values. There was only a 1% survival rate of E. coli (4.5×107 cells/mL) in the presence of 24mg/L nanoparticulate iron oxide at pH 4. S. cerevisiae were less affected by the presence of the nanoparticulate iron oxide. The extent of iron oxide nanoparticle coverage on the surface of the microorganisms appears to be related to electrostatic interaction forces. Furthermore, the toxic effect of the nanoparticle concentration follows the sorption isotherm for E. coli. Based on the resulting hydrodynamic size distributions in the supernatant after sorption experiments, it could be shown that predominantly smaller particle aggregates oxide were sorbed onto E. coli. This was evident by a shift in the particle size distribution towards a larger mean particle size. The effect was observed to a lower extent for S. cerevisiae. The extent of iron oxide nanoparticle sorption on E. coli quickly reached a maximum and remained constant during a 24h period compared to S. cerevisiae where sorption increased over time. [Copyright &y& Elsevier]

Abstract: Cholesterol present in the plasma membrane of target cells has been shown to be important for the infection by SARS-CoV. We show that cholesterol depletion by treatment with methyl-β-cyclodextrin (mβCD) affects infection by SARS-CoV to the same extent as infection by vesicular stomatitis virus-based pseudotypes containing the surface glycoprotein S of SARS-CoV (VSV-ΔG-S). Therefore, the role of cholesterol for SARS-CoV infection can be assigned to the S protein and is unaffected by other coronavirus proteins. There have been contradictory reports whether or not angiotensin-converting enzyme 2 (ACE2), the cellular receptor for SARS-CoV, is present in detergent-resistant membrane domains. We found that ACE2 of both Vero E6 and Caco-2 cells co-purifies with marker proteins of detergent-resistant membranes supporting the notion that cholesterol-rich microdomains provide a platform facilitating the efficient interaction of the S protein with the cellular receptor ACE2. To understand the involvement of cholesterol in the initial steps of the viral life cycle, we applied a cell-based binding assay with cells expressing the S protein and cells containing angiotensin-converting enzyme 2 (ACE2). Alternatively, we used a soluble S protein as interaction partner. Depletion of cholesterol from the ACE2-expressing cells reduced the binding of S-expressing cells by 50% whereas the binding of soluble S protein was not affected. This result suggests that optimal infection requires a multivalent interaction between viral attachment protein and cellular receptors. [Copyright &y& Elsevier]

African trypanosomes are extracellular parasitic protozoa, predominantly transmitted by the bite of the haematophagic tsetse fly. The main mechanism considered to mediate parasitemia control in a mammalian host is the continuous interaction between antibodies and the parasite surface, covered by variant-specific surface glycoproteins. Early experimental studies have shown that B-cell responses can be strongly protective but are limited by their VSG-specificity. We have used B-cell (μMT) and IgM-deficient (IgM-/-) mice to investigate the role of B-cells and IgM antibodies in parasitemia control and the in vivo induction of trypanosomiasis-associated anemia. These infection studies revealed that that the initial setting of peak levels of parasitemia in Trypanosoma brucei-infected mMT and IgM-/- mice occurred independent of the presence of B-cells. However, B-cells helped to periodically reduce circulating parasites levels and were required for long term survival, while IgM antibodies played only a limited role in this process. Infection-associated anemia, hypothesized to be mediated by B-cell responses, was induced during infection in mMT mice as well as in IgM2/2 mice, and as such occurred independently from the infection-induced host antibody response. Antigenic variation, the main immune evasion mechanism of African trypanosomes, occurred independently from host antibody responses against the parasite's ever-changing antigenic glycoprotein coat. Collectively, these results demonstrated that in murine experimental T. brucei trypanosomiasis, B-cells were crucial for periodic peak parasitemia clearance, whereas parasite-induced IgM antibodies played only a limited role in the outcome of the infection. [ABSTRACT FROM AUTHOR]

Activation of macrophages and subsequent ‘killing’ effector functions against infectious pathogens are essential for the establishment of protective immunity. NF-lL6 is a transcription factor downstream of IFN-γ and TNF in the macrophage activation pathway required for bacterial killing. Comparison of microarray expression profiles of Listeria monocytogenes (LM)-infected macrophages from WT and NF-IL6-deficient mice enabled us to identify candidate genes downstream of NF-IL6 involved in the unknown pathways of LM killing independent of reactive oxygen intermediates and reactive nitrogen intermediates. One differentially expressed gene, PKCδ, had higher mRNA levels in the LM-infected NF-1L6-deficient macrophages as compared with WT. To define the role of PKCδ during listeriosis, we infected PKCδ-deficient mice with LM. PKCδ-deficient mice were highly susceptible to LM infection with increased bacterial burden and enhanced histopathology despite enhanced NF-IL6 mRNA expression. Subsequent studies in PKCδ-deficient macrophages demonstrated that, despite elevated levels of proinflammatory cytokines and NO production, increased escape of LM from the phagosome into the cytoplasm and uncontrolled bacterial growth occurred. Taken together these data identified PKCδ as a critical factor for confinement of LM within macrophage phagosomes. [ABSTRACT FROM AUTHOR]

Abstract: The focus of this paper is to present the concepts for a new dynamic, i.e. database-driven information system for the International Earth Rotation and Reference Systems Service (IERS). Its goal is to coordinate the data and information flow between the participating institutions. Therefore, all relevant data and products of the IERS should be archived to guarantee their long-term availability. At the same time the consistency and timeliness of the contents of the information system can be guaranteed by managing the information in a database. Additionally, the metadata of all products (including the IERS publications) should be modelled in a database to allow the users to search for specific data or topics with respect to space, time and content. In order to be able to link related data the heterogeneous formats of the products have to be transformed into a common format. The eXtensible Markup Language (XML) is going to be used, to perform this ambitious task. The usage of XML not only allows to link related data but also to easily exchange data and to present them in different ways like Web pages, pdf, etc. The system will be completed by several tools to manage and coordinate the tasks of the Central Bureau (CB) and by an information system with respect to IERS-related topics. The standardised database tools will also allow an easy exchange of information with and links to other databases within the IGGOS project to realize a powerful instrument to serve the Earth observing system. [Copyright &y& Elsevier]

The synthesis and binding properties of a new tricationic guanidiniocarbonyl pyrrole receptor 7 are described. Receptor 7 binds citrate 9 and other tricarboxylates such as trimesic acid tricarboxylate 8 with unprecedented high association constants of Kassoc > 105 M-1 in water as determined by UV and fluorescence tritration studies. According to NOESY experiments and molecular modeling calculations, the tricarboxylates are bound within the inner cavity of receptor 7 by ion pairing between the carboxylate groups and the guanidiniocarbonyl pyrrole moieties, favored by the nonpolar microenvironment of the cavity. Hence, receptor 7 can be regarded as a molecular flytrap. In the case of the aromatic tricarboxylate 8, additional aromatic interactions further strengthen the complex. The complexes with the tricarboxylates are so strong that even the presence of a large excess of competing anions or buffer salts does not significantly affect the association constant. For example, the association constant for citrate changes only from Kassoc = 1.6 × 105 M-1 in pure water to Kassoc = 8.6 × 104 M-1in the presence of a 170-fold excess of bis-tris buffer and a 1000-fold excess of chloride. This makes 7 one of the most efficient receptors for the binding of citrate in aqueous solvents reported thus far. [ABSTRACT FROM AUTHOR]

Coronaviruses (CoV) mature by a budding process at intracellular membranes. Here we showed that the major surface protein S of a porcine CoV (transmissible gastroenteritis virus) is not transported to the cell surface but is retained intracellularly. Site-directed mutagenesis indicated that a tyrosine-dependent signal (YXXI) in the cytoplasmic tail is essential for intracellular localization of the S protein. Surface expression of mutant proteins was evident by immunofluorescence analysis and surface biotinylation. Intracellularly retained S proteins only contained endoglycosidase H-sensitive N-glycans, whereas mutant proteins that migrated to the plasma membrane acquired N-linked oligosaccha-rides of the complex type. Corresponding tyrosine residues are present in the cytoplasmic tails of the S proteins of other animal CoV but not in the tail portion of the S protein of severe acute respiratory syndrome (SARS)-CoV. Changing the SEPV tetrapeptide in the cytoplasmic tail to YEPI resulted in intracellular retention of the S protein of SARS-CoV. As the S proteins of CoV have receptor binding and fusion activities and are the main target of neutralizing antibodies, the differences in the transport behavior of the S proteins suggest different strategies in the virus host interactions between SARS-CoV and other coronaviruses. [ABSTRACT FROM AUTHOR]

Transmissible gastroenteritis coronavirus (TGEV) is a porcine pathogen causing enteric infections that are lethal for suckling piglets. The enterotropism of TGEV is connected with the sialic acid binding activity of the viral surface protein S. Here we show that, among porcine intestinal brush border membrane proteins, TGEV recognizes a mucin-type glycoprotein designated MGP in a sialic acid-dependent fashion. Virus binding assays with cryosections of the small intestine from a suckling piglet revealed the binding of TGEV to mucinproducing goblet cells. A nonenteropathogenic mutant virus that lacked a sialic acid binding activity was unable to bind to MGP and to attach to goblet cells. Our results suggest a role of MGP in the enteropathogenicity of TGEV. [ABSTRACT FROM AUTHOR]

TGEV is a coronavirus that is still widely spread in pig farming. On molecular level this virus has been studied in detail. However, studying TGEV infection within the complexity of the porcine intestinal epithelium reveals difficulties due to limiting infection models. Here we established a new ex vivo model to analyze the enterotropism of TGEV in porcine intestinal tissue. Precision cut intestinal slices (PCIS) were produced and ATP level was measured to proof vitality of the slices. ATP measurements and HE staining revealed living tissue in culture for up to 24 h. PCIS were infected with three different TGEV strains. TGEV PUR 46-MAD is a commonly used TGEV strain that is known to be attenuated. TGEV Miller was passaged in piglets several times to reveal high infection. Finally, TGEV GFP is a recombinant strain that obtained its main body from TGEV PUR 46-MAD, but its spike protein from TGEV PUR-C11 that showed high mortality in piglets in vivo . Our results were in complete consensus of these statements. TGEV Miller mildly and TGEV GFP extensively infected the cells in the jejunum based on the amount of positive stained epithelial cells. However, for TGEV PUR 46-MAD no nucleocapsid protein was detected in the epithelial cells of the tissue. This shows that differences in TGEV strains and their infectious potential are highly dependent on their S protein. [ABSTRACT FROM AUTHOR]

BODIPYs (boron dipyrromethenes) are fluorescent dyes which show high stability and quantum yields. They feature the possibility of selective 18 F-fluorination at the boron-core. Attached to a bioactive molecule and labeled with [ 18 F]fluorine, the resulting compounds are promising tracers for multimodal imaging in vivo and can be used for PET and fluorescence imaging. A BODIPY containing a phenyl and a hydroxy substituent on boron was synthesized and characterized. Fluorinated and hydroxy substituted dyes were coupled to an isatin-based caspase inhibitor via cycloaddition and the resulting compounds were evaluated in vitro in caspase inhibition assays. The metabolic stability and the formed metabolites were investigated by incubation with mouse liver microsomes and LC-MS analysis. Subsequently the fluorophores were labeled with [ 18 F]fluorine and an in vivo biodistribution study using dynamic PET was performed. [ABSTRACT FROM AUTHOR]

The Respiratory syncytial virus (RSV) is a member of the family of Paramyxoviridae. It infects humans (HRSV) and cattle (BRSV). The cellular receptor of both viruses is unknown. Among the viral proteins, the F protein mediates the fusion of the viral envelope with the cell membrane and is involved in the binding to permissive cells. The glycoprotein (G) enhances attachment of the virion to the host cell by binding to glycosaminoglycans. During maturation the F protein is cleaved into two subunits, F1 and F2. The F1 subunit contains the fusion peptide. The F2 subunit is crucial for the tropism of the virus with respect to epithelial cells of the respiratory tract. We want to characterize the determinant for the host tropism in the F protein in more detail. For this purpose we generated a soluble F protein with a Flag-tag at the C-terminus. This protein is efficiently secreted from transfected cells. The released protein is trimeric like the native F protein. This construct is a valuable tool to analyze the species-specific interaction of the F protein with target cells. [ABSTRACT FROM AUTHOR]

We have analyzed the intracellular transport of the spike (S) protein of infectious bronchitis virus (IBV), an avian coronavirus. Surface expression was analyzed by immunofluorescence microscopy, by surface biotinylation, and by syncytium formation by S-expressing cells. By applying these methods, the S protein was found to be retained intracellularly. Tyr1143 in the cytoplasmic tail was shown to be a crucial component of the retention signal. Deletion of a dilysine motif that has previously been suggested to function as a retrieval signal did not abolish intracellular retention. Treatment of the S proteins with endoglycosidases did not reveal any differences between the parental and the mutant proteins. Furthermore, all S proteins analyzed were posttranslationally cleaved into the subunits S1 and S2. In coexpression experiments, the S protein was found to colocalize with a Golgi marker. Taken together, these results indicate that the S protein of IBV is retained at a late Golgi compartment. Therefore, this viral surface protein differs from the S proteins of transmissible gastroenteritis virus and severe acute respiratory syndrome coronavirus, which are retained at a pre-Golgi compartment or transported to the cell surface, respectively. The implications of these differences are discussed. [ABSTRACT FROM AUTHOR]

Introduction: Dysregulated activity of matrix metalloproteinases (MMPs) drives a variety of pathophysiological conditions. Non-invasive imaging of MMP activity in vivo promises diagnostic and prognostic value. However, current targeting strategies by small molecules are typically limited with respect to the bioavailability of the labeled MMP binders in vivo. To this end, we here introduce and compare three chemical modifications of a recently developed barbiturate-based radiotracer with respect to bioavailability and potential to image MMP activity in vivo.Methods: Barbiturate-based MMP inhibitors with an identical targeting unit but varying hydrophilicity were synthesized, labeled with technetium-99m, and evaluated in vitro and in vivo. Biodistribution and radiotracer elimination were determined in C57/BL6 mice by serial SPECT imaging. MMP activity was imaged in a MMP-positive subcutaneous xenograft model of human K1 papillary thyroid tumors. In vivo data were validated by scintillation counting, autoradiography, and MMP immunohistochemistry.Results: We prepared three new 99mTc-labeled MMP inhibitors, bearing either a glycine ([99mTc]MEA39), lysine ([99mTc]MEA61), or the ligand HYNIC with the ionic co-ligand TPPTS ([99mTc]MEA223) yielding gradually increasing hydrophilicity. [99mTc]MEA39 and [99mTc]MEA61 were rapidly eliminated via hepatobiliary pathways. In contrast, [99mTc]MEA223 showed delayed in vivo clearance and primary renal elimination. In a thyroid tumor xenograft model, only [99mTc]MEA223 exhibited a high tumor-to-blood ratio that could easily be delineated in SPECT images.Conclusion: Introduction of HYNIC/TPPTS into the barbiturate lead structure ([99mTc]MEA223) results in delayed renal elimination and allows non-invasive MMP imaging with high signal-to-noise ratios in a papillary thyroid tumor xenograft model. [ABSTRACT FROM AUTHOR]

Abstract: Cholesterol is a major constituent of detergent-resistant membrane microdomains (DRMs). We localized transmissible gastroenteritis virus (TGEV) spike (S) protein in DRMs in the viral envelope. Though S protein was not solubilized by cold non-ionic detergents, this behavior was unchanged when cholesterol was depleted from viral membrane by methyl-β-cyclodextrin (MβCD) and the protein did not comigrate with cellular DRM marker proteins in flotation analyses. Therefore, the S protein is not anchored in the viral membrane DRMs as they are known to occur in the plasma membrane. Cholesterol depletion from viral membrane may not affect the adsorption process as neither the sialic acid binding activity nor the binding to aminopeptidase N was reduced post-MβCD treatment. Reduced infectivity of cholesterol-depleted TGEV was observed only when the adsorption process occurred at 37°C but not when the virus was applied at 4°C. Cholesterol is important for a post-adsorption step, allowing membrane rearrangements that facilitate virus entry. [ABSTRACT FROM AUTHOR]

To address the initiation of virus infection in the respiratory tract, we established two culture systems for differentiated bovine airway epithelial cells (BAEC). Filter-grown BAEC differentiated under air-liquid interface (ALI) conditions to generate a pseudo-stratified mucociliary epithelium. Alternatively, precision-cut lung slices (PCLS) from the bovine airways were generated that retained the original composition and distribution of differentiated epithelial cells. With both systems, epithelial cells were readily infected by bovine parainfluenza virus 3 (BPIV3). Ciliated cells were the most prominent cell type affected by BPIV3. Surprisingly, differentiated BAEC were resistant to infection by bovine respiratory syncytial virus (BRSV), when the virus was applied at the same multiplicity of infection that was sufficient for infection by BPIV3. In the case of PCLS, infection by BRSV was observed in cells located in lower cell layers but not in epithelial cells facing the lumen of the airways. The identity of the infected cells could not be determined because of a lack of specific antibodies. Increasing the virus titer 30-fold resulted in infection of the ALI cultures of BAEC, whereas in PCLS the ciliated epithelium was still refractory to infection by BRSV. These results indicate that differentiated BAEC are readily infected by BPIV3 but rather resistant to infection by BRSV. Disease caused by BRSV may require that calves encounter environmental stimuli that render BAEC susceptible to infection. [ABSTRACT FROM AUTHOR]

In Germany the coronavirus disease 2019 (COVID-19) pandemic situation is unique among large European countries in that incidence and case fatality rate are distinctly lower. We describe the clinical course and examine factors associated with outcomes among patients hospitalized with COVID-19 in Germany. In this retrospective cohort study we included patients with COVID-19 admitted to a national network of German hospitals between February 12 and June 12, 2020. We examined demographic characteristics, comorbidities and clinical outcomes. We included 1904 patients with a median age of 73 years, 48.5% (924/1904) of whom were female. The mortality rate was 17% (317/1835; 95% confidence interval (95%CI) 16–19), the rate of admission to the intensive care unit (ICU) was 21% (399/1860; 95%CI 20–23), and the rate of invasive mechanical ventilation was 14% (250/1850: 95%CI 12–15). The most prominent risk factors for death were male sex (hazard ratio (HR) 1.45; 95%CI 1.15–1.83), pre-existing lung disease (HR 1.61; 95%CI 1.20–2.16), and increased patient age (HR 4.11 (95%CI 2.57–6.58) for age >79 years versus <60 years). Among patients admitted to the ICU, the mortality rate was 29% (109/374; 95%CI 25–34) and higher in ventilated (33% [77/235; 95%CI 27–39]) than in non-ventilated ICU patients (23%, 32/139; 95%CI 16–30; p < 0.05). In this nationwide series of patients hospitalized with COVID-19 in Germany, in-hospital and ICU mortality rates were substantial. The most prominent risk factors for death were male sex, pre-existing lung disease, and greater patient age. [ABSTRACT FROM AUTHOR]

Many serious diseases such as cancer, atherosclerosis and arthritis are characterized by upregulation of activated matrix metalloproteinases (MMPs). For this reason imaging and quantification of activated MMPs with the molecular imaging techniques such as single photon emission computed tomography (SPECT) and positron emission tomography (PET) using radiolabeled MMP inhibitors would be a valuable tool for the diagnosis and therapy planning of mentioned diseases. This work aims at the synthesis and preliminary in vitro and in vivo evaluation of positron‐emitter 18F‐fluorine labeled radiotracers based on 2‐{[(4‐phenoxyphenyl)sulfonyl]methyl}thiirane (SB‐3CT), a slow‐binding and mechanism‐based MMP‐2 and −9 inhibitor. Therefore, a library of fluorinated SB‐3CT derivatives were prepared and evaluated in vitro in MMP inhibition assays. From this pool the 18F‐labeled triazole [18F]18d was successfully synthesized in a two‐step procedure. However, this compound was unstable in human and mouse serum and showed a biodistribution behavior in C57BL/6 mice that is not favorable for PET imaging preventing further in vivo evaluations in MMP‐associated mouse models of disease. Imaging of matrix metalloproteinases (MMPs) dysregulated in a multitude of diseases with positron emission tomography (PET) is of great clinical interest. For the first time the thiirane‐based SB‐3CT, exhibiting the unique characteristic of slow‐binding inhibition of MMP‐2 and −9, served as lead structure for 18F‐labeled radiotracer development. The first representative of this compound class ([18F]18d) was evaluated in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Dysregulated levels of activated matrix metalloproteinases (MMPs) are linked to different pathologies, such as cancer, atherosclerosis, neuroinflammation, and arthritis. Therefore, imaging of MMPs with positron-emission tomography (PET) represents a powerful tool for the diagnosis of MMP-associated diseases. Moreover, to distinguish between the distinct functions and roles of individual MMPs in particular pathophysiological processes, their specific imaging must be realized with radiolabeled tracers, such as fluorine-18-labeled MMP inhibitors (MMPIs). Therefore, fluorinated dibenzofuransulfonamide-based MMPIs showing excellent inhibition of MMP-12 and selectivity for MMP-12 over other MMPs were prepared. MMP-12 is a key enzyme in diseases such as chronic obstructive pulmonary disease (COPD) and atherosclerosis. Because of their promising in vitro properties, three candidates (4, 9, and 19) were selected from this library, and radiofluorinated analogues ([18F]4, [18F]9, and [18F]19) were successfully synthesized. Initial in vitro serum stability and in vivo biodistribution studies of the radiolabeled MMPIs with PET demonstrated their potential benefit for preferable MMP-12 imaging. [ABSTRACT FROM AUTHOR]

Background: Disturbance of intestinal wound closure leads to insufficient anastomotic healing and is associated with considerable morbidity following colorectal resections. Matrix metalloproteinases (MMPs) play a crucial role in regulation of wound closure. Here fluorescence endoscopy was evaluated for assessment of MMP-2/-9 expression during failed intestinal anastomotic healing. Methods: Distal colonic anastomoses were performed as a model for disturbed healing in 36 Balb/c mice. Healing was evaluated endoscopically, macroscopically, and histologically after 1, 3 and 5 days. For detection of MMP-2/-9 expression fluorescence endoscopy (FE) was used following i.v.-administration of a Cy5.5-labeled MMP-2/-9 specific tracer. FE was complemented by quantification of the fluorescence signal using the MS-FX PRO Optical Imaging System. An overall leakage score was calculated and correlated with the results of FE. Results: With increasing incidence of anastomotic leakage from POD1 (17%) to POD5 (83%) the uptake of the MMP tracer gradually increased (signal-to-noise ratio (SNR), POD1: 17.91 ± 1.251 vs. POD3: 30.56 ± 3.03 vs. POD5: 44.8 ± 4.473, P<0.0001). Mice with defective anastomotic healing showed significantly higher uptake compared to non-defective (SNR: 37.37± 3.63 vs. 26.16± 3.635, P = 0.0369). White light endoscopy and FE allowed evaluation of anastomotic healing and visualization of mucosal MMPs in vivo. Using FE based detection of MMPs in the anastomosis, an overall positive predictive value of 71.4% and negative predictive value of 66.6% was calculated for detection of anastomotic leakage. Conclusion: During disturbed anastomotic healing increased expression of MMP-2/-9 was observed in the anastomotic tissue. Fluorescence endoscopy for detection of MMP-2/-9 during the healing process might be a promising tool for early identification of anastomotic leakage. [ABSTRACT FROM AUTHOR]

Coronavirus spike proteins mediate host-cell-attachment and virus entry. Virus replication takes place within the host cell cytosol, whereas assembly and budding occur at the endoplasmic reticulum-Golgi intermediate compartment. In this study we demonstrated that the last 39 amino acid stretches of Alphacoronavirus spike cytoplasmic domains of the human coronavirus 229E, NL63, and the porcine transmissible gastroenteritis virus TGEV interact with tubulin alpha and beta chains. In addition, a partial co-localization of TGEV spike proteins with authentic host cell β-tubulin was observed. Furthermore, drug-induced microtubule depolymerization led to changes in spike protein distribution, a reduction in the release of infectious virus particles and less amount of spike protein incorporated into virions. These data demonstrate that interaction of Alphacoronavirus spike proteins with tubulin supports S protein transport and incorporation into virus particles. [ABSTRACT FROM AUTHOR]

Purpose: Resistance to bevacizumab (BEV) in glioblastoma is believed to occur via activation of molecular networks including the mTOR/PI3K pathway. Using an MR/PET molecular imaging biomarker approach, we investigated the response to combining BEV with the mTOR/PI3K inhibitor BEZ235. Methods: Tumours were established by orthotopically implanting U87MG-luc2 cells in mice. Animals were treated with BEZ235 and/or BEV, and imaged using diffusion-weighted-MRI, T2-weighted and T2*-weighted before and after administration of superparamagnetic iron oxide contrast agent. Maps for changes in relaxation rates (Δ R2, Δ R2* and apparent diffusion coefficient) were calculated. Vessel size index and microvessel density index were derived. 3′-Deoxy-3′-[F]fluorothymidine ([F]FLT) PET and O-(2-[F]fluoroethyl)- l-tyrosine ([F]FET) PET were further performed and tumour endothelium/proliferation markers assessed by immunohistochemistry. Results: Treatment with BEV resulted in a pronounced decrease in tumour volume (T2-weighted MRI). No additive effect on tumour volume was observed with the BEV/BEZ235 combination compared with BEV monotherapy. The Ki67 proliferation index and [F]FLT uptake studies were used to support the observations. Using Δ R2* and Δ R2 values, respectively, the BEV/BEZ235 combination significantly reduced tumour microvessel volume in comparison to BEV alone. Decreased microvessel density index was further observed in animals treated with the combination, supported by von Willebrand factor (vWF) immunohistochemistry. [F]FET uptake was decreased following treatment with BEV alone, but was not further reduced following treatment with the combination. vWF immunohistochemistry analysis showed that the mean tumour vessel size was increased in all cohorts. Conclusion: Assessing MR imaging biomarker parameters together with [F]FET and [F]FLT PET provided information on mechanism of action of the drug combination and clues as to potential clinical responses. Following translation to clinical use, treatment with a BEV/BEZ235 combination could reduce peritumoral oedema obviating the requirement for steroids. The use of hypothesis-driven molecular imaging studies facilitates the preclinical evaluation of drug response. Studies of this kind may more accurately predict the clinical potential of the BEV/BEZ235 combination regimen as a novel therapeutic approach in oncology. [ABSTRACT FROM AUTHOR]

Classical activation of macrophages (caMph or M1) is crucial for host protection against Mycobacterium tuberculosis (Mtb) infection. Evidence suggests that IL-4/IL-13 alternatively activated macrophages (aaMph or M2) are exploited by Mtb to divert microbicidal functions of caMph. To define the functions of M2 macrophages during tuberculosis (TB), we infected mice deficient for IL-4 receptor α on macrophages (LysMcreIL-4Rα-/lox) with Mtb. We show that absence of IL-4Rα on macrophages does not play a major role during infection with Mtb H37Rv, or the clinical Beijing strain HN878. This was demonstrated by similar mortality, bacterial burden, histopathology and T cell proliferation between infected wild-type (WT) and LysMcreIL-4Rα-/lox mice. Interestingly, we observed no differences in the lung expression of inducible nitric oxide synthase (iNOS) and Arginase 1 (Arg1), well-established markers for M1/M2 macrophages among the Mtb-infected groups. Kinetic expression studies of IL-4/IL-13 activated bone marrow-derived macrophages (BMDM) infected with HN878, followed by gene set enrichment analysis, revealed that the MyD88 and IL-6, IL-10, G-CSF pathways are significantly enriched, but not the IL-4Rα driven pathway. Together, these results suggest that IL-4Rα-macrophages do not play a central role in TB disease progression. [ABSTRACT FROM AUTHOR]

The spike protein S of coronaviruses contains a highly conserved cytoplasmic cysteine-rich motif adjacent to the transmembrane region. This motif is palmitoylated in the Betacoronaviruses MHV and SARS-CoV. Here, we demonstrate by metabolic labeling with [ 3 H]-palmitic acid that the S protein of transmissible gastroenteritis coronavirus (TGEV), an Alphacoronavirus , is palmitoylated as well. This is relevant for TGEV replication as virus growth was compromised by the general palmitoylation inhibitor 2-bromopalmitate. Mutation of individual cysteine clusters in the cysteine-rich motif of S revealed that all cysteines must be replaced to abolish acylation and incorporation of S into virus-like particles (VLP). Conversely, the interaction of S with the M protein, essential for VLP incorporation of S, was not impaired by lack of palmitoylation. Thus, palmitoylation of the S protein of Alphacoronaviruses is dispensable for S–M interaction, but required for the generation of progeny virions. [ABSTRACT FROM AUTHOR]

Transmissible gastroenteritis virus (TGEV) is an enveloped (+) RNA virus belonging to the family Coronaviridae. Among the viral membrane proteins, the spike (S) protein mediates receptor recognition/attachment to the host cell and fusion of viral and cellular membranes. The cytoplasmic tail of the S protein contains a tyrosine-dependent sorting signal with the consensus sequence YXXΦ. In the context of the S protein of TGEV (1440YEPI1443), this motif acts as a retention signal, preventing surface expression of the protein. Here, we show that a chimeric S protein, containing the six C-terminal amino acids of the glycoprotein G of vesicular stomatitis virus (VSV) is no longer retained intracellularly, despite the presence of the tyrosine tetrapeptide motif. Following transport to the cell surface, the chimeric protein was rapidly endocytosed. Analysis of mutant proteins generated by site-directed mutagenesis revealed that a single amino acid exchange (1445K/M, position: +2 downstream of the tyrosine-based motif) was responsible for the altered sorting behavior. [ABSTRACT FROM AUTHOR]

Abstract: Bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus type 3 (BPIV3) and bovine herpesvirus type 1 (BHV-1) are important pathogens associated with the bovine respiratory disease complex (BRDC). Non-bovine ruminants such as goats may also be infected and serve as a virus reservoir to be considered in the development of control strategies. To evaluate the susceptibility of caprine airway epithelial cells to infection by viruses of BRDC, we established a culture system for differentiated caprine epithelial cells. For this purpose, we generated precision-cut lung slices (PCLS), in which cells are retained in their original structural configuration and remain viable for more than a week. The three bovine viruses were found to preferentially infect different cell types. Ciliated epithelial cells were the major target cells of BPIV3, whereas BHV-1 preferred basal cells. Cells infected by BRSV were detected in submucosal cell layers. This spectrum of susceptible cells is the same as that reported recently for infected bovine PCLS. While infection of caprine cells by BRSV and BPIV3 was as efficient as that reported for bovine cells, infection of caprine cells by BHV-1 required a tenfold higher dose of infectious virus as compared to infection of bovine airway cells. These results support the notion that non-bovine ruminants may serve as a reservoir for viruses of BRDC and introduce a culture system to analyze virus infection of differentiated airway epithelial cells from the caprine lung. [Copyright &y& Elsevier]

Binding of influenza viruses to target cells is mediated by the viral surface protein hemagglutinin. To determine the presence of binding sites for influenza A viruses on cells and tissues, soluble hemagglutinins of the H7 and H9 subtype were generated by connecting the hemagglutinin ectodomain to the Fc portion of human immunoglobulin G (H7Fc and H9Fc). Both chimeric proteins bound to different cells and tissues in a sialic acid-dependent manner. Pronounced differences were observed between H7Fc and H9Fc, in the binding both to different mammalian and avian cultured cells and to cryosections of the respiratory epithelium of different virus host species (turkey, chicken and pig). Binding of the soluble hemagglutinins was similar to the binding of virus particles, but showed differences in the binding pattern when compared to two sialic acid-specific plant lectins. These findings were substantiated by a comparative glycan array analysis revealing a very narrow recognition of sialoglycoconjugates by the plant lectins that does not reflect the glycan structures preferentially recognized by H7Fc and H9Fc. Thus, soluble hemagglutinins may serve as sialic acid-specific lectins and are a more reliable indicator of the presence of binding sites for influenza virus HA than the commonly used plant lectins. [ABSTRACT FROM AUTHOR]

Bats (Chiroptera) host major human pathogenic viruses including corona-, paramyxo, rhabdo- and filoviruses. We analyzed six different cell lines from either Yinpterochiroptera (including African flying foxes and a rhinolophid bat) or Yangochiroptera (genera Carollia and Tadarida) for susceptibility to infection by different enveloped RNA viruses. None of the cells were sensitive to infection by transmissible gastroenteritis virus (TGEV), a porcine coronavirus, or to infection mediated by the Spike (S) protein of SARS-coronavirus (SARS-CoV) incorporated into pseudotypes based on vesicular stomatitis virus (VSV). The resistance to infection was overcome if cells were transfected to express the respective cellular receptor, porcine aminopeptidase N for TGEV or angiotensin-converting enzyme 2 for SARS-CoV. VSV pseudotypes containing the S proteins of two bat SARS-related CoV (Bg08 and Rp3) were unable to infect any of the six tested bat cell lines. By contrast, viral pseudotypes containing the surface protein GP of Marburg virus from the family Filoviridae infected all six cell lines though at different efficiency. Notably, all cells were sensitive to infection by two paramyxoviruses (Sendai virus and bovine respiratory syncytial virus) and three influenza viruses from different subtypes. These results indicate that bat cells are more resistant to infection by coronaviruses than to infection by paramyxoviruses, filoviruses and influenza viruses. Furthermore, these results show a receptor-dependent restriction of the infection of bat cells by CoV. The implications for the isolation of coronaviruses from bats are discussed. [ABSTRACT FROM AUTHOR]

: Addition of temozolomide (TMZ) to radiation therapy is the standard treatment for patients with glioblastoma (GBM). However, there is uncertainty regarding the effectiveness of TMZ. Considering the rapid evolution of the disease, methods to assess TMZ efficacy early during treatment would be of great benefit. Our aim was to monitor early effects of TMZ in a mouse model of GBM using positron emission tomography (PET) with 3′-deoxy-3′-[18F]fluorothymidine ([18F]FLT). Methods: Human glioma cells sensitive to TMZ (Gli36dEGFR-1) were treated with sub-lethal doses of TMZ to obtain cells with lower sensitivity to TMZ (Gli36dEGFR-2), as measured by growth and clonogenic assays. Gli36dEGFR-1 and Gli36dEGFR-2 cells were subcutaneously (s.c.) or intracranially (i.c.) xenografted into nude mice. Mice were treated for 7 days with daily injection of 25 or 50 mg/kg TMZ. Treatment efficacy was measured using [18F]FLT-PET before treatment and after 2 days. Computed Tomography (CT) or Magnetic Resonance Imaging (MRI) were used to determine tumor volumes before treatment and after 7 days. Results: A significant difference was observed between TMZ and DMSO treated tumors in terms of variations of [18F]FLT T/B ratio as soon as day 2 in the i.c. as well as in the s.c. mouse model. Variations of [18F]FLT T/B uptake ratio between days 0 and 2 correlated with variations of tumor size between days 0 and 7 (s.c. model: ntumor = 17 in nmice = 11, P<0.01; i.c. model: ntumor/mice = 9, P<0.01). Conclusions: Our results indicate that [18F]FLT-PET may be useful for an early evaluation of the response of GBM to TMZ chemotherapy in patients with glioma. [ABSTRACT FROM AUTHOR]

Porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PDEV) can cause severe diarrhea in pigs. Development of effective vaccines against TGEV and PEDV is one of important prevention measures. The spike (S) protein is the surface glycoprotein of TGEV and PEDV, which can induce specific neutralization antibodies and is a candidate antigen for vaccination attempts. In this study, the open reading frames of the TGEV S1 protein and in addition of the S or S1 proteins of PEDV were inserted into the eukaryotic expression vector, pIRES, resulting in recombinant plasmids, pIRES-(TGEV-S1-PEDV-S1) and pIRES-(TGEV-S1-PEDV-S). Subsequently, 6–8 weeks old Kunming mice were inoculated with both DNA plasmids. Lymphocyte proliferation assay, virus neutralization assay, IFN-γ assay and CTL activity assay were performed. TGEV/PEDV specific antibody responses as well as kinetic changes of T lymphocyte subgroups of the immunized mice were analyzed. The results showed that the recombinant DNA plasmids increased the proliferation of T lymphocytes and the number of CD4+ and CD8+ T lymphocyte subgroups. In addition, the DNA vaccines induced a high level of IFN-γ in the immunized mice. The specific CTL activity in the pIRES-(TGEV-S1-PEDV-S) group became significant at 42 days post-immunization. At 35 days post-immunization, the recombinant DNA plasmids bearing full-length S genes of TGEV and PEDV stimulated higher levels of specific antibodies and neutralizing antibodies in immunized mice. [ABSTRACT FROM AUTHOR]

Background: Swine are important hosts for influenza A viruses playing a crucial role in the epidemiology and interspecies transmission of these viruses. Respiratory epithelial cells are the primary target cells for influenza viruses. Methodology/Principal Findings: To analyze the infection of porcine airway epithelial cells by influenza viruses, we established precision-cut lung slices as a culture system for differentiated respiratory epithelial cells. Both ciliated and mucus-producing cells were found to be susceptible to infection by swine influenza A virus (H3N2 subtype) with high titers of infectious virus released into the supernatant already one day after infection. By comparison, growth of two avian influenza viruses (subtypes H9N2 and H7N7) was delayed by about 24 h. The two avian viruses differed both in the spectrum of susceptible cells and in the efficiency of replication. As the H9N2 virus grew to titers that were only tenfold lower than that of a porcine H3N2 virus this avian virus is an interesting candidate for interspecies transmission. Lectin staining indicated the presence of both α-2,3- and α-2,6-linked sialic acids on airway epithelial cells. However, their distribution did not correlate with pattern of virus infection indicating that staining by plant lectins is not a reliable indicator for the presence of cellular receptors for influenza viruses. Conclusions/Significance: Differentiated respiratory epithelial cells significantly differ in their susceptibility to infection by avian influenza viruses. We expect that the newly described precision-cut lung slices from the swine lung are an interesting culture system to analyze the infection of differentiated respiratory epithelial cells by different pathogens (viral, bacterial and parasitic ones) of swine. [ABSTRACT FROM AUTHOR]

In preparation of activities planned for the realization of the Global Geodetic Observing System (GGOS), a group of German scientists has carried out a study under the acronym GGOS-D which closely resembles the ideas behind the GGOS initiative. The objective of the GGOS-D project was the investigation of the methodological and information-technological realization of a global geodetic-geophysical observing system and especially the integration and combination of the space geodetic observations. In the course of this project, highly consistent time series of GPS, VLBI, and SLR results were generated based on common state-of-the-art standards for modeling and parameterization. These series were then combined to consistently and accurately compute a Terrestrial Reference Frame (TRF). This TRF was subsequently used as the basis to produce time series of station coordinates, Earth orientation, and troposphere parameters. In this publication, we present results of processing algorithms and strategies for the integration of the space-geodetic observations which had been developed in the project GGOS-D serving as a prototype or a small and limited version of the data handling and processing part of a global geodetic observing system. From a comparison of the GGOS-D terrestrial reference frame results and the ITRF2005, the accuracy of the datum parameters is about 5-7 mm for the positions and 1.0-1.5 mm/year for the rates. The residuals of the station positions are about 3 mm and between 0.5 and 1.0 mm/year for the station velocities. Applying the GGOS-D TRF, the offset of the polar motion time series from GPS and VLBI is reduced to 50 μas (equivalent to 1.5 mm at the Earth's surface). With respect to troposphere parameter time series, the offset of the estimates of total zenith delays from co-located VLBI and GPS observations for most stations in this study is smaller than 1.5 mm. The combined polar motion components show a significantly better WRMS agreement with the IERS 05C04 series (96.0/96.0 μas) than VLBI (109.0/100.7 μas) or GPS (98.0/99.5 μas) alone. The time series of the estimated parameters have not yet been combined and exploited to the extent that would be possible. However, the results presented here demonstrate that the experiences made by the GGOS-D project are very valuable for similar developments on an international level as part of the GGOS development. [ABSTRACT FROM AUTHOR]

IL-4Rα-dependent responses are essential for granuloma formation and host survival during acute schistosomiasis. Previously, we demonstrated that mice deficient for macrophage-specific IL-4Rα (LysMcreIl4ra−/lox) developed increased hepatotoxicity and gut inflammation; whereas inflammation was restricted to the liver of mice lacking T cell-specific IL-4Rα expression (iLckcreIl4ra−/lox). In the study presented here we further investigated their role in liver granulomatous inflammation. Frequencies and numbers of macrophage, lymphocyte or granulocyte populations, as well as Th1/Th2 cytokine responses were similar in Schistosoma mansoni-infected LysMcreIl4ra−/lox liver granulomas, when compared to Il4ra−/lox control mice. In contrast, a shift to Th1 responses with high IFN-γ and low IL-4, IL-10 and IL-13 was observed in the severely disrupted granulomas of iLckcreIl4ra−/lox and Il4ra−/− mice. As expected, alternative macrophage activation was reduced in both LysMcreIl4ra−/lox and iLckcreIl4ra−/lox granulomas with low arginase 1 and heightened nitric oxide synthase RNA expression in granuloma macrophages of both mouse strains. Interestingly, a discrete subpopulation of SSChighCD11b+I-A/I-EhighCD204+ macrophages retained expression of mannose receptor (MMR) and Ym1 in LysMcreIl4ra−/lox but not in iLckcreIl4ra−/lox granulomas. While aaMφ were in close proximity to the parasite eggs in Il4ra−/lox control mice, MMR+Ym1+ macrophages in LysMcreIl4ra−/lox mice were restricted to the periphery of the granuloma, indicating that they might have different functions. In vivo IL-10 neutralisation resulted in the disappearance of MMR+Ym1+ macrophages in LysMcreIl4ra−/lox mice. Together, these results show that IL-4Rα-responsive T cells are essential to drive alternative macrophage activation and to control granulomatous inflammation in the liver. The data further suggest that in the absence of macrophage-specific IL-4Rα signalling, IL-10 is able to drive mannose receptor- and Ym1-positive macrophages, associated with control of hepatic granulomatous inflammation. Author Summary: Schistosomiasis is a tropical disease caused by one of the species of the parasitic worm Schistosoma which infects over 200 million people worldwide. Signalling via the IL-4 receptor alpha (IL-4Rα), which is the common receptor chain for the ligands IL-4 and IL-13, is essential for inducing protective Type 2 immune response and granuloma formation in response to the parasite eggs. In experimental Schistosoma mansoni infection and egg-induced inflammation studies with cell type-specific IL-4Rα deficient mice, the role of IL-4Rα-activated alternative macrophages (aaMφ) and IL-4Rα-responsive T cells was investigated with focus on the control of hepatic inflammation and granuloma formation. Interestingly, aaMφ were not essential for the cellular composition or the Th1/Th2 cytokine profile in liver granulomas. In contrast, IL-4Rα-dependent T cell responses were important for predominant Th2 and IL-10 responses, as well as the presence of aaMφ in the granulomas, avoiding major disruption in the granuloma cell composition. Moreover, a macrophage subpopulation was identified and those cells expressed the two aaMφ markers, mannose receptor- and Ym1 in an IL-4Rα-independent but IL-10-dependent manner. These cells might be involved in the control of inflammation. [ABSTRACT FROM AUTHOR]