Black gate bbs virus

Save my name, email, and website in this browser for the next time I comment. A quote from Sherwood Smith, reviewing Black Gate for SFSite , lets you in on our preferences: For the most part, adventure fantasy is not known for complex characterization or skillful prose….

Our findings suggest that human microglia may be a source of neuronal infection and sustain JEV brain pathogenesis. The online version of this article doi JEV is transmitted by mosquito vectors via a zoonotic cycle involving pigs as amplifying and water birds as reservoir host, the latter do not typically develop illness upon JEV infection [ 2 ].

Humans are considered to be dead-end hosts, since low viremia does not allow further virus transmission [ 1 ].

Markedly, competent vectors for JEV have been recently identified in Germany [ 3 ] and the ability of JEV to persist and transmit between pigs in absence of mosquitos [ 4 ] are increasing risks of virus spread and persistence in regions with more moderate climate and becoming a worldwide public health concern.

In regions at risk, vaccination programs are available [ 5 ]. By an unknown mechanism, JEV invades the CNS infecting and killing neurons with a specific tropism for developing neurons [ 7 , 8 ]. In brain autopsies of fatal JE patients, JEV—infected neurons are localized in the thalamus, in the brainstem, as well as in the hippocampus, which are areas of neuronal turn-over, even in adults [ 9 ]. Accumulating evidences highlight the prominent role of the cells of the macrophage lineage in JEV pathogenesis.

In fact, human blood monocytes survive from productive infection by JEV and can maintain infectious virus for 5 days [ 10 ]. Human blood monocyte-derived dendritic cells MoDC and monocyte-derived macrophages MDM support virus replication in vitro [ 11 , 12 ]. Microglial cells are the unique resident immune cells of the nervous system, which populate the brain during early development, but can also derive from blood monocytes after birth under specific conditions [ 13 ].

Mouse microglia are activated and produce numbers of pro-inflammatory factors upon JEV infection in vivo and in vitro [ 16 ]. Moreover, mouse microglia are productively infected by JEV in vitro and are proposed to play a role in long-lasting infection [ 17 ]. Despite differences between humans and most of animals, rodent models are widely used in the laboratory to characterize and understand JEV pathogenicity.

The present study explores the interactions between human microglia and JEV. To achieve this work, vaccine-containing inactivated JEV and two live JEV strains were applied in-vitro on human primary microglia and human monocyte-derived microglia.

Our results show that human microglia were activated upon JEV exposure, without affecting viability. Finally, human microglia supported viral replication but cell-cell contact was required for viral transmission to other cells. The present study demonstrates the implication of human microglia in inflammatory responses and virus propagation upon exposure to JEV. Authorization number A was obtained from the Federal Office for the Environment FOEN, Bern, Switzerland for collection of human samples and manipulation of the various cells and viruses.

All samples were analysed anonymously. Human primary microglia cells BdMG were isolated from the cortex of brains derived from an anonymous donor of the body donation program of the University of Fribourg according to a newly established protocol from [ 18 ]. Briefly, the brain was excised within 8 h post-mortem. Human blood monocyte-derived microglia M-MG were generated from buffy coats of anonymous healthy donors, alternatively obtained from Don du sang Lausanne, Switzerland and Blutspendedienst Bern, Switzerland , as previously described [ 19 ].

Briefly, human peripheral blood mononuclear cells PBMC were isolated from buffy coat after Ficoll-Paque density gradient centrifugation. After adherence, non-adherent cells and contaminants were washed away. Filtered suspension of aluminium hydroxide 0.

Cells were cultured until cytopathogenic effects appear approx. Viral titres were determined by end-point titration on BHK cells.

Addition of serum abrogated the responsiveness of human microglia data not shown. After treatment, cells and supernatants were separately collected for further analysis. On one hand, human microglia and BHK cells were cultured in cell-cell contact condition by addition of BHK cells on top of human microglia. On another hand, human microglia and BHK cells were separated using transwell insert 0. In these conditions, the lower chamber contained human microglia and the upper chamber contained BHK cells.

Cells were successively fixed with 0. Ultrathin sections 0. Viral particles were detected intracellularly using the pan-immune anti-flavivirus antibody ATCC as primary antibody. Concentrations for use of the antibodies were optimized in our laboratory. Cells were analysed using single colour flow cytometry. Beads were analysed using multi-colour flow cytometry and concentrations of chemokines were determined by comparison with the standard material included in the kit.

Reference dye was ROX Life technologies. A study using mouse microglia suggest that those cells are a possible viral reservoir and consequently contribute substantially to JEV pathogenesis [ 17 ]. In order to investigate the interactions between microglia in humans and JEV, changes in the morphology of the cells were explored using bright field microscopy and flow cytometry.

Under the light microscope, Alum-treated human microglia presented cellular processes with a uniform cytoplasmic content whereas JEV vaccine exposure led to an amoeboid shape and the presence of large intracellular vacuoles of various sizes in human microglia Fig.

Changes in morphology were confirmed by flow cytometry Fig. Then, the viability of cells was measured in order to evaluate whether JEV induced cytotoxicity to human microglia.

Flow cytometry analysis of cell surface externalization of phosphatidylserine indicated by Annexin-V staining was used to measure apoptotic cells in culture. Overall, live JEV does not alter the viability of human microglia in vitro. Morphology and cell death of JEV-treated human microglia.

Cell morphology and cell death were investigated. Black gate delineates microglia cells excluding other cell types and cell debris. Data are of 2 independent experiments with each condition performed in triplicate cultures. The bars represent the mean value; the error bars the standard deviation.

Untreated human microglia demonstrated a constitutive production of chemokines that did not alter upon treatment with Alum or Mock data not shown. Moreover, chemokine responses of live JEV-treated human microglia were viral dose-dependent Fig. To conclude, live JEV is able to modulate the production of inflammatory chemokines in human microglia in a dose-dependent. Chemokine responses of JEV-treated human microglia. The black line represents the mean value; the error bars the standard deviation.

The marker represents the mean value and the error bar the standard deviation. Values are of 2 independent experiments with each condition performed in triplicate cultures. In various flavivirus infections such as for dengue and JEV, chemokine receptors have been shown to be critical in the disease outcome [ 26 , 27 ].

In order to explore the influence of JEV on the expression of chemokine receptors on human microglia, modifications in the expression of chemokine receptors of the CC, CXC and CX 3 C sub-families were investigated using flow cytometry. This supported the validity of the model of in-vitro generated microglia in comparison to primary cells.

Interestingly, an MOI of 0. Human microglia were treated with Alum, JEV vaccine used at a concentration of 1. Expression of indicated chemokine receptor on human microglia gated as in Fig. The black line represents the mean value and the error bars the standard deviation.

Lower panels are histogram bars of fold change in expression for the indicated chemokine receptor in human microglia. The bar represents the mean value; the error bars the standard.

Values are of 3 independent experiments with each condition performed in triplicate cultures. Low and high doses are respectively a concentration of 0. Values are of a representative experiment with each condition performed in triplicate cultures. In the CNS, microglial cells link the innate and the adaptive immunity by presenting MHC molecules loaded with exogenous antigen to T-cells.

Microglia cells have a constitutive, but low expressions of MHC class I and II that increase in response to insults [ 15 ]. Expression of indicated MHC molecules on human microglia gated as in Fig. Oh, and starvation. There is a lot of starvation in this book. The wealthy are simply paranoid and treacherous. Amsden has created a very thorough world, and it is clear that she knows exactly how it functions or not and why. Such a feat is quite impressive, but it was difficult to understand how a culture that creates hovercars and amazing wireless technology has gotten absolutely nowhere with agricultural yields.

If this seems like picking nits, the issue of food and lack thereof drives a great deal of the conflicts and drama in the book. This perspective makes The Immortality Virus a bit exhausting: there is no humor, kindness or humanity to ease a very harsh idea of a possible future. Amsden tries to humanize Grace Harper, but was unable to give enough of a glimpse into her interior life to make her random acts of kindness seem anything but halfhearted and grudging.

Grace Harper loathes the idea of being in a relationship, although flings are certainly acceptable, leaving her suitors to rather swoon after her, in a reversal of several hundred years of great literature. It leaves the men emasculated and Grace without a heart. Well, that and the fact that I thought it was an anthology. Tales of Robin Hood was originally published as a paperback by Baen and pretty much vanished without a trace, until iUniverse reprinted it in under the title Robin Hood and the Beasts of Sherwood.

It caught on with modern readers in the new incarnation, and was warmly reviewed by a new generation of readers. Gabriel Syme was not merely a detective who pretended to be a poet; he was really a poet who had become a detective.

Nor was his hatred of anarchy hypocritical. He was one of those who are driven early in life into too conservative an attitude by the bewildering folly of most revolutionists. He had not attained it by any tame tradition. His respectability was spontaneous and sudden, a rebellion against rebellion. He came of a family of cranks, in which all the oldest people had all the newest notions.

One of his uncles always walked about without a hat, and another had made an unsuccessful attempt to walk about with a hat and nothing else. His father cultivated art and self-realisation; his mother went in for simplicity and hygiene. Hence the child, during his tenderer years, was wholly unacquainted with any drink between the extremes of absinth and cocoa, of both of which he had a healthy dislike. The more his mother preached a more than Puritan abstinence the more did his father expand into a more than pagan latitude; and by the time the former had come to enforcing vegetarianism, the latter had pretty well reached the point of defending cannibalism.

Being surrounded with every conceivable kind of revolt from infancy, Gabriel had to revolt into something, so he revolted into the only thing left—sanity.

Gabriel Syme is a poet who fears the world is destined for destruction under a wave of moral relativism and nihilism. He is recruited to a special anti-anarchist unit of the British police by a mysterious figure who remains hidden in the shadows. Cover artist unknown I have recently been entertained by a Facebook group called View from My Window.

So why am I telling you this and what does it have to do with horror? Zu: Warriors from the Magic Mountain Hong Kong, The worldwide success of the Star Wars movies, followed by that of Conan the Barbarian , opened the funding floodgates for fantasy films, not just in Hollywood and Europe, but in Asia as well.