Tissue Tropism and Target Cells of NSs-Deleted Rift Valley Fever Virus in Live Immunodeficient Mice

Rift Valley fever, caused by a member of the Bunyaviridae family, has spread during recent years to most sub-Saharan African countries, in Egypt and in the Arabian peninsula. The virus can be transmitted by insect vectors or by direct contacts with infectious tissues. The analysis of virus replication and dissemination in laboratory animals has been hampered by the need to euthanize sufficient numbers of animals and to assay appropriate organs at various time points after infection to evaluate the viral replication. By following the bioluminescence and fluorescence of Rift Valley fever viruses expressing light reporters, we were able to track the real-time dissemination of the viruses in live immunodeficient mice. We showed that the first infected organs were the thymus, spleen and liver, but the liver rapidly became the main location of viral replication. Phagocytes also appeared as important targets, and their systemic depletion by use of clodronate liposomes decreased the number of viruses in the blood, delayed the viral dissemination and prolonged the survival of the infected mice

Hypoxia Disruption of Vertebrate CNS Pathfinding through EphrinB2 Is Rescued by Magnesium

The mechanisms of hypoxic injury to the developing human brain are poorly understood, despite being a major cause of chronic neurodevelopmental impairments. Recent work in the invertebrate Caenorhabditis elegans has shown that hypoxia causes discrete axon pathfinding errors in certain interneurons and motorneurons. However, it is unknown whether developmental hypoxia would have similar effects in a vertebrate nervous system. We have found that developmental hypoxic injury disrupts pathfinding of forebrain neurons in zebrafish (Danio rerio), leading to errors in which commissural axons fail to cross the midline. The pathfinding defects result from activation of the hypoxia-inducible transcription factor (hif1) pathway and are mimicked by chemical inducers of the hif1 pathway or by expression of constitutively active hif1α. Further, we found that blocking transcriptional activation by hif1α helped prevent the guidance defects. We identified ephrinB2a as a target of hif1 pathway activation, showed that knock-down of ephrinB2a rescued the guidance errors, and showed that the receptor ephA4a is expressed in a pattern complementary to the misrouting axons. By targeting a constitutively active form of ephrinB2a to specific neurons, we found that ephrinB2a mediates the pathfinding errors via a reverse-signaling mechanism. Finally, magnesium sulfate, used to improve neurodevelopmental outcomes in preterm births, protects against pathfinding errors by preventing upregulation of ephrinB2a. These results demonstrate that evolutionarily conserved genetic pathways regulate connectivity changes in the CNS in response to hypoxia, and they support a potential neuroprotective role for magnesium

The Use of Spinning-Disk Confocal Microscopy for the Intravital Analysis of Platelet Dynamics in Response to Systemic and Local Inflammation

Platelets are central players in inflammation and are an important component of the innate immune response. The ability to visualize platelets within the live host is essential to understanding their role in these processes. Past approaches have involved adoptive transfer of labelled platelets, non-specific dyes, or the use of fluorescent antibodies to tag platelets in vivo. Often, these techniques result in either the activation of the platelet, or blockade of specific platelet receptors. In this report, we describe two new methods for intravital visualization of platelet biology, intravenous administration of labelled anti-CD49b, which labels all platelets, and CD41-YFP transgenic mice, in which a percentage of platelets express YFP. Both approaches label endogenous platelets and allow for their visualization using spinning-disk confocal fluorescent microscopy. Following LPS-induced inflammation, we were able to measure a significant increase in both the number and size of platelet aggregates observed within the vasculature of a number of different tissues. Real-time observation of these platelet aggregates reveals them to be large, dynamic structures that are continually expanding and sloughing-off into circulation. Using these techniques, we describe for the first time, platelet recruitment to, and behaviour within numerous tissues of the mouse, both under control conditions and following LPS induced inflammation

Consensus guidelines for the use and interpretation of angiogenesis assays

The formation of new blood vessels, or angiogenesis, is a complex process that plays important roles in growth and development, tissue and organ regeneration, as well as numerous pathological conditions. Angiogenesis undergoes multiple discrete steps that can be individually evaluated and quantified by a large number of bioassays. These independent assessments hold advantages but also have limitations. This article describes in vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis and highlights critical aspects that are relevant for their execution and proper interpretation. As such, this collaborative work is the first edition of consensus guidelines on angiogenesis bioassays to serve for current and future reference

Bone marrow B cell apoptosis during in vivo influenza virus infection requires TNF-α and lymphotoxin-α

Suppression of bone marrow myeloid and erythroid progenitor cells occurs after infection with a variety of different viruses. In this study, we characterize the alterations in bone marrow (BM) lymphocytes after influenza virus infection in mice. We found a severe loss of BM B cells, particularly CD43Blow/-B220+ pre-B and immature B cells, in influenza virus-infected mice. Depletion of BM B lineage cells resulted primarily from cell cycle arrest and most likely apoptosis within the BM environment, rather than from increased trafficking of BM emigrants to peripheral lymphoid tissues. Use of gene-knockout mice indicates that depletion of BM B cells is dependent on TNF-α lymphotoxin-α, and both TNF receptors, TNFR1-p55 and TNFR2-p75. Thus, TNF-α and lymphotoxin-α are required for loss of BM B lineage cells during respiratory infection with influenza virus

Prevalentie van antibioticaresistente bacteriën in Maas, Rijn en Nieuwe Maas

In de grote Nederlandse rivieren de Maas, de Rijn en de Nieuwe Maas komen bacteriën voor waarvan hoge percentages resistent zijn tegen een of meer soorten antibiotica. Dit blijkt uit verkennend onderzoek van het RIVM. Blootstelling via oppervlaktewater: Als mensen aan verontreinigd oppervlaktewater blootgesteld worden, kunnen zij antibioticaresistente bacteriën binnenkrijgen. Dit kan bijvoorbeeld via recreatiewater of via water dat gebruikt worden om gewassen te besproeien. Dergelijk contact kan risico's voor de volksgezondheid met zich meebrengen, omdat deze antibiotica nodig zijn om infecties te behandelen. Antibioticaresistente bacteriën kunnen op meerdere manieren in oppervlaktewater terechtkomen, bijvoorbeeld doordat mest van dieren die met antibiotica zijn behandeld, afspoelt naar het oppervlaktewater. Een andere oorzaak kan zijn dat gedeeltelijk gezuiverd of ongezuiverd afvalwater in oppervlaktewater wordt geloosd, bijvoorbeeld afkomstig van ziekenhuizen waar mensen zijn behandeld met antibiotica. In totaal waren gemiddeld eenderde tot de helft van alle Escherichia coli en van de intesintale enterococcen resistent tegen een of meer soorten antibiotica. In sommige van de monsters werden antibioticaresistente stammen van Staphylococcus aureus, Campylobacter en Salmonella aangetoond. De meeste van deze bacteriën zijn darmbacteriën; Staphylococcus aureus komt vooral voor op de huid, en in de neus en keel van mensen. Diverse risico's: De risico's kunnen zich op verschillende manieren manifesteren. Op de eerste plaats kunnen mensen die aan antibioticaresistente bacteriën worden blootgesteld, daarvan ziek worden en vervolgens problemen krijgen bij de behandeling ervan. Daarnaast is het mogelijk dat mensen die worden blootgesteld aan de resistente bacteriën zelf niet ziek worden, maar deze overdragen aan mensen met verminderde weerstand, zoals ziekenhuispatiënten en ouderen. Deze groep mensen kan vervolgens wel ziek worden door deze bacteriën. Ten slotte is er het risico dat onschadelijke antibioticaresistente bacteriën zich in de darmen nestelen en daar genen die resistentie veroorzaken doorgeven aan andere, ziekteverwekkende bacteriën. Nader onderzoek naar volksgezondheidsrisico's: Onderzoek naar de mate waarin antibioticaresistente bacteriën in oppervlaktwater voorkomen is van belang om te kunnen inschatten in hoeverre mensen via het milieu worden blootgesteld aan deze bacteriën. Nader RIVMonderzoek zal hierop gericht zijn, en wat dit betekent voor de volksgezondheid.A high percentage of bacteria in large Dutch rivers is resistant to one or more antibiotics. This was demonstrated in an exploratory study performed at the RIVM. Exposure through surface water: If people come into contact with contaminated surface water, they risk exposure to bacteria that are resistant to one or more antibiotics. This may occur for instance during recreation in surface water or through water used for irrigation of crops. Because antibiotics are needed for treatment of infections, such contact may involve public health risks. Antibiotic resistant bacteria can end up in surface water by different routes, for example through run off of manure from animals treated with antibiotics to surface water. Another source may be the discharge of partially treated or untreated waste water, for instance derived from hospitals where people are treated with antibiotics. Overall, one third to half of all Escherichia coli and intestinal enterococci were resistant to one or more antibiotics. In some of the samples antibiotic resistant strains of Staphylococcus aureus, Campylobacter and Salmonella were detected. Most of these bacteria are gut bacteria; Staphylococcus aureus is predominantly present on people's skin and in their noses and throats. Diverse risks: The risks can manifest themselves in different ways. Firstly, after exposure to antibiotic resistant bacteria people may develop disease caused by these bacteria, which may therefore be hard to treat. Additionally, people may not become ill themselves, but transfer the resistant bacteria to people who are more vulnerable, such as hospital patients and the elderly. Subsequently, this category of people can develop disease. Finally, there is a risk that the antibiotic resistant bacteria establish themselves in the gut and transfer resistance genes to disease-causing bacteria if these are subsequently ingested Further research on public health risks: Data on the prevalence of antibiotic resistant bacteria in surface water are necessary to estimate the contribution of the environment to human exposure to these bacteria. Further RIVM research is focused on this type of exposure and the associated possible public health risks.VROM-Inspecti