Hepatitis E virus RNA in commercially available porcine livers in The Netherlands

Hepatitis E virus (HEV) infections caused by genotype 3 are increasingly observed in industrialized countries, without a distinct source. High similarity between human and swine strains of HEV strongly suggest possible zoonotic transmission. It was reported previously that in 55% of Dutch pig farms HEV-excreting fattening pigs were present. In the current study, presence of HEV RNA in commercially available porcine livers was shown. We examined 62 commercially available porcine livers for HEV contamination. Before examination of livers, the most sensitive combination of tissue disruption and RNA-extraction was chosen from four disruption and seven RNA-extraction methods. Four of 62 livers were shown to be positive for HEV RNA by RT-PCR and Southern blot hybridization, and three sequences were obtained. Phylogenetic analysis showed clustering of the sequences with previously published Dutch HEV genotype 3 sequences from humans and swine. To study infectivity of possible virus, three pigs were intravenously inoculated with suspensions from commercially available HEV positive livers. Two other pigs served as high-dose or low-dose controls. The low-dose control received a comparable viral count as animals receiving inocula from commercially available livers, the high dose control received a viral count that was known to generate infection. Faecal shedding of HEV was observed in the high-dose control, indicating that the control virus was infectious. No faecal shedding of HEV was observed for the low-dose control and the three pigs that were administered the commercially available livers extracts. In conclusion, HEV RNA was found in commercially available porcine livers. inoculation of susceptible pigs with extracts from HEV-positive livers did not lead to infection, but this may be a dose-dependent effect. The risk for consumers should be investigated further

Hepatitis E virus sequences in swine related to sequences in humans, The Netherlands.

Hepatitis E virus (HEV), a major cause of viral hepatitis in much of the developing world, has recently been detected in swine in North America and Asia, raising concern about potential for zoonotic transmission. To investigate if HEV is commonly present in swine in the Netherlands, pooled stool samples from 115 swine farms and nine individual pigs with diarrhea were assayed by reverse transcription-polymerase chain reaction (RT-PCR) amplification. HEV RNA was detected by RT-PCR and hybridization in 25 (22%) of the pooled specimens, but in none of the individual samples. RT-PCR amplification products of open reading frames 1 and 2 were sequenced, and the results were compared with published sequences of HEV genotypes from humans and swine. HEV strains from swine in the Netherlands were clustered in at least two groups, together with European and American isolates from swine and humans. Our data show that HEV in swine in the Netherlands are genetically closely related to HEV isolates from humans. Although zoonotic transmission has not been proven, these findings suggest that swine may be reservoir hosts of HEV

Truly reconciled? A dyadic analysis of post-conflict social reintegration in Northern Uganda

In the aftermath of civil war or violent internal conflict, one of the key peacebuilding challenges is the reconciliation of former enemies who are members of the same small-scale societies. A failure of social reintegration may contribute to what is known as a conflict trap. To detect lingering hostile attitudes among a community’s various factions is crucial, but the approaches adopted in previous studies tend to focus on the impact of conflict on one or other aggregated indicator of social cohesion rather than on how violence-affected individuals regard and act towards their fellow community members. Here we demonstrate the value of concentrating on this latter dyadic component of social interactions and we use behavioural experiments and a social tie survey to assess, in an appropriately disaggregated manner, social cohesion in a post-conflict setting in northern Uganda. Whereas in self-reported surveys, ex-combatants appear to be well-connected, active members of their communities, the experiments unveil the continued reluctance of other community members to share or cooperate with them; fewer resources are committed to ex-combatants than to others, which is statistically significant. The dyadic nature of our analysis allows us to detect which groups are more prone to discriminate against ex-combatants, which may help facilitate targeted interventions

Genetic diversity of Brazilian isolates of feline immunodeficiency virus

We isolated Feline immunodeficiency virus (FIV) from three adult domestic cats, originating from two open shelters in Brazil. Viruses were isolated from PBMC following co-cultivation with the feline T-lymphoblastoid cell line MYA-1. All amplified env gene products were cloned directly into pGL8MYA. The nucleic acid sequences of seven clones were determined and then compared with those of previously described isolates. The sequences of all of the Brazilian virus clones were distinct and phylogenetic analysis revealed that all belong to subtype B. Three variants isolated from one cat and two variants were isolated from each of the two other cats, indicating that intrahost diversity has the potential to pose problems for the treatment and diagnosis of FIV infection

The Distances to Open Clusters from Main-Sequence Fitting. III. Improved Accuracy with Empirically Calibrated Isochrones

We continue our series of papers on open cluster distances with a critical assessment of the accuracy of main-sequence fitting using isochrones that employ empirical corrections to the color-temperature relations. We use four nearby open clusters with multicolor photometry and accurate metallicities and present a new metallicity for Praesepe ([Fe/H] = +0.11 +/- 0.03) from high-resolution spectra. The internal precision of distance estimates is about a factor of 5 better than the case without the color calibrations. After taking into account all major systematic errors, we obtain distances accurate to about 2%-3% when there exists a good metallicity estimate. Metallicities accurate to better than 0.1 dex may be obtained from BVIcKs photometry alone. We also derive a helium abundance for the Pleiades of Y = 0.279 +/- 0.015, which is equal within the errors to the Sun's initial helium abundance and that of the Hyades. Our best estimates of distances are (m - M)_0 = 6.33 +/- 0.04, 8.03 +/- 0.04, and 9.61 +/- 0.03 to Praesepe, NGC 2516, and M67, respectively. Our Pleiades distance at the spectroscopic metallicity, (m - M)_0 = 5.66 +/- 0.01 (internal) +/- 0.05 (systematic), is in excellent agreement with several geometric distance measurements. We have made calibrated isochrones for -0.3 <= [Fe/H] <= +0.2 available at http://www.astronomy.ohio-state.edu/iso/ .Comment: 28 pages, 23 figures; accepted for publication in the Ap

Pathogenic mycobacteria achieve cellular persistence by inhibiting the Niemann-Pick Type C disease cellular pathway

Background. Tuberculosis remains a major global health concern. The ability to prevent phagosome-lysosome fusion is a key mechanism by which intracellular mycobacteria, including Mycobacterium tuberculosis, achieve long-term persistence within host cells. The mechanisms underpinning this key intracellular pro-survival strategy remain incompletely understood. Host macrophages infected with intracellular mycobacteria share phenotypic similarities with cells taken from patients suffering from Niemann-Pick Disease Type C (NPC), a rare lysosomal storage disease in which endocytic trafficking defects and lipid accumulation within the lysosome lead to cell dysfunction and cell death. We investigated whether these shared phenotypes reflected an underlying mechanistic connection between mycobacterial intracellular persistence and the host cell pathway dysfunctional in NPC.  Methods. The induction of NPC phenotypes in macrophages from wild-type mice or obtained from healthy human donors was assessed via infection with mycobacteria and subsequent measurement of lipid levels and intracellular calcium homeostasis. The effect of NPC therapeutics on intracellular mycobacterial load was also assessed.  Results. Macrophages infected with intracellular mycobacteria phenocopied NPC cells, exhibiting accumulation of multiple lipid types, reduced lysosomal Ca 2+ levels, and defects in intracellular trafficking. These NPC phenotypes could also be induced using only lipids/glycomycolates from the mycobacterial cell wall. These data suggest that intracellular mycobacteria inhibit the NPC pathway, likely via inhibition of the NPC1 protein, and subsequently induce altered acidic store Ca 2+ homeostasis. Reduced lysosomal calcium levels may provide a mechanistic explanation for the reduced levels of phagosome-lysosome fusion in mycobacterial infection. Treatments capable of correcting defects in NPC mutant cells via modulation of host cell calcium were of benefit in promoting clearance of mycobacteria from infected host cells.  Conclusion. These findings provide a novel mechanistic explanation for mycobacterial intracellular persistence, and suggest that targeting interactions between the mycobacteria and host cell pathways may provide a novel avenue for development of anti-TB therapies

Complement as an Endogenous Adjuvant for Dendritic Cell-Mediated Induction of Retrovirus-Specific CTLs

Previous studies have demonstrated the involvement of complement (C) in induction of efficient CTL responses against different viral infections, but the exact role of complement in this process has not been determined. We now show that C opsonization of retroviral particles enhances the ability of dendritic cells (DCs) to induce CTL responses both in vitro and in vivo. DCs exposed to C-opsonized HIV in vitro were able to stimulate CTLs to elicit antiviral activity significantly better than non-opsonized HIV. Furthermore, experiments using the Friend virus (FV) mouse model illustrated that the enhancing role of complement on DC-mediated CTL induction also occurred in vivo. Our results indicate that complement serves as natural adjuvant for DC-induced expansion and differentiation of specific CTLs against retroviruses