Faecal shedding, alimentary clearance and intestinal spread of prions in hamsters fed with scrapie

Shedding of prions via faeces may be involved in the transmission of contagious prion diseases. Here, we fed hamsters 10 mg of 263K scrapie brain homogenate and examined the faecal excretion of disease-associated prion protein (PrPTSE) during the course of infection. The intestinal fate of ingested PrPTSE was further investigated by monitoring the deposition of the protein in components of the gut wall using immunohistochemistry and paraffin-embedded tissue (PET) blotting. Western blotting of faecal extracts showed shedding of PrPTSE in the excrement at 24–72 h post infection (hpi), but not at 0–24 hpi or at later preclinical or clinical time points. About 5% of the ingested PrPTSE were excreted via the faeces. However, the bulk of PrPTSE was cleared from the alimentary canal, most probably by degradation, while an indiscernible proportion of the inoculum triggered intestinal infection. Components of the gut-associated lymphoid tissue (GALT) and the enteric nervous system (ENS) showed progressing accumulation of PrPTSE from 30 days post infection (dpi) and 60 dpi, respectively. At the clinical stage of disease, substantial deposits of PrPTSE were found in the GALT in close vicinity to the intestinal lumen. Despite an apparent possibility of shedding from Peyer’s patches that may involve the follicle-associated epithelium (FAE), only small amounts of PrPTSE were detected in faeces from clinically infected animals by serial protein misfolding cyclic amplification (sPMCA). Although excrement may thus provide a vehicle for the release of endogenously formed PrPTSE, intestinal clearance mechanisms seem to partially counteract such a mode of prion dissemination

Identification of Epigenetic Signature Associated With Alpha Thalassemia/Mental Retardation X-linked Syndrome

BACKGROUND: Alpha thalassemia/mental retardation X-linked syndrome (ATR-X) is caused by a mutation at the chromatin regulator gene RESULTS: We performed genome-wide DNA methylation assessment of the peripheral blood samples from 18 patients with ATR-X and compared it to 210 controls. We demonstrated the evidence of a unique and highly specific DNA methylation epi-signature in the peripheral blood of ATRX patients, which was corroborated by targeted bisulfite sequencing experiments. Although genomically represented, differentially methylated regions showed evidence of preferential clustering in pericentromeric and telometric chromosomal regions, areas where ATRX has multiple functions related to maintenance of heterochromatin and genomic integrity. CONCLUSION: Most significant methylation changes in the 14 genomic loci provide a unique epigenetic signature for this syndrome that may be used as a highly sensitive and specific diagnostic biomarker to support the diagnosis of ATR-X, particularly in patients with phenotypic complexity and in patients wit

Mechanistically-guided materials chemistry: synthesis of new ternary nitrides, CaZrN2_2 and CaHfN2_2

Recent computational studies have predicted many new ternary nitrides, revealing synthetic opportunities in this underexplored phase space. However, synthesizing new ternary nitrides is difficult, in part because intermediate and product phases often have high cohesive energies that inhibit diffusion. Here, we report the synthesis of two new phases, calcium zirconium nitride (CaZrN$_2$) and calcium hafnium nitride (CaHfN$_2$), by solid state metathesis reactions between Ca$_3$N$_2$ and $M$Cl$_4$ ($M$ = Zr, Hf). Although the reaction nominally proceeds to the target phases in a 1:1 ratio of the precursors via Ca$_3$N$_2$ + $M$Cl$_4$ $\rightarrow$ Ca$M$N$_2$ + 2 CaCl$_2$, reactions prepared this way result in Ca-poor materials (Ca$_xM_{2-x}$N$_2$, $x<1$). A small excess of Ca$_3$N$_2$ (ca. 20 mol\%) is needed to yield stoichiometric Ca$M$N$_2$, as confirmed by high-resolution synchrotron powder X-ray diffraction. In situ synchrotron X-ray diffraction studies reveal that nominally stoichiometric reactions produce Zr$^{3+}$ intermediates early in the reaction pathway, and the excess Ca$_3$N$_2$ is needed to reoxidize Zr$^{3+}$ intermediates back to the Zr$^{4+}$ oxidation state of CaZrN$_2$. Analysis of computationally-derived chemical potential diagrams rationalizes this synthetic approach and its contrast from the synthesis of MgZrN$_2$. These findings additionally highlight the utility of in situ diffraction studies and computational thermochemistry to provide mechanistic guidance for synthesis

Cell-Autonomous Sex Differences in Gene Expression in Chicken Bone Marrow-Derived Macrophages

We have identified differences in gene expression in macrophages grown from the bone marrow of male and female chickens in recombinant chicken M-CSF (CSF1). Cells were profiled with or without treatment with bacterial LPS for 24 h. Approximately 600 transcripts were induced by prolonged LPS stimulation to an equal extent in the male and female macrophages. Many transcripts encoded on the Z chromosome were expressed ∼1.6-fold higher in males, reflecting a lack of dosage compensation in the homogametic sex. A smaller set of W chromosome-specific genes was expressed only in females. LPS signaling in mammals is associated with induction of type 1 IFN-responsive genes. Unexpectedly, because IFNs are encoded on the Z chromosome of chickens, unstimulated macrophages from the female birds expressed a set of known IFN-inducible genes at much higher levels than male cells under the same conditions. To confirm that these differences were not the consequence of the actions of gonadal hormones, we induced gonadal sex reversal to alter the hormonal environment of the developing chick and analyzed macrophages cultured from male, female, and female sex-reversed embryos. Gonadal sex reversal did not alter the sexually dimorphic expression of either sex-linked or IFN-responsive genes. We suggest that female birds compensate for the reduced dose of inducible IFN with a higher basal set point of IFN-responsive genes

Space allowance and the behaviour of captive southern hairy-nosed wombats (Lasiorhinus latifrons)

Captive southern hairy-nosed wombats (Lasiorhinus latifrons) often display indicators of substandard welfare, including aggression and stereotypical pacing. To determine if space availability influences the welfare of wombats, the behaviour of three groups of L. latifrons (n = 3) was studied in three different sized enclosures: small (S) (75.5 m2; the minimum space requirement for three wombats in Queensland, Australia), medium (M) (151 m2, twice the minimum space) and large (L) (224 m2, three times the minimum space) in a Latin Square design. Compared to wombats in larger enclosures, those in the small enclosure were observed to display more biting (S: 1.96; M: 0.42; L: 0.28, SED &plusmn; 0.56 counts / day, P = 0.01), retreat from conspecifics (S: 15.0; M: 9.9; L: 7.1 SED &plusmn; 2.66 counts / day, P = 0.03), and visual scanning (S: 52.8; M: 33.9; L: 28.8, SED &plusmn; 4.62 counts / day, P &lt; 0.001); they also spent more time fenceline digging, which may represent attempts to escape (S: 0.78; M: 0.16; L: 0.24, SED &plusmn; 0.07 min / m / day, P &lt; 0.0001). Those in the largest enclosure showed less self-directed grooming behaviour than those in the two smaller enclosures (S: 23.80; M: 24.08; L: 14.42, SED &plusmn; 3.22 counts / day, P = 0.02). It is concluded that small 2 enclosure size had a negative impact on the behaviour of wombat, and as a consequence, current 30 minimum space requirements for wombats in captivity should be reassessed

HIV DNA subspecies persist in both activated and resting memory CD4+ T cells during antiretroviral therapy

The latent HIV reservoir is a major impediment to curing HIV infection. The contribution of CD4 T cell activation status to the establishment and maintenance of the latent reservoir was investigated by enumerating viral DNA components in a cohort of 12 individuals commencing antiretroviral therapy (ART) containing raltegravir, an integrase inhibitor. Prior to ART, the levels of total HIV DNA were similar across HLA-DR and HLA-DR (HLA-DR) CD38 memory CD4 T cell phenotypes; episomal two-long terminal repeat (2-LTR) HIV DNA levels were higher in resting (HLA-DR CD38) cells, and this phenotype exhibited a significantly higher ratio of 2-LTR to integrated HIV DNA (P = 0.002). After 1 year of ART, there were no significant differences across each of the memory phenotypes of any HIV DNA component. The decay dynamics of integrated HIV DNA were slow within each subset, and integrated HIV DNA in the resting HLA-DR CD38 subset per mm of peripheral blood exhibited no significant decay (half-life of 25 years). Episomal 2-LTR HIV DNA decayed relative to integrated HIV DNA in resting cells with a half-life of 134 days. Surprisingly, from week 12 on, the decay rates of both total and episomal HIV DNA were lower in activated CD38 cells. By weeks 24 and 52, HIV RNA levels in plasma were most significantly correlated with the numbers of resting cells containing integrated HIV DNA. On the other hand, total HIV DNA levels in all subsets were significantly correlated with the numbers of HLA-DR CD38 cells containing integrated HIV DNA. These results provide insights into the interrelatedness of cell activation and reservoir maintenance, with implications for the design of therapeutic strategies targeting HIV persistence

First results from the L3+C experiment at CERN

The L3+C experiment combines the high-precision spectrometer of the L3 detector at LEP, CERN, with a small air shower array. The momenta of cosmic ray induced muons can be measured from 20 to 2000 GeV/c. During the 1999 data taking period 5 billion muon events were recorded in the spectrometer. From April until mid Summer 2000 an additional 3 billion muon events have been recorded as well as 25 million air shower events. Here the first results on the muon momentum spectrum and charge ratio will be presented

The state of the Martian climate

60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

Switching Virally Suppressed, Treatment-Experienced Patients to a Raltegravir-Containing Regimen Does Not Alter Levels of HIV-1 DNA

Background: Current HIV-1 antiretroviral therapy (ART) greatly reduces virus replication but does not significantly affect the viral reservoir. Raltegravir, a recently introduced integrase inhibitor, could, at least theoretically, reduce residual viremia in patients on ART and affect the viral reservoir size. The aim of this study was to assess whether switching therapy in treatment-experienced patients that were virally suppressed to a raltegravir-containing regimen reduces the size of the viral reservoir, and if such treatment leads to a change in levels of HIV 2-LTR circles in this patient group. Methods: 14 ART experienced individuals with a suppressed viral load (,50 HIV-1 RNA copies/mL plasma) at baseline (for at least 2 months) were switched to a raltegravir-containing regimen. Blood samples were taken at baseline and at $2 timepoints up to 4866 weeks. Levels of total HIV-1 DNA and 2-LTR circles in peripheral blood mononuclear cells (PBMCs) were measured using real-time PCR assays. Results: There was no significant change in HIV-1 total DNA levels over the study duration (p = 0.808), median slope 0.24 (conservative nonparametric 95 % CI: 211.78, 26.23). Low levels of 2-LTR circles were detected in 2 patients. One had 16 copies/10 6 PBMCs at baseline and the other had 34 copies/10 6 PBMCs at week 51. Conclusions: The switch to a raltegravir containing regimen was not associated with a significant change in HIV-1 total DNA levels in this cohort. There were no observed changes in the levels of HIV-1 2-LTR circles associated with raltegravi

Progressive Activation of CD127+132− Recent Thymic Emigrants into Terminally Differentiated CD127−132+ T-Cells in HIV-1 Infection

AIM: HIV infection is associated with distortion of T-cell homeostasis and the IL-7/IL7R axis. Progressive infection results in loss of CD127+132- and gains in CD127-132+ CD4+ and CD8+ T-cells. We investigated the correlates of loss of CD127 from the T-cell surface to understand mechanisms underlying this homeostatic dysregulation. METHODS: Peripheral and cord blood mononuclear cells (PBMCs; CBMC) from healthy volunteers and PBMC from patients with HIV infection were studied. CD127+132-, CD127+132+ and CD127-132+ T-cells were phenotyped by activation, differentiation, proliferation and survival markers. Cellular HIV-DNA content and signal-joint T-cell receptor excision circles (sjTRECs) were measured. RESULTS: CD127+132- T-cells were enriched for naïve cells while CD127-132+ T-cells were enriched for activated/terminally differentiated T-cells in CD4+ and CD8+ subsets in health and HIV infection. HIV was associated with increased proportions of activated/terminally differentiated CD127-132+ T-cells. In contrast to CD127+132- T-cells, CD127-132+ T-cells were Ki-67+Bcl-2(low) and contained increased levels of HIV-DNA. Naïve CD127+132- T-cells contained a higher proportion of sjTRECs. CONCLUSION: The loss of CD127 from the T-cell surface in HIV infection is driven by activation of CD127+132- recent thymic emigrants into CD127-132+ activated/terminally differentiated cells. This process likely results in an irreversible loss of CD127 and permanent distortion of T-cell homeostasis