Complete Genomic Sequences of Three Salmonella enterica subsp. enterica Serovar Muenchen Strains from an Orchard in San Joaquin County, California.

We present here the complete genome sequences of three Salmonella enterica subsp. enterica serovar Muenchen strains, LG24, LG25, and LG26. All three strains were isolated from almond drupes grown in an orchard in San Joaquin County, California, in 2016. These genomic sequences are nonidentical and will contribute to our understanding of S. enterica genomics

Inoculation of Scrapie with the Self-Assembling RADA-Peptide Disrupts Prion Accumulation and Extends Hamster Survival

Intracerebral inoculation of 263K Scrapie brain homogenate (PrPsc) with a self-assembling RADA-peptide (RADA) significantly delayed disease onset and increased hamster survival. Time of survival was dependent on the dose of RADA and pre-incubation with PrPsc prior to inoculation. RADA treatment resulted in the absence of detectable PrPsc at 40 d followed by an increased rate of PrPsc accumulation at 75 d up to sacrifice. In all PrPsc inoculated animals, clinical symptoms were observed ∼10 d prior to sacrifice and brains showed spongiform degeneration with Congo red positive plaques. A time-dependent increase in reactive gliosis was observed in both groups with more GFAP detected in RADA treated animals at all time points. The PrP protein showed dose-dependent binding to RADA and this binding was competitively inhibited by Congo Red. We conclude that RADA disrupts the efficacy of prion transmission by altering the rate of PrPsc accumulation. This is the first demonstration that a self-assembling biomolecular peptide can interact with PrPsc, disrupt the course of Scrapie disease process, and extend survival

Construction of a Global Pain Systems Network Highlights Phospholipid Signaling as a Regulator of Heat Nociception

The ability to perceive noxious stimuli is critical for an animal's survival in the face of environmental danger, and thus pain perception is likely to be under stringent evolutionary pressure. Using a neuronal-specific RNAi knock-down strategy in adult Drosophila, we recently completed a genome-wide functional annotation of heat nociception that allowed us to identify α2δ3 as a novel pain gene. Here we report construction of an evolutionary-conserved, system-level, global molecular pain network map. Our systems map is markedly enriched for multiple genes associated with human pain and predicts a plethora of novel candidate pain pathways. One central node of this pain network is phospholipid signaling, which has been implicated before in pain processing. To further investigate the role of phospholipid signaling in mammalian heat pain perception, we analysed the phenotype of PIP5Kα and PI3Kγ mutant mice. Intriguingly, both of these mice exhibit pronounced hypersensitivity to noxious heat and capsaicin-induced pain, which directly mapped through PI3Kγ kinase-dead knock-in mice to PI3Kγ lipid kinase activity. Using single primary sensory neuron recording, PI3Kγ function was mechanistically linked to a negative regulation of TRPV1 channel transduction. Our data provide a systems map for heat nociception and reinforces the extraordinary conservation of molecular mechanisms of nociception across different species. © 2012 Neely et al

RADA Promotes Survival.

<p>Survival data following intracerebral inoculation of hamsters with 1% Scrapie brain homogenates (PrPsc) and 0.9% w/v RADA after 24 h. Four independent groups consisting of PrPsc brain homogenate combined with RADA (PrPsc+RADA) were evaluated for survival in days (Mean±SEM). The average survival time in days of all PrPsc+RADA (Groups 1–4) inoculated hamsters is compared to survival of those inoculated with equivalent doses of PrPsc alone. N = number of animals.</p

Effect of RADA on Mortality within 24 h.

<p>Mortality data at <24 h following single unilateral intracerebral (ic) inoculation of brain homogenates and RADA. Normal (PrPc) or Scrapie (PrPsc) infected hamster brain homogenates were inoculated alone (−) or with 0.9% w/v RADA (+). Low melt Agarose (0.9% w/v) combined with PrPsc served as an additional control. RADA alone or combined with PrPc brain homogenate resulted in no lethality. Combined RADA with PrPsc resulted in a rapid mortality within 24 h in ∼20% of the animals inoculated. N = number of hamsters inoculated.</p

Scrapie induced reactive gliosis and PrP aggregation is increased by RADA.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004440#pone-0004440-g005" target="_blank"><i>Fig. 5A</i></a> shows dual labeled immunofluorescent GFAP (red) and PrP (green) hamster brain micrographs. Limited staining of GFAP and no detectable PrP was observed in PrPc+RADA treated brain after 115 d (top right panel). PrPsc treatment alone results in an increased detection of GFAP-positive astrocytes (red) and weak PrP protein (green, white arrows) after 75 d (middle panel). Combined PrPsc+RADA treatment (right panel) results in abundant and strong detection of both GFAP-positive astrocytes and large PrP deposits (green, arrows). V = ventricle. Bv = blood vessel. DAPI nuclei (blue). Bars = 50 µm. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004440#pone-0004440-g005" target="_blank"><i>Fig. 5B</i></a> shows micrographs of hamster brain sections labeled with GFAP (red) and IBA1 (green). Increased IBA1-positive immunoreactive microglia are detected in brain following inoculation of PrPsc alone after 75 d (left panel; white arrows) and PrPsc+RADA after 115 d (middle panel; white arrows) as compared to PrPc+RADA after 115 d (right panel; white arrows). IBA1 and GFAP immunoreactivity do not co-localize in dual labeled sections from PrPsc and PrPsc+RADA brain. V = ventricle. DAPI nuclei (blue). Bars = 50 µm.</p

Effect of RADA on Secondary Passage of Scrapie Brain Homogenate.

<p>Survival data following intracerebral inoculation of hamsters with 263K Scrapie brain homogenate following primary passage with RADA (PrPsc/RADA). Survival (Mean±SEM) of hamsters inoculated with 1% Scrapie alone (PrPsc) is compared to those inoculated with 1% PrPsc/RADA (homogenate derived from a 1% PrPsc+0.9% w/v RADA infected brain) and 1% PrPsc/RADA combined a second time with 0.9% w/v RADA. N = number of animals.</p

RADA promotes increased survival of hamsters inoculated with 263K Scrapie.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004440#pone-0004440-g001" target="_blank"><i>Fig. 1A</i></a> compares hamster survival in days (mean±SEM) following intracerebral inoculation with 1% Scrapie alone (PrPsc), Scrapie pre-incubated with 0.9% w/v RADA (PrPsc+RADA) or Scrapie agarose plugs (PrPsc+agarose). <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004440#pone-0004440-g001" target="_blank"><i>Fig. 1B</i></a> compares hamster survival in days (mean±SEM) with increased dilution of Scrapie brain homogenate alone (open squares) to equivalent doses of Scrapie pre-incubated with 0.9% RADA (open circles). <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004440#pone-0004440-g001" target="_blank"><i>Fig. 1C</i></a> depicts a hamster survival curve in days (N = 6; mean±SEM) with increasing dilution of Scrapie inoculant (open circles). Plotted on the curve (open square) is the mean survival in days of hamsters inoculated with 10⁻² Scrapie combined with 0.9% RADA depicting equivalent titer of Scrapie inoculum. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004440#pone-0004440-g001" target="_blank"><i>Fig. 1D</i></a> shows a dose-dependent increase in hamster survival in days (mean±SEM) with increasing concentration of RADA (open circles) inoculated with 1% Scrapie. A synthetic 16-mer RADA-peptide was hydrated combined with 1% Scrapie and inoculated. Survival in days was plotted (grey triangle) and used to estimate RADA concentration.</p