Cytosolic chaperones influence the fate of a toxin dislocated from the endoplasmic reticulum

The plant cytotoxin ricin enters target mammalian cells by receptor-mediated endocytosis and undergoes retrograde transport to the endoplasmic reticulum (ER). Here, its catalytic A chain (RTA) is reductively separated from the cell-binding B chain, and free RTA enters the cytosol where it inactivates ribosomes. Cytosolic entry requires unfolding of RTA and dislocation across the ER membrane such that it arrives in the cytosol in a vulnerable, nonnative conformation. Clearly, for such a dislocated toxin to become active, it must avoid degradation and fold to a catalytic conformation. Here, we show that, in vitro, Hsc70 prevents aggregation of heat-treated RTA, and that RTA catalytic activity is recovered after chaperone treatment. A combination of pharmacological inhibition and cochaperone expression reveals that, in vivo, cytosolic RTA is scrutinized sequentially by the Hsc70 and Hsp90 cytosolic chaperone machineries, and that its eventual fate is determined by the balance of activities of cochaperones that regulate Hsc70 and Hsp90 functions. Cytotoxic activity follows Hsc70-mediated escape of RTA from an otherwise destructive pathway facilitated by Hsp90. We demonstrate a role for cytosolic chaperones, proteins typically associated with folding nascent proteins, assembling multimolecular protein complexes and degrading cytosolic and stalled, cotranslocational clients, in a toxin triage, in which both toxin folding and degradation are initiated from chaperone-bound states

Cytosolic chaperones influence the fate of a toxin dislocated from the endoplasmic reticulum

The plant cytotoxin ricin enters target mammalian cells by receptor-mediated endocytosis and undergoes retrograde transport to the endoplasmic reticulum (ER). Here, its catalytic A chain (RTA) is reductively separated from the cell-binding B chain, and free RTA enters the cytosol where it inactivates ribosomes. Cytosolic entry requires unfolding of RTA and dislocation across the ER membrane such that it arrives in the cytosol in a vulnerable, nonnative conformation. Clearly, for such a dislocated toxin to become active, it must avoid degradation and fold to a catalytic conformation. Here, we show that, in vitro, Hsc70 prevents aggregation of heat-treated RTA, and that RTA catalytic activity is recovered after chaperone treatment. A combination of pharmacological inhibition and cochaperone expression reveals that, in vivo, cytosolic RTA is scrutinized sequentially by the Hsc70 and Hsp90 cytosolic chaperone machineries, and that its eventual fate is determined by the balance of activities of cochaperones that regulate Hsc70 and Hsp90 functions. Cytotoxic activity follows Hsc70-mediated escape of RTA from an otherwise destructive pathway facilitated by Hsp90. We demonstrate a role for cytosolic chaperones, proteins typically associated with folding nascent proteins, assembling multimolecular protein complexes and degrading cytosolic and stalled, cotranslocational clients, in a toxin triage, in which both toxin folding and degradation are initiated from chaperone-bound states

Measurement of the quenching factor of Na recoils in NaI(Tl)

Measurements of the quenching factor for sodium recoils in a 5 cm diameter NaI(Tl) crystal at room temperature have been made at a dedicated neutron facility at the University of Sheffield. The crystal has been exposed to 2.45 MeV mono-energetic neutrons generated by a Sodern GENIE 16 neutron generator, yielding nuclear recoils of energies between 10 and 100 keVnr. A cylindrical BC501A detector has been used to tag neutrons that scatter off sodium nuclei in the crystal. Cuts on pulse shape and time of flight have been performed on pulses recorded by an Acqiris DC265 digitiser with a 2 ns sampling time. Measured quenching factors of Na nuclei range from 19% to 26% in good agreement with other experiments, and a value of 25.2 \pm 6.4% has been determined for 10 keV sodium recoils. From pulse shape analysis, the mean times of pulses from electron and nuclear recoils have been compared down to 2 keVee. The experimental results are compared to those predicted by Lindhard theory, simulated by the SRIM Monte Carlo code, and a preliminary curve calculated by Prof. Akira Hitachi.Comment: 21 pages, 13 figure

Folding-competent and folding-defective forms of Ricin A chain have different fates following retrotranslocation from the endoplasmic reticulum

We report that a toxic polypeptide retaining the potential to refold upon dislocation from the endoplasmic reticulum (ER) to the cytosol (ricin A chain; RTA) and a misfolded version that cannot (termed RTAΔ), follow ER-associated degradation (ERAD) pathways in Saccharomyces cerevisiae that substantially diverge in the cytosol. Both polypeptides are dislocated in a step mediated by the transmembrane Hrd1p ubiquitin ligase complex and subsequently degraded. Canonical polyubiquitylation is not a prerequisite for this interaction because a catalytically inactive Hrd1p E3 ubiquitin ligase retains the ability to retrotranslocate RTA, and variants lacking one or both endogenous lysyl residues also require the Hrd1p complex. In the case of native RTA, we established that dislocation also depends on other components of the classical ERAD-L pathway as well as an ongoing ER–Golgi transport. However, the dislocation pathways deviate strikingly upon entry into the cytosol. Here, the CDC48 complex is required only for RTAΔ, although the involvement of individual ATPases (Rpt proteins) in the 19S regulatory particle (RP) of the proteasome, and the 20S catalytic chamber itself, is very different for the two RTA variants. We conclude that cytosolic ERAD components, particularly the proteasome RP, can discriminate between structural features of the same substrate

A comparison of the effects of manual hyperinflation and ventilator hyperinflation on restoring end-expiratory lung volume after endotracheal suctioning: a pilot physiologic study

Purpose: Endotracheal suctioning (ES) of mechanically ventilated patients decreases end-expiratory lung volume (EELV). Manual hyperinflation (MHI) and ventilator hyperinflation (VHI) may restore EELV post-ES but it remains unknown which method is most effective. The primary aim was to compare the efficacy of MHI and VHI in restoring EELV post-ES. Materials and methods: ES was performed on mechanically ventilated intensive care patients, followed by MHI or VHI, in a randomised crossover design. The washout period between interventions was 1 h. End-expiratory lung impedance (EELI), measured by electrical impedance tomography, was recorded at baseline, during ES, during hyperinflation and 1, 5, 15 and 30 min post-hyperinflation. Results: Nine participants were studied. ES decreased EELI by 1672z (95% CI, 1204 to 2140) from baseline. From baseline, MHI increased EELI by 1154z (95% CI, 977 to 1330) while VHI increased EELI by 769z (95% CI, 457 to 1080). Five minutes post-VHI, EELI remained 528z (95% CI, 4 to 1053) above baseline. Fifteen minutes post-MHI, EELI remained 351z (95% CI, 111 to 592) above baseline. At subsequent time-points, EELI returned to baseline. Conclusions: MHI and VHI effectively restore EELV above baseline post-ES and should be considered post suctioning

Listado anotado de Solanum L. (Solanaceae) en el Peru.

The genus Solanum is among the most species-rich genera both of the Peruvian flora and of the tropical Andes in general. The present revised checklist treats 276 species of Solanum L., of which 253 are native, while 23 are introduced and/or cultivated. A total of 74 Solanum species (29% of native species) are endemic to Peru. Additional 58 species occur only in small number of populations outside Peru, and these species are here labelled as near-endemics to highlight the role Peru playes in their future protection. Species diversity is observed to peak between 2500 – 3000 m elevation, but endemic species diversity is highest between 3000 – 3500 m elevation. Cajamarca has the highest number of endemic (29 spp.) and total species (130 spp.), even when considering the effect of area. Centers of endemic species diversity are observed in provinces of Cajamarca (Cajamarca), Huaraz and Carhuaz (Ancash), and Canta and Huarochirí (Lima). Secondary centres of endemism with high concentrations of both endemics and near-endemics are found in San Ignacio and Cutervo (Cajamarca), Santiago de Chuco (La Libertad), Oxapampa (Pasco), and Cusco (Cusco). Current diversity patterns are highly correlated with collection densities, and further collecting is needed across all areas, especially from Arequipa, Ayacucho, Puno, Ancash, Huánuco, Amazonas and Cajamarca, where high levels of species diversity and endemism are indicated but only a few collections of many species are known.Solanum L. es uno de los géneros que posee una alta riqueza de especies dentro de la flora peruana y dentro de los Andes tropicales en general. Presentamos una lista revisada de 276 especies de Solanum para el Perú, de estas 253 son nativas, mientras que 23 son introducidas y/o cultivadas. Un total de 74 especies de Solanum (29% de las especies nativas) son endémicas de Perú. Además 58 especies se encuentran solamente en pequeñas poblaciones fuera del Perú, y estas especies están designadas aquí como casi endémicas para destacar el rol importante del Perú en la futura protección de estas especies. El pico de diversidad de especies es observado entre 2500 – 3000 m de elevación, pero la diversidad de especies endémicas es más alta entre 3000 – 3500 m. Cajamarca tiene el más alto número de especies (130 spp.) y de especies endémicas (29 spp.), incluso si se considera el efecto del área. Centros de diversidad de especies endémicas se localizan en las provincias de Cajamarca (Cajamarca), Huaraz y Carhuaz (Ancash), Canta y Huarochirí (Lima). Centros de endemismos secundarios con una alta concentración tanto de especies endémicas y de casi endémicas se encuentran en San Ignacio y Cutervo (Cajamarca), Santiago de Chuco (La Libertad), Oxapampa (Pasco), y Cusco (Cusco): Los actuales patrones de diversidad están altamente correlacionados con la densidad de colecciones, por lo que es necesario una mayor colecta en todas las regiones, especialmente en Arequipa, Ayacucho, Puno, Ancash, Huánuco, Amazonas y Cajamarca, donde se indican altos niveles de diversidad y endemismo de especies, pero de las cuales existen pocas colecciones

Neutralino Dark Matter beyond CMSSM Universality

We study the effect of departures from SUSY GUT universality on the neutralino relic density and both its direct detection and indirect detection, especially by neutrino telescopes. We find that the most interesting models are those with a value of $M_3|_{GUT}$ lower than the universal case.Comment: 20 pages, 12 figures, JHEP format. Figures improved for B&W, references added, typos and english correcte