A structural basis for prion strain diversity

Recent cryogenic electron microscopy (cryo-EM) studies of infectious, ex vivo, prion fibrils from hamster 263K and mouse RML prion strains revealed a similar, parallel in-register intermolecular β-sheet (PIRIBS) amyloid architecture. Rungs of the fibrils are composed of individual prion protein (PrP) monomers that fold to create distinct N-terminal and C-terminal lobes. However, disparity in the hamster/mouse PrP sequence precludes understanding of how divergent prion strains emerge from an identical PrP substrate. In this study, we determined the near-atomic resolution cryo-EM structure of infectious, ex vivo mouse prion fibrils from the ME7 prion strain and compared this with the RML fibril structure. This structural comparison of two biologically distinct mouse-adapted prion strains suggests defined folding subdomains of PrP rungs and the way in which they are interrelated, providing a structural definition of intra-species prion strain-specific conformations

2014 Behavioral Health and Performance Standing Review Panel

The 2014 Behavioral Health and Performance (BHP) Standing Review Panel (from here on referred to as the SRP) met for a site visit in Houston, TX on December 17 - 18, 2014. The SRP reviewed the updated research plan for the Risk of Performance Errors Due to Fatigue Resulting from Sleep Loss, Circadian Desynchronization, Extended Wakefulness, and Work Overload (Sleep Risk) and also received a status update on the Risk of Adverse Behavioral Conditions and Psychiatric Disorders (BMed Risk) and the Risk of Performance Decrements Due to Inadequate Cooperation, Coordination, Communication, and Psychosocial Adaptation within a Team (Team Risk)

Structural features distinguishing infectious ex vivo mammalian prions from non-infectious fibrillar assemblies generated in vitro

Seeded polymerisation of proteins forming amyloid fibres and their spread in tissues has been implicated in the pathogenesis of multiple neurodegenerative diseases: so called "prion-like" mechanisms. While ex vivo mammalian prions, composed of multichain assemblies of misfolded host-encoded prion protein (PrP), act as lethal infectious agents, PrP amyloid fibrils produced in vitro generally do not. The high-resolution structure of authentic infectious prions and the structural basis of prion strain diversity remain unknown. Here we use cryo-electron microscopy and atomic force microscopy to examine the structure of highly infectious PrP rods isolated from mouse brain in comparison to non-infectious recombinant PrP fibrils generated in vitro. Non-infectious recombinant PrP fibrils are 10 nm wide single fibres, with a double helical repeating substructure displaying small variations in adhesive force interactions across their width. In contrast, infectious PrP rods are 20 nm wide and contain two fibres, each with a double helical repeating substructure, separated by a central gap of 8-10 nm in width. This gap contains an irregularly structured material whose adhesive force properties are strikingly different to that of the fibres, suggestive of a distinct composition. The structure of the infectious PrP rods, which cause lethal neurodegeneration, readily differentiates them from all other protein assemblies so far characterised in other neurodegenerative diseases

A Cloud-Ozone Data Product from Aura OMI and MLS Satellite Measurements

Ozone within deep convective clouds is controlled by several factors involving photochemical reactions and transport. Gas-phase photochemical reactions and heterogeneous surface chemical reactions involving ice, water particles, and aerosols inside the clouds all contribute to the distribution and net production and loss of ozone. Ozone in clouds is also dependent on convective transport that carries low troposphereboundary layer ozone and ozone precursors upward into the clouds. Characterizing ozone in thick clouds is an important step for quantifying relationships of ozone with tropospheric H2O, OH production, and cloud microphysicstransport properties. Although measuring ozone in deep convective clouds from either aircraft or balloon ozonesondes is largely impossible due to extreme meteorological conditions associated with these clouds, it is possible to estimate ozone in thick clouds using backscattered solar UV radiation measured by satellite instruments. Our study combines Aura Ozone Monitoring Instrument (OMI) and Microwave Limb Sounder (MLS) satellite measurements to generate a new research product of monthly-mean ozone concentrations in deep convective clouds between 30oS to 30oN for October 2004 April 2016. These measurements represent mean ozone concentration primarily in the upper levels of thick clouds and reveal key features of cloud ozone including: persistent low ozone concentrations in the tropical Pacific of 10 ppbv or less; concentrations of up to 60 pphv or greater over landmass regions of South America, southern Africa, Australia, and Indiaeast Asia; connections with tropical ENSO events; and intra-seasonalMadden-Julian Oscillation variability. Analysis of OMI aerosol measurements suggests a cause and effect relation between boundary layer pollution and elevated ozone inside thick clouds over land-mass regions including southern Africa and Indiaeast Asia

Prion 2016 poster abstracts

Until now, the 3-dimensional structure of infectious mammalian prions and how this differs from non-infectious amyloid fibrils remained unknown. Mammalian prions are hypothesized to be fibrillar or amyloid forms of prion protein (PrP), but structures observed to date have not been definitively correlated with infectivity. One of the major challenges has been the production of highly homogeneous material of demonstrable high specific infectivity to allow direct correlation of particle structure with infectivity. We have recently developed novel methods to obtain exceptionally pure preparations of prions from prion-infected murine brain and have shown that pathogenic PrP in these high-titer preparations is assembled into rod-like assemblies (Wenborn et al. 2015. Sci. Rep. 10062). Our preparations contain very high titres of infectious prions which faithfully transmit prion strain-specific phenotypes when inoculated into mice making them eminently suitable for detailed structural analysis. We are now undertaking structural characterization of prion assemblies and comparing these to the structure of non-infectious PrP fibrils generated from recombinant Pr

Evaluating the causality of novel sequence variants in the prion protein gene by example

The estimation of pathogenicity and penetrance of novel prion protein gene (PRNP) variants presents significant challenges, particularly in the absence of family history, which precludes the application of Mendelian segregation. Moreover, the ambiguities of prion disease pathophysiology renders conventional in silico predictions inconclusive. Here, we describe 2 patients with rapid cognitive decline progressing to akinetic mutism and death within 10 weeks of symptom onset, both of whom possessed the novel T201S variant in PRNP. Clinically, both satisfied diagnostic criteria for probable sporadic Creutzfeldt-Jakob disease and in one, the diagnosis was confirmed by neuropathology. While computational analyses predicted that T201S was possibly deleterious, molecular strain typing, prion protein structural considerations, and calculations leveraging large-scale population data (gnomAD) indicate that T201S is at best either of low penetrance or nonpathogenic. Thus, we illustrate the utility of harnessing multiple lines of prion disease-specific evidence in the evaluation of the T201S variant, which may be similarly applied to assess other novel variants in PRNP

Prevalência de depressão e fatores associados em homens

El objetivo del presente estudio fue evaluar la prevalencia de depresión, detectar el riesgo suicida e identificar los factores sociodemográficos y personales asociados a este trastorno. La muestra no aleatorizada estuvo conformada por 1525 hombres colombianos con edades entre 18 y 83 años procedentes de 22 departamentos y de distintos niveles educativos. Para evaluar la depresión se usó el Cuestionario de Depresión para Hombres (Álvarez y Londoño, 2012); para evaluar la comorbilidad con ansiedad se usó la Escala de Ansiedad HADS (Zigmond y Snaith, 1983) y el IMAFE (Lara, 1991); y para recolectar información acerca de los factores personales y sociodemográficos se usó una ficha de registro. Se analizaron los datos para calcular la prevalencia de corte, el riesgo suicida, la comorbilidad a través del uso del paquete estadístico SPSS. Se concluye que la prevalencia real reportada y el riesgo suicida en la población estudiada son más altos que los detectados usando un instrumento no sensible al género.O objetivo deste estudo foi avaliar a prevalência de depressão, detectar o risco suicida e identificar os fatores sociodemográficos e pessoais associados com esse transtorno. A amostra não aleatorizada foi conformada por 1525 homens colombianos com idade entre 18 e 83 anos, procedentes de 22 estados e de diferentes níveis de escolaridade. Para avaliar a depressão, foi utilizado o Teste de Depressão para Homens (Álvarez e Londoño, 2012); para avaliar a comorbilidade com ansiedade, usou-se a Escala Hospitalar de Ansiedade e Depressão (Hads – Zigmond e Snaith, 1983) e o Inventário de Masculinidade e Feminilidade (Imafe – Lara, 1991); para coletar informação sobre os fatores pessoais e sociodemográficos, empregou-se um formulário de registro. Analisaram-se os dados para calcular a prevalência de corte, o risco suicida, a comorbilidade por meio do uso do SPSS. Concluise que a prevalência real relatada e o risco de suicídio na população estudada são mais altos do que os detectados usando um instrumento não sensível ao gênero.This epidemiologic study aimed to evaluate the prevalence of depression, the suicide risk and the demographic and personal factors associated with depression severity in men. The non-randomized sample of participants was formed by 1525 Colombian men aged 18 to 83 years old, from 22 departments and different educational levels. The instruments used to evaluate the above factors were the Men’s Depression Questionnaire (Alvarez and Londoño, 2012), the Anxiety Scale HADS (Zigmond y Snaith, 1983) and the IMAFE (Lara 1991), and in order to collect data about personal and socio-demographic factors, a registration card was used. Data were analyzed to calculate the prevalence, suicide risk and comorbidity with anxiety through the use of SPSS. It was concluded that the prevalence of depression and suicide risk in the population object of study is higher than the one identified in previous studies when a non- gender sensitive questionnaire was used

A Dynamic Pathway for Calcium-Independent Activation of CaMKII by Methionine Oxidation

SummaryCalcium/calmodulin (Ca2+/CaM)-dependent protein kinase II (CaMKII) couples increases in cellular Ca2+ to fundamental responses in excitable cells. CaMKII was identified over 20 years ago by activation dependence on Ca2+/CaM, but recent evidence shows that CaMKII activity is also enhanced by pro-oxidant conditions. Here we show that oxidation of paired regulatory domain methionine residues sustains CaMKII activity in the absence of Ca2+/CaM. CaMKII is activated by angiotensin II (AngII)-induced oxidation, leading to apoptosis in cardiomyocytes both in vitro and in vivo. CaMKII oxidation is reversed by methionine sulfoxide reductase A (MsrA), and MsrA−/− mice show exaggerated CaMKII oxidation and myocardial apoptosis, impaired cardiac function, and increased mortality after myocardial infarction. Our data demonstrate a dynamic mechanism for CaMKII activation by oxidation and highlight the critical importance of oxidation-dependent CaMKII activation to AngII and ischemic myocardial apoptosis

Effect of fixation on brain and lymphoreticular vCJD prions and bioassay of key positive specimens from a retrospective vCJD prevalence study

We analyse the process of conversion of near-field thermal radiation into usable work by considering the radiation emitted between two planar sources supporting surface phonon-polaritons. The maximum work flux that can be extracted from the radiation is obtained taking into account that the spectral flux of modes is mainly dominated by these surface modes. The thermodynamic efficiencies are discussed and an upper bound for the first law efficiency is obtained for this process

Isolation of Proteinase K-Sensitive Prions Using Pronase E and Phosphotungstic Acid

Disease-related prion protein, PrPSc, is classically distinguished from its normal cellular precursor, PrPC, by its detergent insolubility and partial resistance to proteolysis. Molecular diagnosis of prion disease typically relies upon detection of protease-resistant fragments of PrPSc using proteinase K, however it is now apparent that the majority of disease-related PrP and indeed prion infectivity may be destroyed by this treatment. Here we report that digestion of RML prion-infected mouse brain with pronase E, followed by precipitation with sodium phosphotungstic acid, eliminates the large majority of brain proteins, including PrPC, while preserving >70% of infectious prion titre. This procedure now allows characterization of proteinase K-sensitive prions and investigation of their clinical relevance in human and animal prion disease without being confounded by contaminating PrPC