Fusion of DARPin to Aldolase Enables Visualization of Small Protein by Cryo-EM

Solving protein structures by single-particle cryoelectron microscopy (cryo-EM) has become a crucial tool in structural biology. While exciting progress is being made toward the visualization of small macromolecules, the median protein size in both eukaryotes and bacteria is still beyond the reach of cryo-EM. To overcome this problem, we implemented a platform strategy in which a small protein target was rigidly attached to a large, symmetric base via a selectable adapter. Of our seven designs, the best construct used a designed ankyrin repeat protein (DARPin) rigidly fused to tetrameric rabbit muscle aldolase through a helical linker. The DARPin retained its ability to bind its target: GFP. We solved the structure of this complex to 3.0 Å resolution overall, with 5–8 Å resolution in the GFP region. As flexibility in the DARPin position limited the overall resolution of the target, we describe strategies to rigidify this element

Fusion of DARPin to Aldolase Enables Visualization of Small Protein by Cryo-EM

Solving protein structures by single-particle cryoelectron microscopy (cryo-EM) has become a crucial tool in structural biology. While exciting progress is being made toward the visualization of small macromolecules, the median protein size in both eukaryotes and bacteria is still beyond the reach of cryo-EM. To overcome this problem, we implemented a platform strategy in which a small protein target was rigidly attached to a large, symmetric base via a selectable adapter. Of our seven designs, the best construct used a designed ankyrin repeat protein (DARPin) rigidly fused to tetrameric rabbit muscle aldolase through a helical linker. The DARPin retained its ability to bind its target: GFP. We solved the structure of this complex to 3.0 Å resolution overall, with 5–8 Å resolution in the GFP region. As flexibility in the DARPin position limited the overall resolution of the target, we describe strategies to rigidify this element

Late-life falling and depressive symptoms associated with the risk of Parkinsons disease: a nationwide cohort data analysis

Abstract Background This study aimed to evaluate the relationship between the history of late-life falling and the development of Parkinsons disease (PD) and investigate whether depressive symptoms interact with falling to increase PD risk. Methods We identified 1,223,726 subjects without PD who underwent the National Screening Program for Transitional Age at 66 years between 2009 and 2013 using the National Health Cohort database. In this program, every participant was assessed whether they experienced falling for the past six months. Depressive symptoms were evaluated with a three-item questionnaire extracted from the Geriatric Depression Scale. Incident PD was traced for a mean 4.23 ± 1.50 years. Cox proportional hazard regression models were used to assess the risk of PD by falling history with and without depressive symptoms after adjusting for other confounding variables. Results In this cohort, the PD incidence rate was 1.30 and 1.03 cases per 1000 person-years in groups with and without falling and 1.34 and 1.00 cases per 1000 person-years in groups with and without depressive symptoms. The predictive risk of PD was increased by either a history of falling (HR = 1.24; 95% CI 1.11–1.40) or the presence of depressive symptoms (HR = 1.31; 95% CI 1.21–1.42) after adjusting for possible confounding variables. For individuals with both falling and depressive symptoms, PD risk increased further (HR = 1.66; 95% CI 1.40–1.97), but with sex-differences. The two factors increased PD risk in a sub-additive manner in men, whereas synergistically in women. Conclusions This national cohort database shows that late-life depressive symptoms accompanied by a falling event pose an increase in the risk of PD in older adults

Global Gene Expression Profiling in Whole-Blood Samples from Individuals Exposed to Metal Fumes

Accumulating evidence demonstrates that particulate air pollutants can cause both pulmonary and airway inflammation. However, few data show that particulates can induce systemic inflammatory responses. We conducted an exploratory study using microarray techniques to analyze whole-blood total RNA in boilermakers before and after occupational exposure to metal fumes. A self-controlled study design was used to overcome the problems of larger between-individual variation interferences with observations of relatively smaller changes caused by environmental exposure. Moreover, we incorporated the dichotomous data of absolute gene expression status in the microarray analyses. Compared with nonexposed controls, we observed that genes with altered expression in response to particulate exposure were clustered in biologic processes related to inflammatory response, oxidative stress, intracellular signal transduction, cell cycle, and programmed cell death. In particular, the preinflammatory cytokine interleukin 8 and one of its receptors, chemokine receptor 4, seemed to play important roles in early-stage response to heavy metal exposure and were down-regulated. Furthermore, most observed expression variations were from nonsmoking exposed individuals, suggesting that smoking profoundly affects whole-blood expression profiles. Our study is the first to demonstrate that with a paired sampling study design of pre- and postexposed individuals, small changes in gene expression profiling can be measured in whole-blood total RNA from a population-based study. This technique can be applied to evaluate the host response to other forms of environmental exposures

Spurious Shear in Weak Lensing with LSST

The complete 10-year survey from the Large Synoptic Survey Telescope (LSST) will image $\sim$ 20,000 square degrees of sky in six filter bands every few nights, bringing the final survey depth to $r\sim27.5$, with over 4 billion well measured galaxies. To take full advantage of this unprecedented statistical power, the systematic errors associated with weak lensing measurements need to be controlled to a level similar to the statistical errors. This work is the first attempt to quantitatively estimate the absolute level and statistical properties of the systematic errors on weak lensing shear measurements due to the most important physical effects in the LSST system via high fidelity ray-tracing simulations. We identify and isolate the different sources of algorithm-independent, \textit{additive} systematic errors on shear measurements for LSST and predict their impact on the final cosmic shear measurements using conventional weak lensing analysis techniques. We find that the main source of the errors comes from an inability to adequately characterise the atmospheric point spread function (PSF) due to its high frequency spatial variation on angular scales smaller than $\sim10'$ in the single short exposures, which propagates into a spurious shear correlation function at the $10^{-4}$--$10^{-3}$ level on these scales. With the large multi-epoch dataset that will be acquired by LSST, the stochastic errors average out, bringing the final spurious shear correlation function to a level very close to the statistical errors. Our results imply that the cosmological constraints from LSST will not be severely limited by these algorithm-independent, additive systematic effects.Comment: 22 pages, 12 figures, accepted by MNRA

GALEX Ultraviolet Photometry of Globular Clusters in M31

We present ultraviolet photometry for globular clusters (GCs) in M31 from 15 square deg of imaging using the Galaxy Evolution Explorer (GALEX). We detect 200 and 94 GCs with certainty in the near-ultraviolet (NUV; 1750 - 2750 Angstroms) and far-ultraviolet (FUV; 1350 - 1750 Angstroms) bandpasses, respectively. Our rate of detection is about 50% in the NUV and 23% in the FUV, to an approximate limiting V magnitude of 19. Out of six clusters with [Fe/H]>-1 seen in the NUV, none is detected in the FUV bandpass. Furthermore, we find no candidate metal-rich clusters with significant FUV flux, because of the contribution of blue horizontal-branch (HB) stars, such as NGC 6388 and NGC 6441, which are metal-rich Galactic GCs with hot HB stars. We show that our GALEX photometry follows the general color trends established in previous UV studies of GCs in M31 and the Galaxy. Comparing our data with Galactic GCs in the UV and with population synthesis models, we suggest that the age range of M31 and Galactic halo GCs are similar.Comment: This paper will be published as part of the Galaxy Evolution Explorer (GALEX) Astrophysical Journal Letters Special Issue. Links to the full set of papers will be available at http://www.galex.caltech.edu/PUBLICATIONS/ after November 22, 200

Quantitative Profiling of the Lymph Node Clearance Capacity

Transport of tissue-derived lymphatic fluid and clearance by draining lymph nodes are pivotal for maintenance of fluid homeostasis in the body and for immune-surveillance of the self- and non-self-proteomes. Yet a quantitative analysis of nodal filtration of the tissue-derived proteome present in lymphatic fluid has not been reported. Here we quantified the efficiency of nodal clearance of the composite proteomic load using label-free and isotope-labeling proteomic analysis of pre-nodal and post-nodal samples collected by direct cannulation. These results were extended by quantitation of the filtration efficiency of fluorophore-labeled proteins, bacteria, and beads infused at physiological flow rates into pre-nodal lymphatic collectors and collected by post-nodal cannulation. We developed a linear model of nodal filtration efficiency dependent on pre-nodal protein concentrations and molecular weight, and uncovered criteria for disposing the proteome incoming from defined anatomical districts under physiological conditions. These findings are pivotal to understanding the maximal antigenic load sustainable by a draining node, and promote understanding of pathogen spreading and nodal filtration of tumor metastasis, potentially helping to improve design of vaccination protocols, immunization strategies and drug delivery

Glucocorticoids activate a synapse weakening pathway culminating in tau phosphorylation in the hippocampus

Evidence suggests that the stress hormones glucocorticoids (GCs) can cause cognitive deficits and neurodegeneration. Previous studies have found GCs facilitate physiological synapse weakening, termed long-term depression (LTD), though the precise mechanisms underlying this are poorly understood. Here we show that GCs activate glycogen synthase kinase-3 (GSK-3), a kinase crucial to synapse weakening signals. Critically, this ultimately leads to phosphorylation of the microtubule associated protein tau, specifically at the serine 396 residue, and this is a causal factor in the GC-mediated impairment of synaptic function. These findings reveal the link between GCs and synapse weakening signals, and the potential for stress-induced priming of neurodegeneration. This could have important implications for our understanding of how stress can lead to neurodegenerative disease

H3N2 Influenza Virus Transmission from Swine to Turkeys, United States

Swinelike H3N2 influenza viruses were isolated from two geographically distinct turkey farms in the United States

CSF total tau/α-synuclein ratio improved the diagnostic performance for Alzheimers disease as an indicator of tau phosphorylation

Abstract Background Recently, several studies suggested potential involvements of α-synuclein in Alzheimers disease (AD) pathophysiology. Higher concentrations of α-synuclein were reported in cerebrospinal fluid (CSF) of AD patients with a positive correlation towards CSF tau, indicating its possible role in AD. We analyzed the CSF biomarkers to verify whether α-synuclein could be an additional supported biomarker in AD diagnosis. Methods In this cross-sectional study, CSF samples of 71 early-onset AD, 34 late-onset AD, 11 mild cognitive impairment, 17 subjective cognitive decline, 45 Parkinsons disease, and 32 healthy control (HC) were collected. CSF amyloid-β1-42 (A), total tau (N), and phosphorylated tau181 (T) were measured by commercial ELISA kits, and in-house ELISA kit was developed to quantify α-synuclein. The cognitive assessments and amyloid-PET imaging were also performed. Results CSF α-synuclein manifested a tendency to increase in AD and to decreased in Parkinsons disease compared to HC. The equilibrium states of total tau and α-synuclein concentrations were changed significantly in AD, and the ratio of total tau/α-synuclein (N/αS) was dramatically increased in AD than HC. Remarkably, N/αS revealed a strong positive correlation with tau phosphorylation rate. Also, the combination of N/αS with amyloid-β1-42/phosphorylated tau181ratio had the best diagnosis performance (AUC = 0.956, sensitivity = 96%, specificity = 87%). In concordance analysis, N/αS showed the higher diagnostic agreement with amyloid-β1-42 and amyloid-PET. Analysis of biomarker profiling with N/αS had distinctive characteristics and clustering of each group. Especially, among the group of suspected non-Alzheimers disease pathophysiology, all A−T+N+ patients with N/αS+ were reintegrated into AD. Conclusions The high correlation of α-synuclein with tau and the elevated N/αS in AD supported the involvement of α-synuclein in AD pathophysiology. Importantly, N/αS improved the diagnostic performance, confirming the needs of incorporating α-synuclein as a biomarker for neurodegenerative disorders. The incorporation of a biomarker group [N/αS] could contribute to provide better understanding and diagnosis of neurodegenerative disorders