19 research outputs found
Altered proteostasis in aging and heat shock response in C. elegans revealed by analysis of the global and de novo synthesized proteome
COVID-19 symptoms at hospital admission vary with age and sex: results from the ISARIC prospective multinational observational study
Background:
The ISARIC prospective multinational observational study is the largest cohort of hospitalized patients with COVID-19. We present relationships of age, sex, and nationality to presenting symptoms.
Methods:
International, prospective observational study of 60â109 hospitalized symptomatic patients with laboratory-confirmed COVID-19 recruited from 43 countries between 30 January and 3 August 2020. Logistic regression was performed to evaluate relationships of age and sex to published COVID-19 case definitions and the most commonly reported symptoms.
Results:
âTypicalâ symptoms of fever (69%), cough (68%) and shortness of breath (66%) were the most commonly reported. 92% of patients experienced at least one of these. Prevalence of typical symptoms was greatest in 30- to 60-year-olds (respectively 80, 79, 69%; at least one 95%). They were reported less frequently in children (â€â18 years: 69, 48, 23; 85%), older adults (â„â70 years: 61, 62, 65; 90%), and women (66, 66, 64; 90%; vs. men 71, 70, 67; 93%, each Pâ<â0.001). The most common atypical presentations under 60 years of age were nausea and vomiting and abdominal pain, and over 60 years was confusion. Regression models showed significant differences in symptoms with sex, age and country.
Interpretation:
This international collaboration has allowed us to report reliable symptom data from the largest cohort of patients admitted to hospital with COVID-19. Adults over 60 and children admitted to hospital with COVID-19 are less likely to present with typical symptoms. Nausea and vomiting are common atypical presentations under 30 years. Confusion is a frequent atypical presentation of COVID-19 in adults over 60 years. Women are less likely to experience typical symptoms than men
Etablierung von bio-orthogonaler Markierung und Click-Chemie in Caenorhabditis elegans zur Identifizierung neu synthetisierter Proteine
In this protocol we describe the incorporation of bio-orthogonal amino acids
as a versatile method for visualizing and identifying de novoâsynthesized
proteins in the roundworm Caenorhabditis elegans. This protocol contains
directions on implementing three complementary types of analysis: âclick
chemistryâ followed by western blotting, click chemistry followed by
immunofluorescence, and isobaric tags for relative and absolute quantification
(iTRAQ) quantitative mass spectrometry. The detailed instructions provided
herein enable researchers to investigate the de novo proteome, an analysis
that is complicated by the fact that protein molecules are chemically
identical to each other, regardless of the timing of their synthesis. Our
protocol circumvents this limitation by identifying de novoâsynthesized
proteins via the incorporation of the chemically modifiable azidohomoalanine
instead of the natural amino acid methionine in the nascent protein, followed
by facilitating the visualization of the resulting labeled proteins in situ.
It will therefore be an ideal tool for studying de novo protein synthesis in
physiological and pathological processes including learning and memory. The
protocol requires 10 d for worm growth, liquid culture and synchronization;
1â2 d for bio-orthogonal labeling; and, with regard to analysis, 3â4 d for
western blotting, 5â6 d for immunofluorescence or ~3 weeks for mass
spectrometry. This abstract is reproduced with the permission of Nature
Publishing Group. It is part of the following publication: Ullrich M, Liang V,
Chew YL, Banister S, Song X, Zaw T, Lam H, Berber S, Kassiou M, Nicholas HR,
Gotz J. Bio-orthogonal labeling as a tool to visualize and identify newly
synthesized proteins in Caenorhabditis elegans. Nat Protoc, 2014. 9(9): p.
2237-55.In diesem Versuchsprotokoll beschreiben wir die Inkorporation von bio-
orthogonalen AminosÀuren als eine vielfÀltig anwendbare Methode um neu
synthetisierte Proteine im Rundwurm Caenorhabditis elegans zu visualisieren
und identifizieren. Unsere Methode ermöglicht die Analyse des de novo Proteoms
mittels drei verschiedener, sich ergÀnzender Verfahren: Click Chemistry,
gefolgt von Western Blotting, Immunfluoreszenz oder quantitativer
Massenspektrometrie durch isobare Markierung fĂŒr relative und absolute
Quantifizierung (iTRAQ). Die Analyse des de novo Proteoms war bislang nur in
begrenztem Umfang möglich, weil die chemischen Eigenschaften von Proteinen
unabhÀngig vom Zeitpunkt ihrer Synthese sind. Unser Protokoll umgeht dieses
Hindernis, indem es die Aufnahme von chemisch modifiziertem Azidohomoalanin
anstelle der natĂŒrlichen AminosĂ€ure Methionin in neu entstehende Proteine
ermöglicht. So kann das markierte de novo Proteom identifiziert und in situ
visualisiert werden. Die Methode ist daher ein Ă€uĂerst geeignetes Instrument
um die de novo Proteinbiosynthese in physiologischen und pathologischen
Prozessen, wie etwa Lernen und GedÀchtnisbildung, zu untersuchen. Es werden 10
Tage fĂŒr die Anzucht und Synchronisation der WĂŒrmer in flĂŒssigem Medium
benötigt; 1-2 Tage fĂŒr die bio-orthogonale Markierung; sowie 3-4 Tage fĂŒr die
Analyse mittels Western Blotting, bzw. 5-6 Tage fĂŒr Immunfluoreszenz oder ~3
Wochen fĂŒr Massenspektrometrie
Postural Stability in Parkinsonâs Disease Patients Is Improved after Stochastic Resonance Therapy
Background. Postural instability in Parkinsonâs disease (PD) increases the risk of falls and is not improved by pharmacological therapy. Objective. We performed a double-blind, randomized sham-controlled study to test the effects of stochastic resonance (whole body vibration) therapy on postural stability in PD. Methods. Fifty-six PD participants were allocated to either experimental or sham groups. The experimental group received four series of vibration over eight days, with each series consisting of six stimulus trains of 60-second duration using a randomized whole body vibration. Participants allocated to the control group received a sham treatment. Results. Within-group analysis revealed that postural stability in the experimental group improved by 17.5% (p=0.005) comparing experimental and sham groups. The between-group analysis of change after treatment comparing both groups also showed a significant improvement of postural stability (p=0.03). Only in the within-group analysis several items were improved after Bonferroni correction, too, rigor 41.6% (p=0.001), bradykinesia 23.7% (p=0.001), tremor 30.8% (p=0.006), and UPDRSIII sum score 23.9% (p=0.000), but did not reach the level of significance in the between-group analysis. Conclusions. Stochastic resonance therapy significantly enhanced postural stability even in individuals with increased risk of falling. Thus it offers a potential supplementation to canonical treatments of PD
Altered proteostasis in aging and heat shock response in C. elegans revealed by analysis of the global and de novo synthesized proteome
Protein misfolding and aggregation as a consequence of impaired protein homeostasis (proteostasis) not only characterizes numerous age-related diseases but also the aging process itself. Functionally related to the aging process are, among others, ribosomal proteins, suggesting an intimate link between proteostasis and aging. We determined by iTRAQ quantitative proteomic analysis in C. elegans how the proteome changes with age and in response to heat shock. Levels of ribosomal proteins and mitochondrial chaperones were decreased in aged animals, supporting the notion that proteostasis is altered during aging. Mitochondrial enzymes of the tricarboxylic acid cycle and the electron transport chain were also reduced, consistent with an age-associated energy impairment. Moreover, we observed an age-associated decline in the heat shock response. In order to determine how protein synthesis is altered in aging and in response to heat shock, we complemented our global analysis by determining the de novo proteome. For that, we established a novel method that enables both the visualization and identification of de novo synthesized proteins, by incorporating the non-canonical methionine analogue, azidohomoalanine (AHA), into the nascent polypeptides, followed by reacting the azide group of AHA by 'click chemistry' with an alkyne-labeled tag. Our analysis of AHA-tagged peptides demonstrated that the decreased abundance of, for example, ribosomal proteins in aged animals is not solely due to degradation but also reflects a relative decrease in their synthesis. Interestingly, although the net rate of protein synthesis is reduced in aged animals, our analyses indicate that the synthesis of certain proteins such as the vitellogenins increases with age
Bio-orthogonal labeling as a tool to visualize and identify newly synthesized proteins in Caenorhabditis elegans
In this protocol we describe the incorporation of bio-orthogonal amino acids as a versatile method for visualizing and identifying de novo-synthesized proteins in the roundworm Caenorhabditis elegans. This protocol contains directions on implementing three complementary types of analysis: 'click chemistry' followed by western blotting, click chemistry followed by immunofluorescence, and isobaric tags for relative and absolute quantification (iTRAQ) quantitative mass spectrometry. The detailed instructions provided herein enable researchers to investigate the de novo proteome, an analysis that is complicated by the fact that protein molecules are chemically identical to each other, regardless of the timing of their synthesis. Our protocol circumvents this limitation by identifying de novo-synthesized proteins via the incorporation of the chemically modifiable azidohomoalanine instead of the natural amino acid methionine in the nascent protein, followed by facilitating the visualization of the resulting labeled proteins in situ. It will therefore be an ideal tool for studying de novo protein synthesis in physiological and pathological processes including learning and memory. The protocol requires 10 d for worm growth, liquid culture and synchronization; 1-2 d for bio-orthogonal labeling; and, with regard to analysis, 3-4 d for western blotting, 5-6 d for immunofluorescence or âŒ3 weeks for mass spectrometry
Erratum: Corrigendum: Bio-orthogonal labeling as a tool to visualize and identify newly synthesized proteins in Caenorhabditis elegans
Distinct single-component adjuvants steer human DC-mediated T-cell polarization via Toll-like receptor signaling toward a potent antiviral immune response
The COVID-19 pandemic highlights the importance of efficient and safe vaccine development. Vaccine adjuvants are essential to boost and tailor the immune response to the corresponding pathogen. To allow for an educated selection, we assessed the effect of different adjuvants on human monocyte-derived dendritic cells (DCs) and their ability to polarize innate and adaptive immune responses. In contrast to commonly used adjuvants, such as aluminum hydroxide, Toll-like receptor (TLR) agonists induced robust phenotypic and functional DC maturation. In a DC-lymphocyte coculture system, we investigated the ensuing immune reactions. While monophosphoryl lipid A synthetic, a TLR4 ligand, induced checkpoint inhibitors indicative for immune exhaustion, the TLR7/8 agonist Resiquimod (R848) induced prominent type-1 interferon and interleukin 6 responses and robust CTL, B-cell, and NK-cell proliferation, which is particularly suited for antiviral immune responses. The recently licensed COVID-19 vaccines, BNT162b and mRNA-1273, are both based on single-stranded RNA. Indeed, we could confirm that the cytokine profile induced by lipid-complexed RNA was almost identical to the pattern induced by R848. Although this awaits further investigation, our results suggest that their efficacy involves the highly efficient antiviral response pattern stimulated by the RNAsâ TLR7/8 activation
The DOE E3SM Coupled Model Version 1: Description and Results at High Resolution
This study provides an overview of the coupled high-resolution Version 1 of the Energy Exascale Earth System Model (E3SMv1) and documents the characteristics of a 50-year-long high-resolution control simulation with time-invariant 1950 forcings following the HighResMIP protocol. In terms of global root-mean-squared error metrics, this high-resolution simulation is generally superior to results from the low-resolution configuration of E3SMv1 (due to resolution, tuning changes, and possibly initialization procedure) and compares favorably to models in the CMIP5 ensemble. Ocean and sea ice simulation is particularly improved, due to better resolution of bathymetry, the ability to capture more variability and extremes in winds and currents, and the ability to resolve mesoscale ocean eddies. The largest improvement in this regard is an ice-free Labrador Sea, which is a major problem at low resolution. Interestingly, several features found to improve with resolution in previous studies are insensitive to resolution or even degrade in E3SMv1. Most notable in this regard are warm bias and associated stratocumulus deficiency in eastern subtropical oceans and lack of improvement in El Niño. Another major finding of this study is that resolution increase had negligible impact on climate sensitivity (measured by net feedback determined through uniform +4K prescribed sea surface temperature increase) and aerosol sensitivity. Cloud response to resolution increase consisted of very minor decrease at all levels. Large-scale patterns of precipitation bias were also relatively unaffected by grid spacing