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 60 109 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: ‘Typical’ 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 (≤ 18 years: 69, 48, 23; 85%), older adults (≥ 70 years: 61, 62, 65; 90%), and women (66, 66, 64; 90%; vs. men 71, 70, 67; 93%, each P < 0.001). 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

Comparative therapeutic strategies for preventing aortic rupture in a mouse model of vascular Ehlers-Danlos syndrome

International audienceVascular Ehlers-Danlos syndrome is a rare inherited disorder caused by genetic variants in type III collagen. Its prognosis is especially hampered by unpredictable arterial ruptures and there is no therapeutic consensus. We created a knock-in Col3a1 +/G182R mouse model and performed a complete genetic, molecular and biochemical characterization. Several therapeutic strategies were also tested. Col3a1 +/G182R mice showed a spontaneous mortality caused by thoracic aortic rupture that recapitulates the vascular Ehlers-Danlos syndrome with a lower survival rate in males, thin non-inflammatory arteries and an altered arterial collagen. Transcriptomic analysis of aortas showed upregulation of genes related to inflammation and cell stress response. Compared to water, survival rate of Col3a1 +/G182R mice was not affected by beta-blockers (propranolol or celiprolol). Two other vasodilating anti-hypertensive agents (hydralazine, amlodipine) gave opposite results on aortic rupture and mortality rate. There was a spectacular beneficial effect of losartan, reversed by the cessation of its administration, and a marked deleterious effect of exogenous angiotensin II. These results suggest that blockade of the renin angiotensin system should be tested as a first-line medical therapy in patients with vascular Ehlers-Danlos syndrome

Expression of N protein in GS115, KM71 and SMD1168 <i>P. pastoris</i> strains.

<p>Concentrations of Geneticin in selection plates for the specific clones, and clone numbers, are indicated. Yeast lysates were diluted 1/600 before loading on western blot.</p

Expression of N-PbCS in <i>P. pastoris</i>.

<p>(A) Schematic representation of N-PbCS fusion protein. MV-N (dark grey) is composed of a core domain in N-terminal and a unstructured tail domain in C-terminal <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086658#pone.0086658-Longhi1" target="_blank">[69]</a>. The GAAGAGA linker is in black. PbCS (light grey) corresponds the central repeat region flanked by major portions of the N-terminal and C-terminal domains of the protein <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086658#pone.0086658-Plassmeyer1" target="_blank">[39]</a>. Amino acids numbering are given according to N from the MV Schwarz vaccine strain and PbCS from the <i>Pb</i> ANKA strain. For sequence details, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086658#pone.0086658.s001" target="_blank">Figure S1</a>. (B) Quantitative western blot analysis of SMD1168 expressing N-PbCS or (C) N. In (B) and (C), yeast lysates were diluted as indicated, the MV-N protein was used as a standard with increasing concentrations and western blots were probed with an anti-N antibody.</p

ELISA quantification of N or PbCS proteins in ultracentrifugation (U) fractions and pellets of SMD1168 lysates expressing N alone at 871 ng/YU (A) or N-PbCS at 12 ng/YU (B).

<p>Yeast cultures, lysates and ultracentrifugations were performed in duplicate (3 U and 4 U for N expressing yeast, and 5 U and 6 U for N-PbCS yeast). Values correspond to optical densities at OD_{450 nm} (taking OD_{620 nm} as reference) multiplied by sample dilutions. SB: suspension buffer.</p

Experimental challenge of immunized mice.

<p>(<b>A</b>) Mean and standard deviations log₁₀ values of parasitemia in mice immunized by N-PbCS, PbCS, N or WT yeast, and in non-immunized mice following infection with 6,000 GFP⁺<i>Pb</i> sporozoites. Blood parasitemia is expressed in log₁₀ scale as the percentage of infected red blood cells (iRBCs) out of total RBCs along the first 7 days follow up. Asterisks (*) indicate the significance level of the Mann-Whitney nonparametric test: two symbols correspond to p<0.005 and three to p<0.0005. (<b>B</b>) Parasitemia at day 5 post-challenge. Bars correspond to medians. Asterisks (*) indicate significant median differences (one symbol for p<0.05 and two for p<0.005; Mann-Whitney nonparametric test). (<b>C</b>) Inverse correlation between the day of death (x axis) and the percentage of iRBCs per total RBCs (y axis; arithmetic scale) per mouse. The cause of death is given in the upper part of the graph. (<b>D</b>) Survival curves of immunized mice after challenge with 6,000 GFP⁺<i>Pb</i> sporozoites.</p

Humoral responses elicited in mice after immunization.

<p>(<b>A</b>) Kinetics of anti-PbCS IgG responses in mice immunized with N-PbCS yeasts. OD_{450 nm} are expressed in log₁₀ scale. Black arrows indicate immunization schedule. (<b>B</b>) Isotyping of humoral IgG responses at day 42 in mice immunized with N-PbCS. The bars correspond to median values per group. Asterisks (*) indicate significant median differences (p<0.05; Mann-Whitney nonparametric test). (<b>C</b>) Anti-N IgG titers in mice serums collected at day 42 after immunization with WT yeast or yeasts expressing N, PbCS or N-PbCS. Median values were compared by the Wicoxon Two Sample Test (p = 0.4558). (<b>D</b>) Antibody titers of N-PbCS mice are compared (see panels B and C). The lines associate titers from the same mouse. (<b>E</b>) Anti-<i>P. pastoris</i> IgG responses towards whole yeast. (<b>F</b>) Anti-<i>P. pastoris</i> IgG responses towards lysed yeast. The bars in (E) and (F) correspond to mean values per group. Hash sign (#) indicates anti-N antibody-negative mice from the N-PbCS group (see panel C).</p

Schematic representation of the immunization protocol.

<p>Immunization and challenge schedule is given in days (d) and bleeding time points in weeks.</p

Immunofluorescence analysis of N or N-PbCS expression in yeasts (N staining in green, PbCS staining in red and nuclei in blue).

<p>Each image is the maximal intensity projection of three consecutive focal planes spaced 0.5 µm apart.</p