Urinary Peptidomics to Address Age-Related Disabilities (preprint)

Background: The Global Burden Disease 2019 report called for innovation in addressing age-related disabilities. The current study aimed at identifying and validating a urinary peptidomic profile (UPP) differentiating healthy from unhealthy ageing in the general population, to test the UPP predictor in patients, and to search for targetable molecular pathways. Methods: In a Flemish population study (n=778; 50·8% women; age, 16·2-82·1 years), 559 participants were examined twice and made up the derivation and internal validation datasets; 219 were examined once and constituted the independent validation dataset. The UPP was assessed by capillary electrophoresis coupled with mass spectrometry. Statistical methods included linear and proportional hazard regression. Pathway exploration rested on the Reactome and KEGG databases. The multidimensional UPP signature reflecting ageing was further validated in patients with diabetes, COVID-19 or chronic kidney disease. Findings: With correction for multiple testing and multivariable adjustment, chronological age (C‑age) was associated with 210 sequenced peptides mainly showing downregulation of collagen fragments. The trained model relating C‑age to UPP, derived by elastic net regression, included 54 peptides from 17 proteins. In the derivation and the internal and independent validation datasets, the trained model explained 76·3%, 54·4% and 65·3% of C‑age. Compared with the derivation data, the UPP-predicted C‑age was greater (p<0·0001) in age-matched patients with diabetes (n=1575), COVID‑19 infection (n=110) or chronic kidney disease (n=202): 50·3 vs 56·9 vs 58·5 vs 62·3 years. In the population, risk carrying biomarkers were associated (p≤0·037) with UPP‑age, independent of C‑age. Over 12·8‑year (median), the incidence of total and cardiovascular mortality and osteoporosis in the population was associated with UPP‑age, independent of C‑age, with hazard ratios per 10‑year higher UPP‑age of 1·54, 1·72 and 1·40, respectively (p≤0.018). The overrepresented proteins were key nodes in collagen and extracellular-matrix (ECM) turnover. Interpretation: Ageing is associated with a specific UPP signature, reflecting fibrosis and ECM remodelling. UPP‑age was associated with risk factors and adverse health outcomes in the population and with accelerated ageing in patients. Innovation in addressing disability should overcome the ontology of diseases and focus on shared disease mechanisms, in particular the bodywide ageing associated fibrosis and ECM remodelling.Funding: European Research Council, Ministry of the Flemish Community, OMRON Healthcare. Declaration of Interest: HM is the co-founder and co-owner of Mosaiques-Diagnostics GmbH, Hannover, Germany, and AL is an employee of Mosaiques Diagnostics. All other authors declare no conflict of interestEthical Approval: The Flemish Study on Environment, Genes and Health Outcomes (FLEMENGHO) complies with the Helsinki declaration and is registered at the Belgian Data Protection Authority (reference number III 11/1234/13; 22 August 2013). The ethics committee of the University Hospital Leuven, Belgium, approved the secondary use of FLEMENGHO data (national registration number, B32220083510).

Clinical Characteristics of Recurrent-positive Coronavirus Disease 2019 after Curative Discharge: a retrospective analysis of 15 cases in Wuhan China (preprint)

In China, the patients with previously negative RT-PCR results again test positive during the post-discharge isolation period. We aimed to determine the clinical characteristics of these recurrent-positive patients. We retrospectively reviewed the data of 15 recurrent-positive patients and 107 control patients with non-recurrent, moderate COVID-19 treated in Wuhan, China. Clinical data and laboratory results were comparatively analyzed. We found that recurrent-positive patients had moderate disease. The rate of recurrent-positive disease in our hospital was 1.87%. Recurrent-positive patients were significantly younger (43(35-54) years) than control patients (60(43-69) years) (P=0.011). The early LOS (length of stay in hospital before recurrence) was significantly longer in recurrent-positive patients (36(34-45) days) than in control patients (15(7-30) days) (P =0.001). The time required for the first conversion of RT-PCR results from positive to negative was significantly longer in recurrent-positive patients (14(10-17) days) than in control patients (6(3-9) days) (P =0.011). Serum COVID-19 antibody levels were significantly lower in recurrent-positive patients than in control patients (IgM: 13.69 {+/-} 4.38 vs. 68.10 {+/-} 20.85 AU/mL, P = 0.015; IgG: 78.53 {+/-} 9.30 vs. 147.85 {+/-} 13.33 AU/mL, P < 0.0001). Recurrent-positive patients were younger than control patients. The early LOS (length of stay in hospital before recurrence) was significantly longer in recurrent-positive group than that in control group. COVID-19 IgM/IgG antibody levels were significantly lower in recurrent-positive group than those in control group, which might explain why the virus RNA RT-PCR was positive after the initial clinical cure(with three times of virus RNA RT-PCR negative). The virus might not be fully eliminated because of the lower IgG level and their later replicating might result in recurrent-positive virus RNA RT-PCR.

Association between lung injury and cytokine profile in COVID-19 pneumonia (preprint)

Background: Coronavirus disease 2019 (COVID-19) is a systemic disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The purpose of the present study was to investigate the association between lung injury and cytokine profile in COVID-19 pneumonia.Methods: This retrospective study was conducted in COVID-19 patients. Demographic characteristics, symptoms, signs, underlying diseases, and laboratory data were collected. The patients were divided into COVID-19 with pneumonia and without pneumonia. CT severity score and PaO2/FiO2 ratio and were used to assess lung injury.Results: 106 patients with 12 COVID-19 without pneumonia and 94 COVID-19 with pneumonia were included. Compared with COVID-19 without pneumonia, COVID-19 with pneumonia had significant higher serum interleukin (IL)-2R, IL-6, and tumor necrosis factor (TNF)-α. Correlation analysis showed that CT severity score and PaO2/FiO2 were significantly correlated with age, presence of any coexisting disorder, lymphocyte count, procalcitonin, IL-2R, and IL-6. In multivariate analysis, log IL6 was only independent explanatory variables for CT severity score (β=0.397, p<0.001) and PaO2/FiO2 (β=-0.434, p=0.003).Conclusions: Elevation of circulating cytokines was significantly associated with presence of pneumonia in COVID-19 and the severity of lung injury in COVID-19 pneumonia. Circulating IL-6 independently predicted the severity of lung injury in COVID-19 pneumonia.

Prognostic value of neutrophil-to-lymphocyte ratio, lactate dehydrogenase, D-dimer and CT score in patients with COVID-19 (preprint)

Background To explore the significance of neutrophil-to-lymphocyte ratio (NLR), lactate dehydrogenase (LDH), D-dimer and CT score in evaluating the severity and prognosis of coronavirus disease – 2019 (COVID-19).Methods Patients with laboratory confirmed COVID-19 were retrospectively enrolled. The baseline data, laboratory findings, chest computed tomography (CT) results evaluating by CT score on admission, and clinical outcomes were collected and compared. The logistic regression was used to assess the independent relationship between the baseline level of four indicators (NLR, LDH, D-dimer, CT score) and the severity of COVID-19.Results Among 432 patients, 125 (28.94%) cases were divided into severe group, the remaining (n = 307, 71.06%) were in non-severe group. In multivariate logistic regression, high level of NLR, LDH were independent predictor of the severe group in COVID-19 (OR = 2.163; 95%CI = 1.162–4.026; p = 0.015 for NLR > 3.82; OR = 2.298; 95%CI = 1.327–3.979; p = 0.003 for LDH > 246U/L). Combining NLR > 3.82 and LDH > 246U/L increased the sensitivity of diagnosis in severe patients (NLR > 3.82 [50.40%] vs. Combined diagnosis [72.80%]; p = 0.0007; LDH > 246 [59.2%] vs. Combined diagnosis [72.80%]; p < 0.0001).Conclusions High levels of NLR and LDH in serum have potential value in the early identification of severe patients with COVID-19. The combination of LDH and NLR can improve the sensitivity of diagnosis.

Efficacy and safety of chloroquine or hydroxychloroquine in moderate type of COVID-19: a prospective open-label randomized controlled study (preprint)

The outbreak of novel coronavirus disease 2019 (COVID-19) has become a pandemic. Drug repurposing may represent a rapid way to fill the urgent need for effective treatment. We evaluated the clinical utility of chloroquine and hydroxychloroquine in treating COVID-19. Forty-eight patients with moderate COVID-19 were randomized to oral treatment with chloroquine (1000 mg QD on Day 1, then 500 mg QD for 9 days; n=18), hydroxychloroquine (200 mg BID for 10 days; n=18), or control treatment (n=12). Adverse events were mild, except for one case of Grade 2 ALT elevation. Adverse events were more commonly observed in the chloroquine group (44.44%) and the hydroxychloroquine group (50.00%) than in the control group (16.67%). The chloroquine group achieved shorter time to clinical recovery (TTCR) than the control group (P=0.019). There was a trend toward reduced TTCR in the hydroxychloroquine group (P=0.049). The time to reach viral RNA negativity was significantly faster in the chloroquine group and the hydroxychloroquine group than in the control group (P=0.006 and P=0.010, respectively). The median numbers of days to reach RNA negativity in the chloroquine, hydroxychloroquine, and control groups was 2.5 (IQR: 2.0-3.8) days, 2.0 (IQR: 2.0-3.5) days, and 7.0 (IQR: 3.0-10.0) days, respectively. The chloroquine and hydroxychloroquine groups also showed trends toward improvement in the duration of hospitalization and findings on lung computerized tomography (CT). This study provides evidence that (hydroxy)chloroquine may be used effectively in treating moderate COVID-19 and supports larger trials.

RT-PCR Status and Antibody Response in Patients with Coronavirus Disease 2019 (preprint)

Background: Novel coronavirus (COVID-19) is a new viral species that causes pneumonia. Currently, RT-PCR and IgM/IgG antibody assays have been recommended for the diagnosis of COVID-19 infection. However, the correlation between RT-PCR status and antibody (IgG, IgM) response remains unknown. Methods: Consecutive COVID-19 patients admitted to our department between February 10, 2020 and March 10, 2020, were diagnosed by guidelines issued by the World Health Organization (WHO) and included in this study. RT-PCR and antibody (IgM/IgG) assays for COVID-19 infection were performed for all patients according to the manufactures’ protocols. Other data, such as demographic, clinical, laboratory, as well as treatment and outcome, were collected using data collection tables from electronic medical records.Results: During the study period, a total of 103 patients were diagnosed as having a moderate type of COVID-19 at our department, including 55 males and 48 females, with an average age of 57.53 ± 1.65 years old (range 23 to 90 years old). The peak level of SARS-CoV-2 IgM antibody (243.10 ± 89.84 AU/ml) was reported 4 days after the negative RT-PCR (-) (all P < 0.05). Subsequently, the IgM decreased to 42.69 ± 22.39 AU/ml 21 days after RT-PCR (-). However, the IgG was maintained at a high level 4 days before RT-PCR (-) and later. The lymphocyte count was at the lowest level on day7 before the RT-PCR(-) result (P<0.05), and then elevated after RT-PCR conversion (viral clearance).Conclusions: SARS-CoV-2 IgM/IgG levels did not correlate with RT-PCR status in our study sample. We found that SARS-CoV-2 IgM/IgG could be a potential biomarker to monitor clinical course, determine discharge, and assess recovery of those infected patients with the novel coronavirus. Trial registration: A prospective, open label, randomized, control trial for chloroquine or hydroxychloroquine in patients with mild and common novel coronavirus pulmonary (COVIP-19). ChiCTR2000030054. Registered 18 Feb,2020. http://www.chictr.org.cn/edit.aspx?pid=49869&htm=4

Prognostic Value of Neutrophil-To-Lymphocyte Ratio, Lactate Dehydrogenase, D-Dimer and CT Score in Patients with COVID-19 (preprint)

Background: To explore the significance of neutrophil-to-lymphocyte ratio (NLR), lactate dehydrogenase (LDH), D-dimer and CT score in evaluating the severity and prognosis of coronavirus disease – 2019 (COVID-19) in two centers of Hubei, China.Methods: A total of 432 patients with laboratory confirmed COVID-19 were retrospectively enrolled and divided into non-severe and severe groups. The baseline data, laboratory findings, chest computed tomography (CT) results evaluating by CT score on admission, and clinical outcomes were collected and compared. The logistic regression was used to assess the independent relationship between the baseline level of four indicators (NLR, LDH, D-dimer, CT score) on admission and the severity of COVID-19, respectively.Results: Among 432 patients, 125 (28.94%) cases were divided into severe group, the remaining (n = 307, 71.06%) were in non-severe group. In multivariate logistic regression, the high level of NLR, LDH were independent predictor in the early classification of patients with COVID-19 (OR = 2.163; 95%CI = 1.162–4.026; p = 0.015 for NLR > 3.82; OR = 2.298; 95%CI = 1.327–3.979; p = 0.003 for LDH > 246U/L). Furthermore, combining NLR > 3.82 and LDH > 246U/L could increase the sensitivity of diagnosis in severe patients (NLR > 3.82 [50.40%] vs. Combined diagnosis [72.80%]; p = 0.0007; LDH > 246 [59.2%] vs. Combined diagnosis [72.80%]; p < 0.0001).Conclusions: The high levels of NLR and LDH in serum have potential value in the early identification of severe patients with COVID-19. The combination of LDH and NLR can improve the sensitivity of diagnosis.Importance: COVID-19 has been a global pandemic. The mortality rate is range from 3.5-6.0%. In order to predict the risk factors of severity of COVID-19. we explore the significance of neutrophil-to-lymphocyte ratio (NLR), lactate dehydrogenase (LDH), D-dimer and CT score in evaluating the severity and prognosis of coronavirus disease – 2019 (COVID-19) in two centers of Hubei, China. We found that the high levels of NLR and LDH in serum have potential value in the early identification of severe patients with COVID-19. The combination of LDH and NLR can improve the sensitivity of diagnosis.