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1.
Blood ; 138:1022, 2021.
Artigo em Inglês | EMBASE | ID: covidwho-1582180

RESUMO

[Formula presented] Introduction: Patients (pts) with immune thrombotic thrombocytopenic purpura (iTTP) are at high risk of severe COVID-19, therefore protection from SARS-CoV-2 by vaccination is particularly relevant in this setting, although concerns may exist on possible adverse reactions or disease relapse after vaccination. In this study, in a group of iTTP pts who received in-hospital COVID-19 vaccination in a special program for ‘fragile patients’, we prospectively evaluated over time the antibody response, the clinical and laboratory disease parameters and hemostatic biomarker levels. Methods: Twelve iTTP pts in clinical remission and regularly followed-up in our Center were enrolled in April 2021, all of them received 2 doses of BNT162b2 vaccine (Pfizer-BioNTech) over 21 days, and were followed-up for clinical and laboratory testing for 60 days. Blood samples were collected at enrollment (day 0, D0) before the 1 st vaccine dose;on day 21 (D21) before the 2 nd dose;and on day 60 (D60) after the 1 st dose. Blood cell counts, anti-Spike receptor-binding-domain protein (anti-S/RBD) IgG, ADAMTS-13 activity, and anti-ADAMTS-13 IgG (chromogenic assay and ELISA), were measured at each time point. Additionally, an extensive study of hemostatic markers (i.e. FVIII, von Willebrand Factor (vWF) antigen and activity, fibrinogen, D-dimer, tPA, PAI, and F1+2) was performed. Follow up is currently continuing. Results: Median age of our cohort was 65 years with M/F ratio of 4/8. Median time since last acute iTTP episode was 40 months, median follow up of the cohort was 71 months (95% CI 30-126). All pts were in clinical remission, except one patient (P1) who had an iTTP relapse after contracting SARS-CoV-2 infection, in Dec 2020, and was on low-dose steroids on D0. One patient (P2) had an ADAMTS-13 relapse in Jan 2021, and received pre-emptive rituximab. No other pts were on immunosuppressive therapy. Concerning the status of ADAMTS-13 activity on D0, 6 pts showed normal levels (>50%), while 5 had a moderate (50-20%) and 1 a complete (<10%) ADAMTS-13 deficiency. This latter patient (P3) had normal ADAMTS-13 activity before the pandemic. All patients were negative for anti-ADAMTS-13 inhibitor. Further, on D0, the anti-S/RBD IgG testing was positive in 3/12 pts (median 704,1 AU/mL), due to symptomatic infection in 1 case (P1), and asymptomatic in 2 (P3 and 1 pt with ADAMTS-13 activity of 54%, P4). The study of hemostatic markers on D0 showed an increase in median levels of FVIII and vWF antigen and activity. These parameters were altered in 7/12, 11/12 and 8/12 pts, respectively. Fibrinogen and D-dimer were increased in 3/12 and 2/12, respectively. Notably, P1, P3 and P4 presented the highest levels of FVIII and vWF antigen, associated with high levels of vWF activity in P1 and P3 (mean 233%);moreover, P3 showed higher levels of D-dimer (708 ng/mL) and tPA (13 ng/ml). After the 2 doses of BNT162b2, no significant clinical side effects were reported, and no changes in platelet counts. ADAMTS-13 activity and inhibitors did not significantly change on D21 and D60. A complete ADAMTS-13 activity deficiency persisted in P3 on D21 and D60, associated with anti-ADAMTS-13 IgG titer >15 U/ml, despite clinical remission. Overall, a significant increase in anti-S/RBD IgG level was observed on D21 (p = 0.0005) and D60 (p = 0.0005). Remarkably, only P2 did not show an increase in anti-S/RBD IgG titer after both doses of BNT162b2. Median levels of FVIII and vWF antigen did not significantly change during follow up, while increased vWF activity was seen on D60 (p = 0.05). Fibrinogen levels were stable, and an increase in D-dimer (>1000 ng/mL both on D21 and D60) was seen in P3. There were no changes in the other hemostatic parameters, and no thromboses were observed. Conclusions: In our cohort of iTTP pts, COVID-19 was associated with 1 clinical and 1 ADAMTS-13 relapse. Our data show that SARS-CoV-2 vaccination was effective in inducing an antibody response in all but one patient who received rituximab within 3 months before vaccinat on, confirming recent findings. Overall, vaccination had no relevant impact on the hemostatic profile of our pts, and did not appear to be a driver of iTTP relapses. However, anti-SARS-CoV-2 antibodies monitoring in iTTP pts may be useful after vaccination, as currently it is unknown how long the antibody titer may persist. Although small, this study is in favor of efficacy and safety of mRNA vaccines in pts with iTTP. Disclosures: Falanga: Bayer: Honoraria;Sanofi: Honoraria;Leo Pharma: Honoraria;Pfizer: Honoraria.

2.
European Heart Journal ; 42(SUPPL 1):1288, 2021.
Artigo em Inglês | EMBASE | ID: covidwho-1554493

RESUMO

Background: Hospitalised COVID-19 pneumonia patients are characterised by the occurrence of a hypercoagulable state associated to a high risk of thromboembolic events. The main laboratory findings of this coagulopathy include D-dimer increase, mild thrombocytopenia, prolonged PT, and increase endothelial activation biomarkers (vWF, thrombomodulin). No data are available about coagulation profile in patients presenting with an acute coronary syndrome (ACS) combined with SARS-CoV-2 infection. Purpose: In this prospective study, we aimed to evaluate the contribute of concomitant SARS-CoV-2 infection to the haemostatic system derangement (i.e., from endothelial cell activation to fibrinolytic phase) observed in patients presenting with ACS. Further, the role of haemostatic biomarkers (HB) for in-hospital mortality risk prediction was also explored. Methods: Consecutive patients admitted to our hospital for ACS at peak intensity of local pandemia were enrolled into this study. At admission, all patients underwent routine blood examinations with blood count, serum biochemical tests and an extensive coagulation profiling. Data from coronary angiography and percutaneous coronary intervention (PCI), when performed, were collected. In-hospital major adverse cardio and cerebrovascular events -MACCEs- (total and cardiovascular death, stroke, systemic or pulmonary embolism, re-MI and bleedings) are reported. Results: A total of 99 (76M/23F) consecutive patients with a median age of 66.7 (±12.1) were enrolled. According to nasal swab, 24 patients were SARS-CoV-2 positive and 75 negative. The two groups, similar in age, sex and cardiovascular risk factors, significantly differed in presenting symptoms (p<.001) and radiological signs of pneumonia (p<.0001). At admission, there were no differences in routine laboratory values between groups. Differently, analysis of the HB showed significantly higher values of D-dimer, vWF antigen, vWF activity and vWF;RiCof, t-PA and PAI-1 and lower levels of ADAMTS-13 in the positive group. Furthermore, among ACS patients, both STEMI and NSTEMI subjects, positive for SARS-CoV-2, had significantly higher plasma values of all the HB compared to the respective negative counterparts, with SARS-CoV-2 positive STEMI subjects displaying the highest values. When performed, PCI finished more frequently with a final TIMI flow <3 (p=.004) in positive patients. The in-hospital rate of MACCEs was 24% (24/99 patients) with a higher (p<.0001) prevalence in SARS-Co-V2 positive group. Cardiovascular mortality accounted for the majority of deaths (8/10;p=.019). At multivariable analysis, we identified dyspnoea at presentation, vWF antigen and leukocyte values as independent risk factors for in-hospital death. Conclusions: In patients presenting with ACS combined with SARS-Cov- 2 infection an additional HB asset derangement with stronger endothelial cell activation occurs which negatively impact the outcome, regardless of the invasive treatment.

3.
Research and Practice in Thrombosis and Haemostasis ; 5(SUPPL 2), 2021.
Artigo em Inglês | EMBASE | ID: covidwho-1509186

RESUMO

Background: Hypercoagulability, complement activation and endothelial perturbation characterize sever COVID-19. After disease remission, a proportion of convalescent subjects still experience post-COVID-19 symptoms. No information is available on persistence of hemostatic alterations in this setting. Bergamo city, represents one of the first and most affected area by SARS-CoV-2 infection in the world. For this reason, since the beginning we were actively involved in recruiting CCP donors. Aims: In a large cohort of CCP donors, we aimed to characterize biomarkers of hypercoagulability and of endothelial perturbation in order to find associations with disease severity, demographic characteristics, and anti-SARS-CoV-2 antibody levels. Methods : Candidate CCP donors were tested for the anti-SARSCoV-2 antibodies, including anti-S IgG antibodies (Anti-S Ab) and anti-N IgG antibodies (Anti-N Ab). In addition, the following plasma biomarkers were assessed: fibrinogen, protein C, protein S, factor V, factor VIII, factor XIII, D-dimer, and von Willebrand factor (vWF). Results: 425 CCP candidates (275M/150F, age range 19-67 years) were admitted to donation. Male gender, age > 40 years, and severe form of COVID-19 were identified as independent predictors of high levels of both anti-S and anti-N Ab ( p <0.001). Among hemostatic parameters, levels of vWF antigen, vWF activity and protein C were significantly higher in CCP donors who had severe COVID-19 compared to donors who had non-severe COVID-19 ( p <0.001). Furthermore, vWF levels positively correlated with anti-S Ab levels (vWF-antigen r=0.216 vWF-activity r=0.257 and vWFRiCof r=0.226, p <0.01). Conclusions: Our data show that gender, age and severe disease can be predictors of an increased immunological response. Furthermore, convalescent subjects show a persistently high vWF levels, suggesting a persistence of the endothelial activation, despite of clinical disease remission.

4.
Research and Practice in Thrombosis and Haemostasis ; 5(SUPPL 2), 2021.
Artigo em Inglês | EMBASE | ID: covidwho-1509058

RESUMO

Background : Endothelial damage and hypercoagulability are major players behind the hemostatic derangement in SARS-CoV-2 infection. Aims : In this prospective cohort study of COVID-19 patients, we aimed to assess the role of circulating endothelial activation/damage biomarkers in predicting in-hospital mortality. Methods : Clinical data of COVID-19 patients hospitalized in intensive care (ICU) and non-ICU units at 2 Bergamo (Italy) hospitals from March 23 to May 30, 2020, were analyzed. Markers of endothelium activation including von-Willebrand factor (vWF), soluble thrombomodulin (sTM), and fibrinolytic proteins (t-PA and PAI-1) were measured. Additionally, D-dimer, Fibrinogen, FVIII, nucleosomes, CRP and procalcitonin were assessed. Results : Sixty-three (45 ICU, and 18 non-ICU) patients, with a median age of 62 years were analyzed. Increased plasma levels of Ddimer, FVIII, fibrinogen, nucleosomes, CRP, and procalcitonin were observed in the whole cohort. Extremely elevated vWF levels characterized all patients (highest values in ICU-subjects). Patients with a moderate and severe ARDS (i.e. PaO2/FiO2 ≤200%) have considerably higher vWF and sTM levels, and lower t-PA/PAI-1 values compared to patients in the mild ARDS group (i.e. PaO2/FiO2 >200%). After a median time of 30 days, death occurred in 13 (21%) patients. By multivariable analysis, vWF-activity, neutrophil-count and PaO2/ FiO2 were significantly associated with death. Using these variables, we generated a linear score with 3-risk groups (AUC 0.903) that provided a cumulative incidence of death of 0 % in the low-, 32% in the intermediate-, and 78% in the high-risk group ( P < 0.001). Conclusions : In conclusion, our study provides an extensive overview of the endothelial damage induced by SARSCoV-2 infection in hospitalized patients with virus-induced pneumonia and different degrees of disease severity. In addition, despite the small sample size and the need for the external validation, we could generate an accurate score based on circulating vWF to predicting mortality in severe COVID-19 patients.

5.
HemaSphere ; 5(SUPPL 2):105, 2021.
Artigo em Inglês | EMBASE | ID: covidwho-1393468

RESUMO

Background: Severe COVID-19 is associated with a profound derangement of the hemostatic system characterized by hypercoagulability, complement activation and endothelial cell perturbation. After disease resolution, some convalescent subjects still experience post-COVID-19 symptoms. No information is available on persistence of hemostatic alterations in this setting. Bergamo city, represents one of the first and most affected area by SARS-CoV-2 infection in the world. For this reason, since the beginning we were actively involved in hyperimmune plasma collection from COVID-19 convalescent subjects. Aims: In this study, in a large cohort of convalescent donors of hyperimmune plasma, we aimed to characterize select hemostatic parameters of hypercoagulability and endothelial cell perturbation and their association with disease severity, demographic characteristics, and antibody levels. Methods: Recovered COVID-19 patients eligible to plasma donation were tested for the SARS-CoV-2 antibodies by the anti-N IgG SARSCoV- 2 antibodies (Abbott Laboratories, IL, USA, Anti-N Abs), and/or the anti-S IgG SARS-CoV-2 antibodies (Liaison-Diasorin, Sallugia-VC, Italy, Anti-S Abs), according to the manufacturer's instructions. Fibrinogen, protein C, protein S, factor V, factor VIII, factor XIII, D-dimer, and von Willebrand factor (vWF) were assessed. Results: 425 subjects have been included (275M/150F) with a median age of 48 years (range: 19-67 years). Among convalescent subjects admitted to the donation, male gender, age > 40 years, and previous hospitalization for COVID-19, were identified as independent predictive factors for significantly (p<0.001) higher levels of SARS-CoV-2 IgG (both anti-S and anti-N). Hemostatic parameters including fibrinogen, protein S, factor V, factor VIII, factor XIII, and D-dimer were not different between severe and non-severe COVID-19. Differently, convalescent subjects with previous severe COVID-19 showed significantly higher levels of vWF (124±40 vs 121±41 %, p<0.001) and PC (119±19 vs 109±19 %, p<0.001) compared with non-severe COVID-19 subjects. In addition, significant positive correlations were found between vWF levels and anti-S Abs (vWF antigen r=0.216;vWF activity r=0.257 and vWF RiCof r=0.226, p<0.01). Summary/Conclusion: Our data show that gender, age and severe disease can be potential predictors of an increased immunological response. Furthermore, convalescent subjects show a persistently high vWF levels, suggesting a persistence of the endothelial activation, despite of clinical disease remission.

6.
Vaccines (Basel) ; 9(4):15, 2021.
Artigo em Inglês | MEDLINE | ID: covidwho-1210082

RESUMO

COVID-19 is an ongoing pandemic caused by the highly infectious coronavirus SARS-CoV-2 that is engaging worldwide scientific research to find a timely and effective eradication strategy. Great efforts have been put into anti-COVID-19 vaccine generation in an effort to protect the world population and block SARS-CoV-2 spread. To validate the protective efficacy of the vaccination campaign and effectively control the pandemic, it is necessary to quantify the induction of neutralizing antibodies by vaccination, as they have been established to be a correlate of protection. In this work, a SARS-CoV-2 pseudovirus neutralization assay, based on a replication-incompetent lentivirus expressing an adapted form of CoV-2 S protein and an ACE2/TMPRSS2 stably expressing cell line, has been minimized in terms of protocol steps without loss of accuracy. The goal of the present simplified neutralization system is to improve SARS-CoV-2 vaccination campaign by means of an easy and accessible approach to be performed in any medical laboratory, maintaining the sensitivity and quantitative reliability of classical serum neutralization assays. Further, this assay can be easily adapted to different coronavirus variants by simply modifying the pseudotyping vector.

7.
International Journal of Environmental Research & Public Health [Electronic Resource] ; 18(7):04, 2021.
Artigo em Inglês | MEDLINE | ID: covidwho-1209448

RESUMO

Italy was the first country in Europe to face the coronavirus pandemic. The aim of the study was to analyze healthcare workers' (HCWs) level of information, practice, and risk perception towards COVID-19. We set up a cross-sectional study through SurveyMonkey<sup> R</sup> and distributed the link through Facebook and Whatsapp closed groups. The research instrument was a 31 items questionnaire distributed using Facebook and Whatsapp. It was conducted in Italy from February to May 2020. The study participants were general practitioners, pediatricians and other health professionals. A total of 958 participants were included: 320 (33.4%) general practitioners, 248 (25.9%) pediatricians and 390 (40.7%) other health professionals. The highest response rate was from Northern Italy (48.1%), followed by Central Italy (29.9%) and Southern Italy (22.0%). Less than a half (46%) of respondents felt they had a good level of information of COVID-19 case definition and of national prevention guidelines. Respondents reported to have changed their clinical practice;particularly, they increased the use of masks (87.1%, p < 0.001), disinfection and sanitization of doctors' offices (75.8%, p < 0.001), the use of protective glasses (71.2%, p < 0.001), alcoholic hand solution (71.2%, p < 0.001), and hand washing (31.8%, p = 0.028). HCWs are at high risk of infection;less than a half of them felt adequately prepared to face COVID-19 pandemic, so they need extensive information and awareness of the disease to take adequate precautionary measures, and they are crucial to disseminate good practices.

9.
Biochimica Clinica ; 44(SUPPL 2):S92, 2020.
Artigo em Inglês | EMBASE | ID: covidwho-984401

RESUMO

Focusing attention on cancer patients' frailty, Cancer Institute-IRCCS "Fondazione Pascale" in Naples has planned a tightened program of health surveillance for patients (PTs) and healthcare providers (HPs), in order to early detect and promptly quarantine subjects with SARSCoV-2 infection. This program requires a multidisciplinary asset, within which Laboratory plays a crucial role. In our experience, the attention has focused on application of biosafety conditions, as stated by World Health Organization's guidance. A validated internal protocol was derived from it. Laboratory diagnostic workflow provided 3 steps: 1) rapid immunochromatographic assays (ICAs) to detect SARS-CoV-2 IgM and IgG from plasma samples of PTs and HPs;2) automated qualitative electrochemiluminescence immunoassays (ECLIAs) to detect SARS-CoV-2 antibodies from serum samples of HPs;3) Real-time PCR detection of SARS-CoV-2 RNA from nasopharyngeal swabs (NS) of PTs and HPs. Health surveillance program planned an initial evaluation of PTs and HPs with rapid ICAs (automated tests being not available yet) and a further screening with molecular tests on NS sent to external COVID-19 reference Laboratories. Subsequently, our Laboratory was included in CORONET network, allowing us to perform molecular tests for SARSCoV-2. At the same time, availability of automated ECLIAs allowed us to screen HPs periodically, continuing to perform ICAs to all PTs afferent to hospital triage. At the beginning of the surveillance, rapid ICAs allowed to screen 1920 PTs (89 of which resulted positive, 4.63%) and 1050 HPs (48 positive, 4.57%). Subsequently, HPs' screening with ECLIAs revealed 25 positive subjects out of 1018 tested (2.46%). Early detection and quarantine of positive cases allowed to find very low percentages of positive SARS-CoV-2 subjects to molecular tests from NS: 1 positive out 2215 NS from HPs (0.05%) and 1 positive out of 742 NS from PTs (0.13%). All Laboratory staff efforts have been directed to guarantee an adequate turnaround time. The element "time" has been crucial to subject promptly to quarantine personnel identified as positive and to isolate patients affected, thus allowing health surveillance program to ensure an adequate protection for cancer patients afferent to our Institute.

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