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1.
Critical Care Medicine ; 51(1 Supplement):451, 2023.
Article in English | EMBASE | ID: covidwho-2190635

ABSTRACT

INTRODUCTION: Fluid stewardship is a pillar in the management of ARDS. Previous studies evaluating conservative fluid management had difficulty achieving daily fluid targets and evaluations of adjunctive albumin have found conflicting results. METHOD(S): This was a retrospective study conducted at a large community hospital. Patients were included if >17 years old, admitted to an ICU from January 2016 to September 2021, had a diagnosis of ARDS, and received loop diuretic and albumin within one hour of each other. Exclusion criteria included liver failure, ESRD, and positive test for SARSCOV- 2. Patients were divided into two groups by baseline serum albumin, where low albumin was defined as <= 3.5g/ dL. The primary outcome was the percentage of patients with a positive response to combination therapy, defined as UOP >= 600mL within 6 hours of administration. Secondary outcomes included UOP at 6 hours, and change in PF ratio and serum albumin at 24 hours. Nominal data are presented as percentage, whereas continuous variables are presented as median and were analyzed using Chi-square and Mann Whitney U tests, respectively. Alpha of < 0.05 was deemed statistically significant. RESULT(S): 108 patients were in the low-albumin (LA) group and 67 in the normal-albumin (NA) group. The NA group were statistically heavier (99 vs. 88kg), had higher total protein (7.2 vs. 5.1 g/dL), had diagnosis of sepsis (59 vs. 6%) and had less prior diuretic use (48 vs. 74%). Key baseline similarities included PF ratio (158 vs. 160, p=0.7), creatinine (1.22 vs. 1.20 mg/dL), and presence of shock (27 vs. 33%). Statistically, more patients in the NA group received 5% albumin, (63 vs. 34%), received less albumin (12.5 vs. 25 grams), and similar loop diuretic dose expressed as furosemide equivalents (40 vs. 40 mg). 43 (64%) were positive responders in the NA group compared to 61 (57%) in the LA group (p=0.48). UOP at 6 hours (670 vs. 653mL) and change in PF ratio at 24 hours (+23 vs. +13) were statistically similar. The change in albumin was different between the Normal and Low groups (-0.6 vs. +0.2 g/dL, p< 0.05). CONCLUSION(S): Low albumin did not affect the urinary response in patients with ARDS receiving albumin and loop diuretics. These findings are limited by heterogeneity in baseline characteristics and components of the intervention.

2.
Nucleic acids research ; 05, 2023.
Article in English | EMBASE | ID: covidwho-2189416

ABSTRACT

The number of genetic variations in the SARS-CoV-2 genome has been increasing primarily due to continuous viral mutations. Here, we report that the human APOBEC3A (A3A) cytidine deaminase plays a critical role in the induction of C-to-U substitutions in the SARS-CoV-2 genome. Bioinformatic analysis of the chronological genetic changes in a sequence database indicated that the largest UC-to-UU mutation signature, consistent with APOBEC-recognized nucleotide motifs, was predominant in single-stranded RNA regions of the viral genome. In SARS-CoV-2-infected cells, exogenous expression of A3A but not expression of other APOBEC proteins induced UC-to-UU mutations in viral RNA (vRNA). Additionally, the mutated C bases were often located at the tips in bulge or loop regions in the vRNA secondary structure. Interestingly, A3A mRNA expression was drastically increased by interferons (IFNs) and tumour necrosis factor-alpha (TNF-alpha) in epithelial cells derived from the respiratory system, a site of efficient SARS-CoV-2 replication. Moreover, the UC-to-UU mutation rate was increased in SARS-CoV-2 produced from lung epithelial cells treated with IFN-s and TNF-alpha, but not from CRISPR/Cas9-based A3A knockout cells. Collectively, these findings demonstrate that A3A is a primary host factor that drives mutations in the SARS-CoV-2 RNA genome via RNA editing. Copyright © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

3.
New Journal of Chemistry. ; 2023.
Article in English | EMBASE | ID: covidwho-2186153

ABSTRACT

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), is a highly infectious disease with a significant impact on global public health security, and the development of effective antiviral drugs is warranted. In this study, based on HEK293 membrane chromatography (CMC) model that overexpresses angiotensin-converting enzyme 2 (ACE2), we screened six compounds with long retention time on ACE2h/CMC, namely BU-1 to BU-6, from the biphenyl furanocoumarin compounds previously synthesized by our team. The binding properties of the screened compounds to ACE2 were investigated by frontier analysis. Cytotoxicity assay, virtual molecular docking assay and pseudo-viral invasion assay were used to examine the affinity and potential antiviral activity of the selected compounds towards ACE2 protein. The virtual molecular docking results showed that BU-1, BU-2 and BU-5 could form significant hydrogen bonds with hotspot amino acid residues on the ACE2 receptor. And BU-1, BU-2 and BU-5 significantly inhibited the ability of SARS-COV-2 pseudovirus to enter ACE2h cells. Therefore, BU-1, BU-2 and BU-5 have the potential to be used as lead compounds for further modification to develop more effective anti-SARS-CoV-2 drugs. Copyright © 2023 The Royal Society of Chemistry.

4.
Neurological Sciences ; 43(Supplement 1):S462-S463, 2022.
Article in English | EMBASE | ID: covidwho-2174295

ABSTRACT

Background: Vaccination campaign to contrast the spread of SARSCoV- 2 has raised the issue of vaccine immunogenicity in frail populations, especially multiple sclerosis (MS) patients on disease modifying therapies (DMTs). Material(s) and Method(s): Before (T0) and after 2 months from booster dose of mRNABNT162b2 vaccine (T1), MS patients under DMTs and healthy donors (HDs) were enrolled. For both T0 and T1, anti-Spike (S) antibody titer as well as IFNg, IL2 and TNFa T cells production upon S peptide libraries stimulation were assessed. According to DMTs mechanism of action, MS patients were stratified into immunosuppressive (such as fingolimod, cladribine, ocrelizumab, and alemtuzumab) and immunomodulating (natalizumab) groups [1]. "Activated" cells were defined as T cells producing any of IFNg or IL2 or TNFa while polyfunctional T cells were defined as those simultaneously producing all 3 cytokines. All possible combinations of intracellular expression of IFNg, IL2, and TNFa in cytokine-producing T cells were evaluated. Result(s): Sixteen MS patients (11 females/5 males, median age [IQR] 41.5 [34.3-48.8] years) and nine HDs (7 females/2 males) were enrolled. An increase of anti-S antibody titers at T1 compared to T0 in bothMSand HDs was seen (1930 [85.75-5895] and 198.5 [80.73-1140] BAU/ml, respectively, p=0.0017;3590 [1575-10850] and 320 [124.1- 662.0], respectively, p=0.0039). Reduced percentage of CD4 and CD8 "activated" and CD4 polyfunctional T cells were observed inMS compared toHDs at T0 (CD4: 1.025 [0.795-1.275] and 1.640 [1.325-2.245], respectively, p=0.0111;CD8 1.0 [0.603-1.328] and 1.65 [1.315-2.360], respectively, p=0.0135;CD4: 0.045 [0.029-0.089] and 0.10 [0.10-0.125], respectively, p=0.0211). Stratifying MS population, only immunomodulating showed an increase in anti-S antibody titers production at T1 (5410 [2655-9893] and 871 [175.3-1360], respectively, p=0.0313), while a reduced production was seen in immunosuppressive compared to immunomodulating and HDs (369.5 [49.8-1975] and 5410 [2655-9893], respectively, p=0.0172;369.5 [49.8-1975] and 3590 [1575-10850], respectively, p=0.0431]. At T0 in immunosuppressive patients a reduced percentage of "activated" CD4 and CD8 T cells was observed when compared to HDs (0.875 [0.658-1.025] and 1.64[1.325-2.245], respectively, p=0.0020;0.91[0.53-1,293] and 1.65[1.315-2.36], respectively, p=0.019). While, at T1 a reduced percentage of CD8 polyfunctional T cells was seen in immunosuppressive patients when compared to HDs (0.036[0.019-0.065] and 0.1[0.048- 0.1291], respectively, p=0.0232). Conclusion(s): Both humoral and T cell specific response to vaccination in MS patients seems to be significantly influenced by different DMTs mechanism. Moreover, a higher percentage of TNFa and a reduced IFNg production was observed, mainly in immunosoppressive group.

5.
Springer Protocols Handbooks ; : 33-42, 2022.
Article in English | EMBASE | ID: covidwho-2173501

ABSTRACT

Expression and purification of turkey coronavirus (TCoV) nucleocapsid (N) protein from a prokaryotic expression system as histidine-tagged fusion protein are presented in this chapter. Expression of histidine-tagged fusion N protein with a molecular mass of 57 kDa is induced with isopropyl beta-D-1-thiogalactopyranoside (IPTG). The expressed N protein inclusion body is extracted and purified by chromatography on nickel-agarose column to near homogeneity. The protein recovery can be 10 mg from 100 ml of bacterial culture. The purified N protein is a superior source of TCoV antigen for antibody-capture ELISA for detection of antibodies to TCoV. Copyright © Springer Science+Business Media New York 2016.

6.
Small ; : e2206153, 2023.
Article in English | PubMed | ID: covidwho-2173458

ABSTRACT

Natural membrane vesicles, including extracellular vesicles and enveloped viruses, participate in various events in vivo. To study and manipulate these events, biomembrane-coated nanoparticles inspired by natural membrane vesicles are developed. Herein, an efficient method is presented to prepare organic-inorganic hybrid materials in high yields that can accommodate various lipid compositions and particle sizes. To demonstrate this method, silica nanoparticles are passed through concentrated lipid layers prepared using density gradient centrifugation, followed by purification, to obtain lipid membrane-coated nanoparticles. Various lipids, including neutral, anionic, and cationic lipids, are used to prepare concentrated lipid layers. Single-particle analysis by imaging flow cytometry determines that silica nanoparticles are uniformly coated with a single lipid bilayer. Moreover, cellular uptake of silica nanoparticles is enhanced when covered with a lipid membrane containing cationic lipids. Finally, cell-free protein expression is applied to embed a membrane protein, namely the Spike protein of severe acute respiratory syndrome coronavirus 2, into the coating of the nanoparticles, with the correct orientation. Therefore, this method can be used to develop organic-inorganic hybrid nanomaterials with an inorganic core and a virus-like coating, serving as carriers for targeted delivery of cargos such as proteins, DNA, and drugs.

7.
The Egyptian journal of immunology ; 30(1):73-86, 2023.
Article in English | EMBASE | ID: covidwho-2168476

ABSTRACT

The worldwide medical systems are still being severely impacted by the coronavirus disease-2019 (COVID-19) pandemic, which is responsible for catastrophic mortality and morbidity. It becomes more and more obvious that this unique respiratory virus's impacts go beyond the respiratory system as time goes on and our comprehension of it deepens. The transmembrane serine protease 2 (TMPRSS2) protein is necessary for the severe acute respiratory syndrome coronavirus 2, which is the cause of COVID-19, to gain cellular entry through the angiotensin-converting enzyme 2 (ACE2) receptor. Most endocrine glands exhibit high levels of expression for ACE2 and TMPRSS2. This pays the attention to the effect of COVID-19 on the endocrine system. Besides its capability to pass to the central nervous system especially the hypothalamus inducing a lot of functional disorders in COVID-19 individuals. Although effective vaccines became widely available, and mortality declined but attention is shifting more and more to the lengthy health impacts on COVID-19 survivors. To inform suitable research and effective management, this review provides an overview of the data examining the impacts of COVID-19 on the endocrine glands besides the hypothalamus. In addition, we reported if the endocrinal and thalamic disorders could affect the incidence and progress of COVID-19. Copyright© by the Egyptian Association of Immunologists.

8.
Chinese Journal of Clinical Pharmacology and Therapeutics ; 27(11):1292-1298, 2022.
Article in Chinese | EMBASE | ID: covidwho-2203682

ABSTRACT

The spread of COVID-19 has greatly threatened human health and economic growth. Angiotensin-converting enzyme 2 (ACE2) is a receptor for severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). By attaching to ACE2, SARS-COV-2 reduces its expression and induces lung injury. Vitamin D can inhibit the progression of COVID-19 by inhibiting the activity of ROCK pathway, up-regulating ACE2 expression and bio-availability, and slowing down the adverse reactions caused by Ang II accumulation. This study explored a novel mechanism, i.e., vitamin D protects against COVID-19-induced injury by upregulating ACE2 expression. It provides theoretical guidance for the role of Vitamin D in the prevention and treatment of COVID-19. Copyright © 2022 Chinese Journal of Clinical Pharmacology and Therapeutics. All rights reserved.

9.
Anatolian Journal of Cardiology ; 26(Supplement 1):S115-S116, 2022.
Article in English | EMBASE | ID: covidwho-2202576

ABSTRACT

Background and Aim: Apart from the direct and immediate invasion of coronavirus to vital tissues such as the heart, the virus is also capable of damaging these tissues based on the host's genetic susceptibility. The present bioinformatic- based study aimed to determine the genes and related microRNAs that most likely to be associated with susceptibility rates for virus-induced cardiovascular vulnerabilities. Method(s): A deep search was scheduled in databases including Pubmed (Medline), Google Scholar, Web of Science, and Scopus databases to assess all microRNAs and targeted genes related to cardiovascular defects induced by the coronavirus. The bioinformatic professional software systems were employed to assess gene-microRNAs interactions and mechanisms involved in cardiovascular injury. Result(s): The coronavirus can induce cardiovascular defects by the three mechanisms of inducing cardiac fibrosis (by up-regulating miR-367-3p and down-regulating hsa-miR- 5692a), inducing hypertension (by up-regulating miR-18b- 5p), and inhibiting microvascular angiogenesis (by up-regulation of miR-18b-5p and down-regulating hsa-miR-5692a). Such processes can be triggered by the effects on NFAT5, CD69, and HGF expression. Conclusion(s): Considering the central role of the revealed microRNAs and their targeted genes in cardiovascular injuries induced by coronavirus, such microRNAs can be applied for finding a way to stabilize the host against virus attacks as well as genetically based treatment for the affected host. (Figure Presented).

10.
Journal for ImmunoTherapy of Cancer ; 10(Supplement 2):A1276, 2022.
Article in English | EMBASE | ID: covidwho-2161958

ABSTRACT

Background Cell-specific transduction remains one of the next frontiers for virally-delivered gene therapy. Efforts to achieve cell-specific transduction have largely been limited to borrowing of preexisting viral glycoproteins and pseudotyping viral surface envelopes to result in altered tropism. VSVG is derived from vesicular stomatitis virus (VSV) and is one of the most commonly used lentiviral (LV) pseudotype glycoproteins as its cognate receptor (LDLR) is present on nearly all dividing and non-dividing cells, enabling broad tropism of VSVG-pseudotyped LVs. Methods Our lab recently developed a receptor-blinded version of VSVG, in which point mutations (K47Q, R354A) of this glycoprotein results in a mutated VSVG with inability to bind and infect LDLR-expressing cells. This mutant viral glycoprotein, which we call VSVGmut, thereby loses its broad tropism, but critically retains its fusogen ability, enabling codisplay of a new LV pseudotype ligand to drive LV tropism. Results Initial experiments displaying high-affinity anti-CD19 scFvs alongside VSVGmut on the LV surface demonstrated infection of CD19+, but not CD19- cells. Subsequent work using endogenous ligands (CD80), Fabs (a-CD3e), cytokines (IL-13), viral glycoproteins (SARS-CoV-2 RBD), and peptide MHCs (pMHCs) revealed the modularity of this platform for achieving potent transduction of on-target cells, with minimal infection of bystander cells, across a range of affinities (pM to uM) and at frequencies as low as 1 in 100,000. This technology allowed for library on library screening of 96 viral pMHC-displaying LVs against a repertoire of >450,000 TCRs in pool, which accurately uncovered EBV- and Flu-specific TCRs through scRNA sequencing. Conclusions The VSVGmut platform has resulted in our lab's ability to pair pMHCs with cognate TCRs and viral surface antigens with cognate BCRs, in addition to achieving lineagespecific transduction of T and B cell subsets, setting the stage for cell-specific gene therapy.

11.
Journal of Pharmaceutical Negative Results ; 13:1082-1087, 2022.
Article in English | EMBASE | ID: covidwho-2156323

ABSTRACT

This study was established to detect some of the immune parameters changes that accompany Patients with SARS-CoV-2 who have varying degrees of illness severity, by targeting three groups of mild, moderate and severe cases of patients, while the fourth group was healthy volunteers as well as unvaccinated and it represents the group we took 20 samples from each of these four groups, and we performed the following tests for them: IL-1beta, IL-17A and IL-18. In mild case: According to the study's findings, there has been a considerable rise, (p<=0.01) in the levels of IL1beta, IL-17A and IL-18 in SARSCoV-2 patients group when compared with healthy group. In moderate case: The results of this study found a high significant increase (p<=0.01) in levels of IL1beta, IL-17A and IL-18 in SARS-CoV-2 patients group when compared with healthy group, healthy group, but IL-17A level lower than in moderate case compared with mild case. In severe case: The results of this study found a high significant increase (p<=0.01) levels of IL1beta, IL-17A and IL-18 in SARS-CoV-2 patients group when compared with healthy group, but IL-17A level lower than in moderate case compared with mild case and moderate case. Copyright © 2022 Wolters Kluwer Medknow Publications. All rights reserved.

12.
Biochemical and Biophysical Research Communications ; 2022.
Article in English | ScienceDirect | ID: covidwho-2149376

ABSTRACT

Several SARS-CoV-2 variants of interest (VOI) have emerged since this virus was first identified as the etiologic agent responsible for COVID-19. Some of these variants have demonstrated differences in both virulence and transmissibility, as well as in evasion of immune responses in hosts vaccinated against the original strain of SARS-CoV-2. There remains a lack of definitive evidence that identifies the genetic elements that are responsible for the differences in transmissibility among these variants. One factor affecting transmissibility is the initial binding of the surface spike protein (SP) of SARS-CoV-2 to human angiotensin converting enzyme-2 (hACE2), the widely accepted receptor for SP. This step in the viral replication process is mediated by the receptor binding domain (RBD) of SP that is located on the surface of the virus. This current study was conducted with the aim of assessing potential differences in binding affinity between recombinant hACE2 and the RBDs of emergent SARS-CoV-2 WHO VOIs. Mutations that affect the binding affinity of SP play a dominant initial role in the infectivity of the virus.

13.
Medical Journal of Malaysia ; 77(Supplement 4):49, 2022.
Article in English | EMBASE | ID: covidwho-2147171

ABSTRACT

Introduction: Increase of cytokines is often associated with inflammatory condition during infections and in autoimmune diseases. A sudden increase of pro-inflammatory cytokines, also known as 'cytokine storm', can cause devastating damage on host cells as well as organs and has been frequently described in the recent COVID-19 infection. Objective(s): This study aimed to evaluate the levels of proinflammatory cytokines (IL-6, IL-8, TNF-alpha and IL-1alpha) among COVID-19 patients in comparison with non-COVID-19 patients in Malaysia. Material(s) and Method(s): We retrospectively analysed the data of samples sent for cytokine panel test (IL-6, IL-8, TNF-alpha and IL-1alpha) at Autoimmune Laboratory, Institute for Medical Research, Malaysia, from September 2021 until April 2022. A total of 58 samples were included in this study and were categorised into two groups: COVID-19 (37 samples) and non-COVID-19 (21 samples). Levels of cytokines were determined by microfluidic immunoassay system. Result(s) and Conclusion(s): Majority of the samples (75.9%) were from patients aged 17 and below, with the overall median age of 8 years. Between the two groups, COVID-19 patients had significantly higher cytokines levels (median IL-6: 25.2pg/ml;IL-8: 64.8pg/ml and TNF-alpha: 28.8pg/ml) compared with patients from non-COVID-19 (median IL-6: 9.5pg/ml;IL-8: 33.9pg/ml and TNF-alpha: 18.9pg/ml). As for IL-1beta, the levels were comparable between the two groups. The results of this study showed that higher levels of proinflammatory cytokines were released during COVID-19 infection as compared with non-COVID-19 cases.

14.
Glycobiology ; 32(11):1043, 2022.
Article in English | EMBASE | ID: covidwho-2135204

ABSTRACT

The COVID-19 pandemic has become a major human health concern. The pathogen responsible for COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), invades its host through the interaction of its spike (S) protein with a host cell receptor, angiotensin-converting enzyme 2 (ACE2). In addition to ACE2, heparan sulfate (HS) on the surface of host cells also plays a significant role as a co-receptor. In this study, SARS-CoV-2 SP receptor binding domain (RBD) wild type (WT), Delta and Omicron variants were expressed in Expi293F cells and used in the kinetic and structural analysis on their interactions with heparin by Surface plasminogen resonance (SPR). Detailed kinetic and structural analysis of the interactions of SARS-CoV-2 SP RBDs with heparin provides important information for designing anti-SARS-CoV-2 molecules.

15.
Research and Practice in Thrombosis and Haemostasis Conference ; 6(Supplement 1), 2022.
Article in English | EMBASE | ID: covidwho-2128293

ABSTRACT

Background: As COVID-19 is associated with a prothrombotic condition and some critically ill patients may even undergo extracorporeal oxygenation treatment, heparin is the essential for treatment in these situations. The prevalence of anti-platelet factor 4 (PF4)/heparin antibodies associated with thrombotic tendency may be present in patients hospitalized for COVID-19 without heparin therapy. Lupus anticoagulant (LA) included in the diagnostic criteria for antiphospholipid syndrome, which is one of the common causes of thrombophilia, is also commonly detected during SARS-CoV- 2 infection. Most patients hospitalized for COVID-19 showed elevated D-dimer and prolonged activated partial thromboplastin time (aPTT), However, there were no studies on the association of SARS-CoV- 2 infection with LA and anti-PF4/ heparin antibodies. Aim(s): The purpose of this study was to analyze the expression of LA and anti-PF4/ heparin antibodies associated with thrombotic tendency in COVID-19 patients. Method(s): We performed LA test (Instrumentation Laboratory, Bedford, MA) and LIFECODES PF4 IgG assay (Immucor, Norcross, GA) on 46 COVID-19 patients admitted to Asan Medical Center and analyzed their frequency. Result(s): Of a total of 46 COVID-19 patients, 26 patients (56.5%) were positive for LA test, 3 patients (6.5%) for anti-PF4/ heparin antibodies. In particular, anti-PF4/ heparin antibodies was detected only in LA-negative patients and showed low optical density values (3 out of 20 LA-negative patients, 15.0%). All three patients positive for anti-PF4/ heparin antibodies had no history of heparin treatment. Conclusion(s): In COVID-19- patients, anti-PF4/ heparin antibody test does not predict clinically relevant HIT antibodies. Anti-PF4/ heparin antibodies appear in LA-negative COVID-19 patients, so they are carefully expected to serve as LA-independent indicators. (Table Presented).

16.
Research and Practice in Thrombosis and Haemostasis Conference ; 6(Supplement 1), 2022.
Article in English | EMBASE | ID: covidwho-2128279

ABSTRACT

Background: SARS-CoV- 2 tropism for the ACE2 receptor, along with the multifaceted inflammatory reaction, is likely to drive the generalized hypercoagulable state seen in patients with COVID-19. Aim(s): According to our previously published bioinformatics analysis, we aimed to analyze the diagnostic and predictive utility of miRNAs regulating ACE2 network (miR-26b- 5p, miR-10b- 5p, miR-302c- 5p, hsa-miR- 200b- 3p, hsa-miR- 124- 3p) in patients with COVID-19 Methods: We determined the expressions of ACE2-related- miRNAs in 79 hospitalized COVID-19 patients and 32 healthy volunteers by PCR and monitored miRNAs patterns during the acute phase of COVID-19, as well as the prognostic potential of these miRNAs as biomarkers. Result(s): The expression levels of miR-26b- 5p in COVID-19 patients were found lower at the baseline, 7 and 21-days after admission. compared to the healthy controls (p < 0.0001 for all time points). Similarly, miR-10b- 5p expression levels were lower at the baseline and 21-days post admission in COVID-19 patients when compared to the healthy individuals (p = 0.001 in both time points). Moreover, expression levels of this miRNA were higher 7-days post-admission when compared to the baseline (p = 0.003). According to the ROC curve analysis, low miR-200b- 3p expression presents predictive utility in assessment of the hospital length of stay and/or death (AUC:0.730, p = 0.002). According to the multivariable logistic regression model, low delta miR-200p expression, together with diabetes mellitus (DM), are independent predictors of increased hospital length of stay and/or death (OR: 5.775;95% CI, 1.572-21.214;p = 0.008 and OR: 4.888;95% CI, 1.001-23.858;p = 0.050, respectively). Conclusion(s): This study enabled us to better characterize changes in gene expression and signaling pathways related to COVID-19 thrombosis. In this study we identified, characterized and validated miRNAs which could serve as novel, thrombosis-related biomarkers of the COVID-19, can be used for early stratification of patients and prediction of severity of infection development in an individual. (Figure Presented).

17.
Research and Practice in Thrombosis and Haemostasis Conference ; 6(Supplement 1), 2022.
Article in English | EMBASE | ID: covidwho-2128198

ABSTRACT

Background: Platelets are transfused therapeutically for hemostasis, and are an integral part of hemorrhage management. However, transfusions can be ineffective in the most severe cases of hemorrhage. Platelets are also a potential cell therapy in other applications, but development has been hindered by inadequate methods to control which proteins are expressed by platelets. Currently, there are no methods to express exogenous proteins in transfusable platelets, which would expand their use to help treat the diseases they modulate. A method is therefore needed to modify transfusable platelets, and thus enhance their protein composition for specific applications. Aim(s): To produce engineered, transfusable platelets to enhance their natural coagulability and functional repertoire by directly transfecting donor-derived platelets with mRNA via lipid nanoparticle (LNP)-mediated delivery. The recent advances through the COVID-19 mRNA vaccines demonstrates the clinical safety and efficacy of LNP-mediated gene therapy, and thus offers a promising strategy to effectively engineer modified platelets. Method(s): Donor-derived platelets were washed and subsequently incubated with a systematic array of LNPs encapsulating Cy5-labeled mRNA encoding for nanoluciferase in comparison to commercial transfection reagents. LNP uptake and platelet activation via CD62p levels was assessed following 4 hours by flow cytometry, while luciferase expression was assessed by normalizing the luminescence intensity to the total protein content. Result(s): Platelets took up the mRNA through all conditions tested;nanoluciferase was only expressed, however, in platelets treated with LNPs and not commercial reagents. Systematically optimizing LNPs increased nanoluciferase expression nine-fold relative to pre-optimized LNPs. Exogenous protein expression did not appear to correlate with mRNA uptake nor platelet activation. Conclusion(s): Platelets transfected with LNPs can express exogenous protein. Further optimization can eventually lead to the creation of a platform technology that in the long-term will allow platelets to deliver therapeutic proteins and yield more effective platelet products.

18.
Research and Practice in Thrombosis and Haemostasis Conference ; 6(Supplement 1), 2022.
Article in English | EMBASE | ID: covidwho-2128134

ABSTRACT

Background: Blood product in therapeutic transfusion are now commonly acknowledged to present biologically active constituents during processes of preparation. In the midst of worldwide COVI-19 pandemic, preliminary evidence, suggest that convalescent plasma may lessen the severity of COVID-19, particularly concerning patients with profound B-cell lymphopenia and prolonged COVID-19 symptoms. Aim(s): This study examined the influence of photochemical pathogen reduction treatment (PRT) using amotosalen-HCl and UVA light vs untreated control convalescent plasma (n = 72 -paired samples) -cFFP. Method(s): This study investigated the soluble inflammatory factors: SCD40L, IFN-alpha, IFN-beta, IFN-gamma, IL-1 beta, IL-6, IL-8, IL-10, IL-18, TNF-alpha and ex-vivo inflammatory bioactivity on endothelial cells. Result(s): We observed that IL-8 concentrations were significantly decreased in cFFP w PRT, whereas IL-18 concentration was increased. We observed after activation with cFFP w PRT and w/o PRT no significant modulation of IL-6 released by endothelial cells. CD54 and CD31 expression in the presence of cFFP (w or w/o PRT) is close to negative controls, even if CD54 and CD31 were significant decreased in presence of cFFP w vs w/o PRT. Conclusion(s): It appears valuable to carry on investigations, of IL-18 and IL-8, on both the physiopathology of PRT convalescent plasma treated and post marketing clinical trials. Further research, including a careful clinical evaluation of CCP-treated patients, will be required to further define the clinical relevance of these findings.

19.
Research and Practice in Thrombosis and Haemostasis Conference ; 6(Supplement 1), 2022.
Article in English | EMBASE | ID: covidwho-2128124

ABSTRACT

Background: Thrombotic thrombocytopenic purpura (TTP) has occasionally been described after vaccination. Since the availability of anti-SARS- CoV- 2 vaccines, 12 cases have been described on a possible association with TTP onset. Aim(s): This study aims to evaluate the relapse rates in patients affected by TTP undergoing anti-SARS- CoV- 2 vaccination. Method(s): All consecutive TTP patients undergoing anti-SARS- CoV- 2 vaccination from March to May 2021 were enrolled. Blood samples were collected before vaccination (T0), 2 weeks after the first (T1) and the second dose (T2) to evaluate ADAMTS13 activity and anti-ADAMTS13 antibody titer. Result(s): A total of 49 TTP patients were enrolled (48 acquired and 1 congenital), all vaccinated with an mRNA vaccine. No patients had a clinical TTP relapse, with an ADAMTS13 relapse rate of 1.36% per month. Mean levels of ADAMTS13 activity were stable among the three timepoints (Figure). In only two patients a significant drop in ADAMTS13 levels occurred after the first dose (from 28% to <3% and from 101% to 82%), and both remained stable after the second dose, with negative anti-ADAMTS13 antibodies. Due to a stable undetectable ADAMTS13, the first patient was treated with 4 doses of weekly 375 mg/m2 rituximab with a rapid ADAMTS13 response. One patient had positive basal anti-ADAMTS13 antibodies with a titer remaining stable after the two vaccine doses, while in another patient anti-ADAMTS13 antibodies became detectable after the first dose, with no corresponding drop in ADAMTS13 levels and a stable titer after the second dose. Conclusion(s): The result of our study prospectively evaluating the effect of anti-SARS- CoV- 2 vaccination on the risk of relapse in a large cohort of patients with TTP in Milan showed a lower than reported relapse rate (1.36% vs 2.6%) with an observed to expected incidence rate ratio of 0.52, confirming the safety of mRNA-based anti-SARS- CoV- 2 vaccination in TTP patients.

20.
Research and Practice in Thrombosis and Haemostasis Conference ; 6(Supplement 1), 2022.
Article in English | EMBASE | ID: covidwho-2128119

ABSTRACT

Background: Venovenous extracorporeal membrane oxygenation (ECMO) is a cornerstone in the management of severe acute respiratory distress syndrome (ARDS) and causes hemostatic system activation and inflammation. COVID-19 is known to cause thromboinflammation. Elucidation of the underlying pathomechanisms is of great importance. Aim(s): To evaluate markers of NET formation in COVID-19 and non-COVID- 19 associated ARDS and ECMO and to explore the role of different NET parameters as markers of inflammation and coagulopathy. Method(s): We studied 31 adult COVID-19 patients and 23 adult non-COVID- 19 patients with severe ARDS requiring ECMO and 47 sex-and age-matched healthy controls. Blood was collected at time point A (ECMO day 0-4) and at time point B (ECMO day 7-17). Citrullinated histone H3 (citH3), cell-free DNA (cfDNA), and plasma myeloperoxidase DNA (mpoDNA), as well as d-Dimer, were evaluated. Values are given as median (25th, 75th percentile). Statistical testing was performed using unpaired t-tests of logarithmized parameters. Result(s): COVID-19 and non-COVID- 19 patients exhibited significantly higher levels of citH3, cfDNA, and mpoDNA than healthy controls (Table 1). Levels of citH3 decreased significantly from time point A to B in COVID-19 patients. No comparable effect was identified for cfDNA and mpoDNA (Table 1). In non-COVID- 19 patients, no differences in levels of citH3, cfDNA, and mpoDNA were found between timepoints A and B (Table 1). d-Dimer increased from time point A to timepoint B in both groups (Table 1). Conclusion(s): NET formation is comparably increased in patients on ECMO because of COVID-19 and non-COVID- 19 ARDS. Markers of NET formation could add information on immunothrombosis and the complex interplay of coagulation and inflammation in the context of ECMO.

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