COVID-19-associated pulmonary aspergillosis (CAPA) is a frequent and severe complication among critically ill patients with COVID-19. The absence of a clear diagnostic gold standard and the multiplicity of proposed definitions (EORTC/MSG, AspICU, and ISHAM) complicate its diagnosis. This study aimed to compare the performance of the EORTC/MSG, AspICU, and ISHAM classifications in diagnosing CAPA among mechanically ventilated COVID-19 patients and to assess their correlations with clinical outcomes. We conducted a retrospective, monocentric study including all adult COVID-19 patients requiring invasive mechanical ventilation admitted to the ICUs of CHU-Charleroi Chimay between March 2020 and December 2021. Patients were classified according to EORTC/MSG, AspICU, and ISHAM criteria. Demographics, comorbidities, management, and outcomes were compared across groups. In total, 405 patients were included during the four waves. The incidence of probable or possible CAPA varied widely: 6.1% with EORTC/MSG, 9.7% with AspICU, and 15.1% with ISHAM criteria. ICU mortality reached approximately 76% among patients with probable CAPA versus 43% in patients without aspergillosis. The frequency of CAPA diagnosis increased across COVID-19 waves, possibly correlating with changes in management, particularly corticosteroid use. The choice of CAPA diagnostic criteria has a major impact on incidence estimates and patient management. Prospective validation of CAPA definitions, integrating multiple mycological criteria, is urgently needed to guide clinical decision-making and antifungal therapy.

OBJECTIVES:During the SARS-CoV-2 pandemic, there has been significant increased use of vvECMO as rescue therapy. Patients with COVID-19 as anticoagulation is needed for vvECMO support, may develop bleeding complications requiring an increased number of RBC transfusions. We would like to report the RBC transfusion needs following the implementation of an ECMO program. Data on blood usage in this population is important in view of the decline in donations due to the pandemic. STUDY DESIGN AND METHODS:We analyzed data on RBC transfusions in patients who required vvECMO for COVID-19 related ARDS in a Belgian ICU from March 2020 to March 2022. The primary end point was RBC transfusion requirements. and the relationship to outcome. We also analyzed the evolution of this requirement during the four waves. RESULTS:We admitted 538 patients for hypoxemic ARDS due to COVID-19. Sixty patients (11%) required vvECMO, of whom 27 (45%) died. Forty-seven (78%) of the vvECMO patients were transfused a total of 403 packed RBC units. Sites of hemorrhagic complications were ECMO cannulation sites and lungs. RBC transfusion per patient per day on vvECMO was 0.50 [0.30-0.67] units. There were no differences in hemorrhagic complications in vvECMO survivors and non-survivors. The percentage of vvECMO patients receiving a RBC transfusion increased slightly during the last COVID-19 wave, with 92 % of patients transfused. CONCLUSIONS:vvECMO program is associated with a major need for RBC transfusions. These data are important when blood availability is decreased due to a pandemic and illustrates the need for studies on optimizing blood management including therapeutic anticoagulation target, threshold for RBC transfusion or alternatives to RBC transfusion.

ABSTRACT:Background : Diabetic ketoacidosis (DKA) is a life-threatening emergency. Microvascular hyporeactivity has been reported in these patients and is completely reversible when acidosis is corrected with aggressive treatment. The shape of the red blood cell (RBC), a sensor of local hypoxia and a component of the microcirculation, is altered in diabetic patients, but no data are available concerning RBC deformability in DKA during treatment. Methods : In this prospective observational study, we included all adult patients admitted with DKA to a 32-bed medico-surgical intensive care unit (ICU) over a 6-month period. We excluded patients with infection. We measured RBC deformability in the DKA patients and compared results with those from patients with type 1 diabetes and from a group of healthy volunteers (HV). RBC deformability was assessed using ektacytometry. In the DKA patients, it was assessed at ICU admission, 8 and 24 h after admission, and prior to ICU discharge (48-72 h). The RBC elongation index (EI) was determined based on the laser diffraction pattern changes. A higher EI indicates greater RBC deformability. Results : A total of 46 diabetic patients (15 DKA and 31 type 1 diabetes patients) and 20 HV were included. RBC deformability was more altered at ICU admission in DKA patients, with significantly lower EI values than in the other groups, and these alterations persisted during the ICU stay despite treatment. There were no correlations between these alterations and the quantity of fluids or insulin received. Conclusions : In contrast to the reversible microvascular hyporeactivity observed in DKA, RBC deformability was already altered at ICU admission in patients with DKA and remained altered despite treatment. These alterations may contribute to the blood flow abnormalities observed in these patients.

Introduction Severe hypoxemia is the leading cause of admission in intensive care (ICU) in patients with COVID-19 related acute respiratory distress syndrome (ARDS). In these patients, several studies reported a left shift of the oxyhemoglobin dissociation curve associated with a lower mortality. However, these results are conflicting, as these studies include few patients and often no control groups. Moreover, the calculation of P50, representing the PaO2 value at which 50% of hemoglobin is saturated, is not corrected for factors known to influence it (pH, PaCO2 or temperature). For all of these reasons, we compared the corrected P50 between ICU patients with severe COVID-19 related ARDS on mechanical ventilation or not, and ARDS from other causes. We investigated the evolution of the corrected P50 during the first 3 days of ICU and its relationship with ICU mortality. Methods and Patients We retrospectively calculated the corrected P50 in three groups of patients: intubated and mechanically ventilated COVID-19 related ARDS, non-intubated COVID-19 related ARDS and intubated patients with ARDS due to other infectious causes. The corrected P50 was calculated, on the worst blood gas analysis on days 1 and 3 of ICU admission, by the formula of Hill but modified by Dash et al., controlled for pH, PaCO2 and temperature. We collected ICU mortality. Results 463 blood gas analysis at days 1 and 3 from 214 ICU COVID-19 related ARDS patients (114 with intubation and 100 without) and 35 ICU patients with ARDS from other causes were analyzed. All patients were severely hypoxemic: PaO2/FiO2 of 76 [58-108] mmHg for intubated COVID-19, 79 [60-108] mmHg for non-intubated COVID-19 and 142 [78-197] mmHg for the third group (p < 0.001). The mortality rate was higher in intubated COVID-19 related ARDS patients (44.7 versus 14 versus 37% in ARDS from other causes; p < 0.001). The corrected P50 was significantly lower in COVID-19 patients, especially in non- intubated patients (21.2 [18.8-25.2] mmHg vs. 25.5 [19.2-30.3] mmHg in intubated patients; compared to ARDS from other causes: 27.2 [23.3-35.4] mmHg; p < 0.001. The corrected P50 does not change over the first 3 days, except for the non intubated COVID-19 related ARDS and is not correlated with ICU mortality (odds ratio = 0.98 [0.95-1.03]; p = 0.51), in contrast of PaO2/FiO2 and ICU gravity scores. Conclusion The oxyhemoglobin dissociation curve at ICU admission was left shifting in severe COVID-19 related ARDS patients regardless of the type of ventilation. This deviation increases the third day only in non-intubated COVID-19 related ARDS and was not related to the outcome.

Introduction: Red blood cells (RBC) are one of the key elements of the microcirculation. Their ability to pass through capillaries and to deliver oxygen to cells is due to their large degree of deformability linked to the characteristics of the RBC membrane. Alterations in RBC deformability as a result of membrane damage, linked in part to increased synthesis of reactive oxygen species (ROS), can be observed in several diseases, such as sepsis, and may contribute to the altered microcirculation observed in these pathologies. Hyperbaric oxygen therapy (HBOT), with inhalation of 100 % oxygen, has been proposed in several acute or chronic pathologies, including carbon monoxide poisoning. Objective: We investigated the effects of HBOT on oxidative stress from ROS produced by myeloperoxidase (MPO) and on RBC deformability in patients with acute or chronic inflammation (n = 10), in patients with acute carbon monoxide poisoning (n = 10), and in healthy volunteers (n = 10). Methods: RBC deformability was evaluated before and after HBOT in the various populations using the ektacytometry technique (Laser-assisted Optical Rotational Red Cell Analyzer - LORRCA). Deformability was determined by the elongation index (EI) in relation to the shear stress (SS) over a range of 0.3 to 50 Pa. Oxidative stress was estimated through changes in proteins (chlorotyrosine and homocitrulline) induced by MPO activity measured by liquid chromatography-tandem mass spectrometry analysis. Results: Before HBOT, EI was significantly lower in patients with acute or chronic inflammation than in healthy volunteers and patients with acute carbon monoxide poisoning for the majority of SS values studied. After one session of HBOT, the EI was significantly higher than before HBOT for SS values of 1.93 Pa or higher in patients with acute or chronic inflammation. This effect remains constant after 10 sessions. There were no differences before and after HBOT in protein or amino acid oxidation due to ROS generation mediated by MPO in the three populations. Conclusions: Our results confirm altered RBC deformability in patients with acute and chronic conditions associated with an underlying inflammatory process. HBOT improves deformability only after one session and therefore may improve microcirculation in this population. According to our results, this improvement does not seem mediated by the ROS pathway via MPO. These results need to be confirmed in a larger population.

Guidelines for acute respiratory distress syndrome (ARDS) management include sedation, neuromuscular blockade for protective ventilation and the need for the prone position. This technique can lead to neurologic sequelae like the Tapia syndrome following the patient position. We reported two ARDS COVD-19 patients with Tapia syndrome due to prone position. Both patients, a white European 57 year-old female and a Black African 50 year-old male, were intensive care unit (ICU) admitted for ARDS due to COVID-19. They were orotracheal intubated and mechanical ventilated for respectively 23 and 68 days with 7 and 4 sessions of prone position. Both were tracheotomized during their ICU stay. We observed during the respiratory weaning, swallowing difficulties associated with a tongue deviation during protusion and paralysis of the ipsilateral vocal cord suggesting a Tapia syndrome. Prone position for the treatment of hypoxic ARDS COVID-19 patients with lateral flexion of the head could induce a compression on the lower cranial nerves and their collateral branches and lead to the Tapia syndrome with long neurologic sequelae: swallowing disorder, dysarthria and dysphagia due to difficulty to move the tongue with paralysis of the vocal cord. Because mechanical ventilation on prone position is a cornerstone of the treatment of hypoxic COVID-19 patients, the number of patients with Tapia syndrome may increase in the future.

Inflammatory processes are common in intensive care (ICU) patients and can induce multiple changes in metabolism, leading to increased risks of morbidity and mortality. Metabolomics enables these modifications to be studied and identifies a patient's metabolic profile. The objective is to precise if the use of metabolomics at ICU admission can help in prognostication. This is a prospective ex-vivo study, realized in a university laboratory and a medico-surgical ICU. Metabolic profiles were analyzed by proton nuclear magnetic resonance. Using multivariable analysis, we compared metabolic profiles of volunteers and ICU patients divided into predefined subgroups: sepsis, septic shock, other shock and ICU controls. We also assessed possible correlations between metabolites and mortality. One hundred and eleven patients were included within 24 h of ICU admission, and 19 healthy volunteers. The ICU mortality rate was 15%. Metabolic profiles were different in ICU patients compared to healthy volunteers (p < 0.001). Among the ICU patients, only the subgroup of patients with septic shock had significant differences compared to the ICU control patients in several metabolites: pyruvate, lactate, carnitine, phenylalanine, urea, creatine, creatinine and myo-inositol. However, there was no correlation between these metabolite profiles and mortality. On the first day of ICU admission, we observed changes in some metabolic products in patients with septic shock, suggesting increased anaerobic glycolysis, proteolysis, lipolysis and gluconeogenesis. These changes were not correlated with prognosis.

OBJECTIVES: RBCs from critically ill patients have depressed deformability, especially in sepsis. Prolonged exposure of RBCs from healthy volunteers to physiologic shear stress (the preconditioning technique) has been associated with improved deformability, but the effect of preconditioning on RBCs from critically ill patients with or without sepsis has never been studied. DESIGN: Prospective study. SETTING: A 32-bed medico-surgical ICU and a university-affiliated cell biology laboratory. SUBJECTS: RBCs from 26 healthy volunteers and 40 critically ill patients (20 with and 20 without sepsis). INTERVENTIONS: RBC deformability was measured using the elongation index (EI) with an ektacytometer, at shear stress levels ranging from 0.3 to 50 Pa. To assess the effects of preconditioning in the three groups, we measured EI after first applying a shear stress of 5 Pa for 300 seconds. To study the potential mechanisms involved in preconditioning, we looked at deformability after incubation of an RBC solution from the healthy volunteers with glutaraldehyde, a membrane-stabilizing protein, and neuraminidase, an enzyme that releases membrane sialic acid. MEASUREMENTS AND MAIN RESULTS: Baseline RBC deformability was significantly depressed in the septic patients compared with the volunteers at all shear stress levels greater than or equal to 4.89 Pa. Preconditioning improved deformability only in the volunteers (at shear stress levels of 0.48 and 0.76 Pa). Among the critically ill patients, preconditioning worsened RBC deformability at higher shear stress levels. After incubation (with glutaraldehyde or neuraminidase) of RBCs from five volunteers in whom preconditioning had significantly improved deformability, the positive effect of preconditioning was lost with glutaraldehyde. CONCLUSIONS: RBC deformability is depressed in septic patients. There was a deleterious effect of preconditioning on RBC deformability in septic patients, unlike the positive effect on RBCs from healthy volunteers. The effect of preconditioning may be associated with elasticity of the cell membrane.

Introduction: Red blood cells (RBC) are one of the key elements of the microcirculation. Their ability to pass through capillaries and to deliver oxygen to cells is due to their large degree of deformability linked to the characteristics of the RBC membrane. Alterations in RBC deformability as a result of membrane damage, linked in part to increased synthesis of reactive oxygen species (ROS), can be observed in several diseases, such as sepsis, and may contribute to the altered microcirculation observed in these pathologies. Hyperbaric oxygen therapy (HBOT), with inhalation of 100% oxygen, has been proposed in several acute or chronic pathologies, including carbon monoxide poisoning. Objective: We investigated the effects of HBOT on oxidative stress from ROS produced by myeloperoxidase (MPO) and on RBC deformability in patients with acute or chronic inflammation (n=10), in patients with acute carbon monoxide poisoning (n=10) , and in healthy volunteers (n=10).Methods: RBC deformability was evaluated before and after HBOT in the various populations using the ektacytometry technique (Laser-assisted Optical Rotational Red Cell Analyzer – LORRCA). Deformability was determined by the elongation index (EI) in relation to the shear stress (SS) over a range of 0.3 to 50 Pa. Oxidative stress was estimated through changes in proteins (chlorotyrosine and homocitrulline) induced by MPO activity measured by liquid chromatography-tandem mass spectrometry analysis.Results: Before HBOT, EI was significantly lower in patients with acute or chronic inflammation than in healthy volunteers and patients with acute carbon monoxide poisoning for the majority of SS values studied. After one session of HBOT, the EI was significantly higher than before HBOT for SS values of 1.93 Pa or higher in patients with acute or chronic inflammation. This effect remains constant after 10 sessions. There were no differences before and after HBOT in protein or amino acid oxidation due to ROS generation mediated by MPO in the three populations. Conclusions: Our results confirm altered RBC deformability in patients with acute and chronic conditions associated with an underlying inflammatory process. HBOT improves deformability only after one session and therefore may improve microcirculation in this population. According to our results, this improvement does not seem mediated by the ROS pathway via MPO. These results needs to be confirmed in a larger population.

BackgroundDuring sepsis, red blood cell (RBC) deformability is altered. Persistence of these alterations is associated with poor outcome. Activation of the complement system is enhanced during sepsis and RBCs are protected by membrane surface proteins like CD35, CD55 and CD59. In malaria characterized by severe anemia, a study reported links between the modifications of the expression of these RBCs membrane proteins and erythrophagocytosis. We studied the evolution of RBCs deformability and the expression of RBC membrane surface IgG and regulatory proteins in septic patients.MethodsBy flow cytometry technics, we measured at ICU admission and at day 3–5, the RBC membrane expression of IgG and complement proteins (CD35, 55, 59) in septic patients compared to RBCs from healthy volunteers. Results were expressed in percentage of RBCs positive for the protein. RBC shape was assessed using Pearson's second coefficient of dissymmetry (PCD) on the histogram obtained with a flow cytometer technique. A null value represents a perfect spherical shape. RBC deformability was determined using ektacytometry by the elongation index in relation to the shear stress (0.3–50 Pa) applied to the RBC membrane. A higher elongation index indicates greater RBC deformability.ResultsRBCs from 11 septic patients were compared to RBCs from 21 volunteers. At ICU admission, RBCs from septic patients were significantly more spherical and RBC deformability was significantly lower in septic patients for all shear stress ≥1.93 Pa. These alterations of shape and deformability persists at day 3–5. We observed a significant decrease at ICU admission only in CD35 expression on RBCs from septic patients. This low expression remained at day 3–5.ConclusionsWe observed in RBCs from septic patients a rapid decrease expression of CD35 membrane protein protecting against complement activation. These modifications associated with altered RBC deformability and shape could facilitate erythrophagocytosis, contributing to anemia observed in sepsis. Other studies with a large number of patients and assessment of erythrophagocytosis were needed to confirm these preliminary data.

BackgroundAcute respiratory distress syndrome due to coronavirus disease 2019 (COVID-19) is associated with high mortality. Several studies have reported that the microcirculation responds adequately to hypoxia in COVID-19 patients by increasing oxygen availability, in contrast to the inadequate response observed in patients with bacterial sepsis. Red blood cells (RBCs), the key cells for oxygen transport, and notably their rheology, are altered during bacterial sepsis, but few data are available in patients with COVID-19.MethodsIn this prospective, non-interventional study, shape was assessed on admission (or inclusion for the volunteers) using Pearson’s second coefficient of dissymmetry (PCD) on the histogram obtained with a flow cytometer technique. A null value represents a perfect spherical shape. RBC deformability was determined using ektacytometry by the elongation index in relation to the shear stress (0.3 to 50 Pa) applied to the RBC membrane. A higher elongation index indicates greater RBC deformability. Results were compared across groups. Scanning electronic microscopy was performed on RBCs from COVID-19 patients. RBC shape and deformability were also assessed on days 3 and 7 in COVID-19 patients.ResultsForty-nine ICU patients were included (30 with COVID-19 ARDS and 19 with bacterial sepsis). ARDS was more severe in patients with COVID-19 than in those with sepsis (PaO2/FiO2 99 [73–154] vs. 270 [239–295] mmHg p < 0.001) and mechanical ventilation was more frequently required (87 vs. 21%; p < 0.001). Mortality was significantly higher in COVID-19 patients (15/30 [50%] vs. 4/19 [21%], p = 0.046). RBCs were significantly more spherical in septic patients (PCD −0.40 [−0.56; −0.18]) than in healthy volunteers (PCD −0.54 [−0.66; −0.49]) but not than in COVID-19 patients (−0.48 [−0.55; −0.43]). In COVID-19 non-survivors (n = 11), sphericity was more marked on day 7 (PCD −0.40 [−0.47; −0.28]) than on day 1 (PCD vs. −0.49 [−0.59; −0.44]); p = 0.045. At ICU admission, RBC deformability was altered for all shear stress values studied in septic patients compared to COVID-19 patients and healthy volunteers (maximum elongation index for septic patients: 0.600 [0.594–0.630] vs. 0.646 [0.637–0.653] for COVID-19 patients and 0.640 [0.635–0.650] for healthy volunteers; both p < 0.001). In the 18 COVID-19 patients studied for 7 days, RBC deformability did not change over time and was not related to outcome. At day 1, RBCs from COVID-19 patients showed a normal structure on scanning electronic microscopy.ConclusionIn contrast to the significantly altered shape and decreased deformability in patients with bacterial sepsis, RBCs from severely hypoxemic COVID-19 patients had normal deformability on admission, and this pattern did not change over the first week despite a more spherical shape in non-survivors. As RBCs are the key cell for oxygen transport, this maintenance of normal deformability may contribute to the adequate microcirculatory response to severe hypoxia of the microcirculation that has been observed in these patients.

Data were obtained at ICU admission and are presented as n (%) or median [IQR

BACKGROUND:Consensus for transfusion in intensive care unit (ICU) patients recommends a restrictive strategy, based on a hemoglobin (Hb) concentration of 7 g/dL. Red blood cell (RBC) transfusion is used to prevent tissue hypoxia by improving oxygen transport (DO2 ) and therefore oxygen consumption (VO2 ). We studied the effects of RBC transfusion on systemic oxygenation parameters reflecting systemic oxygen extraction (EO2 = DO2 /VO2 ): S(c)vO2 , lactate level, venous-to-arterial carbon dioxide difference (Pv-aCO2 ), and cardiac index/EO2 (CI/EO2 ) and evaluated their usefulness in guiding transfusion decisions in ICU patients. STUDY DESIGN AND METHODS:Prospectively, all adult patients transfused were included except those with active bleeding or without a jugular or subclavian catheter. We measured O2 parameters before and after transfusion. Patients were a priori grouped according to their initial S(c)vO2 (< or ≥70%), treatment with vasopressors, cardiac function, and septic status. RESULTS:A total of 62 patients received 105 RBC transfusions. For all, mean arterial pressure (77 [69-88] to 81 [73-91] mm Hg), Hb concentration (7.4 [7.0-7.8] to 8.4 [7.7-8.9] g/dL) and S(c)vO2 (65% [59%-73%] to 69% [62%-75%]) increased after transfusion (all P < .001). S(c)vO2 improved after transfusion only when initial S(c)vO2 was less than 70% (62% [56%-65%] to 66% [61%-71%]; P < .001). In this group, Pv-aCO2 , lactate concentrations, and CI/EO2 did not change after transfusion. Cardiac function, sepsis, or vasopressor therapy did not affect these results. CONCLUSIONS:Among systemic O2 parameters, only a S(c)vO2 < 70% in anemic ICU patients improves after transfusion. As S(c)vO2 can reflect a DO2 /VO2 imbalance, it could be helpful when combined with the Hb concentration to decide whether to transfuse. However, the benefit on outcome should be further studied.

It is now reported that coronavirus disease 2019 ICU patients are at increased risk of thrombosis. Expert opinion and scientific societies recommend a higher dose of low-molecular-weight heparin, but definitive data is lacking. We report our adapted thromboprophylaxis practice of low-molecular-weight heparin administration in coronavirus disease 2019 ICU patients. One-hundred six measurements in 19 ICU patients were collected. Despite enoxaparin 60 mg once daily, only two measurements of the trough anti-Xa were in the upper end of prophylactic range. Anti-Xa activity peaks increased significantly after administration, but all measurements were under the optimal prophylactic ranges. Despite an adapted protocol, three of the 19 patients (16%) developed venous thromboembolism. We show in coronavirus disease 2019 ICU patients, despite higher prophylactic low-molecular-weight heparin administration due to body mass index, anti-Xa activity was well below peak serum levels in our cohort of critically ill coronavirus disease 2019 patients. This evaluation suggests the need for rapid studies on adequate thromboprophylaxis in these patients.

Purpose: To investigate age-related differences in outcomes of critically ill patients with sepsis around the world. Methods: We performed a secondary analysis of data from the prospective ICON audit, in which all adult ( >16 years ) patients admitted to participating ICUs between May 8 and 18, 2012, were included, except admissions for routine postoperative observation. For this sub-analysis, the 10,012 patients with completed age data were included. They were divided into five age groups - <= 50, 51-60, 61-70, 71-80, >80 years. Sepsis was defined as infection plus at least one organ failure. Results: A total of 2963 patients had sepsis, with similar proportions across the age groups (<= 50 = 25.2%: 51-60 = 30.3%; 61-70 = 32.8%; 71-80 = 30.7%; >80 = 30.9%). Hospital mortality increased with age and in patients >80 years was almost twice that of patients <= 50 years (493% vs 25.2%, p < .05). The maximum rate of increase in mortality was about 0.75% per year, occurring between the ages of 71 and 77 years. In multilevel analysis, age > 70 years was independently associated with increased risk of dying. Conclusions: The odds for death in ICU patients with sepsis increased with age with the maximal rate of increase occurring between the ages of 71 and 77 years. (C) 2019 Elsevier Inc. All rights reserved.

BACKGROUND AND AIMS:Endothelial cells are main actors in vascular homeostasis as they regulate vascular pressure and permeability as well as hemostasis and inflammation. Disturbed stimuli delivered to and by endothelial cells correlate with the so-called endothelial dysfunction and disrupt this homeostasis. As constituents of the inner layer of blood vessels, endothelial cells are also involved in angiogenesis. Apolipoprotein Ls (APOL) comprise a family of newly discovered apolipoproteins with yet poorly understood function, and are suggested to be involved in inflammatory processes and cell death mechanisms. Here we investigate the role of APOLs in endothelial cells stimulated with factors known to be involved in atherogenesis and their possible contribution to endothelial dysfunction with an emphasis on inflammation driven-angiogenesis in vitro.METHODS:Using the CRISPR/Cas9 technique, we analyzed the effect of APOL3 gene knock out in HMEC-1 endothelial cells on cell migration, tubulogenesis, endothelial permeability, intracellular signal transduction as assessed by kinase phosphorylation, and angiogenesis gene expression (measured by qRT-PCR).RESULTS:Our results indicate that among the family, APOL3 was the only member induced by myeloperoxidase, oxidized LDL, VEGF and FGF treatments. APOL3 invalidation increased endothelial permeability, reduced wound repair and tubule formation in vitro, the latter only in MPO and VEGF-induced conditions. Accordingly, some pro-angiogenic signaling pathways (ERK1/2 and FAK but not Akt) and some pro-angiogenic genes were partially inhibited in APOL3 knock out cells.CONCLUSIONS:These findings suggest the involvement of APOL3 in angiogenesis in vitro and as a modulator of MAPK and FAK signaling in endothelial cells.