Coagulation Potential Research Articles

ROTEM could be useful for lupus anticoagulant hypoprothrombinemia syndrome.

Lupus anticoagulant-hypoprothrombinemia syndrome (LAHPS) is a rare disease caused by acquired factor II (FII) deficiency and lupus anticoagulant. Patients with LAHPS typically present with thrombosis and bleeding. However, little information is available on the evaluation of coagulation potential in patients with LAHPS. We examined global coagulation potentials in patients with LAHPS during the clinical course in this study. Coagulation potentials in two pediatric patients with LAHPS were assessed by measuring clotting time (CT) and clot formation time using Ca2+-triggered rotational thromboelastometry (ROTEM), CT and maximum coagulation velocity using clot waveform analysis (CWA), and lag time and peak thrombin using the thrombin generation assay (TGA). The day of admission was defined as day 0. In case 1, the bleeding symptoms disappeared by day 5. However, the TGA and CWA results were markedly lower than normal, although FII activity (FII:C) returned to within the normal range by day 14. In contrast, ROTEM revealed a recovery to near-normal levels (day 14). All coagulation parameters (day 80) were within normal ranges. In case 2, coagulation potential was severely depressed until day 12, although FII:C returned to normal levels. Bleeding symptoms disappeared on day 19, and the ROTEM data revealed that the parameters were close to the normal range. The coagulation parameters in all assays were normalized on day 75. Recovery of coagulation potential in patients with LAHPS was slower than the recovery of FII:C. Moreover, ROTEM appeared to be clinically useful for assessing coagulation potential in patients with LAHPS.

Insighting the interaction mechanism of zirconium xerogel coagulants with humic acid: the protective effect of acetylacetone

The use of efficient modification methods determines the wide application potential of zirconium salt coagulants.

Fresh Frozen Plasma Versus Solvent Detergent Plasma for Cardiopulmonary Bypass Priming in Neonates and Infants Undergoing Cardiac Surgery: A Retrospective Cohort Study

ObjectiveCompared with Fresh Frozen Plasma (FFP), Omniplasma has been attributed to an increased coagulation potential and an increased fibrinolytic potential. This study aimed to compare Omniplasma and FFP used for cardiopulmonary bypass (CPB) priming regarding the incidence of post-operative thrombotic-or haemorrhagic complications and outcome in paediatric patients undergoing cardiac surgery. DesignA retrospective observational cohort study SettingThis single-center study was performed in the University Medical Center Groningen ParticipantAll paediatric patients up to 10 kg undergoing cardiac surgery with CPB. InterventionsProcedures where FFP was used for CPB priming were compared with those where Omniplasma was used. Measurements and main resultsThe primary outcome parameter was a composite endpoint consisting of 1) Paediatric Intensive Care Unit (PICU) mortality, 2) thromboembolic complications, and 3) haemorrhagic complications during PICU stay. We included 143 procedures in the analyses, 90 (63%) in the FFP- and 53 (37%) in the Omniplasma-group. The occurrence of the combined primary endpoint (FFP 20% vs omniplasma 11%, p=0.18 and its components did not differ between the used CPB priming agent). Omniplasma for CPB priming was associated with decreased Unfractionated Heparin administration per kg bodyweight (585 IU vs 510 IU, p=0.03), a higher pre- and post-operative Activated Clotting Time (ACT) discrepancy (90% vs 94%, p=0.03), a lower post-operative ACT value (125 seconds vs 118 seconds, p=0.01), and less Red Blood Cell transfusion per kilogram bodyweight (78 ml vs. 55 ml, p=0.02). However, none of the variables differed statistically significant in the multivariate logistic regression analyses. ConclusionsWe did not find an association between the used plasma for CPB priming and thromboembolic, haemorrhagic complication, and death in neonates and infants undergoing cardiac surgery. Omniplasma seems to be safe to use in this population.

Changes in coagulogram parameters and acid-base composition of blood in pregnant women with moderate and severe preeclampsia

Objective: to conduct a comparative analysis of coagulogram and acid-base composition of blood in pregnant women with moderate and severe preeclampsia (PE) and to assess the impact of the detected disorders on pregnancy outcomes. Material and methods. Group 1 consisted of patients with moderate PE (n=33), group 2 — with severe PE (n=34), and group 3 —women with physiological gestation (n=39). Parameters of coagulogram, acid-base composition of capillary blood of pregnant women (pH, pC02, p02, HC03-act) were determined on the RAPIDLAB 1265 device. Results. An increase in the level of fibrinogen was revealed in group 1 — by 9.9% relative to the indicator of group 3 (p=0.10), in group 2 — by 28.2% (p=0.002) against the background of a shortening of the APTT, respectively by 6.8% (p=0.003) and 9.9% (p<0.001) and a decrease in the number of platelets — by 11.9% (p=0.03) and 27.3% (p<0.001). Changes in the acid-base composition of capillary blood in PE were established: a decrease in HC03-act and p02 with an increase in pC02, which reflect the tendency to develop metabolic acidosis and respiratory alkalosis, more pronounced in group 2. Conclusions. PE is associated with disorders of blood coagulation potential and changes in blood gas composition, more pronounced in severe PE, which contributes to an increase in the frequency of premature birth (with moderate PE — up to 75.8%, severe — up to 100%) and cesarean section (respectively, by 7.7 and 18 times). The measurement of pC02 and p02 levels should be classified as informative and promising tests in terms of differential diagnosis of the severity of PE.

Emicizumab-mediated hemostatic function assessed by thrombin generation assay in an in vitro model of factor VIII-depleted thrombophilia plasma.

Patients with hemophilia A (PwHA) may have concurrent deficiency of representative anticoagulant proteins, protein (P)C, PS, and antithrombin (AT), which reduces bleeding frequency. However, emicizumab-driven hemostasis in PwHA with such thrombophilic potential remains unclarified. This study investigated the influence of natural anticoagulants on emicizumab-driven coagulation in HA model plasma. Various concentrations of PS and AT were added to PS-deficient plasma and AT-deficient plasma in the presence of anti-FVIII antibody (FVIIIAb; 10BU/mL). PC-deficient plasma was mixed with normal plasma at various concentrations in the presence of FVIIIAb. Emicizumab (50µg/mL) was added to these thrombophilic HA model plasmas, prior to tissue factor/ellagic acid-triggered thrombin generation assays. Co-presence of emicizumab increased peak thrombin values (PeakTh) dependent on PS, AT, and PC concentrations. Maximum coagulation potentials in the PS-reduced HA model plasmas remained normal in the presence of emicizumab. PeakTh were close to normal in the presence of 50%AT irrespective of emicizumab, but were higher than normal in the presence of 25%AT. Addition of recombinant FVIIa (corresponding to an administered dose of 90μg/kg) enhanced coagulation potential to normal levels. Our findings provide novel information on hemostatic regulation in emicizumab-treated PwHA with a possible thrombophilic disposition.

Hemostatic imbalance underlying preterm delivery in COVID-19 convalescent patients

Aim: to study the role of the hemostatic system in pretem delivery in pregnant women who have had COVID-19 in the gestation period from 14 to 16 weeks.Materials and Methods. A prospective single-center observational study was conducted by enrolling 63 pregnant women with verified COVID-19 at 14–16 weeks of gestation. The main group consisted of 37 patients with preterm birth (PB), comparison group – 26 patients labour activity that occurred at least at gestational age of 37 weeks. Clinical and anamnestic data and dynamic changes in fibrinogen and D-dimer level, activity of tissue factor (TF), tissue factor pathway inhibitor (TFPI), plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) were analyzed; thrombin generation assay (TGA) was performed.Results. It was found that severity of COVID-19 infection did not determine the timing of delivery that depended on patient comorbid condition. All PB observations (37 out of 63, 58.7 %) were caused by decompensated placental function manifested by acute obstetrical complications: increasing intrauterine fetal hypoxia (64.9 %) along with intrauterine growth retardation (51.4 %), severe preeclampsia (13.5 %) and premature abruption of the normally located placenta (5.0 %). In both study groups, COVID-19 experienced at 14–16 weeks of pregnancy was associated with coagulation and fibrinolytic imbalances. At the same time, at least 6 weeks post-COVID-19 infection, patients with PB had higher level of the “Peak thrombin” vs. comparison group (3050 vs. 2527 pmol/L; p = 0.0433). Also, patients with term vs. preterm delivery had TF activity decreased significantly: by 47.1% and 28.1%, respectively (p = 0.0546). Patients in preterm delivery group were characterized by fibrinolytic imbalance. At the first time point, suppressed fibrinolysis (PAI-1 level – 18.4 vs. 12.5 ng/ml in the comparison group; p = 0.0209) was concomitant with elevated level of u-PA (1.5 vs. 0.55 ng/ml in comparison group, p = 0.0015), which suggests a potential prolonged immunoinflammatory response in patients with PB. Magnitude of fibrinogen concentration and D-dimer level during post-COVID-19 follow-up study was within the reference values specific to gestational age.Conclusion. A significant increase in coagulation potential was found and verified by elevated activity of tissue factor and potential to thrombin generation in COVID-19 convalescent patients. In the case of preterm delivery, there was an imbalance in fibrinolysis system revealed by decreased blood fibrinolytic activity elevating along with increasing gestational age.

Successful Management of Intracranial Hemorrhage with Emicizumab in a Newborn with Severe Hemophilia a: A Case Report

Introduction Intracranial hemorrhage (ICH) is the second most common first hemorrhage in neonates with congenital Hemophilia, with an incidence of 3.4-4.0%. Nearly half (41%) of ICHs occur within the first week of life. ICH is the most feared complication of hemophilia with a high rate of morbidity and up to 20% of mortality. Timely diagnosis and prompt treatment are critical, even though the clinical signs of ICH in neonates are difficult to assess. Most studies recommend infusion of Factor VIII (FVIII) rising its plasma level to 100% as soon as possible. Unfortunately, approximately 25% to 40% of neonates develop inhibitors against FVIII after substitutive treatment; the percentage may be even higher in neonates with ICH requiring prolonged continuous treatment over days to weeks. We report a case of ICH in a newborn with severe haemophilia A (HA) treated with an initial-single dose of FVIII concentrate and then only with emicizumab. Case report A male neonate, born at 38+4 weeks of gestation by cesarean delivery, presented abnormal bleeding on his third day of life after heel blood sampling. Activated Partial Thromboplastin test resulted 147 seconds and FVIII was less then 1%. Family history for bleeding disorders was negative. A cerebral ultrasound scan documented a brain area suspicious for intracranial hemorrhage. Before transferring the neonate to the neonatal intensive care unit (NICU), he received an additional intravenous dose of vitamin K and 10 ml/kg fresh frozen plasma. On admission in NICU, the neonate presented alert and responsive with stable hemodynamics and vital signs. A second brain ultrasound and head MRI confirmed an intracerebral hemorrhage in the left parieto-occipital area near the splenium of the corpus callosum associated with subdural hematoma in the left temporo-occipital, occipito-mesial area, in falx cerebri and left tentorium cerebelli. Neonate received tranexamic acid infusions 40 mg/kg die and 50 IU/Kg of intravenous recombinant factor VIII concentrate (octocog alfa). Then, it was decided to start treatment with Emicizumab, a bispecific antibody mimicking FVIII, at a dose of 3 mg/kg subcutaneously once a week for 4 weeks, continuing prophylaxis at 1.5 mg/kg once a week.Serial brain ultrasound revealed progressive reduction of the ICH extension over the following weeks; one month later, brain MRI confirmed the improvement. The neonate's clinical condition remained always stable; he never showed seizures or neurological signs of impaired consciousness and his neurodevelopment is appropriate for age. MRI performed at three months of age documented complete resolution of the cerebral hemorrhage. Discussion Intensive FVIII replacement therapy for ICH in pups is associated with a high risk of inhibitor formation.Emicizumab, a humanised bispecific monoclonal antibody, is indicated for routine prophilaxis in patients with severe HA with and without antibodies to FVIII. Emicizumab is attractive due to its weekly or even longer dosing intervals and subcutaneous route of administration but, despite the baseline hemostasis achieved with emicizumab, acute bleeding events may still require aggressive therapy. Theoretically, it needs few days to be hemostatically effective but its in vitro coagulant potential in neonate is heterogeneous, thus we can not completely explain the therapeutic success with only one dose of FVIII. Conclusions The combined use of emicizumab and antifibrinolytics was successful in treating acute intracranial hemorrhage. To the best of our knowledge, this is the first pediatric case of successful management of intracranial hemorrhage with emicizumab in a newborn with severe HA.

NXT007-mediated hemostatic potential is suppressed by activated protein C-catalyzed inactivation of activated factor V

BackgroundActivated protein C (APC) inactivates activated factor (F) V (FVa) and FVIIIa. NXT007, an emicizumab-based engineered therapeutic bispecific antibody, enhances the coagulation potential of FVIII-deficient plasma (FVIIIdef-plasma) to near normal levels. However, little is known about the effect of APC-induced inactivation in NXT007-mediated hemostatic function. ObjectivesTo investigate the contribution of APC-mediated reactions to NXT007-driven hemostasis. MethodsIn pooled normal plasma (PNP) or FVIIIdef-plasma spiked with NXT007 (10 μg/mL), effects of APC (0-16 nM) were measured using a thrombin generation assay (TGA). The direct effects of APC on cofactor activity of NXT007 or FVIIIa in a FXa generation assay were also measured. The FVdef-plasma and FV Leiden plasma (FVLeiden plasma) were preincubated with 2 anti-FVIII monoclonal antibodies (termed FVIII-depleted), and the APC effect in the presence of NXT007 in FVIII-depleted FVdef-plasma with the addition of exogenous FV (7.5-30 nM) or FVIII-depleted FVLeiden plasma was investigated. ResultsThe APC dose-dependent suppression effect in TGA of FVIIIdef-plasma spiked with NXT007 was similar to that of PNP. FXa generation with NXT007 was not impaired by the addition of APC, suggesting that the APC-induced reaction in TGA with NXT007 was attributed to the direct inactivation of FVa. The addition of APC to FVIII-depleted FVdef-plasma, along with NXT007 and various FV concentrations, showed a similar attenuation to PNP. The NXT007-driven thrombin generation in FVIII-depleted FVLeiden plasma was suppressed by APC, similar to the reaction in native FVLeiden plasma. ConclusionNXT007 did not impair APC-mediated downregulation of FVa in FVIIIdef-plasmas, regardless of the presence of FV mutation with APC resistance.

A bispecific antibody NXT007 exerts a hemostatic activity in hemophilia A monkeys enough to keep a nonhemophilic state

BackgroundEmicizumab, a factor (F) VIIIa-function mimetic bispecific antibody (BsAb) to FIXa and FX, has become an indispensable treatment option for people with hemophilia A (PwHA). However, a small proportion of PwHA still experience bleeds even under emicizumab prophylaxis, as observed in the long-term outcomes of clinical studies. A more potent BsAb may be desirable for such patients. ObjectivesTo identify a potent BsAb to FIXa and FX, NXT007, surpassing emicizumab by in vitro and in vivo evaluation. MethodsNew pairs of light chains for emicizumab’s heavy chains were screened from phage libraries, and subsequent antibody optimization was performed. For in vitro evaluation, thrombin generation assays were performed with hemophilia A plasma. In vivo hemostatic activity was evaluated in a nonhuman primate model of acquired hemophilia A. ResultsNXT007 exhibited an in vitro thrombin generation activity comparable to the international standard activity of FVIII (100 IU/dL), much higher than emicizumab, when triggered by tissue factor. NXT007 also demonstrated a potent in vivo hemostatic activity at approximately 30-fold lower plasma concentrations than emicizumab’s historical data. In terms of dose shift between NXT007 and emicizumab, the in vitro and in vivo results were concordant. Regarding pharmacokinetics, NXT007 showed lower in vivo clearance than those shown by typical monoclonal antibodies, suggesting that the Fc engineering to enhance FcRn binding worked well. ConclusionNXT007, a potent BsAb, was successfully created. Nonclinical results suggest that NXT007 would have a potential to keep a nonhemophilic range of coagulation potential in PwHA or to realize more convenient dosing regimens than emicizumab.

Shear Forces Induced Platelet Clearance Is a New Mechanism of Thrombocytopenia.

Thrombocytopenia has been consistently described in patients with extracorporeal membrane oxygenation (ECMO) and associated with poor outcome. However, the prevalence and underlying mechanisms remain largely unknown, and a device-related role of ECMO in thrombocytopenia has been hypothesized. This study aims to investigate the mechanisms underlying thrombocytopenia in ECMO patients. In a prospective cohort of 107 ECMO patients, we investigated platelet count, functions, and glycoprotein shedding. In an ex vivo mock circulatory ECMO loop, we assessed platelet responses and VWF (von Willebrand factor)-GP Ibα (glycoprotein Ibα) interactions at low- and high-flow rates, in the presence or absence of red blood cells. The clearance of human platelets subjected or not to ex vivo perfusion was studied using an in vivo transfusion model in NOD/SCID (nonobese diabetic/severe combined Immunodeficient) mice. In ECMO patients, we observed a time-dependent decrease in platelet count starting 1 hour after device onset, with a mean drop of 7%, 35%, and 41% at 1, 24, and 48 hours post-ECMO initiation (P=0.00013, P<0.0001, and P<0.0001, respectively), regardless of the type of ECMO. This drop in platelet count was associated with a decrease in platelet GP Ibα expression (before: 47.8±9.1 versus 24 hours post-ECMO: 42.3±8.9 mean fluorescence intensity; P=0.002) and an increase in soluble GP Ibα plasma levels (before: 5.6±3.3 versus 24 hours post-ECMO: 10.8±4.1 µg/mL; P<0.0001). GP Ibα shedding was also observed ex vivo and was unaffected by (1) red blood cells, (2) the coagulation potential, (3) an antibody blocking VWF-GP Ibα interaction, (4) an antibody limiting VWF degradation, and (5) supraphysiological VWF plasma concentrations. In contrast, GP Ibα shedding was dependent on rheological conditions, with a 2.8-fold increase at high- versus low-flow rates. Platelets perfused at high-flow rates before being transfused to immunodeficient mice were eliminated faster in vivo with an accelerated clearance of GP Ibα-negative versus GP Ibα-positive platelets. ECMO-associated shear forces induce GP Ibα shedding and thrombocytopenia due to faster clearance of GP Ibα-negative platelets. Inhibiting GP Ibα shedding could represent an approach to reduce thrombocytopenia during ECMO.

RISK FACTORS FOR THE DEVELOPMENT OF VENOUS THROMBOEMBOLISM IN PATIENTS WITH UNCOMPLICATED ACUTE CALCULOUS CHOLECYSTITIS

The aim is to identify and analyze risk factors for venous thromboembolism in patients diagnosed with uncomplicated acute calculous cholecystitis.&#x0D; Materials and Methods. A prospective analysis of clinical data of 71 patients diagnosed with uncomplicated acute calculous cholecystitis was conducted. Two clinical groups were formed: subgroup 1A (n=6) included patients diagnosed with venous thromboembolism, and subgroup 1B (n=65) included patients without venous thromboembolism.&#x0D; Results and Discussion. During the study, the coagulation potential of the patients was studied. The prothrombin index on the third day of the postoperative period and at the time of discharge was significantly higher in subgroup 1A patients (109.1% and 123.0%) compared to subgroup 1B patients (98.0% and 96.4%). The situation with total fibrinogen is similar: the mean level of this indicator during the entire observation period was significantly higher in patients of subgroup 1A (p&lt;0.05). The mean D-dimer level at the time of hospital discharge in subgroup 1A was 3.6 mg/ml higher compared to the level of this indicator on the first day after surgery, and in subgroup 1B it was only 0.5 mg/ml higher. The risk of venous thromboembolism according to the Caprini score in subgroup 1A was 7.5 [5.5; 9.0] points, compared to 6.0 [4.0; 8.0] points in subgroup 1B. Assessment of the risk of venous thromboembolism according to the Rogers scale showed similar results: 8.0 [6.3; 9.8] points for patients of subgroup 1A and 7.0 [6.0; 8.0] points for subgroup 1B. When assessing the interleukin 1 beta (IL1b) level, its increased value was detected in subgroup 1A - 14.0 [7.0; 33.7] pg/ml, as opposed to subgroup 1B, where it was 9.0 [3.5; 26.2] pg/ml. Similar results were found in the assessment of tumor necrosis factor TNF: the mean values of this cytokine level were also higher in subgroup 1A (13.9 [7.9; 15.8] pg/ml) than in subgroup 1B (8.8 [6.0; 13.1] pg/ml).&#x0D; Conclusions. According to the results of the study, the following factors for the development of venous thromboembolism in patients with uncomplicated acute calculous cholecystitis were determined: elevated levels of indicators of total fibrinogen, D-dimer, interleukin 1 beta (IL1b) and tumor necrosis factor (TNF) at the time of admission. The mean values of the total risk assessment points according to the Caprini score and Rogers scale were significantly higher (p&lt;0.05) in the subgroup of patients with uncomplicated acute calculous cholecystitis who later developed venous thromboembolism.

The in vitro effect of anticoagulant agents on coagulation and fibrinolysis in the presence of emicizumab in the plasmas from patients with haemophilia A.

Emicizumab is used as hemostatic prophylaxis for patients with hemophilia A (PwHA), irrespective of the presence of inhibitors. Although bacterial infection can lead to a procoagulant state, there is limited information on coagulation and fibrinolysis potentials in emicizumab-treated PwHA and on the use of anticoagulants in such cases. We examined whether anticoagulants affect the coagulation and fibrinolysis potentials in plasma from PwHA spiked with emicizumab. Plasma from PwHA was in vitro supplemented with emicizumab (50μg/mL; emi-plasma) and anticoagulants (recombinant thrombomodulin (rTM), nafamostat mesylate (NM), unfractionated heparin (UFH), or low-molecular-weight heparin (LMH)). PwHA plasma spiked with rFVIII (1 IU/mL) was used as a reference (ref-plasma). The coagulation and fibrinolysis potentials in plasma was measured by thrombin and plasmin generation assay (T/P-GA) and clot-fibrinolysis waveform analysis (CFWA). In T/P-GA and CFWA, coagulation potentials (maximum coagulation velocity; |min1|, and peak thrombin; Th-Peak) in plasma rose with increasing concentrations of emicizumab and rFVIII, but fibrinolytic potentials (peak plasmin; Plm-Peak, and maximum fibrinolytic velocity; |FL-min1|) remained unchanged. Adding rTM, NM, and UFH to emi-plasma suppressed coagulation and fibrinolysis potentials, similar to ref-plasma. Regarding the heparin, UFH and LMH inhibited the improved coagulation in emi-plasma. UFH inhibited fibrinolysis as well, but LMH did not. Anticoagulants could exhibit the inhibitory effects on the coagulation and fibrinolysis potentials in plasma from PwHA spiked with emicizumab, similar to those in normal plasma.

No Observed Difference in Inflammatory and Coagulation Markers Following Diets Rich in n-6 Polyunsaturated Fat vs Monounsaturated Fat in Adults With Untreated Hypercholesterolemia: A Randomized Trial

BackgroundInflammatory and prothrombotic responses are hallmark to the progression of cardiovascular disease and may be influenced by the type of dietary fat. Cottonseed oil (CSO) is rich in n-6 polyunsaturated fats and improves traditional cardiovascular disease risk factors such as cholesterol profiles. However, some clinicians are still hesitant to promote n-6 polyunsaturated fats consumption despite growing evidence suggesting they may not be independently pro-inflammatory. ObjectiveTo investigate the inflammatory and coagulation marker responses to an 8-week diet intervention rich in either CSO or olive oil (OO) (OO is rich in monounsaturated fat) in adults with untreated hypercholesterolemia. DesignThis was a secondary analysis of a parallel-arm randomized clinical trial with the main outcome of cholesterol measures. Participants/settingParticipants included in this analysis were 42 sedentary adults aged 30 to 75 years (62% women) in the Athens, GA, area, between May 2018 and June 2021, with untreated hypercholesterolemia or elevated blood lipids and body mass index >18.5. Hypercholesterolemia was defined as at least two blood lipid levels in a borderline undesirable/at risk range (total cholesterol level ≥180 mg/dL, low-density lipoprotein cholesterol level ≥110 mg/dL, high-density lipoprotein cholesterol level <50 mg/dL, or triglyceride level ≥130 mg/dL), or at least one in an undesirable range (total cholesterol level ≥240 mg/dL, low-density lipoprotein cholesterol level ≥160 mg/dL, high-density lipoprotein cholesterol level <40 mg/dL, or triglyceride level ≥200 mg/dL). InterventionParticipants were randomly assigned to either the CSO or OO group in a partial outpatient feeding trial. Meals from the study provided approximately 60% of their energy needs with 30% of energy needs from either CSO or OO for 8 weeks. Participants fulfilled their remaining energy needs with meals of their choosing. Main outcome measuresFasting plasma concentrations of inflammatory markers, including C-reactive protein, tumor necrosis factor-α, interleukin-6, and interleukin-1β were measured at baseline and 8 weeks. Markers of coagulation potential, including plasminogen activator inhibitor-1, and tissue factor were measured at the same time points. Statistical analyses performedRepeated measures linear mixed models were used with treatment and visit in the model for analyses of all biochemical markers. ResultsThere were no significant differences in fasting C-reactive protein (P = 0.70), tumor necrosis factor-α (P = 0.98), interleukin-6 (P = 0.21), interleukin-1β (P = 0.13), plasminogen activator inhibitor-1 (P = 0.29), or tissue factor (P = 0.29) between groups across the intervention. ConclusionsInflammation and coagulation marker responses to diets rich in CSO vs OO were not significantly different between groups, and neither group showed changes in these markers in adults with untreated hypercholesterolemia. This provides additional evidence suggesting that dietary n-6 polyunsaturated fats may not promote inflammation compared with monounsaturated fatty acids, even in adults at increased risk for cardiovascular disease.

Predictors of functional and morphological arterial wall properties in coronary artery disease patients with increased lipoprotein (a) levels before and after treatment with proprotein convertase subtilisin-kexin type 9 inhibitors

BackgroundIn addition to proatherogenic properties, lipoprotein (a) (Lp(a)) has also pro-inflammatory, antifibrinolytic and prothrombogenic features. The aim of the current study was to identify the predictors of functional and morphological properties of the arterial wall in patients after myocardial infarction and increased Lp(a) levels at the beginning and after treatment with proprotein convertase subtilisin-kexin type 9 (PCSK9) inhibitors.MethodsSeventy-six post-myocardial infarction patients with high Lp(a) levels were included in the study. Ultrasound measurements of flow-mediated dilation of brachial artery (FMD), carotid intima-media thickness (c-IMT) and pulse wave velocity (PWV) were performed initially and after 6 months of treatment. At the same time points lipids, Lp(a), inflammatory and hemostasis markers were measured in blood samples.ResultsIn linear regression model FMD significantly correlated with age at first myocardial infarction (β = 0.689; p = 0.022), high-sensitivity C-reactive protein (β = -1.200; p = 0.009), vascular cell adhesion protein 1 (VCAM-1) (β = -0.992; p = 0.006), overall coagulation potential (β = 1.428; p = 0.014) and overall hemostasis potential (β = -1.473; p = 0.008). c-IMT significantly correlated with age at first myocardial infarction (β = 0.574; p = 0.033) and Lp(a) (β = 0.524; p = 0.040). PWV significantly correlated with systolic blood pressure (β = 0.332; p = 0.002), tumor necrosis factor alpha (β = 0.406; p = 0.002), interleukin-8 (β = -0.315; p = 0.015) and plasminogen activator inhibitor 1 (β = 0.229; p = 0.031). After treatment FMD reached statistical significance only in univariant analysis with systolic blood pressure (r = -0.286; p = 0.004) and VCAM-1 (r = -0.229; p = 0.024). PWV and c-IMT correlated with age (r = 0.334; p = 0.001 and r = 0.486; p < 0.0001, respectively) and systolic blood pressure (r = 0.556; p < 0.0001 and r = 0.233; p = 0.021, respectively).ConclusionsOur results suggest that age, systolic blood pressure, Lp(a) levels and other biochemical markers associated with Lp(a) are predictors of functional and morphological properties of the arterial vessel wall in post-myocardial patients with high Lp(a) levels initially. However, after 6 months of treatment with PCSK9 inhibitors only age and systolic blood pressure seem to be predictors of these properties.Trial registrationThe protocol for this study was registered with clinicaltrials.gov on November, 3 2020 under registration number NCT04613167.Graphical

Future Perspective and Technological Innovation in Cheese Making Using Artichoke (Cynara scolymus) as Vegetable Rennet: A Review.

Milk coagulation is a process used for the formulation of different dairy products such as cheese. In this process, milk undergoes changes in its chemical stability thanks to acidification or enzymatic reactions. Traditionally, milk coagulation has been carried out with rennet of animal origin, but recently, the research of new types of rennet such as microbial rennet and vegetable rennet has increased. This study aims to present an organized review of the most relevant information on lactic coagulation, its relationship with vegetable rennets, and the importance of the botanical genus Cynara in the extraction of vegetable rennets, focusing on the coagulant potential of artichoke (Cynara scolymus). We conducted this literature review and found that lactic coagulation and vegetable rennets are linked through the enzymatic activity of the latter. The results of the main studies demonstrated a strong relationship between vegetable rennets and protease enzymes as well as the presence of these enzymes in extracts of cardoon (Cynara scolymus) and artichoke (Cynara scolymus). In addition, studies highlight the presence of thistle extracts in artisanal cheese preparations in the Iberian Peninsula. Based on the results of the studies, a comparison between cheeses made with vegetable rennet and those made with traditional rennet was also carried out. Although the results show that the use of vegetable rennet in the manufacture of cheese can confer undesirable characteristics, the use of extracts from Cynara plants demonstrates that vegetable rennets have an industrial potential, especially the one obtained from artichoke (Cynara scolymus) due to its high availability. Nevertheless, specific studies are required for a better understanding and application of this rennet.

Characteristics in dissolved organic matter and disinfection by-product formation during advanced treatment processes of municipal secondary effluent with Orbitrap mass spectrometry

Dissolved organic matter (DOM) is reported to be a precursor to disinfection by-products (DBPs), which have adverse effects on human health. Therefore, it is crucial to effectively remove DOM before water disinfection. Characteristics of DOM and DBPs formation during advanced treatment processes including coagulation, adsorption, ultraviolet (UV) irradiation, and ozone (O3) oxidation in municipal secondary effluent were investigated in this research. DOM was characterized by Fourier transform infrared spectroscopy (FTIR), excitation-emission matrix fluorescence spectroscopy (EEM), and Orbitrap mass spectrometry (Orbitrap MS). Moreover, DBPs formation potential under different advanced treatment processes was also discussed. FTIR results indicated that various functional groups existing in DOM may react with the disinfectant to form toxic DBPs. EEM analysis indicated that DOM in all water samples was dominated by soluble microbial product-like (SMPs) and humic acid-like (HA) substances. The municipal secondary effluent was abundant with DOM and rich in carbon, hydrogen, oxygen, and nitrogen atoms, contained a certain dosage of phosphorus and sulfur atoms, and the highest proportion is lignin. Most of the precursors (CHO features) had positive double bond equivalent subtracted oxygen per carbon [(DBE-O)/C] and negative carbon oxidation state (Cos) in all four different advanced treatment processes. DBPs formation potential (DBPFP) of coagulation, adsorption, UV irradiation, and O3 oxidation advanced treatment processes were 487 μg L−1, 586 μg L−1, 597 μg L−1, and 308 μg L−1, respectively. And the DBPs precursors removal efficiency of coagulation, adsorption, UV irradiation, and O3 oxidation advanced treatment processes were 50.8%, 40.8%, 39.8%, and 69.0%, respectively. This study provides in-depth insights into the changes of DOM in municipal secondary effluent at the molecular level and the removal efficiency of DBPs precursors during coagulation, adsorption, UV irradiation, and O3 oxidation advanced treatment processes.

Coagulant potentials of emicizumab in the plasmas from infant and toddler patients with hemophilia A.

Emicizumab significantly reduces bleedings in patients with hemophilia A (PwHA). A clinical study (HAVEN 7; NCT04431726) for PwHA aged less than or equal to 12months is ongoing, but emicizumab-driven coagulation potential in PwHA in early childhood remains to be clarified. To investigate the in vitro or in vivo coagulation potential of emicizumab in plasmas obtained from infant and toddler PwHA. Twenty-seven plasma samples from 14 infant/toddler PwHA (aged 0-42months, median 19months) who received emicizumab (n=9), factor (F)VIII products (n=8), or no treatment (n=10) were obtained. FVIII activity in FVIII-treated plasmas was cancelled by the addition of anti-FVIII monoclonal antibody (mAb). Emicizumab-treated plasmas (in vivo) and emicizumab-spiked plasmas (in vitro) were analyzed. Emicizumab-untreated plasma or emicizumab-treated plasma supplemented with two anti-emicizumab mAbs were used as references. Adjusted maximum coagulation velocity (Ad|min1|) by clot waveform analysis and peak thrombin (Peak-Th) by thrombin generation assay was assessed. Ad|min1| values in 24 samples were improved by the presence of emicizumab. Values did not improve in the three remaining samples (aged 1, 23, and 31months). Although the presence of emicizumab showed an age-dependent increase in Peak-Th in 20 samples, this increase was not observed in seven samples (aged 0, 1, 1, 2, 8, 19, and 36months). Emicizumab-dependent increases in both Ad|min1| and Peak-Th were shown in 18 samples, and increases in either parameter were shown in eight samples. One sample (from patient aged 1month) showed no increase in both, however. Emicizumab could improve coagulant potential in plasmas from infant/toddler patients with hemophilia A.

Production and characterization of a novel poly amino acid from a thermophilic bacterium, and preliminary testing of its coagulating potential for imminent wastewater treatment application

The increasing demand for biopolymers across diverse fields, such as food, medicine, cosmetics, and environmental applications, has prompted researchers to explore novel molecules with enhanced functionalities that meet these demands. In this study, a thermophilic strain of Bacillus licheniformis was employed to produce a unique polyamino acid. This thermophilic isolate exhibited rapid growth at 50 °C in a sucrose mineral salts medium, resulting in a biopolymer concentration of 7.4 g/L. Interestingly, the biopolymer produced at different temperatures exhibited varying glass-transition temperatures (ranging from 87.86 °C to 104.11 °C) and viscosities (7.5 cP to 16.3 cP), suggesting that the fermentation temperature significantly influenced the degree of polymerization. Furthermore, the biopolymer was characterized using various techniques, including Thin Layer Chromatography (TLC), Fourier Transform Infrared (FTIR) spectroscopy, Liquid Chromatography-Electrospray Ionization-Mass Spectroscopy (LC-ESI MS), Nuclear Magnetic Resonance (NMR), and Differential Scanning Calorimetry-Thermogravimetric Analysis (DSC-TGA). The results revealed that the obtained biopolymer was a poly amino acid, with poly-γ-glutamic acid as the major monomeric component in the polymer backbone with a few appendages of aspartic acid residues in its side chain. Finally, the biopolymer demonstrated significant coagulation potential for water treatment applications, as evidenced by coagulation studies conducted under varying pH conditions using kaolin-clay as a model precipitant.

Factor VIII mutated with Lys1813Ala within the factor IXa-binding region enhances intrinsic coagulation potential.

Factor VIII (FVIII) functions as a cofactor of FIXa for FX activation in the intrinsic tenase complex. The 1811-1818 region in the FVIII A3 domain was observed to contribute to FIXa binding, and the K1813A/K1818A mutant increased the binding affinity for FIXa. The current study aims to identify mutated FVIII protein(s) that increase FVIIIa cofactor activity in the 1811-1818 region. FVIII mutants with K1813A, K1818A, and K1813A/K1818A were expressed in baby hamster kidney cells and were followed by assessments using purified and global coagulation assays for mouse models with hemophilia A (HA). A surface plasmon resonance-based assay revealed that the Kd value of FVIII-K1813A for FIXa interaction was lower than that of the wild-type (WT) (3.9±0.7/6.3±0.3 nM). However, the Km value of FVIII-K1813A for FIXa on tenase activity was comparable with that of the WT, whereas the kcat of this mutant was significantly greater than that of the WT. Thrombin-catalyzed FVIII-K1813A activation was ∼1.3-fold more enhanced than that of the WT, and the spontaneous decay of activated FVIII-K1813A was ∼2.5-fold slower than that of WT. The heat stability assay revealed that the decay rate of FVIII-K1813A was ∼2.5-fold slower than that of WT. Thrombin generation assay and rotational thromboelastometry using blood samples from patients with HA demonstrated that the addition of FVIII-K1813A (0.5 nM) exhibited a coagulation potential compatible with that of WT (1 nM). In the tail clip assay of HA mice, FVIII-K1813A showed a two- to fourfold higher hemostatic potential than that of the WT. FVIII-K1813A, with higher a FIXa binding affinity, enhances the global coagulation potential because of the stability of FVIII/FVIIIa molecules.

STUDY OF THE ROLE OF HEMOCOAGULATION DISBALANCE AS A KEY PATHOGENETIC PREDICTORS IN THE FORMATION OF SYSTEMIC DISORDERS IN EXPERIMENTAL PERITONITIS

Background. Peritonitis today continues to be one of the most difficult problems of both abdominal surgery and theoretical pathophysiology. One of the leading pathogenetic mechanisms of peritonitis is disorders in the hemostasis system and the development of dissemination vascular syndrome. Blockade of microcirculation caused by the development of dissemination vascular syndrome leads to tissue hypoxia and, as a result, to dysfunction of organs and body systems.&#x0D; Aim: is to study changes in vascular-platelet and coagulation hemostasis under conditions of experimental fecal peritonitis in rats.&#x0D; Materials and methods. Experimental studies were conducted on 24 non-linear laboratory rats, which were divided into 2 groups: 1 group – intact control (animals received distilled water), 2 group – animals of the control pathology group. According to the "Methodological recommendations for preclinical study of medicinal products", experimental peritonitis was studied on the model of V. A. Lazarenko. Indicators of vascular-platelet and coagulation hemostasis were determined according to generally accepted methods.&#x0D; Results. On the first day of experimental peritonitis, a sharp increase in the rate of blood clotting and changes in the quality properties of platelets were noted, affecting their aggregation ability and retraction of an already formed blood clot. On the 4th day of the experiment, changes in the vascular-platelet link were established, indicating the exhaustion of the platelet potential, due to a decrease in blood clotting time, platelet aggregation ability, and blood clot retraction against the background of a probable increase in the number of blood platelets, which probably by now had time "get" to the bloodstream from the depot. Changes in the coagulation hemostasis system against the background of experimental fecal peritonitis indicate the development of hypercoagulation already on the first day, which is replaced by a hypocoagulation state, which indicates the exhaustion of the enzymatic systems of blood coagulation potential (as early as on the 4th day), which lasted until the end of the experiment.&#x0D; Conclusion. In the development of disorders of the hemostasis system in fecal peritonitis, the key role belongs to both the internal and external mechanisms of activation of the blood coagulation system.