Antithrombin III Concentration Research Articles

Meglumine Cyclic Adenylate Improves Coagulation Abnormalities in Endotoxemic Rats

<p id="p00005"><strong>Background:</strong> Sepsis and endotoxemia are linked to the concurrent activation of inflammatory and hemostatic pathways, which contribute to organ dysfunction and death. Meglumine cyclic adenylate (MCA) has been found to inhibit NF-κB activity during endotoxemia in rats. This study aims to investigate the effect of MCA on coagulation abnormalities in endotoxemic rats. <p id="p00010"><strong>Methods:</strong> seventy-two male SD rats were randomly divided into three groups (n=24 per group): Normal saline group (NS), group LPS, and group MCA. Endotoxemia was induced by LPS 10 mg/kg injected via the femoral vein in the LPS and MCA groups. MCA 2mg/kg was injected 20 min before LPS injection in group MCA via the femoral vein. An equal volume of normal saline was given to NS and LPS groups. Prothrombin time (PT), partial activated thromboplastin time (APTT), platelet count, cAMP, plasminogen activator inhibitor-1(PAl-1), D-dimer, TNF-α, antithrombin III (ATIII), t-PA and Tissue factor (TF) were assessed before (baseline) and 2, 4, 6 h after LPS injection. <p id="p00015"><strong>Results:</strong> After LPS administration, PT and APTT were prolonged, ATIII concentrations, cAMP, and platelet count were significantly decreased, while plasma PAI-1, D-dimer, t-PA, TNF-α, and TF levels were significantly increased compared to group NS. However, these alterations induced by LPS were inhibited by MCA treatment. <p id="p00020"><strong>Conclusion:</strong> MCA amelio-rates coagulation abnormalities induced by LPS in endotoxemic rats.

Monitoring of Unfractionated Heparin in Severe COVID-19: An Observational Study of Patients on CRRT and ECMO.

Objective Severe cases of coronavirus disease 2019 (COVID-19) can require continuous renal replacement therapy (CRRT) and/or extracorporeal membrane oxygenation (ECMO). Unfractionated heparin (UFH) to prevent circuit clotting is mandatory but monitoring is complicated by (pseudo)-heparin resistance. In this observational study, we compared two different activated partial thromboplastin time (aPTT) assays and a chromogenic anti-Xa assay in COVID-19 patients on CRRT or ECMO in relation to their UFH dosages and acute phase reactants. Materials and Methods The aPTT (optical [aPTT-CS] and/or mechanical [aPTT-STA] clot detection methods were used), anti-Xa, factor VIII (FVIII), antithrombin III (ATIII), and fibrinogen were measured in 342 samples from 7 COVID-19 patients on CRRT or ECMO during their UFH treatment. Dosage of UFH was primarily based on the aPTT-CS with a heparin therapeutic range (HTR) of 50–80s. Associations between different variables were made using linear regression and Bland–Altman analysis. Results Dosage of UFH was above 35,000IU/24 hours in all patients. aPTT-CS and aPTT-STA were predominantly within the HTR. Anti-Xa was predominantly above the HTR (0.3–0.7 IU/mL) and ATIII concentration was >70% for all patients; mean FVIII and fibrinogen were 606% and 7.5 g/L, respectively. aPTT-CS correlated with aPTT-STA ( r 2 = 0.68) with a bias of 39.3%. Correlation between aPTT and anti-Xa was better for aPTT-CS (0.78 ≤ r 2 ≤ 0.94) than for aPTT-STA (0.34 ≤ r 2 ≤ 0.81). There was no general correlation between the aPTT-CS and ATIII, FVIII, fibrinogen, thrombocytes, C-reactive protein, or ferritin. Conclusion All included COVID-19 patients on CRRT or ECMO conformed to the definition of heparin resistance. A patient-specific association was found between aPTT and anti-Xa. This association could not be explained by FVIII or fibrinogen.

Low levels of coagulation inhibitors: a high-risk thrombotic factor in thalassemic patients

Background and aimThe β-thalassemia major (β-TM) is defined as a hereditary red blood cell (RBC)- related disease. Thrombotic events are associated with thalassemia in adult patients. Thus, the present investigation was aimed to examine some hemostatic parameters, including anti thrombin- III (AT-III), protein-C (PRC) and protein-S (PRS) in β-TM patients. MethodsThirty B-TM patients who referred for routine follow-up admission to the thalassemia clinic of Kerman Special Disease Center alongside with 30 healthy subjects were selected and enrolled in the present study. Further registration, the peripheral blood specimens were collected after 3 weeks of last transfusion and then the plasma concentrations of AT-III, PRC and PRS were measured in them. ResultsWe have observed that the concentrations of natural coagulation inhibitors (PRC and PRS) were significantly attenuated in β-TM patients (P < 0.05), while the plasma level of AT-III was not remarkably differed in β-TM patients in compare to healthy subjects. ConclusionAccording to the findings of present work, significant changes in the PRC, PRS and AT-III which could be observed in multi transfused β-TM patients may attribute as critical risk factors for the development of upcoming thromboembolic events in their future life.

Bajos niveles de inhibidores de la coagulación: un factor de alto riesgo trombogénico en pacientes talasémicos

Background and aimThe β-thalassemia major (β-TM) is defined as a hereditary red blood cell-related disease. Thrombotic events are associated with thalassemia in adult patients. Thus, the present investigation was aimed to examine some hemostatic parameters, including anti thrombin-III (AT-III), protein-C (PRC) and protein-S (PRS) in β-TM patients. MethodsThirty B-TM patients who referred for routine follow-up admission to the thalassemia clinic of Kerman Special Disease Center alongside with 30 healthy subjects were selected and enrolled in the present study. Further registration, the peripheral blood specimens were collected after 3 weeks of last transfusion and then the plasma concentrations of AT-III, PRC and PRS were measured in them. ResultsWe have observed that the concentrations of natural coagulation inhibitors (PRC and PRS) were significantly attenuated in β-TM patients (P<0.05), while the plasma level of AT-III was not remarkably differed in β-TM patients in compare to healthy subjects. ConclusionAccording to the findings of present work, significant changes in the PRC, PRS and AT-III which could be observed in multi transfused β-TM patients may attribute as critical risk factors for the development of upcoming thromboembolic events in their future life.

Comparison of antithrombin III levels in type 2 diabetes mellitus patients with and without ulcers at Haji Adam Malik Hospital Medan Indonesia from may to july 2017

Introduction: People with diabetes mellitus (DM) have an increased risk for thrombosis compared with non-diabetic patients. Several studies showed contradicting in data on levels of antithrombin III (AT-III) in people with type 2 DM and diabetic ulcers.Methods: This is a descriptive and analytical cross-sectional study on AT-III concentration of patients with type 2 DM with and without foot ulcers. A total of 40 subjects are divided equally into the control group, which consist of type 2 DM patients without foot ulcers, and the case group of type 2 DM patients with a diabetic foot ulcer based on the Wagner criteria. Blood samples are then taken after 8 to 10 hours of fasting to check for AT-III concentration and hemostasis examination including platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), fibrinogen levels, and D-dimers.Results: The mean AT-III level of diabetic foot ulcers group was 216.02 ± 71.23 pg/mL, which was lower than the mean AT-III level of the group without diabetic foot ulcers, i.e. 243.05 ± 48.05 pg/mL. The mean AT-III concentration in the diabetic foot ulcers group with hypercoagulation state was 221.52 ± 64.25 pg/mL, which was slightly higher than the mean AT-III concentration of the diabetic foot ulcers group with hypocoagulation state of 203.18 ± 90.92 pg/mL.Conclusion: No statistically significant difference was found in the levels of AT-III between the diabetic foot ulcers group and the non-diabetic foot ulcers group (p&gt; 0.05). There was no statistically significant difference in AT-III levels of diabetic foot ulcers with hypercoagulation compared with no diabetic foot ulcers with hypocoagulation and no statistically significant relationship between grade diabetic foot ulcers and AT-III concentration.

Predictive value of thromboelastography parameters combined with antithrombin III and D-Dimer in patients with recurrent spontaneous abortion.

To investigate the value of thromboelastography (TEG) combined with antithrombin III (AT-III) and D-Dimer in predicting the occurrence of recurrent spontaneous abortion (RSA). One hundred and five RSA patients and 40 fertile women were enrolled. The subjects were subjected into four groups: group 1 (40 fertile women), group 2 (58 women with 2 abortions), group 3 (30 women with 3 abortions), and group 4 (17 women with four abortions). TEG was conducted on all subjects. Clotting time, reaction time, angle degree, coagulation index, and maximum amplitude were measured. The levels AT-III, D-Dimer, platelet counts, and fibrinogen concentration were determined. The ROC curve analysis was done using MedCalc software to analyse the diagnosis accuracy of the parameters of interest and the combined approach. The AT-III level in all group 4 was significantly lower than in fertile women. The D-Dimer concentration, platelet count, and MA in patients with four prior abortions were significantly higher than the other three groups. CI and fibrinogen concentration in patients with four prior pregnancy losses were significantly higher than group 1. The ROC curves suggested that combined use of CI, MA, AT-III, and D-Dimer was with the highest accuracy 92.8%, thus predicting the most accurate diagnosis for RSA. Recurrent spontaneous abortion is associated with abnormal coagulation and anticoagulation. TEG combined with detection of AT-III and D-Dimer levels can distinguish patient with RSA from those with normal fertility and highly possibly predict the occurrence of RSA.

Is tri-iodothyronine a better choice than activated protein C in sepsis treatment?

Sepsis can be defined as a life-threatening organ dysfunction due to a dysregulated host response to infection. In sepsis, the coagulation cascade is activated and the balance shifts to the procoagulant side. Recently, the use of protein C is proposed for the treatment of sepsis. Another therapeutic agent that has been intensively studied is tri-iodothyronine. This study aimed to compare the effects of activated protein C and tri-iodothyronine, which are administered at a single dose to sepsis-induced rats at the late phase. Leukocyte, platelet, hemoglobin and antithrombin-III concentrations and histopathological changes in the small intestine, liver and lung were evaluated at 24 hours. Single-dose intraperitoneal recombinant human APC (activated protein C) has a partial curative effect on hematological parameters in the late phase, while it is possible to state that it has significant therapeutic effects on hepatic and intestinal tissues, but more remarkably on the lung tissue. Tri-iodothyronine is also considered to be used for the treatment and has a strong potential to be a therapeutic agent. We observed that the T3 hormone has significantly limited and reduced the sepsis-related damage to hepatic and intestinal tissues, but especially the lung tissue. Tri-iodothyronine can be a good alternative to APC, which is partially allowed due to high cost and complication of bleeding in the treatment of sepsis.

The evaluation of hemostatic dysfunction and disseminated intravascular coagulation in dairy cows with abomasal displacement.

Disseminated intravascular coagulation (DIC) is a complex, dynamic and hemostatic disorder which develops secondarily to a disease characterized with an imbalance in the pro-coagulant and anti-coagulant components of hemostasis. The aim of the study is to evaluate hemostatic dysfunc- tion and the DIC syndrome in cattle with displaced abomasum (DA), with using the hematologic analyses and an extensive coagulation profile in the 96 hour-period including before and after surgery. The animal material of the study consisted of 12 dairy cows diagnosed with displaced abomasum (9 LDA and 3 RDA without volvulus) in the 2-4 week period after parturation and with no other post-partum disease. In dairy cows diagnosed with DA, hematological, coagulomet- ric (PT, APTT, Fibrinogen) and coagulation factor analyses [D-Dimer, TAT (thrombin-anti- thrombin complex), ATIII (antithrombin III), PAI-1 (plazminogen activator inhibitor-1] were performed in blood samples obtained before the operation as well as 30 minutes, 60 minutes and 2, 5, 10, 24, 48, 72 and 96 hours after the operation. In the DA cases, abnormalities were found in 6 of the 8 coagulation parameters. In the LDA and RDA groups, prolonged PT (sec), PT (INR) and APTT, hypofibrinogenemia, an increase in serum D-Dimer concentration at 72 and 96 hours after the operation and an increase in serum ATIII concentrations before and 30, 60 minutes and 2, 5, 72 and 96 hours after the operation was found (p⟨0.05). Hemostatic dysfunction and the risk of DIC developing in DA cases and continuing in the post-operative period was determined.

Effects of tibolone on fibrinogen and antithrombin III: A systematic review and meta-analysis of controlled trials

Tibolone is a synthetic steroid with estrogenic, androgenic and progestogenic activity, but the evidence regarding its effects on fibrinogen and antithrombin III (ATIII) has not been conclusive. We assessed the impact of tibolone on fibrinogen and ATIII through a systematic review and meta-analysis of available randomized controlled trials (RCTs). The search included PUBMED, Web of Science, Scopus, and Google Scholar (up to January 31st, 2016) to identify controlled clinical studies investigating the effects of oral tibolone treatment on fibrinogen and ATIII. Overall, the impact of tibolone on plasma fibrinogen concentrations was reported in 10 trials comprising 11 treatment arms. Meta-analysis did not suggest a significant reduction of fibrinogen levels following treatment with tibolone (WMD: −5.38%, 95% CI: −11.92, +1.16, p=0.107). This result was robust in the sensitivity analysis and not influenced after omitting each of the included studies from meta-analysis. When the studies were categorized according to the duration of treatment, there was no effect in the subsets of trials lasting either <12months (WMD: −7.64%, 95% CI: −16.58, +1.29, p=0.094) or ≥12months (WMD: −0.62%, 95% CI: −8.40, +7.17, p=0.876). With regard to ATIII, there was no change following treatment with tibolone (WMD: +0.74%, 95% CI: −1.44, +2.93, p=0.505) and this effect was robust in sensitivity analysis. There was no differential effect of tibolone on plasma ATIII concentrations in trials with either <12months (WMD: +2.26%, 95% CI: −3.14, +7.66, p=0.411) or≥12months (WMD: +0.06%, 95% CI: −1.16, +1.28, p=0.926) duration. Consistent with the results of subgroup analysis, meta-regression did not suggest any significant association between the changes in plasma concentrations of fibrinogen (slope: +0.40; 95% CI: −0.39, +1.19; p=0.317) and ATIII (slope: −0.17; 95% CI: −0.54, +0.20; p=0.374) with duration of treatment. In conclusion, meta-analysis did not suggest a significant reduction of fibrinogen and ATIII levels following treatment with tibolone.

Identification of a panel of serum protein markers in early stage of sepsis and its validation in a cohort of patients.

Sepsis is a life-threatening illness with a challenging diagnosis. Current serum biomarkers are not sensitive enough for diagnosis. With the aim of finding proteins associated with sepsis, serum protein profile was compared between patients and healthy donors and serum classical inflammatory proteins were analyzed in both groups. Serum protein profiles were characterized by two-dimensional electrophoresis (2DE). Identification of the proteins was carried out by mass spectrophotometry and their validation was performed by Enzyme-Linked-ImmunoSorbent Assay (ELISA) in a cohort of 85 patients and 67 healthy donors. Seven classical inflammatory proteins were analyzed in the same cohort by ELISA: interleukin-2 receptor α-chain (sCD25), scavenger receptor cysteine-rich-type-1 (sCD163), tumor-necrosis factor receptor superfamily-member-6 (sFas), hemeoxigenase-1 decycling (HO-1), interleukin-6 (IL-6), interleukin-18 (IL-18) and intercellular adhesion-molecule-1 (sICAM-1). After 2DE, 20 significantly differently expressed spots were identified by mass spectrometry analysis, revealing deregulation of six different proteins upon sepsis and 50% were validated by ELISA: Antithrombin-III (AT-III), Clusterin (CLUS) and Serum amyloid A-1 (SAA-1). Serum concentration of AT-III and CLUS was significantly lower in patients' serum, whereas SAA-1 showed higher values in septic patients. Serum concentration of the seven inflammatory proteins was significantly increased in septic patients. Functional analysis of the ten deregulated proteins revealed an enrichment of proteins related mainly to the activation of the immune response. We have identified a panel of ten potential sepsis marker proteins biologically connected and validated in a large number of patients, whose analysis could be considered as a complementary tool for the diagnosis of sepsis.

Antithrombin III, but not C1 esterase inhibitor reduces inflammatory response in lipopolysaccharide-stimulated human monocytes in an ex-vivo whole blood setting

In order to examine the immunomodulatory effects of antithrombin III (AT-III) and C1 esterase inhibitor (C1-INH) in human monocytes, we investigated the intracellular expression of interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α in an ex-vivo laboratory study in a whole blood setting.Heparinized whole blood samples from 23 healthy male and female volunteers (mean age: 27±7years) were pre-incubated with clinically relevant concentrations of AT-III (n=11) and C1-INH (n=12), then stimulated with 0.2ng/mL lipopolysaccharide (LPS) for 3h. After phenotyping CD14+ monocytes, intracellular expression of IL-6, IL-8, and TNF-α was assessed using flow cytometry. In addition, 12 whole blood samples (AT-III and C1-INH, n=6 each) were examined using hirudin for anticoagulation; all samples were processed in the same way. To exclude cytotoxicity effects, 7-amino-actinomycin D and Nonidet P40 staining were used to investigate probes.This study is the first to demonstrate the influence of C1-INH and AT-III on the monocytic inflammatory response in a whole blood setting, which mimics the optimal physiological setting. Cells treated with AT-III exhibited significant downregulation of the proportion of gated CD14+ monocytes for IL-6 and IL-8, in a dose-dependent manner; downregulation for TNF-α did not reach statistical significance. There were no significant effects on mean fluorescence intensity (MFI). In contrast, C1-INH did not significantly reduce the proportion of gated CD14+ monocytes or the MFI regarding IL-6, TNF-α, and IL-8. When using hirudin for anticoagulation, no difference in the anti-inflammatory properties of AT-III and C1-INH in monocytes occurs.Taken together, in contrast to TNF-α, IL-6 and IL-8 were significantly downregulated in monocytes in an ex-vivo setting of human whole blood when treated with AT-III. This finding implicates monocytes as an important point of action regarding the anti-inflammatory properties of AT-III in sepsis. C1-INH was unable to attenuate the monocytic response, which supports the hypothesis that other cellular components in whole blood (e.g., neutrophils) might be responsible for the known effects of C1-INH in inflammation.

Hepatic Ischemia/Reperfusion Injury Is Diminished by Atorvastatin in Wistar Rats

Temporal occlusion of the hepatoduodenal ligament (HDL) is often used during liver surgeries in order to reduce blood loss, resulting in ischemia/reperfusion injury (I/R). The aim of the study was to investigate the effects of atorvastatin (ATOR) on hepatic I/R injury and on serum levels of tumor necrosis factor-alpha (TNF-α), endothelin-1 (ET-1), antithrombin III (ATIII) and intracellular adhesion molecule-1 (ICAM-1). Liver ischemia was induced in Wistar rats by clamping the HDL for 60 min, followed by either 60 or 180 min reperfusion. Rats received either vehicle or 10 mg/kg ATOR before hepatic I/R. Control group received sham surgery. Livers were examined for histological damage and serum AST, ALT, TNF-α, ET-1, ATIII and ICAM-1 concentrations were measured. After I/R, AST and ALT were significantly elevated, ATIII levels were significantly depleted, both TNF-α and ICAM-1 levels increased and ET-1 was significantly elevated (at 180 min). ATOR pretreatment attenuated these alterations and diminished histological injury scores. Our results show that ATOR protects the liver from I/R injury.

Alternations of Maternal and Cord Plasma Hemostasis in Preeclampsia Before and After Delivery

Objective: The aim of this study was to investigate the role of hemostatic factors in the pathogenesis of preeclampsia. Materials and Methods: Maternal and cord plasma concentrations of tissue factor (TF), tissue factor pathway inhibitor (TFPI), von willebrand factor (vWF), soluble P-selectin (sP-selectin), fibrinopeptide A (FPA), D-dimer, and antithrombin III (AT-III) were measured by enzyme-linked immunosorbent assay (ELISA) in 46 women with preeclampsia and 40 normotensive pregnant women before and after delivery. Results: The maternal plasma concentrations of TF, vWF, and sP‐selectin were higher, but lower concentrations of TFPI, AT-III, and D-dimer were observed in women with preeclampsia compared to normotensive pregnant women before and after delivery. Compared with maternal plasma, fetal plasma concentrations of TF concentrations were increased significantly in both groups, whereas vWF, FPA, TFPI, AT-III, and D-dimer were decreased. Compared with normotensive pregnancy, fetal plasma concentrations of TF were markedly increased in preeclampsia, accompanied with a higher vWF and a lower sP-selectin and D-dimer levels. Furthermore, fetal plasma TF concentrations were more significantly increased in women with high blood pressure and severe proteinuria. Conclusions: Imbalance in the coagulation/fibrinolysis equilibrium, especially alterations in the extrinsic pathway of coagulation and anticoagulation, may play an important role in the pathogenesis of preeclampsia. In addition, fetal alteration of TF may be involved in the pathogenesis of fetal complications of preeclampsia.

Abstract 2728: Multivariate analysis of cerebrospinal fluid protein biomarkers in central nervous system lymphoma patients and controls

Abstract Establishing the diagnosis of primary central nervous system lymphoma (PCNSL) is often difficult as the clinical presentation and radiographic features are nonspecific. The gold standards for diagnosis are cytologic analysis of the CSF, which has poor sensitivity, or brain biopsy, which may be associated with significant morbidity and a high rate of diagnostic failure. We are testing the hypothesis that the cerebrospinal fluid (CSF) proteome contains information which may facilitate early and noninvasive diagnosis of CNS lymphoma as well as provide prognostic information. We evaluated the CSF concentrations of three candidate CNS lymphoma protein biomarkers: CXCL-13, a B-cell homing chemokine, Interleukin-10 (IL-10), an autocrine B-cell growth factor, and antithrombin-III (ATIII), a candidate tumor biomarker which we previously identified by mass spectrometric analysis of the CSF. Determinations were performed by ELISA of CSF diagnostic specimens from 228 patients: 75 specimens were from patients with a confirmed diagnosis of non-Hodgkin's lymphoma of the brain and 153 controls including 15 specimens from patients with astrocytic neoplasms, 16 specimens from secondary brain tumors including metastases from breast and lung cancer, and 122 other controls including patients with established systemic lymphoma who staged negative for CNS involvement. Mean concentrations of CXCL-13, IL-10 and ATIII were each significantly elevated in the CSF of CNS lymphoma patients compared to the controls (p&amp;lt;0.0003). By contrast, mean CSF albumin concentration, measured to control for disruption of the blood-brain barrier in the two groups, was similar in CNS lymphoma versus controls (p=0.17). The potential utility of each of these candidate biomarkers for CNS lymphoma was evaluated by means of receiver operating characteristic (ROC) curve analysis, both individually as well as in combination. Bivariate determination of CXCL-13 and IL-10 concentrations yield the best potential discrimination of CNS lymphoma versus all other controls. Detection of IL-10 in combination with CXCL-13 in CSF identified CNS lymphoma with the highest accuracy with area under the ROC curve equal to 0.858 (79% sensitivity and 92.2% specificity), (p&amp;lt;0.0001). In addition, we noted a potential relationship between high CSF concentration of CXCL-13 and short survival at first relapse. These preliminary data support a role for CXCL-13 and IL-10 in the pathogenesis of B-cell non-Hodgkin's lymphoma involving the brain and support our hypothesis that molecular signals within the CSF may be rapidly evaluated in combination to facilitate early and relatively non-invasive diagnosis. This approach may also be applied with other diagnostic modalities such as neuroimaging and flow-cytometry. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2728.

Effects of pregnancy and lactation on warfarin-induced changes in blood coagulation-related parameters in rats

Effects of pregnancy and lactation on warfarin-induced changes in blood coagulation-related parameters were examined in rats. Warfarin (0.5 mg/kg/day) was given orally to pregnant and non-pregnant rats for 3 days from gestation day (GD) 17 to 19 or to lactating and non-pregnant rats for 3 days from post partum day (PPD) 10 to 12. Blood samples were collected from the rats on the day following the last administration (GD 20 or PPD 13) to measure prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombotest (TBT), factor VII and X activities and anti-thrombin III concentration (ATIII). Administration of warfarin to non-pregnant rats resulted in significant prolongation of APTT and TBT and significant decreases in factor VII and X activities. On the other hand, similar but not significant changes were observed in pregnant rats and similar significant but less prominent changes were observed in lactating rats. The reduction of the anticoagulant effects of warfarin may partially be related to high plasma 17beta-estradiol concentration in pregnant rats and to high plasma prolactin concentration in lactating rats, respectively.

Resolution of Preoperative Portal Vein Thrombosis After Administration of Antithrombin III in Living Donor Liver Transplantation: Case Report

A 59-year-old man with hepatitis C virus–associated liver cirrhosis was transferred to our hospital to undergo living donor liver transplantation. Coagulation was impaired (prothrombin time [International Normalized Ratio], 3.27), and antithrombin III (AT-III) activity was 23% (normal, 87%–115%). Contrast-enhanced computed tomography scans revealed portal vein thrombosis (PVT) from the junction between the splenic and superior mesenteric vein to the porta hepatica; the portal vein was completely obstructed (PVT). To prevent further development of PVT, 1500 U of AT-III was administered for 3 days, elevating the AT-III activity to 50%. A contrast-enhanced computed tomography scan obtained 9 days after AT-III administration showed resolution of PVT. Living donor liver transplantation was safely performed without portal vein grafting. Thus, a low AT-III concentration may have an important role in the pathogenesis of PVT in patients with cirrhosis.

Overexpression of GADD34 Enhances Production of Recombinant Human Antithrombin III in Chinese Hamster Ovary Cells

To improve the production of recombinant human antithrombin III (AT-III) in Chinese hamster ovary (CHO) cells, the gene encoding growth arrest and DNA damage inducible protein 34 (GADD34), which is a transcription factor involved in the unfolded protein response (UPR), was cloned from CHO-K1 cells. Overexpression of GADD34 significantly enhanced the production of recombinant AT-III in CHO 13D-35D cells. The specific rate of AT-III production in the GADD34-overexpressing CHO 13D-35D cells reached approximately 28 pg/cell/d. After 144 h of incubation, the AT-III concentration in the culture supernatant was approximately 40% higher than that observed in the case of the parental CHO 13D-35D cells. The mRNA expression, specific activity, and fucosylation of AT-III were not affected by GADD34 overexpression. Overexpression of GADD34 is a promising method of improving the production of secreted protein pharmaceuticals in CHO cells.

Improved production of recombinant human antithrombin III in Chinese hamster ovary cells by ATF4 overexpression

To improve the production of recombinant human antithrombin III (AT-III) in Chinese hamster ovary (CHO) cells, the genes encoding transcription factors, ATF4 (activating transcription factor 4) and XBP-1s (the spliced form of X-box binding protein 1), which were involved in the mammalian unfolded protein response (UPR), were cloned from CHO-K1 cells. Overexpression of ATF4 significantly enhanced the production of recombinant AT-III in CHO 13D-35D cells. The specific rate of AT-III production in the ATF4-overexpressed CHO 13D-35D cells reached approximately 23 pg/cell/day. After 144 h of incubation, the AT-III concentration in the culture supernatant was twofold greater compared to that observed with parental CHO 13D-35D cells. In contrast, ectopic expression of XBP-1s failed to enhance the production of recombinant AT-III in CHO 13D-35D cells. RT-PCR analysis revealed that high levels of XBP-1s mRNA were present in the CHO cells, regardless of ectopic expression of XBP-1s. Our results indicate that overexpression of the UPR transcription factor ATF4 is a promising means for improving the production of secreted protein pharmaceuticals in CHO cells.

Serum protease inhibitor concentrations and total antitrypsin activity in diabetic and non-diabetic children during adolescence

The aim of the study was the assessment of the concentrations and establishment of mutual relationships between three main protease inhibitors: alpha-1-antitrypsin (AAT), alpha-2-macroglobulin (alpha-2-M) and antithrombin-III (AT-III), and of the total trypsin inhibitory capacity (TIC) in the serum of diabetic and non-diabetic children during adolescence. Forty-nine children (24 girls and 25 boys) with type 1 diabetes mellitus and 24 non-diabetic children (13 girls and 11 boys) were divided according to the Tanner scale into three groups: pre-, peri- and post-pubertal. The concentrations of AAT, alpha-2-M and AT-III were determined by the radial immunodiffusion method on NOR-Partigen plates (Dade-Behring), while TIC was determined by the method using BAPNA as substrate. Means and medians of serum AAT [1.55 g/l, 1.40 (95% confidence interval, 1.42-1.68), respectively] and TIC [10.6 mg trypsin/100 ml, 10.3 (95% CI, 9.5-11.7)] in diabetic children were lower than means and medians of AAT [1.81 g/l, 1.60 (95% CI 1.55-2.07), respectively] and TIC [12.5 mg trypsin/100 ml, 13.2 (95% CI, 10.9-14.1)] in non-diabetic children. A comparison of variables between Tanner groups shows an increasing trend of AAT concentration in diabetic children and a decreasing trend of TIC in non-diabetic subjects. In contrast to pre- and peri-puberty, no correlations were found in the postpubertal period between the studied parameters, either in diabetic or non-diabetic patients. Hyperglycaemia and the duration of diabetes were found to have a significant association with alpha-2-M and AT-III concentrations, but not with AAT serum concentrations. The concentrations and correlations between serum protease inhibitors in diabetic children during adolescence are disrupted compared with non-diabetic children. Taking into account the unfavourable consequences of vascular complications resulting from serum trypsin inhibitor changes and protease- antiprotease imbalance, diabetic children are at greater risk of this occurring during adolescence.

Plasma antithrombin III and protein C levels in early recognition of late-onset sepsis in newborns

The aim of this prospective, observational study was to establish the prognostic values of plasma antithrombin III (ATIII) and protein C (PC) functional levels, determined in 150 neonates with suspected late-onset sepsis. Both the ATIII and PC concentrations were significantly lower in neonates with sepsis, either confirmed or not confirmed by blood culture, than data obtained in infants with excluded disease. The lowest values of plasma ATIII and PC were observed in neonates who had died in the course of sepsis. The differences between survivors and non-survivors were statistically significant (respectively, for ATIII P=0.003 and for PC P=0.00002). A highly statistically significant correlation (P=0.0016) between plasma PC functional level and risk of death was found in patients with sepsis. However, plasma ATIII concentration, evaluated in the same group of patients, did not correlate with the occurrence of death. We concluded that the measurement of plasma ATIII and, especially, PC levels may facilitate the recognition of sepsis and, with respect to PC, may have a prognostic value. Plasma PC functional concentration below 10% might be the indication for supplementation of PC concentrate.