Newly Developed Recombinant Antithrombin Protects the Endothelial Glycocalyx in an Endotoxin-Induced Rat Model of Sepsis.

(1) Background: The endothelial glycocalyx is a primary target during the early phase of sepsis. We previously reported a newly developed recombinant non-fucosylated antithrombin has protective effects in vitro. We further evaluated the effects of this recombinant antithrombin on the glycocalyx damage in an animal model of sepsis. (2) Methods: Following endotoxin injection, in Wistar rats, circulating levels of hyaluronan, syndecan-1 and other biomarkers were evaluated in low-dose or high-dose recombinant antithrombin-treated animals and a control group (n = 7 per group). Leukocyte adhesion and blood flow were evaluated with intravital microscopy. The glycocalyx was also examined using side-stream dark-field imaging. (3) Results: The activation of coagulation was inhibited by recombinant antithrombin, leukocyte adhesion was significantly decreased, and flow was better maintained in the high-dose group (both p < 0.05). Circulating levels of syndecan-1 (p < 0.01, high-dose group) and hyaluronan (p < 0.05, low-dose group; p < 0.01, high-dose group) were significantly reduced by recombinant antithrombin treatment. Increases in lactate and decreases in albumin levels were significantly attenuated in the high-dose group (p < 0.05, respectively). The glycocalyx thickness was reduced over time in control animals, but the derangement was attenuated and microvascular perfusion was better maintained in the high-dose group recombinant antithrombin group (p < 0.05). (4) Conclusions: Recombinant antithrombin maintained vascular integrity and the microcirculation by preserving the glycocalyx in this sepsis model, effects that were more prominent with high-dose therapy.

[Case of dyspnea due to toilet cleaner containing nitric acid for household use].

A 40-year-old male tried to clean a urinal at his home storing 900 mL of a toilet cleaner containing 9.8% nitric acid to remove calcium deposit, and clean the toilet floor for twenty minutes. Immediately after using the cleaner, he experienced eye irritation. He washed out the toilet cleaner. However, he thereafter experienced dyspnea, a compressive sensation in his chest, and chest and back pain about 40 minutes after the cleaning the toilet. He monitored his symptoms overnight and found them to gradually improve. However, the symptoms still remained the next morning and therefore he came to our department on foot. He had no particular past or family history. On arrival, his physiological findings and chest computed tomography scan were negative for any abnormalities. His arterial blood gas analysis revealed a mild abnormality of oxygenation. Observation without any drugs revealed that a complete remission of his symptoms occurred after approximately 4 weeks. Based on the results of the experiments, contact with the mucosal membrane and nitric acid gas produced by any accidentally coexisting metals or contact with moisture, including nitric acid produced by a reaction between CaCO3 and cleaner, may have been the mechanism of occurrence for the symptoms observed in this case. This is the first reported case of nitric acid poisoning due to the use of a toilet cleanser intended for household use.

A case of cardiac arrest with ST elevation induced by contrast medium.

A 62-year-old man with local recurrence of pancreatic cancer underwent his 17th infusion of contrast medium. He had no history of allergy and had not experienced any side effects from the contrast medium during any of the previous examinations. During infusion, he complained of nausea, followed by a loss of consciousness. He was injected intramuscularly with 0.3 mg adrenalin; however, he temporally went into cardiopulmonary arrest. He was therefore injected with 100 mg hydrocortisone and the continuous infusion of dopamine for shock. His electrocardiogram revealed ST elevation. An urgent cardiac echo evaluation revealed hyperkinetic wall motion. As his blood pressure increased after the initiation of the treatment, the ST elevation started to normalize. After transportation to an intensive care unit, the patient did not show chest pain, ST elevation on cardiograms, or any increase in the levels of cardiac markers. Based on his clinical course, the cause of the patient's ST elevation was considered to be coronary vasospasm. Kounis syndrome is the concurrence of acute coronary syndromes with conditions associated with mast cell activation, including allergic or hypersensitivity and anaphylactic or anaphylactoid insults. In cases of coronary vasospasm with shock due to contrast medium, supportive therapy using catecholamine, which has coronary vasodilator activity, and a steroid might be effective to treat the coronary vasospasm. Attention should therefore be paid to the patient's complaints, the findings of real-time cardiosonography, electrocardiograms, and the levels of cardiac markers to ensure a correct diagnosis and to achieve a good treatment outcome.

Comparison of prothrombin time tests used in the monitoring of edoxaban and their evaluation as indicators of the reversal effect.

Clinical demand for the prompt assessment of the activity of direct-acting factor Xa inhibitors in the emergency care setting is increasing. In the present study, we examined whether prothrombin time (PT) tests can serve as a clinically useful indicator of anti-factor Xa activity. In the first series, the in vitro effect of edoxaban on PT was evaluated by spiking human plasma with edoxaban and measuring PT using three different commercial PT tests. In the second series, the reversal effect of prothrombin complex concentrates (PCC) and activated PCC (aPCC) in edoxaban-spiked plasma was evaluated. In the third series, PT of plasma samples from patients administered either 15 or 30 mg/day of edoxaban was assessed, and the results were compared with edoxaban concentrations determined by a calibrated anti-factor Xa activity assay. The spike test revealed that all PT reagents positively correlated with edoxaban. The sensitivity to edoxaban varied among the three reagents and Triniclot(®) Excel S showed the best performance. Prolonged PT by edoxaban was reversed by PCC and aPCC in a dose-dependent manner; however, complete reversal was not achieved. Positive correlation between anti-factor Xa activity and PT was shown in the clinical samples at the edoxaban range from 0 to >300 ng/mL.

Rivaroxaban attenuates leukocyte adhesion in the microvasculature and thrombus formation in an experimental mouse model of type 2 diabetes mellitus.

INTRODUCTION: Thrombosis is a major complication in diabetes mellitus. Since Factor Xa inhibitors are not only inhibit the coagulation system but also attenuate the leukocyte-endothelial interaction in acute inflammation models, the purpose of this study is to confirm the similar effects of rivaroxaban in a mouse model of type 2 diabetes mellitus. MATERIALS AND METHODS: In the treatment groups, either 5 or 10mg/kg of rivaroxaban dissolved in DMSO was orally given to KK-A(y) mice for 7 weeks (n=6 in each group). KK-A(y) mice fed by chow containing DMSO without rivaroxaban for 7 weeks were served for the control group (n=6). Following clamping of the mesenteric vein for 20 minutes, intravital microscopic observation of the intestinal microcirculation and the measurement of bleeding time after the needle puncture were carried-out. In another series, the calculation for blood cell counts and the measurement of blood fluidity using micro channel array flow analyzer (MC-FAN) were performed. RESULTS: The initial event in the microvasculature is the leukocyte adhesion on endothelium. Then, the leukocytes make clusters and the platelets are involved in. These leukocyte-platelet conjugates aggregate and form thrombus. The leukocyte adherence and the microthrombus formation was significantly suppressed with the treatment of 10 mg/kg of rivaroxaban compared to the control group (P<0.05). While, the bleeding time was significantly extended with the treatment with 10mg/kg of rivaroxaban (P<0.01). The blood fluidity was maintained best with the treatment of 10 mg/kg rivaroxaban. CONCLUSIONS: Rivaroxaban attenuates the leukocyte-platelet-endothelial interaction, which leads to the attenuation of microthrombus formation in a mouse model of diabetes mellitus.

Analysis of non-traumatic truncal back pain in patients who visited an emergency room.

AIM: To investigate epidemiology of acute non-traumatic back pain using modern diagnostic methods in patients who visited an emergency room. METHODS: The medical charts were retrospectively reviewed for all patients with back pain who were treated in our hospital. In addition, the patients were divided into two groups based on whether they were treated at the hospital or as outpatients. RESULTS: There were 95 patients with non-traumatic acute back pain. Leading cause of back pain was ureterolithiasis (53 cases), followed by pyelonephritis (10), orthopedic disease including two cases of purulent spondylitis (24), aortic disease (3), pancreatitis (1), renal bleeding (1), adrenal bleeding (1), psoas abscess (1), and torsion of an ovarian tumor (1). All cases of pyelonephritis, aortic disease, purulent spondylitis, renal bleeding, adrenal bleeding, psoas abscess, and torsion of an ovarian tumor were treated in admission. Using a multiple logistic regression analysis, blood pressure, age, and body temperature were the only factors that were independently associated with whether the patient was admitted or treated as an outpatient. CONCLUSION: This study showed that urological diseases are the most common cause of back pain in patients who visit the emergency room, followed by orthopedic disease. Older age, low blood pressure, and high body temperature were independently associated with the decision to admit the patient who might have lethal disease.

Recombinant thrombomodulin improves the visceral microcirculation by attenuating the leukocyte-endothelial interaction in a rat LPS model.

INTRODUCTION: Abnormalities in vascular endothelial function play an important role in the development of septic organ dysfunction. The aim of this study was to examine the effect of recombinant thrombomodulin (rTM) on leukocyte-endothelial interaction and subsequent malcirculation in endotoxemia. MATERIALS AND METHODS: Wistar rats were administered with either low, medium or high dose of rTM (n=7 each) 2hours after lipopolysaccharide (LPS) infusion. Mesenteric microcirculation after the treatment was observed under the intravital microscopy. In another series (n=5 each), plasma levels of high-mobility group box 1 (HMGB1) levels were measured at 5hours after LPS infusion. RESULTS: Microscopic findings revealed suppression in leukocyte adhesion, thrombus formation and endothelial damage with the treatment by rTM. However, high-dose rTM tended to increase the bleeding events. Thus, blood flow was better maintained with medium-dose rTM (P<0.05). The increase in HMGB1 level was significantly suppressed by medium and high-dose rTM (P<0.05, respectively). CONCLUSIONS: rTM demonstrated a protective effect on microcirculation through the inhibition of leukocyte-endothelial interaction and suppression of HMGB1.

Factor Xa inhibitor attenuates leukocyte adhesion and thrombus formation in an experimental mouse model of the metabolic syndrome.

AIMS: In a model of acute inflammation, Factor Xa inhibitors have been reported not only to suppress the coagulation system but also to exert anti-inflammatory effects. However, this has not been experimentally demonstrated in a model of chronic inflammation. Recent studies demonstrated that vascular inflammation in the metabolic syndrome plays major roles in the development of thrombotic diseases. Therefore, we examined the anti-inflammatory effects of fondaparinux, a Factor Xa inhibitor, in a mouse model of the metabolic syndrome, looking at both leukocyte adhesion on the vascular endothelium and thrombus formation. METHODS: Following clamping of the mesenteric vein for 20 min in the KK-A(y) mouse, mice were administered by subcutaneous injection either low-dose or high-dose fondaparinux or placebo (n = 10 in each group. Microscopic observation of the intestinal microcirculation was carried out. In another series, blood samples were taken and measured for blood cell counts and organ damage markers (n = 6 in each). RESULTS: Both leukocyte adherence and thrombus formation were inhibited by treatment with fondaparinux. Red blood cell and white blood cell counts were maintained better in high-dose group. Levels of alanine aminotransferase (ALT) were significantly reduced in both low-dose and high-dose groups (P < 0.05 and 0.01, compared with control, respectively). CONCLUSIONS: Factor Xa inhibitor attenuates leukocyte adhesion and leukocyte-platelet conjugate formation in a mouse model of the metabolic syndrome. These effects appeared to be related to both inhibition of thrombus formation and reduction in markers of organ damage.

Enoxaparin and fondaparinux attenuates endothelial damage in endotoxemic rats.

BACKGROUND: Prophylactic use of anticoagulants for septic patients in intensive care unit is a standard therapy for the prevention of venous thrombosis. Moreover, recent studies have demonstrated the anti-inflammatory effects of anticoagulants such as Factor Xa inhibitors and heparins. However, there have been no studies to examine the effects of fondaparinux and enoxaparin when applied in a sepsis model. Therefore, we examined the anti-inflammatory effects and bleeding events when these agents are applied in a lipopolysaccharide challenge model. METHODS: Wistar rats received lipopolysaccharides followed by a bolus infusion of fondaparinux, enoxaparin, or placebo. Microscopic observation of the mesenteric microcirculation for endothelial damage and measurement of bleeding area after vascular puncture was performed (n = 6 in each group). In another series, blood samples were taken, and blood cell counts, coagulation markers, and organ damage markers were measured (n = 6 in each). RESULTS: Both leukocyte adherence to vascular endothelium and endothelial damage were reduced in fondaparinux and enoxaparin groups. The bleeding area was markedly increased in the fondaparinux group. Coagulation markers were maintained better in the enoxaparin group. Levels of organ damage markers were significantly suppressed in both fondaparinux and enoxaparin groups (p < 0.01, compared with control, each). CONCLUSIONS: Fondaparinux and enoxaparin reduce organ dysfunction by decreasing endothelial damage. However, bleeding was more prominent in the fondaparinux group compared with the enoxaparin group at an equipotent dose for anti-Xa activity. Because the setting of this experiment is different from the clinical use, further study is required for the comparison of both pharmaceuticals.

Formation of the venous thrombus after venous occlusion in the experimental mouse model of metabolic syndrome.

INTRODUCTION: The metabolic syndrome is considered to be a risk factor for the venous thromboembolism (VTE) as well as arterial thrombosis. Although obesity, hyperglycemia and dyslipidemia are considered to be important triggering factors, it is difficult to evaluate the relationship between VTE and the metabolic syndrome in a clinical study. Furthermore the mechanism of venous thrombosis initiation still remains elusive. MATERIALS AND METHODS: 20 min clamp of superior mesenteric vein was applied to 7 w, 16 w-old KK-A(y) mouse and 16 w-old B6J mouse (n = 6 in each group), after de-clamp, the view of the mesenteric vein and intestinal submucosal venule were observed by the intravital microscopy. RESULTS: Massive thrombi formed in the mesenteric vein in 16 w-old KK-A(y) mice, moderate thrombi formation was observed in 7 w-old KK-A(y) mice, while very few thrombi were observed in B6 J mice. The first event in submucosal venule after de-clamp was the adhesion of leukocytes to the endothelium. Subsequently, leukocytes assembled and platelets covered the leukocyte cluster. These leukocyte-platelet aggregates move from the venule to the vein and finally formed a venous thrombus. CONCLUSION: Metabolic syndrome is a risk factor for venous thrombosis. Intravital microscopic examination revealed leukocyte and platelet recruitment to the venule in the early stages of venous thrombosis formation.

Estimated glomerular filtration rate is an independent predictor for mortality of patients with acute heart failure.

BACKGROUND: Heart failure is a major public health problem in developed countries including Japan, therefore it is important to estimate the future risk in patients with heart failure. Recently, it has been reported that chronic kidney disease (CKD) is an independent predictor for mortality in chronic heart failure. However, it is unknown whether CKD is an independent predictor for mortality in acute heart failure. We retrospectively investigated the relationship between estimated glomerular filtration rate (eGFR) on admission for acute heart failure and long-term mortality. METHODS: We analyzed 194 patients who were admitted for acute heart failure from January, 2002 to February, 2005. Patients were divided into two groups, high-eGFR group (eGFR <60 ml/min, n=75) and low-eGFR group (eGFR > or =60 ml/min, n=119). eGFR was calculated by equation of MDRD (modification of the diet in renal disease) study. eGFR level <60 ml/min/1.73 m(2) is impaired renal function according to the guidelines of the Japanese Society of Nephrology and of the National Kidney Foundation. Serum B-type natriuretic peptide (BNP) level and left ventricular ejection fraction (LVEF), anemia, age, gender, and etiology of heart failure were also evaluated. Median observation period was 609 days (range: 30-1627). Mean age was 69 years and 138 patients were male. RESULTS: Median eGFR on admission was 74.2 ml/min (range: 5.48-238.7), median BNP level was 840 pg/ml (range: 200-4800), and median LVEF was 36% (range: 11-81%). Forty-two percent of patients had eGFR <60 ml/min of eGFR at the time of coronary care unit admission. Patients with low-eGFR had a significantly lower mortality rate by Kaplan-Meier analysis (log rank test, p=0.013). By Cox's proportional-hazard analysis, eGFR was an independent factor for long-term mortality of acute heart failure (p=0.039). CONCLUSIONS: Lower eGFR at the time of admission could be an independent predictor for mortality of acute heart failure.