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The inhibitory effect of human placenta on ADP-induced platelet aggregation was examined. Placental extracts were purified by chromatography and gel filtration. The eluted fractions were subjected to affinity chromatography with Bestatin AH-Sepharose to obtain a more purified substance. A substance markedly inhibited platelet aggregation induced by ADP, and also exhibited high placental leucine aminopeptidase (P-LAP) activity. Preincubation of sample with ADP abolished the platelet aggregatory effect. Thin-layer chromatography (TLC) analysis revealed that the substance hydrolyzed ADP to AMP. The potent anti-platelet aggregating activity of placental extracts appears to be due to ADPase-like action. This action may be one of the controlling factors of hemostasis in fetoplacental circulation.

Before hemodialysis (HD), plasma levels of tissue factor (TF), free-TF pathway inhibitor (TFPI) and thrombo modulin (TM) were significantly higher in patients with HD than in healthy volunteers. Plasma levels of (T-F) TFPI and plasmin plasmin inhibitor complex (PPIC) were significantly higher in patients with HD than in healthy volunteers. During HD, plasma levels of TF and (T-F) TFPI were not significantly increased, but plasma levels of total TFPI and free TFPI at 1 hour after and at the end of HD were significantly increased, compared with levels before start of HD. Plasma level of PPIC 1 hour after start of HD was significantly higher than before start of HD, and plasma levels of thrombin antithrombin com plex (TAT). PPIC. D-dimer. TM, and protein C (PC) at the end of HD were significantly higher than before start of HD. In patients with thrombosis complications, plasma TF levels were significantly higher than in patients without thrombotic com plications during HD. Plasma levels of PC were significantly lower in patients with thrombotic complications than in patients without thrombotic complications. There was no significant difference between both groups during HD in hemostatic pa rameters, with the exception of TF and PC. Hemostatic abnormalities existed in patients with HD; espe cially, increased TF and decreased PC might cause thrombotic complications.

In this study, plasma concentrations of hemostatic molecular markers were investigated in 118 elderly persons with normal renal function (aged 65 to 97 years) who could manage their activities of daily living (ADL) by themselves, to find a strategy for conservation or elevation of ADL and quality of life (QOL). In all subjects, the frequency by which hemostatic markers exceeded their upper limit of normal range was 35.9% for thrombin-antithrombin HI complex (> or = 3.7 ng/ml), 38.3% for soluble fibrin monomer (> or = 4.0 microg/ml), 41.8% for D-dimer (> or = 1.0 microg/ml), 49.0% for plasmin-alpha2 plasmin inhibitor complex (> or = 1.0 microg/ml), and 53.7% for thrombomodulin (> or = 20 ng/ml). The mean plasma levels of these markers were slightly higher than the upper limit of their normal range. These markers were also investigated in samples of patients with and without cardiovascular risk factors and with and without cardiovascular diseases (ischemic heart disease and/or cerebral infarction). Furthermore, the results were analyzed in relationships between cardiovascular disease and cardiovascular risk factor or aging. The findings suggest that aging exerts a stronger influence on plasma levels of these hemostatic molecular markers than the presence of cardiovascular risk factors and cardiovascular diseases. From the viewpoint for conservation or elevation of ADL and QOL in elderly persons, also other factor, such as drug intake, lifestyle, aging, and so on, must be considered to clarify the relationship between the plasma levels of the hemostatic molecular markers and cardiovascular risk factors or cardiovascular diseases.

Fas, a member of the tumor necrosis receptor superfamily, is 36 kD surface protein containing a single transmembrane region and induces apoptosis by Fas-Fas ligand binding. Soluble Fas (sFas) is produced as the form lacking 21 amino acid residues containing the transmembrane domain by alternative splicing. We found that the plasma sFas levels of 33 patients with thrombotic thrombocytopenic purpura/hemolytic uremic syndrome (TTP/HUS), 19 patients with disseminated intravascular coagulation (DIC), and 10 non-DIC patients with multiple organ failure (MOF) were significantly higher than those of 21 non-DIC patients without organ failure and those of 25 healthy volunteers. The plasma sFas ligand levels of the TTP/HUS patients, the DIC patients, and the non-DIC patients with MOF were significantly higher than those of the non-DIC patients without organ failure and those of the healthy volunteers. The plasma sFas levels were significantly correlated with the plasma sFas ligand levels in all subjects. The plasma thrombomodulin (TM) levels were increased significantly in the TTP/HUS patients, the DIC patients, and the non-DIC patients with MOF compared with the levels of the non-DIC patients without organ failure and the healthy volunteers. The plasma sFas antigen levels were correlated significantly with the plasma TM levels in all subjects. These findings suggest that the increases of sFas and sFas ligand that cause apoptosis might be related to the vascular endothelial cell injuries in TTP and DIC with organ failure.

We retrospectively measured various hemostatic markers in 240 patients with disseminated intravascular coagulation (DIC) before the onset of DIC and in 110 non-DIC patients, and examined their usefulness for the diagnosis of pre-DIC. Changes in prothrombin time ratio and fibrinogen levels were not significant before the onset of DIC. The plasma levels of fibrinogen and fibrin degradation products before the onset of DIC were increased and the platelet count was gradually reduced in nonleukemic patients; these changes were already significant in the non-DIC state. The plasma levels of thrombin-antithrombin complex (TAT), plasmin-plasmin inhibitor complex (PPIC), D-dimer, and soluble fibrin monomer (sFM) were increased before the onset of DIC. In leukemic patients, the plasma levels of sFM on day 5, those of TAT on day 3, and D-dimer on day 1, were significantly increased before the onset of DIC. The levels of most hemostatic markers 7 days before the onset of DIC were not different from those observed in the non-DIC state. In nonleukemic patients, only D-dimer, sFM, and TAT levels were significantly increased 7 days before the onset of DIC compared with values in the non-DIC state. The positive rate of hemostatic markers for the diagnosis of DIC, TAT, and PPIC were high during the pre-DIC and non-DIC groups. The plasma levels of sFM and D-dimer were low in non-DIC and increased gradually during the pre-DIC state. These findings suggest that hemostatic molecular markers such as sFM, D-dimer, and TAT are useful for the diagnosis of pre-DIC, although their cutoff values were different among various diseases.

Hemostatic abnormalities were examined in 55 pa tients during maintenance hemodialysis (HD). Before HD, plasma protein C and protein S antigens were almost within the normal range, while plasma thrombin-antithrombin III complex (TAT III) and plasmin-plasmin inhibitor complex (PPIC) levels in HD patients were increased slightly, and plasminogen acti vator inhibitor 1 level was significantly increased, compared to that in normal volunteers. Plasma activated protein C (APC) and protein C inhibitor (PCI) complex and APC α1 antitrypsin (α1AT) complex were not detected in normal volunteers; how ever, plasma APC-PCI complex was increased in 36 of the patients and plasma APC-α1AT complex was increased in 25 patients. Plasma PCI levels in these patients before HD were significantly decreased. Plasma TAT, PPIC, and tissue type plasminogen activator levels were significantly higher before HD than after 1 hour HD and at the end of HD, while the changes in plasma protein C antigen, protein S antigen, PCI antigen, APC-PCI complex, and APC-α1AT complex were not significant after 1 hour of HD or at the end of HD compared to levels before HD. Plasma PCI levels were correlated with APC- PCI complex, suggesting that decreased PCI levels might be caused by the activation of protein C.

We measured plasma-soluble adhesion molecules (granule membrane protein [GMP]-140, endothelial leukocyte adhesion molecule [ELAM]-1, intracellular adhesion molecule [ICAM]-1, and vascular adhesion molecule [VCAM]-1) in 21 patients with thrombotic thrombocytopenic purpura (TTP), 10 with mitomycin C-induced hemolytic uremic syndrome (MMC-HUS) and 59 with disseminated intravascular coagula tion (DIC) or pre-DIC. The plasma levels of soluble GMP-140 in the three groups were significantly higher than the levels in healthy volunteers, suggesting that platelets and vascular en dothelial cells are activated in these diseases. The plasma- soluble GMP-140 level of the DIC patients with nonhemato poietic malignancy was significantly higher than in the DIC patients with hematopoietic malignancy, perhaps due to the release of GMP-140 mainly from platelets. The plasma levels of soluble VCAM-1 in the TTP and DIC patients were signifi cantly higher than the levels in healthy volunteers. The plasma levels of ICAM-1 in DIC patients were significantly higher than those in healthy volunteers, but the levels in the TTP and MMC-HUS groups were not significantly high. The plasma ELAM-1 levels in the TTP and MMC-HUS groups were sig nificantly lower than those in healthy volunteers and DIC pa tients. Since elevated plasma cytokine levels have been ob served in TTP and DIC patients, the adhesion molecules might be upregulated in these diseases. However, plasma ELAM-1 is thought to be reduced in severe vascular endothelial cell dis eases such as TTP.

To define the clinical significance of plasma thrombomodulin (TM) values in elderly, we examined plasma TM in healthy young subjects, healthy elderly subjects and patients with cerebral infarction sequelae. We also studied the relationship with effective renal plasma flow (ERPF) and with the liver's protein-production ability. The TM values of healthy elderly subjects were higher than those of healthy young subjects. There existed an inverse correlation between TM values and ERPF. Accordingly, high TM values might significantly influence renal arteriosclerosis. From the inverse correlation identified between TM and serum cholinesterase, it was estimated that high TM might appear in conjunction with the liver's protein production ability. Patients with cerebral infarction showed higher plasma TM values. It is thought that angiopathy has been maintained in patients as the anamnesis of cerebral infarction even though it occurred in the past. The TM values of patients with diabetes mellitus (DM) were higher than those without it. Moreover, the TM values of patients with DM complicated by retinopathy were higher than those uncomplicated by retinopathy. It is therefore estimated that increased TM might occur with angiopathy resulting from DM. A possibility thus exists that plasma TM could be utilized as one of the markers for endothelial injury.

Plasma-soluble fibrin monomer (SFM) level in patients with disseminated intravascular coagulation (DIC) was significantly higher than the level in patients with pre-DIC or in non-DIC patients, and the level in patients with pre-DIC was significantly higher than that in non-DIC patients. There was no significant difference in plasma SFM levels among various diseases underlying DIC. Plasma SFM level in patients with good outcome was significantly decreased after treatment for DIC. The sensitivity of fibrin degradation products and platelet number was high for DIC, but not for pre-DIC. The sensitivity of thrombin-antithrombin III complex, plasmin-plasmin inhibitor complex, and SFM was high for both DIC and pre-DIC. The specificity of these markers was also high. Receiver operating characteristic analysis suggests that plasma SFM level could be the most useful marker for the diagnosis of both DIC and pre-DIC.

We examined red cell fragmentation syndrome (RCFS) induced by mitomycin C (MMC) (13 patients), by thrombotic thrombocytopenic purpura (TTP) (17 patients), and by disseminated intravascular coagulation (DIC) (15 patients). Plasma cytokine levels were increased in the TTP and DIC patients, but not in those whose RCFS was induced by MMC, suggesting that the activation of the immune system plays an important role in the pathogenesis of RCFS due to TTP and DIC but did not in RCFS due to MMC. Plasma thrombomodulin, tissue type plasminogen activator, and plasminogen activator inhibitor-I levels were increased in all RCFS patients, suggesting that RCFS, whether MMC induced, or due to TTP or DIC, might be associated with vascular endothelial cell injury. In TTP, von Willebrand factor (vWF) antigen and high molecular weight vWF multimer levels were reduced, possibly as a result of microthrombus consumption. The hemostatic data in this study showed that the TTP patients were in a hypercoagulable state without hyperfibrinolysis, and that DIC patients were in both a hypercoagulable and a hyperfibrinolytic state, whereas hemostatic abnormalities were slight in patients with MMC induced RCFS. These findings suggest that vascular endothelial cell injuries might be associated with RCFS, and that those injuries in MMC-induced RCFS might not be related to microthrombi or an activated immune system.

The plasma tissue factor (TF) antigen level was measured in patients with disseminated intravascular coagulation (DIC). The plasma TF antigen was detected In normal volunteers, and it was significantly higher in DIC patients than in non-DiC patients. However, in some patients with DiC, the plasma TF antigen level was within the normal range. The plasma TF antigen level in patients with DIC significantly decreased after therapy, but it was not correlated with organ failure or outcome. The plasma TF antigen level in patients with DIC was not correlated with other hemostatic markers. The plasma TF antigen level tended to be higher in DIC patients with nonlymphoid leukemia than in those with lymphoid tumor. TF might be implicated in the occurrence and progression of DIC. © 1994 Wiley-Liss, Inc.

Two potent inhibitors of protein phosphatase type 1 (PP1) and type 2A (PP2A), calyculin A (CAL-A) and okadaic acid (OKA), inhibited human platelet aggregation induced by thrombin, collagen and 9,11-epithio-11,12-methano-thromboxane A 2 (STA 2 ). IC 50 values of CAL-A and OKA for STA 2 -induced aggregation were 53 nM and 3.5 μM, respectively. These drugs also inhibited thrombin-induced [ 14 C]serotonin secretion of platelets. CAL-A and OKA elicited phosphorylation of certain proteins with an apparent M r (× 10 −3 ) of 200, 60, 50 and 20 light chain of myosin (MLC). Agonist-induced 47,000 M r protein phosphorylation was strongly inhibited by these compounds, whereas phosphorylation of 20,000 M r MLC was enhanced. The increase in 50,000 M r protein phosphorylation by CAL-A and OKA was observed in the presence of agonists, and the 50,000 M r phosphorylation may be involved in the inhibition of platelet activation by these compoundes. Subcellular analysis of the phosphatase activity in human platelets showed that MLC phosphatase activity was present mainly (approx. 78%) in the cytosolic fraction. Chromatography of human platelet extract on heparin-Sepharose resolved two peaks of MLC phosphatase activity: PP2A in 0.1 M NaCl eluate and PP1 in 0.5 M NaCl eluate. PP2A and PP1 isozymes (PP1α PP1α and PP1δ) have also been identified in human platelets, by cross-reactivity with polyclonal antibodies against (PP2A and PP1 isozymes, respectively. These results suggest that PP1 and/or PP2A may play an important role in the process of platelet activation by regulating levels of phosphorylation of certain proteins.

Plasma tissue factor (TF) antigen level and hemostatic markers were measured in 55 patients with disseminated intravascular coagulation (DIC), 15 patients suspected with DIC and 10 patients without DIC. Plasma TF antigen was detected in normal volunteers and it was significantly higher in patients with DIC than in patients suspected with DIC and without DIC. But in some patients with DIC plasma TF antigen level was within normal range. Plasma TF antigen level in patients with DIC significantly decreased after therapy compared with before. And it was not correlated with organ failure and its prognosis. Plasma TF antigen level in patients with DIC was slightly positively correlated with thrombin-antithrombinIII complex but not correlated with other hemostatic markers. In hematopoietic neoplasma with DIC plasma TF antigen level showed a tendency to increase in nonlymphoid leukemia as compared to lymphoid tumor. It was suggested that TF was implicated in occurrence and progression of DIC.

We report a rare case of complete remission for 32 months with continuous treatment with all-trans retinoic acid (ATRA) alone in a patient with acute promyelocytic leukemia which developed as a second malignancy after the treatment of osteosarcoma after failure of conventional chemotherapy. The adverse effects of ATRA were apparently tolerable.

Within the framework of a multicenter clinical study of the recombinant human factor VIII preparation BAY w 6240, we studied the pharmacokinetics of this factor in blood by using the preparation during tooth extraction in a patient with hemophilia A. The results allowed us to conclude that this preparation exerts a hemostatic effect comparable to that of human-derived concentrated factor VIII preparation and that it can be safely used in patients with hemophilia A who undergo tooth extraction.

Hypercholesterolemia is associated with an increased incidence of vascular complications. To assess the actual degree of activation of coagulation systems and vascular disorders in hypercholesterolemia, plasma levels of vascular endothelial cell markers, such as thrombomodulin (TM), tissue-type plasminogen activator, plasminogen activator inhibitor-I (PAI-I), and von Willebrand factor, were measured in 51 patients with hypercholesterolemia. We also investigated the effects of Pravastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, on plasma lipid, lipoprotein a, and hemostatic markers. The mean plasma levels of thrombin-antithrombin III complex (TAT), fibrinopeptide A (FPA), TM, and PAI-I were significantly elevated in hypercholesterolemia. Of the hemostatic markers, only TM was significantly increased in patients with ischemic heart diseases (IHD). The mean concentration of total cholesterol and levels of TAT, FPA, PAI-I, and TM were significantly reduced after the Pravastatin treatment. The PIC/TAT ratio was significantly increased in non-IHD patients after treatment, this was not the case in IHD patients. These findings suggested the presence of a thrombogenic state and vascular endothelial cell disorders in hypercholesterolemia; such a state might well be related to hypofibrinolysis.

We examined vascular endothelial cell markers, thrombomodulin (TM), plasminogen activator inhibitor-l (PAI-I), tissue plasminogen activator (t-PA), and von Willebrand factor, in 80 patients with disseminated intravascular coagulation (DIC). The levels of thrombin-antithrombin III complex (TAT), plasmin-α2 plasmin inhibitor complex (PIC) and FDP-D-dimer were significantly increased both before and after the onset of DIC, but were not correlated with organ failure or prognosis. However, the PIC/TAT ratio was lower in patients with poor prognosis than in those with good prognosis, and it was also lower in those with organ failure than in those without. Plasma TM, PAI-I, and t-PA levels were increased in DIC patients with organ failure or poor outcome, but were not significantly increased before the onset of DIC. We consider that the prognosis of patients with DIC might be related to organ failure or endothelial cell damage and that plasma levels of TM, PAI-I, and t-PA might be useful in the detection of these disorders and in assessing prognosis. A hypofibrinolytic state might enhance organ failure in patients with DIC. © 1993 Wiley-Liss, Inc.

The effect of 2-methyl-3-(1, 4, 5, 6-tetrahydronicotinoyl) pyrazolo [1, 5-a] pyridine (KC-764) on human platelet aggregation and its mechanism of action were investigated. The inhibitory activity of KC-764 on arachidonic acid-induced platelet aggregation was most potent with IC50 of 4.2×10-8 M and was about 100 times more potent than that of acetylsalicylic acid (ASA). KC-764 weakly inhibited prostaglandin G2-and prostaglandin H2-induced platelet aggregations. Neither KC-764 nor ASA had effect on platelet aggregation induced with STA2 (thromboxane A2 analog). KC-764 inhibited strongly human platelet cyclooxygenase activity. The effect of KC-764 on platelet aggregation, differently from ASA, was reversible. The inhibitory effect of KC-764 on cyclic nucleotide phosphodiesterases was weak. KC-764 had no effect on the concentration of cAMP and cGMP.These results suggest that the reversible inhibitory effects of KC-764 on platelet aggregations are mediated mainly via inhibition of platelet cyclooxygenase.

The effect of a novel compound, 3-isobutyryl-2-isopropylpyrazolo [1, 5-a] pyridine (ibudilast, KC-404), on human platelet aggregation and its mechanism of action were investigated.In vitro, KC-404 inhibited human platelet aggregation induced by ADP, collagen, adrenalin, platelet activating factor and arachidonic acid but not by ristocetin. Together, KC-404 and agents which increased cAMP (prostaglandin I2, prostaglandin E1 (PGE1), forskolin) or cGMP (3-morpholinosydnonimine (SIN-1)) produced synergistic inhibitory effects on platelet aggregation.KC-404 inhibited human platelet cAMP phosphodiesterase (PDE) (IC50: 50μM) and cGMP-PDE (IC50: 5.2μM) activities. cAMP and cGMP concentration of human platelets were not increased by KC-404 itself. PGE1, an adenylate cyclase stimulator, increased cAMP content; KC-404 enhanced the effect of PGE1 on cAMP accumulation. SIN-1, which stimulates guanylate cyclase, increased cGMP content; KC-404 enhanced the effect of SIN-1 on cGMP accumulation.These results suggest that effects of KC-404 on accumulation of cyclic nucleotides and inhibition of platelet aggregation are mediated via inhibition of platelet cyclic nucleotide phosphodiesterase activities.

Plasma levels of thrombomodulin antigen in various diseases were measured by ETA. Patients with renal insufficiency were not included in this study. In normal subjects the thrombomodulin levels were 16.3±4.1ng/ml (mean±SD). Thrombomodulin levels were elevated in patients with carcinoma, thrombotic disease, aneurysm and collagen disease. In patients with hematopoietic malignancy and carcinoma, there was no relationship between thrombomodulin levels and the presence or absence of DIC. In one patient with protein C deficiency complicated by DIC and thrombophlebitis, plasma thrombomodulin levels were in the normal range. In another patient with carcinoma of the cervix and without vascular complication, plasma thrombomodulin levels were very high. Our data suggest that DIC dose not always cause elevated plasma levels of thrombomodulin antigen, and that high level of plasma thrombomodulin dose not always indicate the existence of vascular damage.

Cyclic AMP (cAMP) and cyclic GMP contents were measured in 63 leukemic cell lysate from patients with and without disseminated intravascular coagulation (DIC) and compared with tissue factor (TF) activity, plasminogen activator (PA) and PA inhibitor (PAI). The cAMP level was increased in acute promyelocytic leukemia and was sighificantly higher in the leukemic cell lysate from patients with DIC than in from those without DIC. The cAMP level was well correlated with TF activity in the leukemic cell lysate. There was no difference in the phosphodiesterase level between the leukemic cell lysate from patients with DIC and from those without DIC, but calmodulin and s-100 levels were significantly higher in the former. The calmodulin level was also negatively correlated with the cAMP level and TF activity in leukemic cell lysates.It is speculated that cAMP may not be important in TF synthesis of leukemic cells and that calcium might regulate such synthesis.

The dynamic fluctuation of thrombin-antithrombin III complex (TAT) was studied in blood obtained during the daytime (at 9 AM, noon, 3 PM), during extracorporeal circulation and during the course of disseminated intravascular coagulation (DIC), to certify whether the level of TAT in blood can reflect the generation of thrombin. In 10 healthy male volunteers, the mean values of TAT (micrograms/liter) were 1.74 (+/- 1.36) at 9 AM, 1.22 (+/- 0.47) at noon, and 1.25 (+/- 0.68) at 3 PM. TAT did not show a daytime fluctuation, unlike fibrinolytic factors. The mean values of TAT in 38 hemodialyzed patients were 4.83 (+/- 2.8) before the initiation, 6.59 (+/- 4.39) in the first hour, and 13.42 (+/- 10.96) at the end of a dialysis session. In 20 patients undergoing open heart surgery, the mean value of TAT was increased during cardiopulmonary bypass (CPB) and decreased with time after the end of surgery. The fibrinopeptide A (FPA) value was increased with TAT during CPB but achieved a maximum level immediately after heparin neutralization by protamine. In 20 patients with DIC, the values of TAT varied from 5.8 to 297 micrograms/liter in the blood at the onset of DIC. In seven of eight patients treated with low-molecular-weight heparin (LMW-H), the values of TAT and FPA were lower 24 hr after LMW-H than before the treatment. These results suggest that the level of TAT in blood reflected the formation of thrombin and could serve as a sensitive parameter of activated coagulation in circulating blood.

We measured plasma level of tissue type plasminogen activator (t-PA) antigen in patients with disseminated intravascular coagulation (DIC). Plasma levels of t-PA, PA inhibitor-1 (PAT-1), t-PA-PAI-1 complex, von Willebrand factor, thrombin-antithrombin III Complex (TAT), plasmin-α2 plasmin inhibitor complex (PIC), and FDP-D-dimer were markedly increased at the time of the onset of DIC but decreased after treatment. In most patients without leukemia, as the DIC score increased, plasma t-PA increased to 20 ng/ml or more. Plasma t-PA level before treatment for DIC was slightly higher in those showing poor outcome than it was in those showing good outcome. After treatment, t-PA level increased slightly in those showing poor outcome but decreased significantly in those showing good outcome. Plasma t-PA level was significantly higher in patients with organ failure than in those without. Positive correlation of plasma t-PA antigen level to PAI-1

SummaryThe plasma level of interleukin-1β (IL-1β) was determined in normal individuals, patients with disseminated intravascular coagulation (DIC), patients in the pre-DIC period (within 7 days before the onset of DIC), and non-DIC patients to examine the relationship between DIC and the plasma ILlp level. The plasma IL-1β level was 0-0.085 ng/ml in normal individuals, with little difference being seen according to related age. It was significantly higher in the DIC group (0.19 ± 0.19 ng/ml) than in the pre-DIC group (0.05 ± 0.08 ng/ml) or the non-DIC group (0.09 ± 0.01 ng/ml). The plasma IL-1β level was not markedly elevated in leukemia patients, even in the DIC group, but it was significantly increased in the DIC group of solid cancer patients and was generally elevated in patients with sepsis. It was markedly elevated to 0.39 ± 0.26 ng/ml in patients with organ failure. When mononuclear cells were incubated with lipopolysaccharide, it was found that IL-1β, tumor necrosis factor, and tissue factor (TF) were released into the medium, and there was an increase of TF release from endothelial cells incubated with this medium. These results suggest that the increase in IL-Iβ reflected the activation of monocytes and may be an important factor in DIC and its associated organ failure.

In order to reduce risk factors in the treatment of hemophilia B with prothrombin complex concentrates, a new coagulation factor IX concentrate (GA-1013) was developed by Green Cross Corporation, Osaka, Japan. This product is prepared by immunoaffinity chromatography using a monoclonal antibody (3A6) and virally inactivated by solvent/detergent treatment. GA-1013 has a specific activity of 173 factor IX units/mg of protein, representing a 13, 700 fold purification from cryosupernatant.A multicenter clinical study was conducted in 24 previously treated and 1 previously untreated hemophilia B patients with no inhibitor to evaluate the safety and efficacy of GA-1013. We determined the half life and in vivo recovery of this FIX concentrate.The mean age of the patients was 29.0 years with a range of 9 months 62 years. The mean T1/2 value (n=22) was 20.3±8.93hr and the mean in vivo recovery rate was 69.0±20.12% calculated from the labelled FIX: C content (47.6±14.46% calculated from the actual FIX: C content).A total of one hundred and seventy-six bleeding episodes were assessed during the 6 months treatment, chiefly on a demand basis. Hemostatic effectiveness was confirmed with an efficacy rate of 93.7% (165/176). No adverse reactions attributed to GA-1013 were obserbed in any infusion. Neither FIX inhibitors nor antibodies to mouse or goat protein were detected. No significant changes attributed to GA-1013 were seen in vital signs or in laboratory findings including CBC, blood chemistries, virological parameters and immunological data (CD4 and CD8 counts, lymphocyte blastogenesis index and serum immunoglobulins at 1, 3 and 6 months).These results suggest that monoclonal antibody-purified coagulation FIX concentrate (GA-1013) is safe and efficacious as replacement therapy for hemophilia B.

Twelve patients with hemophagocytic syndrome (HPS), were studied. In these patients, marked hemophagocytosis was observed with fever in 10, hepatosplenomegaly in 11, and organ failure and disseminated intravascular coagulation (DIC) in 10 each. Six of these patients were treated, and only 2 have survived with 10 having died. The time of survival was within 3 months in most cases. Pancytopenia was noted in all patients, and GOT, LDH, total bilirubin, and creatinine were increased in many patients. Coagulation examinations showed a marked prolongation of APTT in 2, prolongation of prothrombin time in 3, and high fibrinogen levels in many patients. Increased FDP and thrombocytopenia were noted in most patients, and antithrombin III activity was reduced in half the patients. Organ failure was more frequent than bleeding tendency. Tissue plasminogen activator antigen and plasminogen activator inhibitor (PAI) -1 antigen levels were markedly increased, and the PAI-2 antigen level was slightly increased. Plasma interleukin (IL) -1β, granulocyte macrophage colony stimulating factor, and IL-6 levels were elevated. Thus, HPS was frequently complicated by multiple organ failure due to activation of monocytes/macrophages, hypercytokinemia, and an increase in PAI. Examination of monocyte-specific markers is considered to be useful.

Plasma levels of thrombomodulin (TM), fibrinogen, antithrombin III (ATIII) and thrombin ATIII complex (TAT) were studied in healthy young subjects (group A), healthy elderly subjects (group B) and patients with level of TM in group B tended to be higher than that in group A. Levels of TM, fibrinogen and TAT in group C suggested the presence of a hypercoagulable state. When group C was further divided into those with and without diabetes mellitus (DM), the TM level in the former tended to be higher than that in the latter. Furthermore, among the patients with DM, those with diabetic retinopathy showed significantly higher levels of TM than those without retinopathy. Thus, high TM levels indicate the presence of endothelial injury. In groups B and C, TM correlated positively with fibrinogen, and negatively with ATIII, which also indicates that a high TM level is related to a hypercoagulable state. In conclusion, the TM level is considered to be a potential marker of the presence of endothelial injury.

The blood level of interleukin-1β (IL-1β) was determined in 100 normal individuals, 10 patients with thrombotic thrombocytopenic purpura (TTP), 41 patients with established disseminated intravascular coagulation (DIC), 15 in pre-DIC period (within 7 days before the onset of DIC), and 23 non-DIC patients. In TTP, the blood IL-1β concentration was increased at onset but decreased at remission. The blood IL-1β concentration was significantly higher (p

A 19-year-old man with common acute lymphoblastic leukemia (ALL) developed necrotizing leukoencephalopathy (NL) after extensive chemotherapy and irradiation. Clinicians should exercise care in the treatment of leukemia, since chemotherapy has shown a trend of becoming more intensive, and the survival time of patients with leukemia is being extended.

Disseminated mycobacterial infection was found at autopsy in a male patient with hemophilia B and acquired immunodeficiency syndrome (AIDS). In May 1986, 23 months before death, the patient had encephalitis for one month and in July he developed a fever, malaise and generalized lymphoadenopathy. Human immunosuppressive virus (HIV) was positive and the CD 4/8 ratio of lymphocyte surface markers was 0.1, but mycobacterium was not detected. In September 1986, he had severe dyspnea due to interstitial pneumonia and he was treated with high-dose methylprednisolone. He died after a 23-month course of fever, severe weight loss and terminal progressive deterioration, although he was treated with antibiotics, antifungal agents, gamma-globulin, steroid and a Azidothymidine.

Plasma levels of tissue factor pathway inhibitor (TFPI)-activated factor Xa (FXa) complex were measured in patients with disseminated intravascular coagulation (DIC), pre-DIC, and DIC. Plasma levels of plasmin-plasmin inhibitor complex (PPIC), D-dimer, and soluble fibrin monomer (SFM) were significantly higher in patients with DIC than in those with pre-DIC or non-DIC; the levels of these hemostatic markers were significantly higher in patients with pre-DIC than in those with non-DIC. Plasma levels of thrombin-antithrombin complex (TAT) were significantly higher in patients with DIC or pre-DIC than in those with non-DIC. Plasma levels of tissue factor (TF), total TFPI, free TFPI, and TFPI-Xa complex were significantly higher in patients with DIC than in those with non-DIC. Plasma levels of TFPI-Xa complex were significantly increased in patients with pre-DIC as compared to those with non-DIC; however, plasma free TFPI levels were significantly decreased in patients with pre-DIC as compared to those with non-DIC. These findings suggest that free TFPI might be consumed in the pre-DIC state, thereby confirming the activation of the extrinsic pathway. Plasma levels of TFPI-Xa complex were significantly correlated with TF, free TFPI, and total TFPI. Increased plasma TFPI-Xa complex levels might be useful for the diagnosis of DIC or pre-DIC, particularly that occurring by activation of the extrinsic pathway of blood coagulation.

To evaluate the relationship between the tissue factor (TF) pathway and lupus anticoagulant (LA), in the pres ent study, we measured the plasma levels of TF antigen and TF pathway inhibitor (TFPI) antigen in patients positive for LA. Plasma TF and TFPI levels in LA-positive patients were sig nificantly higher than levels in healthy volunteers (p < 0.01). In LA-positive patients, there were no significant differences in plasma TF and TFPI levels between patients with and without thrombosis. In patients with thrombosis, there was no signifi cant difference in the plasma TF level between LA-positive and LA-negative patients; however, the plasma TFPI level in LA- positive patients was significantly lower than that in LA- negative patients (p < 0.01). We also examined the TF pathway in human umbilical venous endothelial cells (HUVEC) incu bated with plasma of LA-positive patients, LA-negative pa tients, and healthy volunteers. TF activity was significantly higher (p < .05) in HUVECs incubated with the plasma of LA-positive patients than in cells incubated with the plasma of the other two groups (p < .01). However, there was no signifi cant difference in TFPI antigen levels among the media of HUVECs incubated with the plasma of all groups. The viability of HUVEC incubated with the plasma of LA-positive patients with thromboses, LA-positive patients without thromboses, and LA-negative patients with thromboses were significantly lower than that of HUVECs incubated with the plasma of healthy volunteers (p < .01). These findings suggest that abnormalities of the TF pathway plays an important role in the mechanism of hypercoagulability in LA-positive patients. LA may affect vas cular endothelial cells causing thrombogenesis. Key Words: Lupus anticoagulant—Tissue factor—Tissue factor pathway inhibitor—Vascular endothelial cell—MTT assay.

To evaluate that the relationship between the truncated form of tissue factor pathway inhibitor (TFPI) and the stage of disseminated intravascular coagulation (DIC), we measured the plasma levels of tissue factor (TF) antigen and the intact and truncated forms of TFPI antigens in 41 patients with DIC, 12 with pre-DIC, and 20 with non-DIC. The plasma TF and total TFPI antigen levels were significantly higher in patients with DIC than in non-DIC patients. Plasma levels of intact TFPI antigen in the pre-DIC groups were significantly lower than in the non-DIC and DIC groups. The truncated form of TFPI antigen levels in DIC patients were significantly increased compared with those in non-DIC and pre-DIC patients. The fact that the intact form of TFPI was decreased in pre-DIC patients compared with that in non-DIC patients, suggests that it is consumed in the pre-DIC state and that hypercoagulability occurs in pre-DIC patients. The increased level of the truncated form of TFPI in DIC patients may be attributed to proteolysis of the intact form of TFPI in these patients. The increased level of the truncated form of TFPI may be a useful index for the diagnosis of DIC.

In our medical and welfare facilities, many patients with senile dementia require aid in taking a bath. In most institutions, patients usually take a bath in the daytime within the working hours of the staff. However, most of these patients used to take a bath in the evening or at night at their homes. Some patients even fall asleep after daylight bathing. Thus, we studied the stabilizing effects of night-time spa bathing on symptoms associated with dementia. Ten patients (two male and eight females, aged 75–88) in special nursing institution for the aged, were enrolled in this study. They were all assessed as +4 on the Karasawa's clinical criteria for grading dementia. For 9 weeks, night-time spa bathing was performed at 18:00–19:00 twice a week. Except for the night-time spa bathing period, the bathing hour was 14:00–15:00 as usual. The observations of symptoms including restlessness, wandering and aggression were carried out ten times daily along with those on sleeping condition five times daily, to compare symptoms and conditions during 2 weeks of baseline daytime bathing periods, 9 weeks of night-time bathing periods and 2 weeks of daytime bathing periods, totaling 13 weeks. The results showed that sleeping conditions were ameliorated in more than 60–90% of the subjects. Their sleeping conditions began to improve 2 weeks after the start of night-time spa bathing with a remarkably improvement 4–6 weeks after the start. Restlessness was recognized in six subjects, wandering in eight and aggression in four at baseline, and 75–100% of the subjects with such symptoms improved markedly.

Expression of the tissue factor (TF) and the plasminogen activator inhibitor-II were induced in cultured human monocytes-macrophages by incubation with lipoproteins. Very low-density lipoprotein (VLDL) augmented the TF and PAI-II expression the most, followed by low-density lipoprotein (LDL) and a very weak effect by high-density lipoprotein (HDL). In macrophages pre-cultured for 3 days, oxidized LDL augmented the expression of TF activity in the macrophages to a greater extent than native LDL. These findings indicate that lipoproteins affect both monocytes and macrophages, and that they induce a hypercoagulable-hypofibrinorytic state. Thus hyperlipidemia may be a direct risk factor for thrombotic disease.

The role of the cAMP/A-kinase signaling pathway in G-CSF dependent proliferation of murine myeloblastic NFS-60 cells was investigated. G-CSF treatment resulted in a rapid and transient elevation of cAMP content of NFS-60 cells. G-CSF treatment of NFS-60 cells also resulted in the activation of A-kinase parallel to the increase in cAMP concentration. A low concentration (0.2-10 nM) of forskolin augmented the G-CSF-dependent cell proliferation, although forskolin by itself had no effect on NFS-60 cell growth. Forskolin did not affect the IL-3-induced proliferation of this cell line. Addition of forskolin resulted in further increases in the cAMP level, activation of A-kinase in NFS-60 cells stimulated by G-CSF. Proliferation of NFS-60 cells by G-CSF, but not by IL-3, was blocked by the axial diastereoisomer of adenosine 3',5'-phosphorothioate (Rp-cAMPS), a competitive cAMP antagonist. KT-5720(8R*,9S*,11S*)-(-)-9-hydroxy-9-n-hexyloxy-8-methyl-2, 3, 9, 10-tetrahydro-8, 11-epoxy-1H, 8H, 11H-2,7b, 11a-triazadibenzo(a,g) cycloocta(c,d,e)trinden-1-one), an A-kinase inhibitor, inhibited the G-CSF-dependent proliferation. These findings suggest that activation of the cAMP/A-kinase signaling pathway may be involved in G-CSF-mediated cell proliferation of NFS-60 cells, whereas IL-3-dependent proliferation is not mediated in such a manner.

We found that patients with thrombotic thrombocytopenic purpura (TTP) have significantly elevated plasma thrombin antithrombin III complex (TAT) and FDP-D-dimer levels, while the plasmin-α2 plasmin inhibitor complex (PIC) level was only slightly increased. The tissue-type plasminogen activator (t-PA) level was increased, but it was well correlated with the plasminogen activator inhibitor-1 (PAl-l) level. These findings suggest that hypercoagulable and hypofibrinolytic states coexist in these patients, in contrast to patients with disseminated intravascular coagulation, who exhibit coexisting hypercoagulable and hyperfibrinolytic states. Levels of vascular endothelial cell markers, such as PAl-l, thrombomodulin (TM), and t-PA, were increased at the onset of TTP, but the level of von Willebrand factor (vWF) antigen was not increased. The outcome in TTP patients was correlated with plasma t-PA and TM levels but not with TAT or PIC. These results suggest that vascular endothelial cell markers, such as TM and t-PA, are released from injured or stimulated endothelial cells, reflecting the degree of vascular endothelial damage, and that the main factor in the pathogenesis of TTP is vascular endothelial cell injury. © 1993 Wiley-Liss, Inc.

The authors present a patient with massive liver sarcoidosis, showing only slight hepatomegaly on precontrast computed tomogram and multiple low-density intrahepatic septa on postocontrast computed tomogram. Sarcoidosis frequently involves the liver, but rarely shows abnormal findings other than hepatomegaly on computed tomograms. Only a few cases have demonstrated a low density intrahepatic area on computed tomograms1,2. In this report, we present a patient with massive liver and pulmonary sarcoidosis, showing low density intrahepatic septa and pulmonary fibrosis on computed tomograms.

The effects of spa bathing on blood coagulation and fibrinolysis were studied in 20 patients with chronic cerebral infarction. Blood was obtained before and after a 10-minute period of spa bathing at 41 degrees C. Prothrombin time, activated partial thromboplastin time, fibrinogen, factor VIII activity, von Willebrand factor activity, and antithrombin III activity did not show significant changes after bathing, but euglobulin lysis time was significantly reduced (p < 0.01) and fibrin lysis activity was increased (p < 0.05). These findings suggest that spa bathing activates fibrinolysis without markedly changing blood coagulation in patients with chronic cerebral infarction. It is thought that the activation of fibrinolysis without the activation of coagulation has a favorable effect on blood circulation. The results of fibrin-plate assays using C1 inactivator indicated that tissue-type plasminogen activator was the major contributor to the activation of fibrinolysis during spa bathing.

Early diagnosis is necessary for the treatment of disseminated intravascular coagulation (DIC), but criteria for the stage preceding the diagnosis of DIC (pre-DIC) have not yet been established. To clarify hemostatic abnormalities that occur before the onset of DIC, we performed hemostatic studies in 117 patients within at least a week before the onset of DIC (pre-DIC), in 237 patients with DIC, and in 50 patients without DIC or pre-DIC (non-DIC). Levels of FDP, PT, and fibrinogen, and platelet counts were significantly abnormal after the onset of DIC, but not before. Thrombin-antithrombin III complex (TAT), plasmin-alpha 2 plasmin inhibitor complex (PIC), and FDP-D-dimer levels were significantly higher before the onset of DIC compared to the non-DIC patients. Hemostatic abnormalities were observed within a week before the onset of DIC. Monitoring the plasma levels of TAT, PIC, and FDP-D-dimer might be useful for the diagnosis of a pre-DIC condition.

Plasminogen activator (PA) and PA inhibitor (PAI) were measured in homogenates of leukemia cells. Both PA and PAI levels were higher in non-lymphoblastic leukemia than in lymphoblastic leukemia. The levels were below the sensitivity of determination in chronic myelocytic leukemia (CML) but showed significant increases in blast crisis (CML, bc). The level of the tissue type PA (t-PA) antigen was highest in acute myelobtestic leukemia (AML) and that of the urokinase type PA (u-PA) was highest in acute promyeiocytic leukemia (APL). The PAH antigen showed no marked cell specificity, but the PAI-II antigen was markedly increased in myelomonocytic leukemia and acute monocytic leukemia (AMoL). From these findings, various PAs and PAIs are considered to be present in leukemia cells and to be involved in hemostatic disorders, thus they are of diagnostic value in leukemia. © 1993 Wiley-Liss, Inc.

The excessive and untimely formation of blood clot(thrombus) in blood vessels may discontinue the blood supply to vital organ, and sometimes be followed by the aggravation of diseased state and the danger of life. The word “hypercoagulability” is often taken to mean an increased tendency to thrombose, and is usually used to mean an excess or activation of, clotting factors. Furthermore, as analogous words, prethrombotic state seem to imply the previous state of thrombosis, and prethrombotic state is used to connote a predisposition to thrombose(1). It is very important for the prophylaxis of overt thrombosis to know the behaviors of clotting and fibrinolytic factors in these state.

We report a 31-year-old female who had repeated thrombosis and was diagnosed as having congenital homozygous protein C deficiency based on decreased protein C antigen and activity, and the findings of family history. This patient had shown no symptom of thrombosis until the age of 27 years, when she had cerebral infarction as the first symptom. Low molecular weight heparin was useful for disseminated intravascular coagulation (DIC) that complicated protein C deficiency in this patient.

We examined the involvement of cAMP-dependent protein kinase (A kinase)2 in the inhibition by cilostamide, a specific inhibitor of the low Km cAMP-phosphodiesterase (PDE), on 9,11-epithio-11,12-methanothromboxane A2 (STA2)-induced platelet aggregation. For comparative purposes, the PGE1 analogue, 17S-20-dimethyl-trans-delta 2-PGE1 (OP-1206) was used. OP-1206 (IC50 = 18 +/- 0.55 nM) and cilostamide (IC50 = 40 +/- 4.5 nM) were both potent inhibitors of the platelet aggregation induced by STA2 (1 microM). OP-1206 and cilostamide dose-dependently inhibited elevations in intracellular free Ca2+ ([Ca2+]i) caused by STA2. OP-1206 caused an almost complete inhibition of Ca2+ mobilization, but cilostamide did not prevent the STA2-induced elevation in [Ca2+]i to the same extent as OP-1206, even at a high concentration (greater than 200 nM). Cilostamide did not increase the cAMP level at concentrations (5-100 nm) which affected STA2-induced aggregation. OP-1206 significantly increased cAMP contents in platelets, and the degree of aggregation inhibition by OP-1206 appears to be related to the size of increase in cAMP. OP-1206 increased phosphorylation of the 50,000 mol. wt vasodilator-stimulated phosphoprotein, at concentrations of 7.9-79 nM, which inhibited aggregation induced by STA2. Cilostamide treatment resulted in a marginal increase in the 50,000 mol. wt phosphorylation at concentrations (10-100 nM) which completely inhibited the STA2-induced aggregation. (8R*, 9S*, 11S*)-(-)-9-Hydroxy-9-n-hexyloxy-8-methyl-2,3,9,10- tetrahydro-8,11-epoxy-1H, 8H, 11H-2, 7b, 11a-triazadibenzo(a,g)-cycloocta(c,d,e)trinden-1-one (KT-5720), a specific inhibitor of A kinase, not only reversed the inhibition by OP-1206 of STA2-induced platelet aggregation, but also inhibited the OP-1206-induced protein phosphorylation. However, the inhibition by cilostamide of STA2-induced aggregation was not prevented by pretreatment with KT-5720. Inhibition of the STA2-induced aggregation by OP-1206 may be associated with cAMP-dependent protein phosphorylation, while cilostamide may have inhibitory effects on STA2-induced platelet activation through mechanisms other than the activation of A kinase.

A 70-year-old man was admitted to our hospital because of paraplegia. A spinal epidural tumor (Th VII-XI) was diagnosed by magnetic resonance imaging and resected. Histological examination of the tumor showed non-Hodgkin's lymphoma of the diffuse large cell type (Lymphoma Study Group classification), with a B cell phenotype. The clinical stage was IEA by the Ann Arbor classification. Radiation therapy of the involved field was performed (total dose : 40Gy), followed by six courses of modified cyclophosphamide, adriamycine, vincristine, prednisolone (CHOP) therapy. He was discharged walking unaided 10 months later and has shown no evidence of relapse in the 26 months since discharge.(Internal Medicine 31 : 513-515, 1992)