The clinical significance of the platelet dysfunction following cardiopulmonary bypass has been studied by many authers. But the pathogenesis is not clear and prevention of platelet dysfunction had not been achieved yet. In this paper, the morphological parameters of the platelet were evaluated quantitatively, before, during and after cardiopulmonary bypass in 20 adult patients. Cardiopulmonary bypass was drived in moderate hypothermia with bubble oxygenator primed with fresh ACD blood.
The morphological parameters of the platelet, PLT (Platelet Count, ×10⁴/mm³), MPV (Mean Platelet Volume, μm³), PDW (Platelet Distribution Width) and Pct (Plateletcrit, %) were calculated by an electronic particle-sizing apparatus, based on the Coulter counter SP. Blood samples were taken before operation, after anesthesia, during perfusion, postperfusion, and postoperative 1st to 7th days.
Results were as follows
1) PLT decreased during the first 30 minutes of perfusion, and then PLT recovered slightly by 120 minutes of perfusion. During perfusion, reduction of MPV was significant but PDW remained unchanged.
2) After surgery, PLT was in the lowest at the second postoperative day, and then recovered gradually, but was still significantly lower than preoperative level during 7 postoperative days. Significantly increasing of MPV and PDW were observed in this period.

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The distribution and mobility of binding sites on the plasma membrane of platelets were studied by using ferritin-conjugated wheat germ agglutinin (F-WGA).
Washed human platelets were incubated with F-WGA (final ferritin concentration: 0.1mg/ml) at 37°C after or before glutaraldehyde fix atio at room temperature.
In incubation with F-WGA after fixation, the plasma membrane of disk shaped, nonactivated platelets was diffusely studded with the particles.
The binding sites of F-WGA were remarkably decreased in number with pretreatment of neuraminidase (f. c. 10 or 20IU/ml) after fixation.
When platelets were incubated with F-WGA and followed by fixation, the binding sites on the cell surface became loose and irregular in arrangement showing lateral migration. With the lapse of time, the particles were taken up into the dilated open canalicular system. After further incubation with phosphate buffered saline, such a distribution became looser and the particles increased in number in the open canalicular system. When platelets were reincubated with F-WGA and fixed after washing, the particles were distributed again on surface of activated platelets.
The native ferritin used in the respective control experiment was never bound on the membrane of platelets.
These findings suggest that WGA generally binds to N-acetylneuraminic acid (sialic acid) of the cell surface. The results are absolutely compatible with those of the mobility of anionic sites on the cell membrane obtained by utilization of cationized ferritin.
In conclusion, the diffuse distribution of sialic acid may contribute to the unstickness of platelets in association with anionic charge on the cell surface. Under circumstance, however, mobility of binding sites may facilitate activation of platelets due to some kinds of stimulation.

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It has been reported that plasma prekallikrein was correlated with the activation of coagulation and fibrinolysis systems, and their changes and significance in various diseases have been widely discussed.
In this report, the plasma prekallikrein and kallikrein inhibitor levels were studied in cases of collagen disease, especially in cases of systemic lupus erythematodes and allergic purpura. The activity of prekallikrein was measured by amidolytic assay with synthetic substrate (S-2302), using plasma kallikrein activator (Daiichi Pure Chemical Co., Ltd.) containing sufficient amount of high molecular weight kininogen and factor XII_a.
The following results were obtained.
1) A significant correlation of 0.96 was found between the prekallikrein levels measured by the Cephotest and plasma kallikrein activator chromogenic assay with the S-2302 substrate.
2) Kallikrein inhibition in plasma was attributed mainly to the action of C₁INH and α₂ macroglobulin, and partially to α₂ PI.
3) The plasma prekallikrein and plasma kallikrein inhibitor levels were increased in the acute SLE phase unaccompanied by nephropathy, but decreased when either lupoid hepatitis or nephropathy was complicated in this phase.
4) Plasma kallikrein inhibitor levels decreased in acute phase of Behcet disease in ocular crisis.
5) Plasma kallikrein and kallikrein inhibitor levels were within upper limits of the normal range in allergic purpura.
6) Both the plasma kallikrein and kallikrein inhibitor levels decreased in PSS, myositis, thrombotic aortic obliterans, aortic sclerotic obliterans, in which there has been a long suffering time.

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The nature of sugar chain of factor VIII related protein (VIIIR) in the plasma of normal subject and patients with type I and IIA of von Willebrand's disease was examined by crossed affinoimmunoelectrophoresis using anti-human factor VIII rabbit serum, with inserted wheat germ agglutinin (WGA) agarose layer. Molecular weight disfribution and affinity for WGA of factor VIII related antigen (VIIIR: AG) were estimated by the electrophoretic patterns.
VIIIR: AG, in normal plasma and in type I of von Willebrand's disease, showed high affinity for WGA at the portion over 3×10⁶ daltons of molecular weight. On the other hand, VIIIR: AG in plasma of type IIA consists only of the multimers smaller than 3×10⁶ daltons, showing a very weak affinity for WGA.
These results suggested that VIIIR-multimers in type IIA may be poor in sialic acid reactive with WGA, in addition to incomplete polymerization.

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There have been reported many cases of acquired immune deficiency syndrome (AIDS) among the groups of homosexuals, drug addicts, Haitian immigrants, prisoners and hemophiliacs under treatment with concentrate of antihemophilic factors in the United States and many European countries, but no case of definite AIDS has been reported in Japan so far yet.
Authors have observed two possible cases of AIDS in hemophiliacs, one of whom had high fever, generalized lymphadenopathy and splenomegaly, and died of sepsis with Campylobacter jejunii. The other surviving one was revealed abnormality of T cell subsets of peripheral lymphocytes. These observations prompted us to investigate the cellular immunity of hemophiliacs to find whether any case of apparent or latent AIDS was included in them.
Twenty one hemophiliacs (17 cases of hemophilia A and 4 cases of hemophilia B) were investigated their cullular immune functions of peripheral lymphocytes to prove that responsiveness of lymphocytes to mitogens (PHA and PWM) was impaired at various degrees, and the number of T₄ (helper/inducer) and T₈ (suppressor/cytotoxic) positive lymphocytes was decreased moderately or markedly, giving significant depression of T₄/T₈ ratio in all cases of hemopilia A and 3 in 4 cases of hemophilia B. These data showed good concordance with the decrease of natural killer (NK) activity and T cell colony forming activity.

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The purpose of the study is to evaluate the relationship of abnormalities in lipid metabolism and the coagulation induced by contraceptives containing estrogen and progesterone. Ten young female volunteers taking contraceptives (contracetive group) and 10 not taking contraceptives (control group) were randomly selected and their lipid and coagulation parameters were measured. (table 1) Serum triglyceride (TG) and total cholesterol (TC) levels were significantly higher in the contraceptive group than in the control group. Hepatic triglyceride lipase (HTGL) and peripheral lipoprotein lipase (LPL) activities were 5.4±1.5 and 7.8±2.4 unit in the contraceptive group, while 10.8±2.6 and 9.4±2.3 unit in the control group, respectively. As for the coagulation parameters, plasma fibrinogen (Fbn), plasminogen (Pln) and antithrombin (AT) III concentrations were 382±62, 16.5±3.9 and 30.3±3.3mg/d<i>l</i> in the contraceptive group, while 298±73, 10.9±1.1 and 29.4±1.9mg/d<i>l</i> in the control group, respectively. Collagen and ADP induced aggregation and spontane aggregation (PAT III) in the contraceptive group showed higher but not significantly than those in the control group. (Fig. 1) Negative correlation between TG and HTGL (r=-0.69, p<0.01) (Fig. 4), as well as TG and LPL (r=-0.63, p>0.01) means that impaired triglyceride catabolism might be induced by contraceptives. There were significant negative correlations between HTGL and Fbn (r=-0.63, p<0.01) or Pln (r=-0.66, p<0.01) (Fig. 5, 6) in whole subjects, and also positive correlation between AT III and LPL (r=0.93, p<0.01) or HTGL (r=0.67, p<0.02) only in the contraceptive group (Fig. 7).
In conclusion, impaired lipid metabolism and the coagulation tendency due to the contraceptives might occure on the same occasion.

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It was reported that glutathione (GSH) improved the survival of endoxin shock experimentally produced in rats. We investigated the effects of GSH in endotoxin shock on the platelet aggregation and blood coagulation.
Our recent study revealed that GSH inhibited the platelet aggregation induced by ADP and collagen, and prolonged the activated partial thromboplastin time (APTT). The APTT prolonged by GSH was approached to nearly normal time with long incubation, and it was similar to the Fletcher trait plasma. Further more, we investigated the effects of GSH on the kinin-kallikrein and on the contact phase of intrinsic blood coagulation.
Kallikrein was measured with amidolytic activities (S-2302), and kinin was measured by the bioassay method of UCHIDA.
It was observed that the production of kallikrein was predominantly reduced by GSH, and the release of kinin from high molecular weight kininogen (HMW-kininogen) was delayed compared with the not-GSH plasma.
In conclusion, we examined the cause of the APTT prolonged by GSH and our result showed that the APTT prolonged by GSH resulted from the inhibition of the contact phase of intrinsic blood coagulation, because the production of kallikrein was reduced and the release of kinin from HMW-kininogen was delayed.

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In order to clarify the characteristic differences between myocardial infarction with and without preinfarction angina (AP (+) vs AP (-)), we examined 401 patients with myocardial infarction discharged from the Department of Cardiology (Juntendo University) in the period of Jan. 1979 to Dec. 1981. We excluded cases of asymptomatic infarction and subendocardial infarction. In 203 cases of infarction with full clinical history, 36 AP (-) cases were significantly younger than 167 AP (+) cases (49.8 y-o vs 54.1 y-o). There were no significant difference in the coronary risk factors between the two groups except for hypertension in AP (+) group. Out of 159 cases whose coronary arteries were examined by cineangiogram and at autopsy most cases (78.7%) in AP (+) had two to three vessel diseases, but in 19 cases out of 32 cases in AP (-), single vessel disease was observed and most of the lesions (89%) were focal. There were no significant difference in the site of infarction between the two groups.
The abscence of preinfarction angina cannot be explained by the elevation of the threshold of chest pain caused by aging and diabetes mellitus and the presence of inferior ischemia. We postulate a possibility of rapid progressive coronary occlusion in AP (-) cases.

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The incidence of platelet activation in myocardial ischemia was evaluated in 13 patients with acute myocardial infarction (AMI); group A, 7 cases 3-12 hours after onset, group B, 6 cases 15-55 hours after onset, 2 patients with impending myocardial infarction and 6 normal controls by measurement of plasma β-thromboglobulin (β-TG). Vasoactive prostanoids, thromboxane A₂ and prostacyclin in peripheral venous blood were measured as the stable metabolites thromboxane B₂ (TXB₂) and 6-keto-prostaglandin F_1α (6-keto-PGF_1α) in the same subjects. Platelet count was significantly decreased in AMI group B as compared with the control (p<0.05) and mean plasma β-TG concentration was elevated in both AMI group A and B (p<0.05 and 0.01 respectively) (Fig. 1) The TXB₂ levels were not different among 4 groups.. The 6-keto-PGF_1α levels were also not different among 4 groups, however, the level of AMI group B was below the lower measurable limit if one case (3.26ng/ml) is excluded (Fig. 2).
Results indicate that platelets are frequently activated with myocardial infarction. However the measurement of β-TG is of limited value and that of TXB₂ is of little value in detecting myocardial ischemia or infarction. Decreased level of 6-keto-PGF_1α in the recovery phase of myocardial infarction may indicate the decreased production or depletion of prostacyclin in the vessel wall, but it certainly needs further investigation.

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We observed that the generation of factor Xa in hemophilia A plasma incubated with ellagic acid, phospholipid and CaCl₂ was inhibited by anti factor VII IgG by our assay of factor X a generation. This fact suggests that VIIa is effective to increase the intrinsic generation of factor Xa without tissue thromboplastin. In this study, the authors applied a chromogenic synthetic peptide substrate (S-2222) for the assay of factor VII and VIIa activity in the condition that the activity of factor IX was inhibited by the addition of anti IX IgG into the reaction mixture. The intrinsic generation of factor Xa in the deficient plasma of factor XI, IX or VIII was increased by the activation with ellagic acid, so we tried to measure the activated factor VII in these deficient plasma by our newly developed technique before and after the activation. We could detect the increase of activated form of factor VII in these deficient plasma, but not in factor XII deficient plasma. Therefore, we considered that the activation of contact factors in plasma, especially factor XII, could be followed by the generation of activated factor VII, then the activated factor VII could acceralate the intrinsic generation of factor Xa without the addition of tissue thromboplastin on the condition that the intrinsic coagulation was suppressed like hemophilia A.

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The effects of platelet, factor XIII inhibitor and plasmin inhibitor on the dynamic rigidity of plasma clot were examined. The dynamic rigidity (G′) of clot for a mixture of factor XIII deficient platelet poor plasma (PPP) and normal platelets obtained from healthy human was much larger than that for factor XIII deficient PPP. The value of G′ for platelet rich plasma (PRP) from patient with Glanzman's disease was smaller than that for normal PRP. A higher value of G′ for PRP was ascribed to the clot retraction. The contractile force of platelets brings about the reorientation or reconstitution of fibrin fibers attached to platelets and caused the elevation of rigidity of clot.
After reaching a maximum value of G′, a relaxation of the clot occured with subsequent decline in G′. The decrease in G′ after attaining a maximum value was almost abolished by adding 0.3mol ε-aminocaproic acid (plasmin inhibitor). A extremly sharp drop in G′ was observed for PRP containg 5 or 10mM glycine ethyl ester (factor XIII inhibitor). From these results, it is assumed that the rate and magnitude of decrease in G′ are determined by the counterbalance between the formation of α-crosslinks and platelet fatiguing, fiber slippage or fibrinolysis.

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Recently, a two-dimensional echography has been for the detection of intracardiac and intravascular thrombi. However, it is not well known whether all of thrombi consisting of various constituents could be detected or not. Thus, this study attempts to make various kinds of thrombi in vitro, and aims to demonstrate those by the echographic technique.
Fibrin thrombi were made of bovine or human fibrinogen (0.5-2.0%) and thrombin. Whole blood thrombi, Erythrocyte-rich thrombi (Ht 20-80%) and platelet-rich thrombi(0.85-2.60×10⁶/cmm) were produced from bovine or human blood and thrombin. Each thrombus (8mm in diameter, 30mm in length) was inserted a vinyl tube filled with whole blood, and was studied in a water tank using Aloca, SSD-250.
All thrombi were clearly detected. The internal echo density of fibrin thrombi was sparse in comparison with the others. The echo-images of platelet-, or erythrocyte-rich or whole blood thrombi were not different each other.
This study was summarized as the follows. Using by a two-demensional echography, it would be easy to detect many kinds of vascular thrombi in vivo when a blood vessel was clearly recognized. But it would be difficult to distinguish the cell kinds consisting thrombi except pure fibrin thrombi.

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In order to evaluate significance and usefulness of parameters for platelet activation in antithrombotic therapy, platelet count ratio, CPA ratio, platelet shape (discoidness index and pseudopod %), plasma platelet factor 4 or β-thromboglobulin/initial platelet count, serum PF-4, aggregated platelets and plasma thromboxane B₂ levels were exmined in experimental arterial thrombosis in vascular grafts in chaired baboons before and 30min after placing them between femoral arteriovenous shunts.
Antithrombotic therapy was done by heparin (H), PG I₂ (P), ancrod (A), heparin+PG I₂ (HP) or ancrod+PG I₂ (AP). As for preservation of blood flow, A, HP and AP were effective. HP completely inhibited thrombus formation whereas A and AP inhibited only fibrin formation (to be mixed type reported in detail elsewhere).
None of the parameters reflected the blood flow but changes or findings of platelet count ratio, platelet shape parameters, plasma PF-4 or βTG and PF-4/βTG in control were significantly suppressed in HP. Those of these parameters in H, P and AP were similar to those in control or in A rather enhanced.
In conclusion platelet parameters described above are useful only for platelet participation in thrombus, and suppresion of both blood coagulation and platelet activation may be essential for inhibition of mixed type thrombus formation.

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The fine structure of vascular permeability was studied on the dorsal skin of rabbits by a single intradermal injection of the extract, cationic protein, obtained from α-granules of bovine platelets after separation as subcellular fractions.
As early as 1-0min., the ferritin particle, used as a tracer, frequently leaked out through the junctional gaps of postcapillary venular endothelium into the perivascular connective tissue. At this period, however, mild exudation also occurred even in the sites of small veins and capillaries.
At 3-5min., inflammatory cells already infiltrated into the vessel wall or surrounding tissue. With the lapse of time, the inflammatory cells increased in number and strands of fibrin appeared in the connective tissue by 3 hours.
Neither exudation of ferritin particles nor cellular migration appeared in the vessels of control rabbits given isotonic phosphate buffered saline. Such ferritin exudation and cell migration from the vessels were scarcely inhibited in the animals pretreated with histamine H₁- and H₂-receptor antagonists before the injection of extract.
In conclusion, the α-granule extract from bovine platelets has vascular permeability factor(s) as described in that from human platelets and causes early permeability response.

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Exogenous prostaglandin(PG)E₂, 5μg/kg, was orally administered to DONRYU rats. Then, the animals were immersed in water to induce gastric ulcers. Postmortal macroscopic examinations of the stomach revealed that exogenous PGE₂ markedly prevented the occurrence of ulcers. In contrast to the control (immersed in water without PGE₂ pretreatment), the experimental group showed (1) a slight increase in endogenous PGE at Stages II and III, (2) a marked decrease in glandular kallikrein (GK) at all stages and (3) no change in tissue plasminogen activator (TPA) at any stage. On the other hand, endogenous PGE in the group untreated with PGE₂ showed a marked increase at Stage I and a moderate decrease at Stages II and III compared with findings obtained from the corresponding control (neither treated with PGE₂ nor immersed in water). Our previous studies demonstrated that GK gradually decreased with the progress of stage, and that TPA markedly increased at Stage I and gradually decreased at Stages II and III. These findings suggest that PGE may interact with GK and TPA, leading to the mprovement of blood flow in the gastric tissue.

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There has been a discrepancy in several investigators whether prostacyclin-like substance (PGI₃) might be produced from eicosapentaenoic acid (EPA) in the vessel wall. Production of PGI₃ from EPA in 3T3 fibroblasts, which had an ability to produce prostacyclin (PGI₂) from arachidonic acid (AA), was studied. After various times of incubation with EPA and 3T3 fibroblasts, aliquots of the culture media were transferred to platelet-rich plasma, to which ADP was added to induce aggregation. Inhibition of platelet aggregation was observed with samples removed after short minutes incubation, but not from longer periods incubation. The addition of tranylcypromine, an inhibitor of prostacyclin synthetase, to the culture media blocked the formation of a labile inhibitor of platelet aggregation. These data demonstrated sufficient synthesis of PGI₃ like substance to account for the inhibition of platelet aggregation. But the inhibitory effiect of platelet aggregation by PGI₃ like substance from EPA was weaker than by PGI₂ like substance from AA. The formations of PGI₃ and PGI₂ produced from EPA and AA by the culture of 3T3 fibroblasts were measured as their stable metabolites ¹⁷Δ-6-keto-PGF_1α and 6-keto-PGF_1α respectively by gas chromatography-mass spectrometry. Unexpectedly, the amount of ¹⁷Δ-6-keto-PGF_1α was higher than that of 6-keto-PGF_1α. These observations suggest that PGI₃ is inferior to PGI₂ in the ability to inhibit platelet aggregation.

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Arteriosclerosis was induced to rabbits by feeding with 1% cholesterol diet for various terms. After killing the rabbits, the surface of the thoracic aorta was measured and spotaneously released PGI₂ from the thoracic aorta during 10 minutes incubation was determined by assay using platelet aggregation, which was demonstrated as pg/cm₂/min. Plasma concentration of 6-keto PGF_1α thromboxane B₂ (TXB₂) and TXB₂ release from the washed platelets in the aggregatory response were determined by the radioimmunoassay.
Arteriosclerotic plaque coverage in thoracic aorta was quantitatively determined by the planimetric procedure and they were divided into three groups, normal group (group A, n=16), arteriosclerotic group with less than 50% changes (group B, n=9) and arteriosclerotic group with more than 80% changes (group C, n=7). The PGI₂ production of the aortic wall were 54±21 (A), 46±19 (B) and 41±22 (C)pg/cm²/min in each groups (statistically not significant). The microscopic examination of the aorta revealed preservation of the endothelium even at the site of arteriosclerotic lesion. The amount of released TXB₂ from platelets showed no significant differences among three groups.
PGI₂ production was still preserved in advanced arteriosclerotic aortic wall of rabbits. Since the aortic endothelium is the main layer of the PGI₂ production, arteriosclerotic lesion per se might not reduce PGI₂ production of the aorta unless endothelium is damaged.

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Platelet thromboxane A₂ synthetizing activity was examined in patients with myocardial infarction. Platelets were isolated from plasma by centrifugation and suspended in Ca⁺⁺, Mg⁺⁺ free Tyrode solution (pH 7.4). After an addition of arachidonic acid (25μM) or thrombin (0.1U/ml) to the platelet suspension, the production of TXA₂ from platelets was measured, as determined via radio-immunoassay of its stable metabolite TXB₂. In platelets collected from healthy controls, the production of TXB₂ increased from 30 to 60 seconds after the addition of AA or thrombin lineally. At 5 minutes after the addition of both the agents, the production reached to the maximum level.
The production of TXB₂ in the platelets of myocardial infarction was significantly decreased (p<0.001) than in the healthy control at 30, 60 and 300 seconds after the addition of AA. On the other hand, it was significantly increased in the myocardial infarction (p<0.01 and 0.001) than in the control at 30 and 60 seconds after the addition of thrombin. At 5 minutes after, the production was almost the same compared to the control. The results suggest a presence of selective consumption of hyperreactive platelets in myocardial infarction. At the same time, an enhancement of release of AA from platelet membrane related to activity of phospholipase might be present in myocardial infarction.

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It is known that aspirin acts as one of platelet antiaggregatory agents and also inhibits production of prostacyclin which has antiagrregatory effect. Thus administration of small dose of aspirin is recommended. In this point of view, we determined plasma aspirin levels and platelet antiaggregatory effects after administration of small dose of aspirin. Our experimentation includes eight old persons. Aggregation of plateletes was evaluated by Born's method and plasma aspirin and salicylate levels by high pressure liquid chromatography. In five cases with maximum level of aspirin above 1.30μg/ml, excellent antiaggregatory effect was seen. But in three cases, maximum aspirin levels less than 1.01μg/ml revealed no effect. According to our data, it is suggested that a fixed aspirin plasma level is necessary for platelet antiaggregatory effect, but minimum dose for oral administration differs person to person.

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The possible enhancement of prostaglandin I₂ (PGI₂) synthesis in whole blood by the inhibition of thromboxane A₂ (TXA₂) synthesis was investigated, applying OKYs, the specific TXA₂ synthetase inhibitors, to platelet aggregation and PG formation in whole blood and platelet rich plasma (PRP) by arachidonic acid (AA). Platelet aggregation was estimated from the decrease in platelet count in the supernatant of centrifuged blood or PRP. The amount of PGs formed was determined by radioimmunoassay. Platelet count both in whole blood and PRP was decreased following addition of AA. The decrease in platelet count at 3min. after addition of AA both in blood and PRP was slightly inhibited in the presence of OKYs. At 15min. after the addition of AA, however, OKYs inhibited the decrease considerably in blood and not in PRP. The amount of TXB₂ was increased in blood and PRP following addition of AA, which was markedly inhibited by OKY-046 or aspirin. The amount of 6-keto-PGF_1α was also ircreased in the same samples and this increase was significantly potentiated in the presence of OKY-046 in blood but not in PRP. On the other hand, the formation of PGE₂ and PGF_2α was potentiated in the presence of OKY-046 both in blood and PRP. Aspirin did not potentiate the formation of PGs both in blood and PRP. These results suggest that OKYs exert an inhibitory effect on platelet activation by inhibition of TXA₂ synthesis in platelets and by potentiation of PGI₂ synthesis in white blood cells, utilizing PG endoperoxides from platelets.

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The outcome of metastasis depends on the interaction of tumor cells with their host in the different metastatic process. Blood coagulation-fibrinolysis-platelet system is a great important host factor. Our previous studies have shown that intravascular coagulation in the target organ was a very important mechanism at an early stage of hematogenous metastasis.
In order to clarify the role of platelets on cancer metastasis, we studied the effects of various antiplatelet agents with different acting mode, such as ticlopidine, diltiazem, dipyridamole and trapidil on the intravenously induced or spontaneous pulmonary metastases. B16 melanoma or Lewis lung carcinoma cells were inoculated via a tail vein or subcutaneouly implanted into foot pad. Pulmonary metastases were assayed grossly by counting the numbers of pulmonary metastatic nodules 2 to 3 weeks after intravenous inoculation, or 3 to 5 weeks after subcutaneous inoculation of tumor cells.
Optimal inhibitory dose in ADP-induced platelet aggregation was 60-120mg/kg in ticlopidine, 180mg/kg in trapidil, 60mg/kg in dipyridamole or 2mg/kg in diltiazem, respectively. Marked inhibition of intravenously induced pulmonary metastases by B16 melanoma was obtained by the treatment of all these antiplatelet agents with dose described above. Significant reduction of spontaneous metastases by these substances was also observed only after removal of the primary tumor. Role of platelets was discussed with respect to the thrombus formation in the lodgement of tumor cells and participation of platelet-derived tumor growth factor in the growth of micrometastases in the lung.

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It is sometimes difficult to distinguish endoplasmic reticulum (ER) from platelet demarcation membranes (PDM) in megakaryocyte. We utilized the glucoes-6-phosphatase (G-6-Pase) reaction to mark ER at the ultrastrural level. Aspirated bone marrow fragments of an idiopathic thrombocytopenic purpura case were exmined. The modified Wachatein-Method was used for the detection of G-6-Pase activity. The bone marrow fragments were fixed in a 2% glutaraldehyde solution with 0.1M cacodylate buffer (pH7.4) for 10min. at 0-4°C. After rinsing in 0.1M cacodylate buffer, the fragments were incubated in a medium containing 0.375% G-6-Pate, 20ml; 0.2M tris-maleate buffer (pH6.7), 20ml; 2% lead nitrate, 3ml saccharose, 3.0g and H₂O 7ml (G-6-P medium), for 1 hour at room temperature. After rinsing again, the fragments were post-fixed. Ultrathin sections were unstained with lead citrate, or double stained with uranyl acetate and lead citrate prior to examination. Lead phosphate deposits, specifically indicating sites of G-6-Pase activity, were present in the perinuclear space and in the saccules of ER. However, deposites did not appear in the PDM or the single-membrane organelles. We could thus easily distinguish ER from PDM.
This is the first time that G-6-Pase activity has been utilized to mark ER in human megakaryocyte. We have demonstrated that G-6-Pase activity can be used to investigate the ultrastructure and function of megakaryocyte.

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In the present study we observed that peritoneal and alveolar macrophages of mice produced non-plasminic fibrinolytic enzyme (NPFE) which was different from plasminogen activator (PA), and the activity of NPFE was enhanced when the macrophages were stimulated. The stimulation of macrophages was carried out by injecting heat killed Corynebacterium anaerobium into the abdominal cavity of mice, and the peritoneal and alveolar macrophages were harvested one week after, and then they were subjedted to cultivation in the medium (RPMI-1640) containing 20% fetal calf serum for up to 96 hours. The fibrinolytic activity in macrophages were observed by the fibrinolysis autograph according to Todd and the released fibrinolytic activity in the medium was analysed by the polyacrylamide gel electrophoresis and the fibrin plate method.
The following results were obtained;
1) The stimulated macrophages clearly demonstrated two types of fibrinolytic enzyme, one of which was NPFE, and the other was PA. The former lysed plasminogen-free fibrin, and its activity was inhibited with a specific non-plasminic fibrinolytic inhibitor, Boc-Try-Leu-Val-CH₂Cl (TLVCK). The later lysed plasminogen-rich fibrin and its activity were inhibited with t-AMCHA.
2) The NPFE in cells became undetectable when cells were kept in culture medium, whereas PA was still present during cultivation.
3) The non-plasminic fibrinolytic enzyme activity was detected in the culture medium at the early cultivation phase, but tended to be undetectable at the late phase. On the other hand, the release of PA into the medium was ebserved to be retarded.
4) Both NPFE and PA in the macrophages were determind to be basic proteins, but they were an electrophoretically different entity.

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Proteinase and proteinase inhibitor activities were investigated in the scale extracts from five patients with psoriasis vulgaris and five with psoriasis pustulosa, as well as normal cornified cell extract. Hydrolytic activities were measured using peptide chromogenic substrates. PA activity was measured with fibrin plates. Inhibitor activities for urokinase, trypsin and papain were examined.
Psoriatic scale extract from each patient hydrolyzed Glu-Gly-Arg-pNA and Ile-Pro-Arg-pNA (2.6 and 5.8nmol/min/mg protein, respectively). PA activity was found in a range between 0.1-0.9 CTA U/mg protein. However, the enzymatic activity was not shown in the normal keratinized cell extract. The psoriasis extract did not show an inhibitor activity for urokinase, whereas normal extract inhibited urokinase. The following separation of PA with Sephacryl S-200 chromatography revealed three peaks with PA activity at Vo, 75k (PAh) and 25k (PAs) in MW. By DEAE Sepharose chromatography, PAs fraction showed three peaks for PA activity and a peak for synthetic substrate hydrolysis. PAh fraction was not separated by this chromatography. Since inhibitor activities for trypsin and papain did not show different levels between psoriatic and normal cell extract, changes in PA-PA inhibitor relationship in psoriatic condition presently demonstrated may have significant roles in the pathogenesis.

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To study the role of complement on DIC, we investigated whether experimental endotoxin-induced DIC was inhibited or not in decomplemented rats treated with purified cobra venom factor (CVF).
Female Wistar rats of 200g body weight were used. To make decomplemented rats, 20units/100g of purified CVF (Cordis Lab., Miami) was once injected intra-peritoneally. Six hr after the injection of CVF, endotoxin (lipopolysaccharide B, E. coli 055: B5, Difco Lab., Detroit) in a does of 100mg/kg was infused for 4hr to induce the experimental DIC in rats.
After the injection of CVF, coagulation parameters such as fibrin-fibrinogen degradation products (FDP), fibrinogen level, prothrombin time (PT), partial thromboplastin time (PTT), platelet counts, and the number of renal glomeruli with fibrin thrombi (% glomerular fibrin deposits; %GFD) remained within normal range. At 2hr after the injection of CVF, CH50 and C3 leveles were markedly decreased, and low levels continued for 72hr. After the infusion of endotoxin, the increase in FDP and % GFD, the fall in fibrinogen level and platelet counts, and the prolongation in PT and PTT were significantly inhibited in rats pretreated with CVF compared with those of the non-pretreated control rats (p<0.001, 0.001, 0.001, 0.001, 0.02, 0.001, respectively).
It is well known that C3a and C5a induces anaphylactic shock, and the release of platelet factors. Endotoxin-induced DIC was inhibited in decomplemented rats treated with purified CVF. These data indicate that the complement plays an important role in the pathogenesis of DIC.

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