Negatively stained platelts from samples of human citrated-platelet rich plasma (PRP) were examined in electron microscope on the carbon-coated grids after exposure to hypotonic condition, termed here as ultramicrodissection. Two canalicular systems were separated as structural elements; one may be correspondent to the dense tubular system (DTS) and the other the open canalicular system (OCS). No definite branch was formed in the DTS, while obvious connections were present between the α-granule and the branched OCS.
On the carbon-coated grids, spread platelets were also examined without fixation and staining. The electron opaque serotonin granule, apparently different from another organelles in the granulomere, protrudes long tails with time. Decrease in number and vacuolation of the granule may suggest the depletion of the serotonin matrix.
Cytochemical approaches were also carried out on thin-sectioned platelets to obtain a further correlation of the separated canalicular elements to those appeared in the transmission electron microscope. The OCS of platelets responded to adenosine diphosphate (2×10^-7M) was traced with lanthanum nitrate as branched elements. The diaminobenzidine oxidative activity is suggested to localize in the DTS.
The selectivity of release through the canalicular system was discussed in relation to the rapid and slow release reactions.

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The results of in vitro studies on Indium-111-oxine uptake by and loss from human platelets as well as function of platelets incubated with Indium-oxine in ethanol-saline (In-Es) or in HEPES buffer with polysorbate 80 (In-PH), are presented.
When subcellular fractions were analysed, more than 60% of the In-111 was located in the cytoplasmic pool of platelets incubated with In-ES. Less than 30% of label was located in cytoplasm of platelets incubated with In-PH.
Although the distribution ratio of free In between platelet water space and the outside medium exceeded unity in 30min incubation with 0.001 to 10nM of In, uptake of label did not appear to be dependent on intact platelet energy metabolism.
The in vitro sensitivity of platelets in response to different aggregating and release inducing stimuli was suppressed when platelets were incubated with In-ES or HEPES buffer containing polysorbate 80. Higher concentration of polysorbate 80 promoted the leakage of LDH and platelet factor 4 by injuring platelet membrane.
Loss of In-111 from platelets labeled with In-ES or In-PH was 23% and 58%, respectively, during 20hrs in vitro incubation in plasma at 22°C.
This study demonstrated that it is better to use ethanol as a solvent of In-oxine complex rather than surfactant to avoid the damage of cell membrane by it.

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The method reported by Lorand et al. for measuring the activity of the plasma factor XIII consists in determining it from fluoresence intensity of the casein-dansylcadaverine complex formed by a transglutaminase activity of factor XIII, using casein and dansylcadaverine as the substrate. We used gel filtra tion (molecular sieving) as a means of separating the casein-dansylcadaverine complex from the free dansylcadaverine. With this method it was possible to achieve separation easily in a short time. This has enabled the measurement on a large number of samples. The amount of plasma is 20μl and the measurement time is approximately 30 minutes for the total measuring procedure. This assay method shows linearity up to 200% and coefficient of variation is 4.9%. The correlation coefficient of this method vs. Lorand's method is 0.965.

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The aim of this report is to investigate the effects of eicosapentaenoic acid (EPA), procursor of prostaglandins of the 3-series, on platelet aggregation (PA), blood coagulation, blood lipid and fatty acid composition of plasma and platelets in man. Twelve volunteers (average age 60.7ys) were treated with ethylester of EPA, 2g/day in 3 divided dosages for the four weeks. ADP-, collagen- and adrenaline- induced PA, blood coagulation, blood lipid and fatty acid composition of platelet poor plasma and platelet phospholipid were examined before and 2 and 4 weeks following EPA treatment.
Platelet aggregation, induced by ADP, collagen or adrenaline, decreased significantly at 2 and 4 weeks following EPA treatment. There were no significant changes of bleeding time, prothrombin time and plasma fibrinogen after EPA administration. Significant decrease in both serum total cholesterol and triglyceride levels was observed at 4 weeks following EPA treatment, while no significant change in HDL cholesterol was found following the treatment. Following the treatment with EPA, significant increase of EPA content in plasma and platelet and no change of arachidonic acid content of plasma were observed. In conclusion, the administration of EPA was effective in decreasing platelet aggregation, possibly by changing the ratio of EPA to arachidonic acid in the platelet.

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The mechanisms of acceleration of the platelet aggregation in the hypercholesterolemic rabbits were investigated. Rabbits were fed with 1% cholesterol-containing standard diet for 3 months.
No apparent increase was found in ADP-induced aggregation, but the marked acceleration of arachidonic acid (AA)-induced aggregation in the PRP (platelet rich plasma) and the slight acceleration in the WPS (washed platelet suspension) were found at end of 3 months' diet.
Analysis of the fatty acid composition of total lipids in the plasma revealed an increase of AA and a decrease of docosahexaenoic acid (DHA). Also the analysis of the fatty acid composition of phospholipids in the platelet revealed the increase of AA. The incorporation of ¹⁴C-AA into thromboxan B₂ (TXB₂) in platelet was increased approximately 2 times of that of the control. It was suggested that the acceleration of platelet aggregation in the hypercholesterolemic rabbits are caused by 1) the increase of AA component in plasma, 2) increased incorporation of AA into platelet and 3) the increase of AA metabolism to TXB₂ within the platelets.

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Platelet taurine and platelet aggregation were measured in an attempt to clarify the physiological function of taurine in the platelet in both healthy volunteers and essential hypertensives. Platelet taurine was significantly low in untreated essential hypertension as compared to normals. Platelet taurine and mean blood pressure exhibited a significantly negative correlation (r=-0.507, p<0.01) between normals and untreated essential hypertension, while mean platelet volume was significantly great in untreated essential hypertension as compared to normals. Platelet aggregation in response to epinephrine (0.2μg/ml) and ADP (2.3μM) was significantly (p<0.01) great in untreated essential hypertension, suggesting a possible taurine defect. Significant positive correlation (r=0.311, p<0.01) was obtained between mean blood pressure and ADP (2.3μM) platelet aggregation in both normals and untreated essential hypertension. The administration of taurine 6g/day per os 5 months normalized epinephrine and ADP platelet aggregation in 10 patients.
These findings suggest that essential hypertension may result from a insufficient metabolism of taurine, which leads to enhanced platelet aggregation possibly resulting in myocardial infarction and cerebral thrombosis, and that taurine could be useful as a platelet antiaggregative agent.

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The effects of sustained handgrip isometric exercise on blood coagulability and platelet function were studied in 10 patients with ischemic heart disease (IHD) and 10 healthy controls. Isometric exercise was performed for 3min at the 50% level of maximal voluntary contraction. Blood sampling was made at rest, and immediately and 6min after exercise.
No significant changes in the platelet sensitivity to ADP-aggregation and β-TG levels were noticed in IHD and control groups after exercise. In IHD fibrinogen levels were higher and antithrombin III activities were lower than those in healthy controls at rest. These two substances increased after exercise in both groups. Prothrombin time and partial thromboplastin time were shortened immediately after exercise in IHD, while these measurements showed no significant changes in healthy controls, The activity of factor VIII increased immediately after exercise in IHD and the activity of factor XII in healthy controls, respectively.
In conclusion, platelet function was not altered in both groups after exercise. However, there was a tendency to an increase in blood coagulability after exercise in IHD. Hemostatic unbalance might be induced after the isometric exercise in IHD patients.

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Changes in hemorheologic properties and platelet function induced by a multistage ergometric stress testing in the supine position were observed in 31 patients with ischemic heart disease (IHD), 12 middle aged (MH) and 14 young normal subjects (YH). Slight but significant increases in hematocrit and platelet count were observed at peak exercise in all groups. Erythrocyte deformabiltiy (nuclepore filtration method) at peak exercise reduced significantly from 0.83±0.14 to 0.76±0.14ml/min in IHD and 0.80±0.10 to 0.75±0.10ml/min in MH, but unchanged in YH. Corresponding to such change, whole blood viscosity at the shear rate of 230 sec^-1 increased significantly from 4.23±0.48 to 4.38±0.54 cP in IHD, while that at the shear rate of 23sec^-1 remained unchanged. Platelet adhesiveness (modified Salzman's method) was enhanced, and platelet aggregability to ADP and epinephrine was increased at peak exercise in IHD. Epinephrine-induced platelet aggregation was also increased in normal subjects with exercise. Aspirin inhibited platelet aggregability both at rest and during exercise in IHD. The data suggest that hemorheologic characteristics and platelet function during exercise are, at least in part, responsible for an acute coronary event in patients with IHD.

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This report shows the presence of an unidentified enzyme zymogen which is activated by thrombin generated accompanied by platelet activation. This enzyme cleaved a synthetic fluorogenic substrate, Boc-Leu-Ser Thr-Arg-MCA, and was not inhibited by antithrombin III/heparin. The activation of the zymogen occured by the addition of Ca²⁺ (5-20mM) into platelet-rich plasma (PRP), but not platelet-poor plasma (PPP). Thrombin (0.4U/ml) facilitated the activation of the zymogen in PRP, but little in PPP. The activation of the zymogen depended on platelet count in plasma. Inhibitors for platelet activation, C-AMP and EDTA inhibited the activation of the zymogen in proportion to its concentration. Hirudin (10U/ml) prolonged the lag time for the appearance of this enzymatic activity after Ca²⁺ addition, and antithrombin III/heparin or DAPA, a specific inhibitor for thrombin, obviated the activation of the zymogen completely. Although both normal and prekallikrein deficient human plasma containing washed platelets could generate the noticed enzymatic activity after addition of Ca²⁺, the physiological buffer containing washed platelets could not. The enzyme was inhibited by benzamidine, DFP, DAPA and partially by hirudin, neither by anti-prothrombin-IgG, anti-protein C-IgG nor protein C inhibitor. These results indicate that the enzyme resistant to antithrombin III/heparin and similar to thrombin in substrate specificity may differ from the already known active coagulation factors, plasma kallikrein and protein Ca, and that this enzyme zymogen could be activated by abundant thrombin generated accompanied by platelet activation.

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Effects of the antiplatelet drug, ticlopidine (600mg/day, p. o.) on the anginal attacks of effort or rest angina pectoris by placebo-controlled single blind randomized-cross over method. In 5 patients with rest angina refractory to the conventional method (Ca-antagonists, beta-blockers, isosorbide dinitrates), during ticlopidine therapy, frequencies of anginal attacks and doses of nitroglycerin were both decreased. However, in 5 patients with effort angina, using the Bruce treadmill test, no beneficial effects were observed.

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In order to search the changes of response between vascular wall and platelets, plasma thromboxane (TXB₂), cyclic AMP, cyclic GMP levels and platelet aggregation were measured during exercise in 31 patients with the complaint of chest pain. The exercise was performed by submaximal treadmill test and blood samples were collected before, immediately and 30min. after test.
Using Hollenberg's treadmill exercise score (TES), 15patients were judged as TES (-) group (positive in conventional test) and others as TES (+) group (negative test). At the immediately after exercise, plasma levels of TXB₂ in TES (-) group were 208.5±20.8pg/ml, a statistically high level as compared with 168.6±14.4pg/ml, in TES (+) group. On the other hand, in TES (+) group levels of cAMP and cGMP (cAMP; 14.5±0.7, cGMP; 5.37±0.46pmol/ml) increased significantly immediately after exercise, as compared with these in TES (-) group (11.3+0.8, 4.44±0.48pmol/ml). No significant changes of platelet aggregation was seen in both TES (+), TES (-) groups, except that induced by adrenaline. In TES (-) group, maximum aggregation rate induced by adrenaline immediately after exercise was 66.5±5. 8%, a statistically high as compared with 45.5±8.2% before exercise. There was a positive correlation between TXB₂ and cAMP in TES (+) group during exercise, however, such a correlation was not found in TES (-) group.
These data suggest that the poor response of vascular wall for hemodynamic changes exists in patients of coronary heart disease which breaks the exquisite balance of the interrelationship between platelet and vascular wall.

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Changes of, β-thromboglobulin (β-TG) concentration in blood drawn from the proximal aorta (Ao) and coronary sinus (CS) during cardiac catheterizaiton were investigated in patients with ischemic heart diseases and the following observations noted: 1) Catheter manipulation did not significantly affect, β-TG concentration.. 2) β-TG level following exercise stress was higher in the left ventricular asynergy group (ASY) than in the group without ASY, suggesting enhanced platelet function in ASY. 3) In the group with non-provoked variant angina, β-TG level during rest did not differ from the normal control and remained at the same level following exercise and ergonovine provocation. 4) Different β-TG levels were observed in an ergonovine provoked anginal attack case (ERP) compared with a spontaneous attack case (SPN). In the ERP case, β-TG level was higher in CS than Ao during an anginal attack. Conversely, reversed results were obtained for SPN. In addition β-TG level remained higher both before and after an anginal attack in SPN. It was concludcd that an intimate relationship between platelet activation and ocronary artery vasospasm was proved by the direct methods.

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Platelet survival time was measured in 46 patients with various cardiovascular diseases and 5 healthy controls based upon ICSH criteria using ¹¹¹Indium labeled platelets, and thrombus formation was investigated by scintiphotography. Platelet count, β-thromboglobulin and coagulation-fibrinolytic system were also investigated.
Significant shortening of mean platelet survival time was observed in patients with arteriosclerosis obliterans and aortic aneurysm comparing with healthy controls. The same tendency was seen in patients with mitral stenosis who had a history of thromboembolism and in patients with left ventricular aneurysm due to myocardial infarction in spite of warfarin therapy.
Correlations were necognized beween platelet survival time and platelet count and between platelet survival time and laboratory tests of coagulation-fibrinolytic system were not significant.
The accumulation of labeled platelets on a thrombus in the left ventricular aneurysm in one patient and in the left atrium in two patients was clearly detected on the scintiphotogram. An accumulation of labeled platelets was detected on an aortic prosthetic graft, especially at the anastomotic sites and also on the thromboend-arterectomised segment in cares with arteriosclerosis obliterans.
Imaging of accumulated platelets and measurment of platelet survival time using ¹¹¹Indium-oxin labeled platelets was considered to be an useful tool for the diagnosis and the evaluation of treatment in cardiovascular disease.

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We analyzed plasma β-thromboglobulin (β-TG) levels during cardiopulmonary bypass to compare the effects of membrane and bubble oxygenators on platelet, because our and other author's comparative studies by the conventional measurements of circulating platelet counts, sensitivities to activating agents and adhessiveness to glass beads were controversial. In bubble oxygenator group (Bentley, BOS 10, n=8.), plasma β-TG level increased from 34.1±15.1ng/ml before perfusion to 1095.6±463.7ng/ml after 60 minutes of bypass and the rate of increase was 18.3ng/ml/min. In membrane oxygenator group (Sci Med Life System Inc. Minneapolis, Minn.), plasma, β-TG level increased from 29.5±15.6ng/ml to 242.9±158.2ng/ml and the rate of increase was 4.0ng/ml/min.
Plasma β-TG levels 60 minutes after bypass were 12.0±7.1ng/ml/perfusion time (min) in bubble oxygenator group and 2.9±1.5ng/ml/min in membrane oxygenator group (p<0.01).
These data indicate that the traumatic effects on platelet of membrane oxygenator is much less than bubble oxygenator.

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Vasodilator drugs have often been applied clinically to the patients with ischemic heart disease or hypertension, and their antiplatelet activity have been evaluated recently. This paper reports the effects of Hydralazine, Budralazine, Verapamil and Nicardipine, on the platelet aggregation and PGI₂ generation from rat aorta.
Effects of the drugs on the platelet aggregation in vitro were evaluated by ADP, arachidonic acid (A. A.) and collagen induced aggregation. Effects of drugs on the PGI₂ generation from rat aorta in vitro and in vivo were examined by a bioassay system. ADP-induced platelet aggregation was inhibited dose dependently by these drugs at the higher concentrations than 10^-4M. A. A. and collagen-induced platelet aggregations were partially inhibited at 10^-4M of these drugs, and completely inhibited at the concentration over 2×10^-4M. These drugs increased PGI₂ generation of rat aorta both in vitro (10^-5-10^-7M) and in vivo (1mg/Kg).
when hydrocortisone, a phospholipase A₂ inhibitor, was added to the incubation system of anrta PGI₂ activity was not increased even in the presence of Verapamil. However when A. A. was added to this incubation medium, PGI₂ generation was recovered and increased with the addition of Verapamil. These results indicate that Verapamil increases the PGI₂ generation in the process of A. A. metabolism.
In the action of these vasodilators, it was confirmed that the prostaglandins, especially PGI₂ and TXA₂, were involed. And it is suspected that Ca-antagonist affects the process after liberation of arachidonic acid.

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Recently ethanol intoxication has been recognized as a risk factor of cerebral thrombosis in the young adults.
The purpose of this study is to elucidate the effects of ethanol on blood coagulability, fibrinolytic activity, platelet aggregability, blood viscosity, and serum protein and lipids. Each comparative study was made before and after the oral administration of beer in nine young, healthy male volunteers. Beer was taken in the total amount of 1899ml containing about 4.5% ethanol (total ethanol volume: about 85g) over 3 hours.
The results of this study were as follows:
(1) Elevated hepaplastintest (p<0.01).
(2) Prolonged euglobulin clot lysis time (p<0.005).
(3) Enhanced platelet aggregability induced by ADP (2μM) (p<0.05) and ristocetin (1.25mg/ml) (p<0.01).
(4) Decreased whole blood viscosity at shear rate of 18.75sec^-1 (p<0.005), 75sec^-1 (p<0.01) and 375sec^-1 (p<0.05) respectively.
(5) Decreased hematocrit value (p<0.005).
(6) Decreased total protein (p<0.05).
(7) Decreased total cholesterol (p<0.005).
(8) Increased triglycerides (p<0.05).
(9) Decreased HDL-cholesterol (p<0.05).
They suggest that excessive beer abuse may decrease the risk of cerebral thrombosis through its effect on whole blood viscosity, whereas it may increase the risk through its effects on blood coagulability, fibrinolytic activity, platelet aggregability and serum lipids.
In conclusion, excessive ethanol abuse may be a risk factor in the development of cerebral thrombosis.

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Activation of Factor XII was examined as the generation of prekallikrein activator (PKA) activity. The incubation mixture of various plasma samples with activator, such as kaolin and bromelain, was further incubated with partially purified human plasma prekallikrein, and then the amidase activity of plasma kallikrein generated was measured with Z-phe-arg-MCA as a substrate.
By kaolin activation normal plasma and XI deficient plasma yielded PKA activity, but prekallikrein or high molecular weight kininogen deficient plasma could not generate the activity. On the contrary, all plasma except Factor XII deficient yielded PKA. The PKA activities of Fletcher plasma and Fujiwara plasma were less than normal value. However, these values corresponded well to those estimated from other assay method. Fibrinolytic activity in rabbit plasma after intravenous injection of bromelain was the same as that from nontreated animal. Either the decrease in plasminogen in the plasma after bromelain could not be detected. Thus the activation of Factor XII by bromelain could be led to the specific activation even in vivo, and could be used for the functional assay of Factor XII.

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For the analysis of the pathophisiological role of the kallikrein system in hematological disorders, the system of bone marrow blood (BM) was examined. Using chromogenic peptide substrate (S-2302), spontaneous activity, prekallikrein activated by activator (KABI Co.) and kallikrein inhibitor to purified plasma kallikrein (KABI Co.), were measured in BM of 15 cases of hematological disorder.
Spontaneous activity was 13.5±4.5 (mean±SD) u/L in peripheral blood (PB) and 36.1±13.4u/L in BM. Prekallikrein was 86.2±19.3% (PB) and 72.6±20.6% (BM). Kallikrein inhibitor was 91.9±30.3% (PB) and 55.4±21.5% (BM). A good correlation between spontaneous activity and kallikrein inhibitor was observed in BM. Another good correlation between kallikrein inhibitor and C₁-inactivator was observed both in PB and BM. The discrepancy between BM and PB was marked in acute leukemia. Prekallikrein and kallikrein inhibitor were depressed both in PB and BM in myeloproliferative diseases.
Compared to PB, high spontaneous activity, depressed prekallekrein and depressed kallikrein inhibitor were obvious in BM. These suggest a marked activation of the kallikrein system in BM and a possible participation of kallikrein in hematopoietic enviroment.

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We studied on the correlation between kallikrein-kinin systems and haemostasis in pancreatitis.
The results were as follows;
1) Plasma prekallikrein levels showed a significant decrease in acute pancreatitis but a significant increase in chronic pancreatitis.
2) Plasma total kininogen levels decreased significantly in acute prancreatitis.
3) The levels of plasma high molecular weight (HMW) kininogen and low molecular weight (LM W) -kininogen decreased in acute pancreatitis. However, LMW-kininogen level alone decreased in chronic pancreatitis.
4) Serum kininase II and trypsin levels increased in pancreatitis.
5) We attempted to study kinin releasing activities of pancreas juice and trypsin on LMW-kininogen and plasma. Kinin formation level which liberated from LMW-kininogen by pancreatic juice was 17ng/BK eq./5min.
From the above results, trypsin and pancreatic glandular kallikrein are released into blood in patients with pancreatitis. These enzymes activate kallikreinkinin systems in patients with pancreatitis.

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A family of inherited antithrombin III (AT III) deficiency was reported. Propositus was a 42-year-old man with 10-year history of deep vein thrombosis and cerebral thromboembolism.
AT III was assayed for his family members in three consecutive generations by single radial immunodiffusion, electroimmunoassay by Laurell, progressive antithrombin activity by thrombin clotting time and chromogenic assay using S-2238. A significant decrease of plasma AT III levels with low activity was found in 8 (47%) out of 17 members investigated. Five (63%) of them had recurrent episodes of deep vein thrombosis of extremities and/or cerebral thromboembolism, mesenterial thrombosis and pulmonary embolism. In two dimensional crossed immunoelectrophoresis, mobility of their AT III in the heparinized agarose gel was identical with that of normal control. Thus, the family is classified as type I by Nagy or type Ia by Sas.

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In various conditions of rheumatic heart diseases (RHD), as we reported previously, decrease of Antithrombin-III (AT-III) was observed, and it was supposed to be a result of hypercoagulable state in these conditions. In this study AT-III and other hemostatic parameters were examined before and after surgical treatments of 110 RHD patients to know their behaviours. And effects of Warfarin to them were also studied.
Although sites of valvular replacements had differences in AT-III and Plasminogen (Plg.) values before operations, their changes after operations seemed to be similar. Their mean values were significantly higher at 1 month after operations than pre-operative values, but they seemed to be decreased again at 2 months. At 1 month after operation, besides AT-III and Plg., inflammatory parameters remained still higher in responce to surgical procedures. Platelet count was also increased at 1 month, but they seemed to be still slightly higher even at 2 month.
Patients with open mitral commissurotomies without Warfarin therapies showed no difference in AT-III values before and after operations. But with postoperative Warfarin therapies, 62.0% of the patients showed increases in AT-III at 1 month after operations. And statistics showed use of Warfarin was related with AT-III behaviour both 1 and 2 months after operation.
The present study showed that post-operative RHD patients might be still in hypercoagulable states and Warfarin might be beneficial in view of laboratory hemostatic findings.

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Effect of warfarin on left atrial thrombus was studied.
53 rabbits were used. A plastic piece of 3×15mm was inserted into the left atrium of rabbit. Rabbits were sacrificed on the 1st, 5th, 10th, and 30th days and thrombus formation were found in all rabbits.
Fourmg/day of warfarin was administered subcutaneously to 33 rabbits from 4 days after insertion of plastic pieces. Apparent decrease of thrombus was noted even on the 10th day after warfarin administration compared with those without warfarin. This tendency was more clearly noted on the 20th and 30th days, and we could not find any thrombus in two of them.

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Nonprecipitating antibodies directed toward human-F VIII R: Ag were found in the plasma of a multitransfused patient with severe von Willebrand's disease (VIII: C 2%, VIII R: Ag<5%, VIII R: RC<5%, B. T.>30min.) during replacement therapy with VIII concentrate for intracranial hemorrhage.
Such antibodies were detected with newly developed semiquantitative assays of anti-VIII R: Ag antibodies by using ELISA. Partially purified AHF made from commercially available F VIII concentrate by Gel filtration was coated as solid phase antigen on the well of plastic microplate. The reacted patient's antibodies with VIII: Ag on the surface of the wells were detected and titrated by using enzyme conjugated anti-human IgG heavy chain, k or λ Light chain. These studies showed that the types of the anti-VIII R: Ag antibodies were IgG heavy chain, mainly k light chain and partially λ light chain. The anti-VIII R: Ag IgG antibody in the patient's plasma diluted (1:10, 000) with PBS was detected 3 weeks after initation of the repalacement therapy by this method. These antibodies were absorbed with normal plasma but not with severe von Willebrand's plasma.
The anti-VIII R: Ag antibodies in patient's plasma blocked platelet retention by glass beads column measured by Bowie's method. But they did not block ristocetin-induced platelet aggulutination at all, and they weakly inhibited procoagulant activity of AHF in non-specific manner.
These data suggest that the site or sites on the AHF complex molecule that are associated with ristocetin-induced platelet agglutination differ qualitatively from those associated with enhancement of platelet retention by glass beads.

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In order to elucidate the relation between intraglomerular fibrin deposition and types of serum and urinary fibrin/fibrinogen degradation products (FDP) in children with various renal diseases, the extent and distribution of fibrinogen related antigen (FRA) deposition in the kidney tissue which was perfesed with monochloacetic acid (MCA) solution was examind by immunofluorescent microscopy in comparison with deposition of factor XIII subunit-A (XIII-a). In addition to the immunofluorescent studies, the typing of FDP was made by SDS polyacryamide gel electrophoresis and crossed immunoelectrophoresis (SDS-PAGE-CIE).
Intrarenal deposition of FRA after MCA treatment was observed mainly within the glomerular capillary walls, mesangium or vessels, and its extent and distribution showed a good agreement with that of XIII-a. Patterns of SDS-PAGE-CIE of MCA insoluble FRA products by plasmin were similar to those seen in cross-linked fibrin. These findings suggest that MCA insoluble FRA or XIII-a deposition reflects the extent and distribution of cross-linked fibrin.
The presence of D-dimer in the urinary FDP was well seen in the children who had intrarenal fibrin deposition.

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Plasma fibrinogen (Fbg) derivatives were selectively obtained by immuno affinity chromatography. Electrophoretically. Fbg derivatives could be devided in to 3 subfractions (Sf): native Fbg (N) (MW: 33.0×10⁴), modified Fbg 1 (M1) (MW: 31.0×10⁴) and modified Fbg 2 (M2) (MW: 28.0×10⁴). In the normal plasma, percent concentrations of each Sf were 62.9-70.7% in N, 26.1-31.9% in M1 and 2.9-5.5% in M2. Analysis with the reduction method revealed that the difference between these Sf's was mainly due to the degree of defect in Aα chain. In almost all cases a large amount of fragment D was noted in their plasma. In the normal adults and the patients with DIC the time course of changes in Fbg-Sf and FDP-E were observed. In the normal adults, both of Fbg-Sf and FDP-E showed small changes within several days. In contrast, the marked changes in Fbg-Sf and FDP-E were noted in the cases of DIC according to the clinical stages; namely, the decrease of N, the marked increase of FDP-E and the appearance of M3 at the onset of DIC; the marked increase of N and FDP-E and the disappearance of M2 and M3 at the peak stage, and the slight increase of N and FDP-E, and the appearance of M2 in the convalescence. From these results, the destruction of Fbg was considered to in the following sequence: N→M1→M2→FgDP.

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Plasma fibronectin levels were determined immuno-turbidimetrically in 95 cerebrovascular patients (61 with cerebral infarction and 34 with cerebral hemorrhage), 42 patients with hypertension, and 45 healthy controls age-matched with three groups of patients. Cerebrovascular patients were those in chronic stage passed more than one month after stroke.
The average value in controls was 349±88.8μg/ml. There was no difference by sex, but a tendency to increase slightly with advancing age was seen. The average values in patients with infarction, hemorrhage, and hypertension were 403±116.1μg/ml, 402±93.6μg/ml, and 403±96.0μg/ml, respectively. All these three values were significantly higher than that in controls. When cerebrovascular patients were divided into two groups by the score in test of activity of daily living (ADL), those scored higher mark than the average were found to have higher fibronectin level as compared to those scored lower ADL. Plasma fibronectin level correlated positively with platelet adhesiveness, but not with other platelet functions.
These results suggest that fibronectin may have some prognostic value especially in relation to the functional ability of cerebrovascular patients. In addition, higher value observed in hypertensives and the relationship with platelet adhesiveness may be noticeable findings suggesting the involvement of fibronectin in the development of vascular diseases.

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This study was undertaken to evaluate the diagnostic value of β-D-N-Acetylglucosaminidase (NAG) in the acute occlusion of superior mesenteric artery of the dog. In eight adult mongrel dogs of both sexes, superior mesenteric artery and superior mesenteric vein were clamped for 2 hours. Blood samples were taken from the portal and femoral vein before clamping, immediately before declamping and 2 and 4 hours after declamping. NAG was measured by NAG test kit (Shionogi, Japan). Aminogram of the blood samlpes was analyzed. The small intestine with ischemia was taken for histological examination.
NAG level significantly increased after declamping in the portal and femoral vein. Aminogram revealed an increase of taurine 4 hours after declamping. Histologiclly intramucosal bleeding and degeneration of epitherium were observed. Significant increase of blood NAG level was observed soon after acute occlusion of SMA and SMV in the present canine experiment. It was suggested that measurement of blood NAG might be valuable for early detection of acute SMA thrombosis.

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In order to clarify a suggested relevance of DIC to ARDS, we designed an experimental model of DIC in dogs by declamping shock.
As soon as the blood circulation was reestablished, the animals fell into a severe shock with marked decrease in platelets, fibrinogen, antithrombin III and plasminogen and increase in FDP. The PaO₂ decreased simultaneously and the A-a Do₂ increased progressively. Marked perivascular edemas and dilated lymphatic channels were principal histological findings in the lung. These findings closely resemble those observed in the early stage or ARDS in man.
An administration of heparin, prostaglandin E₁ or FOY prior to the induction of shock minimized or even prevented these changes in the respiratory functions as well as blood coagulation.
Thus we presume that DIC may trigger ARDS after severe trauma or extensive surgery.

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Effect of ticlopidine, 5-(2-chlorobenzyl)-4, 5, 6, 7-tetrahydro [3, 2-C] pyridine hydrochloride, on experimental disseminated intravascular coagulation (DIC) was studied in rats. The experimental DIC was induced by 4-h sustained infusion of endotoxin at a dose of 100mg/kg in rats. Rats were injected with ticlopidine at 2.0, 20.0, 50.0, 100.0 or 200.0mg/kg intraperitoneally, and thereafter infused continuously with 100mg/kg/4h of endotoxin. The preventive effect against DIC was noted in all the parameters, such as fibrinogen and fibrin degradation products, fibrinogen level, prothrombin time, partial thromboplastin time, platelet count, and the number of renal glomeruli with fibrin thrombi, in rats treated with 20.0, 50.0, 100.0 or 200.0mg/kg of ticlopidine.
From these results, it was shown that ticlopidine inhibited the aggravation of endotoxin-induced experimental DIC in rats.

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The purpose of this study was to detect the possible factors related to the occurrence of DIC in cancer patients.
I) Our own 20 cancer cases accompanied with DIC and the similar 223 autopsy cases reported by the Japanese Association of Pathology were studied.
Results; 1) The cancers most frequently accompanied with DIC were gastric, biliary system, pancreatic, colon, lung, and hepatic cancer, especially those with distant metastases. 2) Pneumonia, UTI, and biliary tract infections, as well as jaundice and hepatic diseases seemed to be the most important triggers of DIC, because these were frequently seen in cancers with DIC (Figure 1). 3) No significant relationship was found between the anti-cancer chemotherapy and the DIC incidence. 4) Endotoxemia was more frequently detected in the cases having received anti-cancer drug administration than in those without that.
II) The influence of anti-cancer chemotherapy on the incidence of endotoxemia was examined in rats using Limulus test (Figure 2).
Results; Higher incidence of endotoxemia was revealed in the groups treated with high dosage of 5-FU or of Cyclophosphamide. These incidences of endotoxemia seemed to run parallel with the incidences of diarrhea and of weight loss in each animal group.
These results may suggest some of the responsible factors for DIC occurrence in solid carcinoma patients.

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Activation of intrinsic pathway of fibrinolysis and its inhibitory system were studied immunologically. Using anti-plasminogen antibody crossed immunoelectrophoresis (CIE) was done in normal plasma on condition that prekallikrein was activated with prekallikrein activator (Pkk. A.; Kabi) which is a mixture of ellagic acid, cephalin, and plasma fraction containing Hageman factor and high molecular weight kininogen. CIE of normal plasma activated with Pkk. A. revealed two peaks, while that of untreated plasma did single peak. It was supposed that newly appeared first peak was plasmin generated by intrinsic pathway and second peak was original plasminogen.
Plasma in a case of hereditary angioneurotic edema (HANE) showed a higher and wider first peak in pattern of CIE on the same condition as normal plasma. On the other hand these findings were not observed, when normal plasma was treated with other contact activating agents.
CIE using anti-C₁^-esterase inhibitor (C₁^-INH) antibody and anti-α₂-plasmin inhibitor (α₂-PI) antibody were also done in normal plasma. A distoi-tion of ascending shoulder of original peak was seen in both CIE patterns after the activation with Pkk. A.. It seems likely that complexes between kallikrein and C₁^-INH or α₂-PI were generated while activation of prekallikrein.

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Tissue plasminogen activator (TPA) was purified either as a single chain or as a two chain form from the culture medium of a human melanoma cell line. The thrombolytic activity of TPA was investigated in beagle dogs with an experimental femoral vein thrombosis and compared with urokinase. The ¹²⁵I-fibrinogen labeled thrombus was formed in the femoral vein, and the thrombolytic agents were infused over a 4 hour period. The degree of thrombolysis was measured as the difference between the injected and recovered ¹²⁵I. In six control animals with a saline infusion, the degree of thrombolysis was 16.3±3.8 percent (mean±S. E. M.), in 5 dogs receiving 100, 000IU urokinase, 17.4±3.7 percent and in 4 dogs with 1, 000, 000IU urokinase, 40.6±4.8 percent. Infusion of 100, 000IU single chain TPA in 5 dogs resulted in 33.5±7.8 percent lysis and of 100, 000IU two chain TPA in 5 dogs in 60.1±10.8 percent. Infusion of 300, 000IU one chain TPA yielded 57.5 percecent lysis and of the same amount of two chain TPA 72.9 percent. Significant activation of plasminogen, consumption of α₂-antiplasmin and fibrinogen breakdown was only observed in the animals receiving the high dosis of urokinase but not in the saline, TPA or the low dose urokinase groups. It is thus concluded that in this thrombosis model, human TPA has a higher specific thrombolytic effect than urokinase, without systemic fibrinolytic activation and fibrinogen breakdown.

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