Bipolar platelets named by us (1977) have two polar projections issuing from opposite sites of the spherical body, which are usually straight, arrange on a line and thicker than usual pseudopods.
In normal subjects (80 males and 80 females of age 16-80), the bipolar form was found in 1.3+1.1 (SD)% (95% range: 0-4%). There was no significant difference in the frequencies between ages (under 20, 20-40, 40-60, over 60) or between sexs. Clinically, the frequencies of the bipolar form increased in several conditions such as malignant lymphoma, idiopathic thrombocytopenic purpura, pernicious anemia, primary thrombocythemia, females after delivery, whereas they were normal in DIC, acute promyelocytic leukemia, and some thrombotic conditions including diabetes mellitus, varices and thrombophlebitis. The highest frequencies were encountered in each case of primary thrombocythemia and idiopathic multiple lung emboli (4-7%).
In in vitro experiment to produce bipolar form, A23187 produced bipolar form (9%) at 20μg/ml, 30sec, whereas thrombi, ADP, adrenaline, trypsin, plasmin, arachidonate and EDTA-2Na failed. Rewarming of chilled (swollen) platelets produced many bipolar forms (around 60%) before discs increased, indicating transformation from bipolar form to discoid form.
Scanning electron microscopical servey on platelets from some normal subjects and those patients with many bipolar forms revealed various intermediate forms between bipolar and disc forms, i. e. curved form, ring form and discoid form with thinned cytoplasma at the central part.
In conclusion, bipolar platelets exist in both normal and pathological conditions and increase probably related to platelet activation in vivo by hemolysis, immune complex, thrombotic process or others. They are actually an intermediate form in a transformation from spherical form to discoid form.

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A cell surface and tissue protein, termed fibronectin, is antigenically identical to a major plasma glycoprotein, cold-insoluble globulin (CIG). Fibronectin on fibroblast cell surfaces contributes to cell spreading and adhesion, and its protein diminishes from these surfaces when transformed by oncogenic viruses. Recently, fibronectin has been found in platelets and is known to be released from them under stimulation by collagen or thrombin. Moreover, platelet fibronectin (P-fibronectin) is suggested to contribute the receptor for collagen in the platelet (Bensusan, H. B., et al.: Proc. Natl. Acad. Sci. USA, 75: 5864, 1978).
The present study aimed to confirm the findings of Bensusan, et al., and to investigate the influences of P-fibronectin and CIG on the collagen binding to platelets. The results obtained were as follows.
(1) Fibronectin was isolated by affinity chromatography on anti-CIG-IgG-agarose. Its protein, different from CIG, showed a single band SDS-gel electrophoresis after reduction and a molecular weight of approximately 230, 000.
(2) The collagen-induced secretion of serotonin from human platelets in plasma was inhibited by the treatment of collagen with both P-fibronectin and CIG. The secretion was stimulated with gelatin and inhibited by CIG.
(3) Purified anti-CIG-IgG stimulated serotonin secretion from the platelets, but the control IgG did not. The secretion from platelets stimulated with anti-CIG-IgG was inhibited by pretreatment of anti-CIG-IgG with both P-fibronectin and CIG.
(4) Collagen preincubated at 37°C for 120min was found to bind to platelets, but the binding of collagen scarecely saturated with unlabelled collagen. Binding of collagen to platelets was inhibited by pretreatment of collagen with both P-fibronectin and CIG.
These evidences imply that fibronectin could constitute the collagen-binding protein of platelets.

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We have investigated the binding of factor VIII to platelets in the presence of platelet aggregating agents.
Factor VIII was purified from cryoprecipitates by gel filtration on Sepharose 2B column and was iodinated by lactoperoxydase technique. To washed human platelets (5×10⁸/ml in 10mM Tris, 0.15M NaCl, pH 7.4), ¹²⁵I-factor VIII (5μg/ml) and an aggregating agent were added. After incubation at 24°C for 3 minutes platelets were separated by centrifugation at 7, 000×g for 1 minute through silicone oil. The radioactivity of the platelet pellets was measured by gamma counter.
Platelets bound 33ng of factor VIII per 10⁸ platelets without addition of aggregating agents. The addition of aggregating agents such as ADP (20μM), collagen (Horm, 4μg/ml) or epinephrine (2μg/ml) did not significantly increase the binding of ¹²⁵I-factor VIII to platelets. The addition of thrombin (1u/ml) markedly increased ¹²⁵I-factor VIII binding (126ng/10⁸ platelets). This thrombin-induced binding was concentration dependent (0.1-2.5u/ml) and was enhanced when Ca⁺⁺ was present in the reaction mixture.
These results suggest that thrombin may play an important role in the early phase of hemostasis through its action upon the interaction between factor VIII and platelets.

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Difference of platelet aggregation between whole blood and platelet-rich plasma (PRP) was studied. Platelet aggregation was estimated by changes of optical transmission of supernatant fraction which was separated from the citrated blood sample (whole blood or PRP). Prior to the separation procedure, the sample was mixed with an aggregation inducer and 3 minutes later, the mixed sample was fixed with formaldehyde.
Resalts were as follows:
1. Platelet aggregation rate induced by epinephrine was higher in whole blood than in PRP. The aggregation rate induced by ADP or collagen was almost the same in both whole blood and PRP.
2. PGE₁ has less inhibitory effect on platelet aggregation induced by ADP or or collagen in PRP than that in whole blood. PGE₁ had inhibitory effect on the aggregation induced by epinephrine in PRP as much as that in whole blood.
Conclusion:
Platelet aggregation in PRP or whole blood was influenced a certain degree by the inducers respectively.

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Human platelet arachidonic acid (AA) metabolism and functions have been studied in fresh and stored (4°C and 22°C) concentrates. Thrombin-induced thiobarbituric acid (TBA) reactive substance produced by the platelet (Thr-TBARS) significantly decreased after 1 day storage at both temperatures. Platelet cyclo-oxygenase (PCO) and lipoxygenase (PLO) activities as estimated by the TBA reaction were well preserved for 1 day of the storage. However, PCO and PLO activities significantly decreased in the concentrates stored for 2 days at both temperatures and at 22°C, respectively. Thr-TBARS was better preserved at 22°C than at 4°C, but no such difference in the effect of storage temperatures was observed on PCO and PLO activities. Thrombin-induced platelet aggregation and ATP release as monitored by the use of lumi-aggregometer decreased during the storage at 4°C for up to 2 days. These data suggested that AA liberation from phospholipids was more vulnerable to storage than the metabolism of liberated AA and that 22°C was better than 4°C for the storage of concentrates. Furthermore, Thr-TBARS seems to be one of the most sensitive indicators in vitro to test function and viability of stored platelets.

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Platelet factor 3 (PF3) activity was determined using chromogenic substrate S-2238 (Kabi).
During aggregation induced by collagen (20μg/ml) PF3 activity up to 10% was appeared in the supernatant concomitant with LDH activity. No close relation was observed between PF3 activity and aggregation rate or the amount of released platelet specific proteins (Fig. 1). PF3 activity in the supernatant increased up to 32% during A23187 (5-10mM) induced aggregation. PF3 activity was always higher than LDH activity which derived from lysed platelets. PF3 activity was appeared in the supernatant prior to release of alpha-granule contents (Fig. 2). During primary or secondary aggregation induced by ADP no PF3 activity was observed in the supernatant (Fig. 3).
These results suggest that PF3 activity appeared in the supernatant during aggregation was derived by various mechanism, namely platelet lysis and/or mobilization from the membrane. The possibility that PF3 was released from alph-granules is less likely.

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1) Activity of serum sialyltransferase or N-acetyl-β-D-glucosaminidase (GlcNA-case) varies according to the time after venopuncture and platelet counts. Its activity reaches a maximum about 60 minutes after venopuncture and keeps a plateu for a while.
2) As to sera separated 1 minute after venopuncture, (1′-S), there is no significant change in sialyltransferase activities between the normal subjects and the patients with hepatic cirrhosis, but in the latter, GlcNAcase shows a higher activity than the former.
3) As to the difference of the enzyme activities between the sera separated 60 minutes and 1 minute after venopuncture, (60′-S)-(1′-S), both sialyltransferase and GlcNAcase shows significantly lower activity in the patients with hepatic cirrhosis than normal subjects.
4) There exists a close positive correlation between platelet counts and activity of sialyltransferase or GlcNAcase.
5) 1′-S looks predominantly to reflect the liver-derived enzyme activity, on the other hand, (60-S)-(1′-S) reflects platelet-derived enzyme activity.
6) Reduced serum sialyltransferase activities in patients with hepatic cirrhosis are mainly due to their thrombocytopenia.

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The inhibitory effect of gliclazide, a hypoglycemic sulfonylurea, on prostaglandin and thromboxane synthesis in guinea-pig platelets was studied in order to explore its mode of inhibition on platelet functions. 1) Gliclazide inhibited platelet adhesiveness to glass bead columns at 300mg/kg and 30mg/kg/day for 7 days, and increased the thrombus-inducing threshold voltage in mesenteric artery at the similar doses to that for platelet adhesiveness. 2) In in vitro experiments, gliclazide inhibited the thrombin-induced release of radioactive arachidonic acid from labeled platelets and collagen-induced thromboxane B₂ formation in platelet-rich plasma. Gliclazide also inhibited the platelet malondialdehyde (MDA) formation induced by collagen but not by arachidonic acid. Gliclazide did not block the conversion of [¹⁴C] arachidonic acid to radioactive thromboxane B₂. Gliclazide showed no influence on the platelet cyclic AMP levels. 3) In ex vivo experiments, the platelet MDA formation induced by collagen but not by arachidonic acid was significantly inhibited by a single administration at 300mg/kg, and this inhibition was enhanced by the repeated administration for 7 days. The inhibitory effect on the MDA formation by collagen was well correlated with that on platelet adhesiveness or the thrombus formation by electrical stimulation in meseteric artery. Thus, gliclazide seems to exert its inhibition in vivo on platelet functions through the inhibition of arachidonate release from platelet phospholipids.

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Activities of platelet superoxide dismutase (SOD), catalase and glutathione peroxidase were measured in the family members of the hereditary heterozygous glutathione reductase deficiency. The SOD activity of four deficient individuals fell in the range from 3.62 to 4.28unit/mg protein, significantly high compared to normal subject (2.52±0.26unit/mg protein). The cyanide insensitive SOD (mitochondrial SOD) of the members had not increased compared to that of normal subject, and the increased activity was found to be due to cyanide sensitive SOD (cytoplasmic SOD). On the other hand both catalase and glutathione peroxidase activities of the deficient members were almost within normal range (35.4±7.0μ mole H₂O₂/min/mg protein for catalase, 1.15±0.23×10^-2μ mole NADPH/min/mg protein for glutathione peroxidase). The platelet aggregation of the proband was also examined and found to be normal; aggregation induced with ADP (10^-5M), epinephrine (2.6×10^-7M), collagen (10μg/ml) or ristocetin (1.5mg/ml). The increased SOD activity might be due to a compensatory effect for the increasing level of active oxygen, that was brought about due to the defect of glutathione system. The compensation might also account for the normal platelet function.

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Blood viscosity at the shear-rate of 225.00sec^-1, erythrocyte deformability (DI), and other variables were determined in 3 healthy subject groups: the young age group including 13 subjects with average age of 21.8 years, the middle age group 15 subjects with 53.0 years, the old age group 19 subjects with 73.7 years.
In the old age group, whole blood viscosity, plasma viscosity and plasma fibrinogen concentration were significantly higher and DI, ATP and albumin level were significantly lower compared to those in the other 2 groups.
Whole blood viscosity was correlated directly to hematocrit, fibrinogen concentration and fibrinogen/albumin ratio, and plasma viscosity was also correlated directly to fibrinogen concentration in the old age group.
Although whole blood viscosity was correlated directly to DI in all subjects, significant correlation could not be detected in separate observation of each age group. To confirm the relationship with erythrocyte deformability, further investigation on whole blood viscosity at different shear-rates would be necessary.
In the old age group, depletion of ATP and increment of fibrinogen concentration must be causative factors of hypervi scosity state.

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Three cases in a family with platelet release mechanism abnormality who had intact arachidonate metabolism and defective response to ionophore A23187 were reported.
The proband was a 3 year old boy who showed prolonged bleeding time, small platelet volume and normal clot retraction. No second wave of aggregation with ADP, epinephrine or ristocetin and poor aggregation with collagen were observed. ATP release, investigated with a lumi-aggregometer (Chrono-log Co.), was not detected during the aggregation. Not only arachidonate (2mM at final concentration) but also A23187 (10, 20μg/ml at final concentration) failed to induce the aggregation or release reaction of his platelets. In normal subjects who ingested either aspirin or indomethacin, A23187-induced aggregation and release were normal. Production of metabolites of ¹⁴C-arachidonic acid assessed by thinlayer radiochromatogram was normal. Platelet lysate prepared from the patient caused the aggregation of normal platelets as well as lysate from normal subjects, and the amount of platelet adenine nucleotides was within the normal range. His sister and mother were revealed to have similar defects to those of the proband.
From these results, it was assumed that abnormality of release mechanism in these patients was not due to defective arachidonate metabolism (either phospholipase A₂, cyclo-oxygenase or thromboxane A₂ deficiency) but was attributable to defective mobilization of Ca⁺⁺ into the cytoplasm and/or abnormal interaction between Ca⁺⁺, thromboxane A₂ and cyclic AMP. These cases and three patients in another family with similar defects previously reported by us (Takahashi, et al., 1978) might be a new type of platelet release mechanism abnormality.

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It has been suggested that antiplatelet antibody may play a vital role in causing thrombocytopenia in cases with ITP. Recently Luiken, et al. (1) reported a quantitative measurement of platelet IgG, utilizing Fab, anti-Fab assay. With a slight modification of this method, platelet associated IgG (PAIgG), free platelet antibody in serum and plasma were studied in normal subjects and patients with ITP. Platelet antibody in plasma and serum was determined by assaying PAIgG of washed normal platelet (blood type O) incubated with plasma or serum from ITP patients, and index was calculated.
PAIgG level in normal subjects was 759±317ng/10⁹ platelets. The level in ITP patients was 6, 509±6, 726ng and a good correlation was observed between PAIgG and the degree of thrombocytopenia. PAIgG level in patients with thrombocytopenia due to hypersplenism was in the same range as in normal subjects. Index of free platelet antibody in normal subjects was 1.9±0.5 in plasma and 1.8±0.6 in serum. Index in ITP patients was 3.5±0.9 in plasma and 2.6±0.8 in serum. No correlation was observed between the index and the severity of thrombocytopenia. The assay of PAIgG level may be very useful in distinguishing ITP from other types of thrombocytopenia.

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Mechanism of maintaining the integrity of the vascular wall is not well understood. Thrombocytopenia is associated with a bleeding tendency which suggests that platelets play a major role in maintaining the integrity of the vascular wall and a supportive role in endothelial function.
Therefore the effect of certain platelet components on the cultured endothelial cell growth in the cases of thrombocytopenia was studied in this paper. Endothelial cells were obtained from the human umbilical veins by a perfusion technique using 0.2% collagenase as described by Maruyama or Jaffe, et al.
Cultured polygonal cells were identified as the endothelial cells by presence of Weibel-Palade bodies using transmission electron microscopy. These cells were also identified as the endothelial cells by immunofluorescent reaction of Factor VIII antigen.
In our experiments, 10% normal-WBS (whole blood serum) was found to enhace the cultured endothelial cell growth and DNA synthesis, while 10% ITP-WBS and 10% aplastic anemia-WBS had no discernible effect on the growth of cultured endothelial cells.
Our results indicated that the vascular repair mechanism was impaired in the patients with thrombocytopenia, resulted in decrease of platelet derived growth factor which was stimulatory of endothelial cell growth to repair the injured vascular surface.

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The platelet function was investigated in 15 patients with nephrotic syndrome.
Platelet counts and bleeding time [duke method and template Ivy method] were normal, but platelet retention was elevated in 58.3% of patients. Platelet factor 3 activity was normal. β-TG, however, was strikingly high and its average value was 142.6ng/ml. Spontaneous platelet aggregation was observed in 46.7% of patients, and platelet aggregation was enhanced in response to low concentration of ADP and collagen. Enhanced platelet aggregation was significantly correlated with serum total protein level and serum albumin level negatively.

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In order to study the role of renal glomerular tissue thromboplastin (TTP) on intraglomerular coagulation, relation between TTP activity of isolated glomeruli and intraglomerular coagulation was examined in rats. As the experimental model of intraglomerular coagulation, liquoid-induced DIC and Masugi Nephritis of Wistar rat were used. TTP activity in isolated glomeruli was assayed by modified Astrup's method. Histological observation was also done in parallel.
Renal glomeruli had high activity in TTP as brain and lung and TTP activity of renal cortex was middle whereas that of medulla was the lowest. These findings may possibly explain the hypercoagulability in glomeruli.
In DIC, TTP activity in isolated glomeruli and the released TTP activity from isolated glomeruli increased after liquoid injection. The development of histological intraglomerular coagulation was paralleled to the increase of released TTP activity.
In Masugi Nephritis, TTP activity in isolated glomeruli decreased in 1st and 2nd phases after nephrotoxin injection. On the other hand, released TTP activity from isolated glomeruli increased in both phases in parallel to the development of histological intraglomerular coagulation after nephrotoxin injection.

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Qualitative changes of platelets in bacterial endocarditis (BE) could result from endocardial damage due to vegetation or from complication of disseminated intravascular coagulation (DIC) induced by sepsis. But there were no cases with DIC in our study.
To evaluate platelet function, platelets aggregation test before and after preincubation of low concentration of collagen and measurement of β-thromboglobulin (β-TG) were carried out.
Platelets aggregation by ADP was shown as the minimum concentration of ADP which could aggregate platelets before and after the pre-incubation at 37°C for 2 minutes, and as the results, high ability of platelets aggregation was detected especially after the pre-incubation of collagen in BE patients.
On the other hand, patients with BE showed high levels of β-TG regardless of activity of inflammation and BE with prosthetic cardiac valves showed higher levels than BE without it. (The mean values of β-TG in cases of healthy controls, active BE, inactive BE and BE with prosthetic valves were 19, 57, 58 and 164ng/ml)
It was concluded that patients with BE had a thrombotic tendency of platelets in vessels from the above findings.

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Platelet aggregation and ATP release with ADP, epinephrine and collagen were investigated in patients with ischemic heart disease.
The blood was obtained from the subclavian vain, pulmonary artery, aorta and brachial artery. Platelet counts in the heparin used blood of aorta were lower than the others but the number of platelets were similar in the venous blood, the blood of pulmonary artery and arterial blood.
Platelet aggregation and ATP release with ADP and collagen were increased in ischemic group with more than 75% narrowing of coronary artery.
Especially, the ATP release is regarded as the sensitive parameter to evaluate the hyperfunction of platelet in ischemic heart disease.

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Intraplatelet serotonin (5-hydroxytryptamine: 5HT) and 5HT in the plasma was assayed spectrophotometrically in 69 healthy volunteers and in 67 patients with cerebrovascular diseases (CVD), including acute cerebral thrombosis within 72 hours of onset (13 cases), chronic cerebral thrombosis during 2 months to 4 years from onset (27), TIA (11), RIND (2), SAH (5), intracerebral hematoma (ICH) (7) and recurrent cerebral thrombosis (2).
In health, the intraplatelet 5HT level decreased significantly with increasing age, and plasma 5HT level showed slight increase with age (Fig. 1).
In acute cerebral thrombosis, intraplatelet 5HT level was decreased significantly and that of plasma was high compaired to the healthy control at any age group. Chronic cerebral thrombosis, however, showed neither significant decrease in the intraplatelet level nor increase in the plasma. The reversible stroke, such as TIA and RIND, had lower intraplatelet 5HT than that of control, but the plasma level was within normal range. In SAH and ICH as hemorrhagic CVD, either the intraplatelet or plasma 5HT was within normal range. Recurrent cerebral thrombosis stroke under treatment revealed very high level of intraplatelet 5HT (Fig. 2).
Under the above sesults, we may conclude that, in acute thrombus forming state, intraplatelet 5HT level was decreased.

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Plama levels of 6-keto prostaglandin F_1α (6-keto PGF_1α) and thromboxane B₂ (TXB₂) derivatives of prostacyclin (PGI₂) and thromboxane A₂, respectively, were measured in female stroke-prone spontaneously hypertensive rat (SHRSP), stroke-resistant SHR (SHRSR) and Wister-Kyoto rats (WKY) of various ages. Gas chromatograph-mass spectrometer was used for the analysis.
The 6-keto PGF_1α level was decreased in female SHRSP before the occurence of stroke, however, the levels reverted to prestroke values following the stroke. The TXB₂ levels were found to be increased only in the female SHRSP with stroke. Thus, the possibility that PGI₂ may play an important pathophysiological role in the SHRSP strain in which the occurrence of stroke is spontaneous warrants further investigation.

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For the assay of gastric glandular kallikrein of the rat, a synthetic substrate was used, the structural formula of which is as follows; L-Pro-L-Phe-L-Arg-α-NE. The hydrolytic activity of sample solution was used as an esterolytic activity. The influence of the blood and peptic fluid mixed into the sample solution could be negated. When low molecular weight kininogen was added to the sample solution from the glandular stomach which exhibited high esterolytic activity, a significant kallidin value was manifested. Such high correlation coefficient as r=0.953 was detected between an esterolytic activity of the sample solution and kallidin value. From this result, the assay of glandular kallikrein came to be possible. The glandular kallikrein activity was demonstrated. During the process of development of the experimental gastric ulcer due water immersion stress, the glandular kallikrein showed an intimate relationship with tissue plasminogen activator. In the group of rats administered orally with 300mg/kg of cetraxate, the glandular kallikrein and tissue plasminogen activator did not show any obvious alteration irrespective of the progress in stage during the development of the experimental gastric ulcer due to water immersion stress.

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It was studied whether the abnormal fibrinogen could be existed in umbilical cord blood as compared with hepatoma associated dysfibrinogen.
The plasma and purified fibrinogen of patients (12 cases of cord blood and 16 cases of hepatoma) were studied as follows,
[1] with plasma
Fibrin monomer polymerization were estimated with Soria's method.
[2] with purified fibrinogen
(1) thrombin time
(2) reptilase time
(3) fibrin monomer polymerization
(4) SDS-PAGE
(5) immunoelectrophoresis
(6) content of sialic acid
Using with plasma, fibrin monomer polymerization of both cord blood group and hepatoma group were markedly decreased. Using with purified fibrinogen, thrombin time and reptilase time were prolonged in hepatoma group, but not in cord blood group.
The content of sialic acid of fibrinogen in hepatoma was increased, but not marked in cord blood.
On mobility of electrophoresis, the difference was no noticed between two groups.

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It is well known that thromboxane A₂ (TxA₂) and prostacyclin (PGI₂) play an important role in development of thrombosis and other cardiovascular disease. In this study, we examined the effect of aspirin and dipyridamole on the generation of TxA₂ and PGI₂ as well as platelet aggregation in control and experimental hyperlipidemic rabbits.
Experimental hyperlipidemic rabbits were fed 1% cholesterol supplemented diet for 25 weeks. PGI₂ was measured by the inhibitory effect of human platelet aggregation. Arachidonic acid metabolites in arteries and platelets were determined by thin-layer radiochromatography.
Dipyridamole inhibited the platelet aggregation induced with collagen and accelerated the release of PGI₂, but there was no difference between control and hyperlipidemic group.
In the control group, aspirin inhibited the platelet aggregation and the generation of TxA₂ in the concentration of 0.1mg/ml, while PGI₂ generation was less inhibited at the same dose. On the other hand, in the hyperlipidemic group, aspirin did not inhibit the platelet aggregation and the generation of TxA₂ in the concentration of 0.1mg/ml in full measure, but the release of PGI₂ was markedly inhibited at the same dose.
These data indicate that, in the hyperlipidemic state, aspirin inhibit the generation of PGI₂ from arteries at the effective dose showing the inhibition of TxA₂ in platelets.

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Fibrinolytic activity and coagulation state of uterine venous blood (UVB) were investigated at the time of histerectomy due to uterine fibroid. Changes of coagulation factors (fibrinogen, Factor V, VII, VIII and XIII) and protease inhibitors such as α₁-Antitrypsin (α₁-AT), α₂-Macroglobulin (α₂-M), α₂-Plasmin inhibitor (α₂-PI) and Antithrombin-III (AT-III) in UVB were also assayed after strong inhibition of fibrinolytic activity by means of t-AMCHA administered into the uterine artery. Fibrinolytic rates of thrombelastgram and F. D. P. revealed higher fibrinolytic activities in UVB than in cubital venous blood (CVB). Both α₂-PI and AT-III were markedly low in UVB after the operation, indicating hypercoagulable state associated with increased fibrinolysis in the uterus.
The level of α₂-PI in UVB was not low when t-AMCHA was given by intraarterial infusion. This results will confirm the acceleration of fibrinolysis in the uterus during the operation.
The level of factor VIII and factor XIII were considerably high in UVB, compared with almost normal levels in CVB. Although these results might indicate partly acute phase reaction during the operation, it will be difficult to explain the general situation, where the level of factor XIII decrease to the normal in the case of administration of t-AMCHA.
For understanding the mechanism of factor VIII and factor XIII in this situation, further studies will be necessary.

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The serum trypsin and plasma β-thromboglobulin concentrations were investigated in patients with pancreatitis. Both of them were measured by radioimmunoassay.
The results were as follows:
1) Serum trypsin showed a significant increase in acute stage in pancreatitis.
2) There were significant correlations between serum trypsin and amylasecreatinine-clearance, and between serum trypsin and β₂-microglobulin-creatinine-clearance.
3) In case of acute pancreatitis serum trypsin showed the change closely related to α₁-antitrypsin.
4) β-thromboglobulin showed a significant increase in acute stage of pancreatitis, and showed normalization in inactive stage.
5) The tissue β-thromboglobulin investigated in cases of pancreatic cancer with pancreatitis revealed a significant increase in pancreae, lung and kidney.
By the present study it was suggested that serum trypsin increases in acute pancreatitis and that it may bring about renal derangement. Furthermore high β-thromboglobulin concentration both in plasma and in tissues clearly revealed thrombogenetic tendency in pancreatitis.

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Endotoxin from E. coli was infused into the distally ligated common bile duct or ureter of rabbits under the static pressure of 25cm H₂O or 50cm H₂O. Fibrinogen, soluble fibrin monomer complexes (SFMC), antithrombin III (AT III) and platelet counts were estimated before, 2, 4, and 6 hours after endotoxin infusion.
Fibrinogen level, AT III and platelet counts decreased, but on the other hand, SFMC increased after endotoxin infusion into common bile duct or ureter of rabbits.
The change of hematological parameters showed a pattern characteristic of disseminated intravascular coagulation (DIC). Histologically, microclots in the glomeruli of the kidneys could be demonstrated in all entotoxin infused animals. The findings suggest that DIC can be produced by endotoxin infusion into the common bile duct or ureter of rabbits, marking these portions as focal origin.

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1. It was suggested that the cytotoxic damage of bone marrow cells due to endotoxin is responsible for the induction of procoagulant activity of the cells, since a) bone marrow cells of mouse given endotoxin (Post-LPS-cells) had a significant procoagulant activity and time course of procoagulant induction in the cells was the same as those of cytotoxicity and decrease of nucleated cell numbers in the marrow, b) there existed a definite correlation between cytotoxicity and procoagulant activity, and c) the strong cytotoxic changes independent of endotoxin, i. e. after death did not induce a significant procoagulant activity.
2. The procoagulant activity of Post-LPS-cells was not considered to be endotoxin contaminated in the cells but to be tissue thromboplastin, since a) a large amount of endotoxin was required for direct activation of plasma clotting, and dose-response-regression line for endotoxin was not parallel to that for Post-LPS-cells, b) parallelism between the two regression lines for Post-LPS-cells and thromboplastin was recognized, and c) Post-LPS-cells did not shorten the clotting time of plasma deficient in factor VII but did in factor XII, as thromboplastin did.
3. While Post-LPS-cells corrected the prolonged clotting time of factor VIII deficient plasma, it was shown, utilizing a parallel line assay, that no factor VIII clotting activity was present in the cells.
It should be mentioned that the bone marrow cells are one of the sources of procoagulant induction, and Post-LPS-cells either in marrow or circulation would cause DIC in endotoxicosis.

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The effects of various drugs on experimental DIC induced by endotoxin infusion in rabbits were studied. The fibrin deposition rate and mortality were 50% and 33% in control (endotoxin alone) group. Heparin and hydrocortisone phosphate reduced both fibrin deposition (25%, 11%) and mortality (14%, 8%). Increase of fibrin deposition was observed by aprotinin (93%) and noradrenaline (91%), however, mortality was not changed (36%, 30%) as compared with control group. Fibrin deposition did not change by conjugated estrogen, however, mortality was reduced significantly (0%).
Theoretically, it seems that increase in fibrin deposition may produce high mortality. However, there was no pararellism between them as in aprotinin, noradenaline and conjugated estrogen groups. These drugs were capable of inhibiting activity on capillary permeability.

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Activities and antigenicities of factors VIII, II and XIII, plasminogen, antithrombin III, α₂-macroglobulin and α₂-plasmin inhibitor were compared in 34 healthy subjects and in 41 cases of DIC.
Procoagulant activity of factor VIII slightly decreased in patients with DIC, whereas factor VIII related antigen increased in these patients. Activity of factor II was directly proportional to antigenicity of this procoagulant, although ratio of the latter to the former was significantly higher in cases of DIC, in whom prothrombin content decreased, than that in healthy subjects. In regard to factor XIII, plasminogen and antithrombin III, there were no discrepancies between antigenicities and activities. There was no significant correlation between α₂-macroglobulin levels assayed immunologically and antithrombin activity of this protein in cases of DIC. Concentrations of α₂-plasmin inhibitor, which decreased markedly in cases of DIC, were not correlated with antiplasmin activity measured using chromogenic substrate (reaction time of plasma and plasmin was 20 seconds at 37°C) in these subjects. However, antigenicities of these inhibitors in plasma were significantly correlated with antithrombin or antiplasmin capacities in healthy subjects, suggesting that some conformation changes of α₂-macroglobulin occured in DIC and that role of α₂-macroglobulin cannot be negligible in determination of antiplasmin, when levels of α₂-plasmin inhibitor fall markedly.

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Pancreatic involvement in DIC is not fully documented. So a retrospective study was done to clarify it. Four cases (13%) out of 30 autopsy cases, clinically diagnosed as DIC in our laboratory for previous 9 years, showed the pathological evidence of pancreatic involvement.
Serum amylase and trypsin levels of which increase is known to indicate the development of pancreatic lesions were measured in 22 cases of DIC for recent 3 years using neo-amylase test (Daiichi) and trypsin radio-immunoassay ki t (Cis-Sorin) respectively. Mean values of amylase and trypsin were 321.1±377.6 Somogi u/dl and 73.1±53.1ng/ml respectively and both increased as compared to normal controls. Furthermore correlation between trypsin levels and biochemical data or hemostatic data were studied. Only BUN and creatinine showed positive correlation with trypsin levels.
A 29 year-old female at 23 weeks' gestation was presented. DIC had occurred immediately after artificial abortion with the symptomes of genital bleeding, G-I bleeding and oliguria. Coagulation study revealed high level of FDP, positive SFMC and decreased platelet counts and fibrinogen level. Despite of intensive cares, heparin treatment and peritoneal dialysis no sign of improvement was obtained and the patient died on the 32th postoperative day. Postmortem examination showed stomach cancer with multiple metastasis of mild degree and pancreatic infarction, besides multiple fibrin thrombi in lungs and kidneys. Retrospective analysis revealed marked increase of serum trypsin level which would show the occurrence of pancreatic involvement since the onset of DIC and might be concerned with the deteriolation of DIC.
High levels of serum trypsin in DIC are related to renal function, but sometimes its marked increase showed the presence of pancreatic involvement which may lead to one of the aggravatig factors of DIC.

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Burn injuries cause dynamic alterations of the coagulative and fibrinolytic activities of the blood. Up to the present, there has been little research on the anti-coagulative therapies for burned patients. Herein we discuss the clinical significance of heparin therapy as viewed from the results of animal experiments and clinical experiences.
For our animal experiments we induced deep burns covering 40% of the total body surface of rabbits by soaking their backs in 80°C water for 20 seconds, and we followed the hematological alterations. Examinations of the rabbits without therapy revealed that the platelet counts, fibrinogen, and the plasminogen decreased gradually after injury, and the FDP increased soon after injury. Examinations of the rabbits administred only the infusion therapy revealed that almost the same alterations took place as in the rabbits without therapy. On the other hand, in the rabbits administered 1, 000U/kg/6hr. heparin in addition to the infusion therapy, there were significant alterations: no decrease in the platlet count at all; a slight decrease of fibrinogen; and no increase in FDP.
Our clinical administration of heparin in doses of 10, 000-20, 000U/day prevented DIG and was proved to be safe. In the severely burned patients, however, the therapeutic effect of heparin was underestimated because of the decrease in antithrombin III concentration; therefore, we recommended that concentrated antithrombin III and heparin be administered syncronously in severly burned patients.

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The radioactivity in blood, feces, urine and main organs such as liver and kidney of mice and rats was measured after intraduodenal administration of ¹²⁵I-urokinase (¹²⁵I-UK) with well type γ-counter (ALOKA).
In mice, the highest radioactivity in blood, liver and kidney was reached at 15min. after intraduodenal administration of ¹²⁵I-UK and the radioactivity continued for 6hr. On the other hand, the highest radioactivity in rats was observed at 30min in blood., at 15min in liver, and at 1hr in kidney. Fifty percent of the highest radioactivity continued until 6hr. after intraduodenal administration of ¹²⁵I-UK. The decrease of radioactivity in rats was slower than in mice. In comparison with the cases of intravenous administration, intraduodenally administration showed slower decrease of radioactivity from blood and organs of mice and rats.
This finding supported the idea that orally administered urokinase should develop slower effect and maintains longer period in the animals although their rates were variable dependency on animal.

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The metabolic fate of ¹⁴C-urokinase (¹⁴C-UK) after intraduodenal administration was studied in mice and rats. In mice, the maximum radioactivity of serum was observed at 15min, and liver and kidney at 30min. Each value indicated 2.7% (serum), 2.3% (liver) and 6.1% (kidney) of administered activity. In rats, the maximum values of serum, liver and kidney were respectively 1.8%, 1.3% and 1.6% of administered activity at 30min after intraduodenal administration.
While the radioactivity came to their highest level at either 15min or 30min, it gradually fell down to approximately 50% of the maximum activity after 2hr or 3hr. This level was continued for 24hr.
The retention of intraduodenally administered radioactivity of ¹⁴C-UK was obviously longer than that of the intravenously administered activity.

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