In order to investigate the dynamic state of blood fibrinolysis system at oral administration of urokinase (UK), double blind test was performed by a crossover method. Twelve healthy adult male volunteers were divided into two groups of true drug and placebo. Before and after single oral administration of 120, 000 IU UK and placebo, their blood fibrinolytic activity and its inhibitors were measured until 24 hours thereafter and the statistical significance of the variation of these parameters and the absorption of UK through intestinal wall were also analysed. It was revealed that the blood fibrinolytic activity reached the maximal value between 3 to 6 hours and the highest absorption of UK was attained at 1 to 3 hours after the administration.
The blood fibrinolytic activity was then gradually decreased but its effect was continued at least until 9 hours after the administration. The level of fibrinolysis inhibitors (α₂-AP, α₂-M, α₁-antitrypsin and AT-III) did not show significant change during this period.
From these data it was concluded that oral administration of UK could increase blood fibrinolytic activiity much greater and more continuously than in the case of venous injection with the same dosage. The effective thrombolytic therapy could be expected by oral administration of UK.

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Factor IX activity (IX: C) and factor IX antigen (IX: AGN) (Laurell method) were measured in 140 full-term newborn infants with no hemorrhagic sympotoms during the first week of life, and crossed immunoelectrophoersis was performed for qualitative analysis of factor IX. IX: C and IX: AGN in the newborn plasma were lowest on the first day of life, i. e., 24.4±8.2 and 23.4±6.3% respectively, and rose gradually during the first week, reaching 52.6±5.8 and 53.9±7.4% respectively. On the crossed immunoelectrophoresis of IX: AGN in the presence of 1mM calcium lactate, a biphasic arc was observed in 16 newborn plasmas, i. e., in 20, 35, 25 and 10% respectively, in the first 13-24 hours, the second day, the third day and the fourth day. IX: C of these plasmas was apparently low in contrast to IX: AGN. One arc lay in normal factor IX position and the other arc shifted faster to the anode, suggesting the presence of abnormal factor IX. This anodal arc and the anodal arc of plasma obtained from dicumarol treated patient or chronic hepatitis were the same mobility on the crossed immunoelectrophoresis. From this study, the abnormal factor IX which occurs in the newborn period was regarded as PIVKA IX (protein induced by Vitamin K absence or antagonist).

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In our series of study on DIC, we found coincidentally that plasma prekallikrein determined chromogenic tripeptide substrate (S-2302) changed with close relation to labor and sexual cycle. In order to confirm the possible role of prekallikrein in the reproduction physiology, the animal experiment was performed. The results obtained were as follows:
(1) Plasma prekallikrein was increased during pregnancy with advance of gestational ages, and reached maximum (180.9±32.6%) at the end of pregnancy. It decreased abruptly with start of labor, and free kallikrein was demonstrated in the circulating blood during labor. In pregnant rats (Wister descent rats), onset of labor was significantly prolonged (520.7±1.3 hours 564.0±0 hours) by FOY (a synthetic broad specturm serine protease inhibitor, 2mg daily intraperitoneal administration, n=4) as compared with saline control group. It can be therefore concluded that prekallikrein may relate to the labor mechanism.
(2) In women (n=5) with normal ovulation cycle, a significant transient fall of plasma prekallikrein was observed 72 hours before ovulation. The same event was also observed 21 hours before ovulation in the experimentally induced ovulation by PMS-HCG in rats. The experimental ovulation was suppressed by aprotinin.

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Simultaneous determination of factor VII activities in Autoplex and FEIBA was performed by amidolytic (VIIam) and one-stage clotting (VIIc) methods, to evaluate the presence and possible role of activated factor VII in these activated PCC preparations. As a tissue thromboplastin, monkey brain (monkey T. P.) and rabbit lung tissue thromboplastins (rabbit T. P.) were employed in both assay systems.
In healthy subjects, the plasma level of VIIam was quite similar to and correlated well with that of VIIc, and was not significantly influenced by the kinds of tissue factor.
The level of VIIc was significantly higher than that of VIIam in both preparations, showing that the ratio of VIIc to VIIam (VIIc/VIIam, factor VII activity ratio) was 13.3 in Autoplex and 4.8 in FEIBA. The difference of the values was more remarkable when monkey T. P. was used as the tissue thromboplastin reagent, showing 92.9 in Autoplex and 23.8 in FEIBA. These results suggest that some activated factor VII is contained in these two preparations, and may be playing an important role in the treatment of inhibitor patients with classic hemophilia.

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Cold-insoluble globulin (CIg) was prepared from rat glasma by means of heat-defibrinogenstion at 56°C for 4min, salting-out at 25% saturation of ammonium sulfate, ion-exchange chromatography on DEAE-cellulose and affinity chromatography using fibrinmonomer-Sepharose. SDS polyacrylamide gel electrophoresis run on the purified rat CIg chowed a major 4.4×10⁵ dalton polypeptide and a minor closely migrating doublet with a molecular weight half as large as the major one under unreducing conditions. When the sample was reduced, a doublet with an approximate molecular weight of 2.2×10⁵ was noted. Other characteristics and properties described on human and bovine CIg's were found in the purified rat CIg as well, including electrophoretic mobility and immunological cross-reactivity with CIg of other species. The overall recovery and purification in a typical run were 33% and 68-fold, respectively.

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In vivo turnover of canine plasminogen-Fraction I (plg) and the plasminmodified-plasminogen (m-plg) labeled with radioiodine and in vitro binding of either plg or m-plg to fibrin were studied. The plg prepared by the method of Deutsch and the m-plg prepared from the purified plg by the method of Sodetz and Castellino were labeled with ¹²⁵I and ¹³¹I, respectively, by the iodine monochloride method of McFarlane.
1) The plasma clearance of radioiodine in 6 dogs injected with ¹²⁵I m-plg was described by a biexponential curve within a observation period of 72 hours. The half-life of the slower component was 0.96±0.06 days and the catabolic rate of m-plg calculated from the analysis of Atencio's two compartment motabolic model was 1.216/day, respectively, whereas those for ¹³¹I-plg were 1.636±0.150 days and 0.602/day, respectively.
2) About 2.5μ Ci of both ¹²⁵I-m-plg and ¹³¹I-plg solution were added to one ml of canine plasma or canine fibrinogen solution, then the mixture was clotted by the addition of 0.1ml of CaCl₂ solution (0.25M) or of thrombin solution (100u/ml). The mixture containing fibrin clot were incubated for 10min., 30min., 1 hour or 2 hours. After completion of the incubation, the clot were squeezed by a glass stick and the unadsorbed radioactivity was washed out by 0.05M phosphate buffer (pH 8.0). Then the radioactivities of ¹²⁵I and ¹³¹I in fibrin were measured. When the plasma was clotted by thrombin, the binding of both ¹²⁵I-m-plg and ¹³¹I-plg to fibrin was about 20.1% and 12.4%, respectively. Whereas the plasma was clotted by CaCl₂, the binding of these to fibrin were 25.5% and 16.5%, respectively. (p<0.05). In case of the canine fibrinogen solution, the binding of the m-plg and the plg to fibrin after addition of thrombin was increased to 27% and 19.3%, respectively.
3) One day after I. V. injection with ¹³¹I-plg to each of 3 dogs, 20×10³ units of urokinase (UK) was injected intravenously. The plasma samples were obtained and were clotted by an addition of CaCl₂, then the radioactivity of fibrin was measured. It was increased to 2.7 times as compared to initial value.
From these data, it may be concluded that the plasma clearance of the m-plg was faster and the m-plg has higher affinity for fibrin than the plg. The I. V. administration of UK resulted in the activation of a part of plg into plasmin, then the plasmin modified ¹³¹I-plg to ¹³¹I-m-plg which can be easily bound to fibrin.

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We've reported the reverse change of fibrinolysis in uterus in the course of pregnancy. The activation of fibrinolysis is dominant due to endometrial plg. act. in non-pregnant uterus, and on the other hand, its inhibition is dominant in pregnant uterus due to urokinase inhibitor (UKI) which exists in decidua or placenta.
In this paper, purification and identification of plg. act. from placenta were performed, which has not been reported so for.
Plg. act. is purified from placenta by extraction, acid treatment, fraction with ammonium sulfate, and chromatographies on CM-Sephadex, Sephadex G-150 gel filtration, DEAE-Sephacel and UK-UKI Sepharose.
Its activity was measured using fibrin plate and chromogenic substrate S-2444. The molecular weight was estimated about 40, 000 by SDS disc electrophoresis. This plg. act. did not react to anti-UK antiserum, therefore, this suggests placental plg. act. is different from UK.
Although plg. act. really exists in placenta, but is combined with its inhibitor, therefore its activity is inhibited in placenta. But this complex is reversible, so can be dissociated easily.
We suggest plg. act. is related to menstruation, abortion and the prevention of toxemia, and on the contrary, its inhibitor is related to the prevention of abortion and hemorrhage during pregnancy, and the occurrence of toxemia.

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To investigate clinical application of Lys-plasminogen, which is prepared from human placenta, phase I study was carried out. From its biochemical properties distinct from native Glu-plasminogen, it is expected that the agent enhances thrombolysis. The study included three single dose tests (15, 30 and 60mg), three multiple dose tests (15, 30 or 60mg, every 24hrs for 4 days), and single administrations in combination with 30, 000 IU of urokinase and multiple combined administraions. In these tests any significant adverse effect was not noted, particularly, bleeding did not occur. As to the effect on fibrinolytic system shortening of ELT which lasted for many hours was consistently observed and it was more remarkable when urokinase was combined. However, there were no substantial changes in plasma levels of inhibitors, such as α₂-AP, α₂-M and α₁-AT. Subsequently, therapeutic trial of this agent was performed on 20 patients with cerebral thrombosis under a regimen of 60mg of Lysplasminoge and 30, 000 IU of urokinase, every 24hrs for 4 days. The results appeared to be promising and further studies were indicated. One patient developed gastrointestinal bleeding. To avoid such a side effect careful assessment of indication is required.

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Contractile proteins of platelet play an important role in the functions of platelet aggregation or platelet adhesion, and the concentration of calcium has an effect on these functions. Therefore it is considered that there is the factor which regulates the activity of contraction by contractile protein.
In this report, we purified the protein from platelets which inhibits the gelation of actin in the presence of calcium.
Homogenate of human platelets was firstly chromatographied by DEAE Sepharose column with linear gradient of KCl (0.1-0.4M), and secondary by actin binding affinity chromatography column applyed in the presence of 1mM CaCl₂ and then eluted with 10mM EGTA containing buffer. This purified protein had remarkable actin gelation inhibitory activity. And this activity appered in the range of 10^-7-10^-6M of free calcium, which is the concentration of intracellular calcium.
This protein exhibited three bands, molecular weight of 8.5×10⁴, 4.5×10⁴, and 4.3×10⁴, by SDS polyacrylamide gel electrophoresis.
The band of 4.3×10⁴ was thought to be of actin of platelets and the remaining to be of actin gelation inhibitory factor.

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It has been proposed that the contractile activity of platelet actomyosin is determined by the level of phosphorylation of myosin light chain. In this report, we show that; (i) the Mg²⁺-ATPase activity of natural actomyosin from platelet is regulated by Ca²⁺ through “native tropomyosin” (which contains light chain kinase activity) from arterial smooth muscle: (ii) the ATPase activity of synthetic skeletal acto-platelet myosin is enhanced by purified arterial tropomyosin (which has no light chain kinase activity). The results suggest that the contractile activity of platelet actomyosin is modulated not only by the level of light chain phosphorylation, but also by the state of actin-tropomyosin interaction.

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Using immunoperoxidase staining, we have localized platelet factor 4 and fibronectin in human platelets and megakaryocytes at the electron microscope level. Platelet factor 4 was purified by affinity chromatography using heparin agarose. Plasma fibronectin was purified by gelatin sepharose affinity chromatography. Antisera against purified materials were raised in rabbits. Fab' fragments of antibody were conjugated to horseradish peroxidase. Platelets from venous blood and buffy coats of bone marrow aspirates were fixed with periodate-lysine-paraformaldehyde. After incubation with anti-fibronectin or anti-platelet factor 4 Fab'-HRPO conjugates, they were then incubated with diaminobenzidine medium containing H₂O₂. The samples were post-flxed with osmium tetroxide, dehydrated and embedded in epon. Ultrathin sections were examined in the Philips EM 201 electron microscope. The positive reaction products of these antigens are localized mainly in α-granules of platelet and megakaryocyte. Fibronectin is also localized in some parts of open canalicular system of platelet and lining surfaces of demarcation membranes of megakaryocyte. These results provide additional evidence that platelet factor 4 and fibronectin are localized in α-granules of platelet and megakaryocyte and suggest that the synthesis occurs in megakaryocytes where these are packaged into granules before release of platelets into the circulating blood.

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The changes in cytoskeletal structures of thrombasthenic platelets were investigated following thrombin activation. Non-stimulated platelets from normal subjects and thrombasthenic patients yielded essentially the same amount of Triton-precipitated cytoskeletal proteins. When platelets were treated with thrombin at various concentrations, the amount of protein in Triton-precipitates was increased both in normal and thrombasthenic subjects; less degree in the latter. The Triton-insoluble materials from non-stimulated, normal and thrombasthenic platelets showed essentially similar polypeptide composition on SDS-polyacrylamide slab gel electrophoresis; three prominent bands of molecular weight 43, 000, 200, 000 and 250, 000 respectively. A distinct difference was, however, observed in polypeptide composition of cytoskeletons between thrombinstimulated normal and thrombasthenic platelets. While polypeptides of molecular weight 43, 000, 52, 000, 56, 000, 200, 000 and 250, 000 were increased after control platelets were stimulated with thrombin, cytoskeletal proteins of thrombinactivated thrombasthenic platelets showed no increase in the amount of polypeptides of molecular weight of 52, 000 and 56, 000. These findings may suggest the altered cytoskeletal structures of thrombasthenic platelets.

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Electronmicroscopic examinations were performed on platelets in three cases of congenital platelet function disorder, which included Bernard-Soulier Syndrome, Hermansky-Pudlak Syndrome, and non-albino type of Storage-pool Disease.
Ultrastructual abnormalities of giant platelets in Bernard-Soulier Syndrome, such as disorganization of microtubules, Swisscheese appearance, large α-granules etc.., reported by other investigators were also noted in our case. In addition, electronmicroscopic examinations revealed some platelets were clearly demarcated into two or three parts by circumferential bundles of microtubules. These findings strongly suggest that giant platelet of Bernard-Soulier Synrome are formed by membrane fusion of two to three platelets while they are circulating in the peripheral blood stream.
Relatively large-sized platelets containing larger number of α-granules and mitochondrion were recognized in Hermansky-Pudlak Syndrome, Storage-pool Disease associated with albinism. The number of dense bodies are decreased predominantly and the decrease in content of storage-pool ATP, ADP and 5HT was also observed.
Non-albino type of Storage-pool Disease, however, have normal sized platelets with normal number of α-granules and mitochondrion. The decrease in the number of typical dense bodies as well as storage-pool ATP, ADP and 5 HT was slightly less compared with that of Hermansky-Pudlak Syndrome, but many atypical uran-affin electron dense organelles considered to be atypical dense bodies were obserbed.

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The characteristics of platelet organelles in various diseases were morphometrically studied with a manual optical picture analyzing system (MOP/AM 03 from Kontron). We have measured the platelet profile area and the number and area of each platelet organelle [α-granule (α-G), dense granule (DG), morphologically atypical α-G and DG (α′-G, D′G), mitochondria, unidentified granule, open canalicular system (OCS), membrane complex, myelin figure]. Parameters of each organelles were expressed as “F” (total number/platelet area 100cm²), “A” (total area/total number) and “O” (total area/platelet area 100cm²).
“F”, “A” and “O” of mitochondria in almost all patients with hematopoietic disorders (7 cases) and with congenital qualitative platelet diseases (4 cases) were within normal range. “A” and “O” of OCS increased in many patients and myelin figures were often observed in OCS of platelet from 2 patients of them, with thrombasthenic syndrome. “A” of membrane complex increased in hematopoietic disorders, especially increased in an AMMoL patient. The platelet from an AMMoL showed low “F” and “O” of α-G. The sum total “F” of DG and D′G was reduced in 2 patients with storage pool deficiency, while increased in the others except for an AMMoL.

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The effects of treadmill exercise stress (up to 85% of the predicted maximum heart rate) on platelet functions and coagulating activities were studied in healthy males. Blood sampling for the measurements were performed from the antecubital vein at rest, 3, 6, 9 and 12 minutes during exercise, and 6 and 30 minutes after exercise.
No significant changes were noticed in platelet sensitivities and maximum and 3 minute percent platelet aggregation to 4 concentrations of ADP between at rest, during exercise and after exercise. Platelet counts increased during and after exercise in platelet rich plasma without significant changes of platelet counts in whole blood. Plasma thromboxane B₂ levels showed a tendency to increase, while plasma 6-keto-prostaglandin F_1α levels to decrease. Plasma epinephrine levels showed a tendency to increase and norepinephrine levels increased during and after exercise. Among coagulating factors, factor VIII activities and fibrinogen levels increased without any significant change in activities of factors IX, XI and XII. Antithrombin III activities also increased during and after exercise. In spite of significant changes in several coagulating factors, prothrombin time and partial thromboplastin time were not influenced by exercise.
In conclusion, moderate exercise produced a significant elevation of factor VIII, fibrinogen, antithrombin III and catecholamine without affecting the hemostatic balance.

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To determine the role of platelets in the pathogenesis of angina pectoris, plasma levels of β-thromboglobulin (β-TG), thromboxane B₂ (TXB₂) and 6-keto-PGF_1α were determined in the coronary sinus (CS) and left ventricle (LV) in 6 patients with effort angina, 6 with old myocarial infarction, 4 with variant angina and 6 with other diseases. Atrial pacing was performed by placement of a 7F Zucker catheter in the CS and LV pressure was monitored by a 8F pig-tail catheter. β-TG, TXB₂ and 6-keto-PGF_1α were determined by radioimmunoassay using β-TG RIA kit (Radiochemical Center, Amersham), anti-TXB₂ antisera, ³[H] TXB₂, anti-6-keto-PGF_1α antisera and ³[H] 6-keto-PGF_1α. Blood was also obtained for the metabolic study. β-TG in CS increased after pacing in both pacing-positive and negative cases (Fig. 1). Coronary arteriography revealed significant coronary stenosis in all pacing positive cases and many of the pacing-negative cases. Four patients with variant angina developed either spontaneous or ergonovine-induced coronary spasm and β-TG in CS significantly increased. β-TG in LV did not change significantly in cases of either pacing or coronary spasm. TXB₂ in CS seems to be increased in pacing-positive cases and cases with coronary spasm. (Fig. 2). 6-keto-PGF_1α also increased in cases with coronary spasm. Negative coronary (a-v) lactate and (a-v) pyruvate were seen only on pacing in cases of positive-pacing test (Fig. 3). Lactate/pyruvate concentration ratio in CS blood was significantly increased after pacing whereas L/P in LV blood stayed constant. These metabolic data suggest myocardial hypoxia developed in pacing-positive cases. The result indicates that platelet activation in the coronary circulation may be responsible for angina pectoris.

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Exercise-induced changes of plasma β-thromboglobulin (β-TG) and coagulofibrinolytic substances were investigated in patients with ischemic heart diseases (IHD). Plasma β-TG was measured at rest and then immediately, 10 and 30 minutes after Master's two step test in the early morning. Further, in variant angina (VAP), it was also measured in the afternoon on the same day. At rest, IHD except effort angina (EAP) showed high levels of β-TG when compared with normal controls. But the exercise caused different changes of β-TG in IHD. Namely, β-TG levels in old myocardial infarction (OMI) with angina and, especially in VAP, were kept higher even at 30 minutes after the exercise. Moreover, the characteristic finding was that in VAP β-TG level in the early morning was significantly higher than in the afternoon. On the other hand β-TG in EAP and OMI without angina showed higher levels at immediately and 10 minutes after the exercise but returned to the initial levels at 30 minutes. As to coagulo-fibrinolysis mild but significant hypercoagulability, based on lowered levels of plasma antithrombin III, was recognized in IHD. It was concluded that the most remarkable change of β-TG was detected in VAP resulted from vasospasm of the coronary artery.

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The effect of ristocetin on platelet electrophoretic mobility (EPM) was studied with the new automatic Laser Zee System 3000 with special reference to the effects of changing the conductance of the platelet suspension medium. Human platelets were extensively washed using several methods until they no longer agglutinated upon addition of ristocetin alone. Washed platelets in plasma-free Zeiller-Hannig's buffer which has a low specific conductance showed an EPM of -1.95±0.05μm/sec/V/cm (M±S. E. M., n=20) and showed high susceptibility towards EPM decrease by 0.075mg/ml ristocetin alone without any change in medium conductance. Washed platelets in Tris-saline buffer showed an EPM of -0.98±0.03μm/sec/V/cm and showed a significant decrease in EPM with 0.5mg/ml ristocetin, while they did not agglutinate on the further addition of 1/10 volume of normal plasma. Since the platelet EPM decreased by addition of ristocetin was restored to the control value after 2 washes with buffer, the change in platelet EPM in the presence of ristocetin was reversible, suggesting a weak attachment of ristocetin to platelets. Treatment of platelets with neuraminidase resulted in irreversible and dose-related parallel decreases in both EPM and sialic acid content. These tiresults suggest that platelet surface negative charge is decreased by ristocetin and this enhances binding of negatively charged von Willebrand factor to the platelet membrane.

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Indium-111-oxine has been employed as a redioactive platelet label for thrombosis imaging and thrombokinetic studies in man. To evaluate it's suitability for platelet survival and turnover, thrombokinetic studies were carried out in hematological normal subjects, in patients with idiopathic thrombocytopenic purpura (ITP) and chronic congestive splenomegaly. For In-111-oxine labelled platelets, platelets were collected by differential centrifugation from 44ml of whole blood drawn into 6ml of acid citrate dextrose solution. Platelet suspension was incubated with In-111-oxine, which was extracted before use by the method of Thakur and co-workers. The survival, recovery and turnover of In-111-labeled platelets were 8.6±0.5 days, 63.0±5.4% and 3.9±0.3×10⁴/μl/day, respectively, which were similar with those of Cr-51 method. Platelet disappearance curves labelled with In-111 and Cr-51 simultaneously were similar in one case. In patients with ITP, platelet survival shortened in the same degree with Cr-51 method. The two simultaneous labeling studies between In-111 and Cr-51 showed no differences. In the patients with congestive splenomegaly, the same results were obtained. Thrombokinetic studies with In-111-oxine labelled platelets offer the advantages of reduced blood requirements, and the ability to perform external imaging of platelet distribution.

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Detection of intracardiac thrombi by scintiphotography using In-111-oxine labelled autologous platelets was done in patients with various heart diseases. Their results were compared with those of two dimensional echocardiography.
1) Left atrial thrombi in 3 patients with mitral valve disease and left ventricular thrombus in a patient with myocardial infarction could be detected by the both methods. In 2 out of above 3 patients with mitral valve disease, 30 and 21g of left atrial thrombus were certified at surgery. In the patient with myocardial infarction, no radioactivity on the thrombus was shown in the second scintiphotogram during antiplatelet therapy.
2) In a case with mitral stenosis, the image of thrombus by scintiphotography was observed, while none of abnormal echo by two dimensional echocardiography was detected. In this case, only 0.3g of left atrial thrombus was confirmed at surgery.
3) In an another case of mitral stenosis, the image of thrombus by scintiphotography was not observed, while abnormal echo by two dimensional echocardiography was detected. The thrombus in this case may be hematologicaly nonactive thrombus.
Scintiphotography with In-111-oxine labelled platelets is considered to be an excellent method for the detection of intracardiac thrombi and for the evaluation of antithrombotic therapy in vivo.

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The availability of hepaplastin test ^®(HPT), thrombotest ^®(TT) and prothrombin time (PT) in the early diagnosis of fluminant hepatitis (FH) has some conditions as well as antithrombin III (AT-III) and α₂-plasmin inhibitor (α₂-PI). In this paper, the significance of AT-III and α₂-PI in the early diagnosis of FH was evaluated.
HPT, TT, PT AT-III and α₂-PI in plasma were determined at the early stage of 14 cases of FH as compared with 6 severe from of acute hepatitis (with severe symptoms, but without hepatic coma, but HPT, TT and PT being less than 40%; AH-S) and 20 acute hepatitis (AH). And the following results were obtained.
(1) HPT, TT and PT showed a significant decrease in FH and AH-S as compared with AH.
(2) HPT, TT and PT showed no significant difference between FH and AH-S.
(3) AT-III and α₂-PI showed a significant decrease in FH as compared with AH-S and AH.
(4) AT-III and α₂-PI showed no significant difference between AH-S and AH.
Therefore, the determination of AT-III and α₂-PI is useful in early diagnosis of FH.

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As biological characterization of ATIII in patients with liver cirrhosis, the discrepancy between it's progressive activity and heparin cofactor activity obtained from biological measurements determined by clotting method with it's neutralized ability to thrombin or FXa. Both of heparin cofactor activities to thrombin and FXa reduced significantly in comparison with that's progressive activities in patients with liver cirrhosis. By crossed immunoelectrophoresis with heparin agar, the fast moving peak formed with ATIII and heparin was markdly smaller than that of normal plasmas. It might be thought that ATIII affinity to heparin was more reduced in plasma from liver cirrhosis than it's affinity in normal plasma.

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The localization of antithrombin III (AT III) in human vessel was investigated by fluorecent antibody method.
The authors found a single layer of AT III on the surface of the intima of the arteries. This fact suggested that a conformational change inducing an increase in antithrombin activity of AT III was caused by heparinoids on the surface of the intima, and that thrombin was regulated by the progressive type of AT III on the surface of the vessels.

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Two patients with the congenital deficiency of antithrombin III (AT-III) assoiated with superior mesenteric venous thrombosis (SMVT) are described and a possible relevance of the AT-III deficiency to SMVT is discussed.
Patient No. 1: A 34 year-old male without a history of thrombosis was found to have SMVT at operation, which was further confirmed histologically. Hematological studies disclosed abnormally low levels of progressive antithrombin activity, heparin-cofactor as well AT-III antigen. The decreased AT-III was also found in his mother, sister and niece.
Patient No. 2: The diagnosis of SMVT was established surgically as well as histologically in a 24 year-old male, whose father had died of recurrent SMVT. Abnormally decreased levels at about 50% of normal of progressive antithrombin. heparin-cofactor and AT-III antigen were also found in this patient and his sister. The familial involvement in their family indicates that the decrease in AT-III is hereditary, most probably autosomally transmitted. Besides the decreased level of AT-III, no abnormal laboratory findings were noted in both patients and their affected family members.
Thus we suggest that the SMVT found in these patients were due to the congenital deficiecy of AT-III.

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The disappearance rate of heparin and the effect of heparin on activated partial thromboplastin time (APTT) were investigated in five newborn infants between one and four days of age. Plasma antithrombin-III (AT-III) levels of five infants were 12%, 22%, 5%, 116% and 56%, respectively. In comparison with plasma heparin half life (T 1/2) from four normal adults (mean±1SD:102±10min.), significantly shorter T 1/2 was observed in all newborn infants (mean±1SD:40±5min.), excluding one infant (97min.) with normal AT-III level. Prolongation of APTT was observed corresponding with increase of plasma heparin concentration, in the newborn infants with normal and decreased AT-III level. In one infant, plasma concentration was reached to 0.8 unit/ml after 30min. of one shot infusion of heparin by 100 units/kg. However, in the same cases, it was reached to 1.4 units/ml after 30min. of one shot infusion of heparin by 100 units/kg and AT-III concentrates by 30 units/kg.
These results indicate that shortening of plasma heparin T 1/2 in newborn infant may be related to decreased plasma AT-III level.

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It was thought that only some part of heparin was complexed with AT III, when it was administered clinically and this would be a reason of indefinite and uncontrollable effect of anticoagulant therapy with heparin.
When heparin was mixed with whole blood, some amount of heparin was adsorbed to RBC, platelets and a fraction was distributed to plasma; 55% of total heparin mixed at the concentration of 2U/ml in whole blood.
When mixtures of normal plasma and various amount of ³H-Heparin were electrophoresed on 7% PAG, most part of heparin migrated free to the cathode and the small amount of radioactivity was noted at the sites of AT III and albumin, irrespective of the added amount of heparin. Purified AT III was mixed with various amount of heparin and electrophoresed on 1.4% agarose gel. It was found that only a part of AT III and heparin formed irreversible complex, and the most of them remained free. As the result, only a fraction of heparin was irreversibly complexed with AT III; i. e. 7.7-9.1% of total heparin mixed with plasma.

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The relationships between the chemical and physicochemical properties and the antithrombin III (AT III)-potentiating activities of heparan sulfates (HS) from various origins were investigated.
HS of bovine kidney or bovine lung showed higher activity than those of pig skin or pig gastric mucin.
These activities increased with the intensities of anionic charges of HS.
Unlike heparin, all HS preferentially potentiated AT III activities for Xa rather than for thrombin.
These tendencies were pronounced in lower molecular weight of HS.
The oligosaccharides obtained by enzymatic degradation of HS showed only anti-Xa activities and these active fragments were entirely adsorbed to AT III-Sepharose column.
Similar oligosaccharides were also obtained from heparin by heparinase digestion.
These results suggest that HS and heparin are carring similar oligosaccharides which are capable of potentiating AT III activity.

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The inhibitory effects of respective DS and heparin on the fibrinogen-fibrin conversion were studied with purified bovine fibrinogen and thrombin as reagents.
1) Both DS (0.05-0.8mg/ml) and heparin (1.0-5.0u/ml) showed inhibitory effects on the whole fibrin polymerization. Further study was done at every step of all the reactions of fibrinogen-fibrin conversion.
2) The fibrinopeptide release which reflects the proteolysis of fibrinogen induced by thrombin was measured. Heparin inhibited the reaction, but DS didn't at this step.
3) Then both accelerated the fibrin monomer polymerization which reflects the reaction from fibrin monomer to fibrin.
These results showed that the fibrinogen-fibrin conversion was eventually inhibited by either DS or heparin, independent of AT III. The inhibition by heparin operated mainly at the step of thrombin-fibrinogen reaction, though fibrin monomer polymerization seemed to be accelerated later. While the inhibitory mechanisms by DS could not be clarified. These results also suggest that in coagulation studies with heparin or DS administered, there is a possible discrepancy between the anti-thrombin activities estimated by clotting time and those by chromogenic substrate.

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