Mechanism of thrombus formation related to blood procoagulants and their inhibitors in blood was studied. In vitro experiments revealed that amount of thrombin adsorbed to fibrin during blood coagulation process was about 35% of thrombin which was neutralized by normal defibrinated plasma in 5 minutes at 37°C and that about 60% of thrombin inhibiting capacity of defibrinated plasma in this condition was attributed to antithrombin III. Effects of changes of concentrations of procoagulants and/or thrombin inhibitors in plasma on intrinsic blood clotting system were investigated by measuring activated partial thromboplastin times (APTT) of mixtures of 0.1ml of normal plasma and 0.1ml of buffered saline, normal plasma, BaSO₄ adsorbed plasma, heated (56°C, 2 minutes) plasma, bentonite adsorbed plasma or fraction of fibrinogen obtained from human plasma. Results were as follows:
1) APTT of mixture of normal plasma and buffered saline was 28.6 seconds; 2) 0.2ml of normal plasma (concentrations of all procoagulants and antithrombins were doubled), 34.6 seconds; 3) mixture of normal plasma and BaSO₄ adsorbate (did not contain factors VII, IX, X and II, however the other procoagulants and inhibitors remained), 38.8 seconds; 4) mixtures of normal plasma and heated plasma (fibrinogen was eliminated and factor V activity was halved, although the other procoagulants and inhibitors were unaffected), 40.7 seconds; 5) mixture of normal plasma and bentonite plasma (only α₂-macroglobulin and α₁-antitrypsin unchanged. α₂-plasmin inhibitor halved. The other procoagulants and inhibitors were removed), 28.6 seconds; 6) addition of fibrinogen solution to normal plasma; 38.2 seconds. In cases of acute myocardial infarction, levels of factor VIII, V and fibirnogen increased and concentrations of antithrombin III decreased, while levels of prekallikrein, which is a kind of procoagulants, and of factor X unchanged. In healthy subjects or cases of acute myocardial infarction, statistically significant positive correlations between factor X activities and antithrombin III levels in plasma were observed. However ratio of antithrombin III to factor X activity was usually lower in the patients with acute myocardial infarction. In many cases in whom thromboembolic complications developed, levels of antithrombin III decreased frequently. From these results, it is stressed that inhibitors of activated blood procoagulants is more important than blood clotting factors in thrombus formation, probably because process of blood coagulation is amplified by repetition of activation (proteolysis) of procoagulants (zymogen) by activated clotting factors (enzyme). Among cases of arteriosclerotic diseases, hypercoagulable state was observed in patients with acute myocardial infarction.

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Pathological study of thrombi was performed in 154 autopsy cases in respect of atherogenesis. One hundred twenty eight of 154 autopsy cases (83%) were found to have histological evidence of thrombi in more than one organ, and 119 (77%) exclusive of tumor emboli. Thrombi were classified into five types, e. g. microplatelet, red, organized, tumor emboli and thromboembolism. Malignant neoplastic group showed higher in frequency of thrombi than non-malignant neoplastic one.
Microplatelet thrombi were observed in the cases of DIC and found, in the descending order of frequency, in the ovarial cancer, pancreas cancer, leukemia, stomach cancer, pneumonia and central nervous disorders. Cases with clinical DIC showed high in frequency (8/9 cases) and wide in spread of thrombi (3.0 organs at mean).
There was no significant correlation between the severity of aortic atherosclerosis and the frequency of the whole thrombi.
It seemed to me that the organization of thrombi could be concerned intensely with the pathogenesis of atherosclerosis but the frequency of its organization would be very low in vivo.
This was a autopsy survey report, so it was necessary further to analyze the thrombi in various points of view.

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The physiological mechanisms of fibrinolysis are regulated by vascular plasminogen activator and α₂-plasmin inhibitor. The vascular plasminogen activator and plasminogen are adsorbed to fibrin, and plasminogen activation takes place on the surface of fibrin. This efficient mechanism of fibrinolysis is inhibited by α₂-plasmin inhibitor which interferes with the adsorption of plasminogen to fibrin. The blood level of α₂-plasmin inhibitor as well as the production and release of vascular plasminogen activator from blood vassels take determinant roles in the whole process of physiological fibrinolysis.

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Importance of endothelial injury and smooth muscle cell proliferation is emphasized as an early key event in atherosclerosis. We studied the effects of fibrin, fibrinogen and fibrinogen degradation products on swine aortic endothelial cells and rabbit aortic smooth muscle cells in culture.
Endothelial cells in the 5th subculture from the thoracic aorta of swine formed a cobblestone monolayer architecture. Normal monolayer morphology of cultured endothelial cells disorganized rapidly in response to contact with a fibrin clot. The cells of a confluent monolayer separated and degenerated after 24 hours of contact of fibrin.
Smooth muscle cells in the 12th subculture from the thoracic aorta of rabbit formed a “hills and valleys” pattern, showing multilayered or monolayered arrangement. Cultured smooth muscle cells proliferated on the agar plate containing plasminogen-free fibrin during the experimental period. Prominent proliferation of cultured smooth muscle cells on the agar plate containing plasminogen-rich fibrin was found after 24 hours, but smooth muscle cells degenerated after 48 hours. Incorporation of [³H] thymidine by cultured smooth muscle cells seemed to be dose-dependent on the concentration of fibrin during 24 hours. Plasminogen-rich fibrin inhibited the incorporation of [³H] thymidine after 48 hours, but plasminogen-free fibrin showed no inhibition.
Previously we elucidated that cultured smooth muscle cells showed fibrinolytic activity by Todd's fibrinolytis autography. Fibrinogen degradation products inhibited the incorporation of [³H] thymidine by cultured smooth muscle cells dose dependently.
These results suggest that fibrin deposition on the surface of endothelium produces disorganization of endothelial monolayer architecture and increased imbibition of plasma components into the arterial wall. Fibrin stimulates the proliferation of the smooth muscle cells of the media. But, the proliferation of the smooth muscle cells is inhibited by fibrinogen degradation products produced by fibrinolytic activity of smooth muscle cells. Metabolism of fibrinogen and fibrin in the arterial wall may play an important role in atherogenesis.

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The discovery of thromboxane A₂ and prostacyclin has clearly indicated the importance of the interrelationship between platelets and the vascular wall in clinical atherogenesis and thrombogenesis. We confirmed that thromboxane A₂ caused acute plasmal inflow into the aortic wall of rabbits, and such was considered to be an important key mechanism in the initiation and progression of atherosclerosis. When we flushed thromboxane A₂ into coronary arteries, myocardial infarction in cholesterol-fed rabbits was immediately apparent death occurred within 48 hours. In the controls, no changes related to myocardial infarction occurred. The plasma levels of TXB₂ in cholesterol-fed rabbits before a flush of thromboxane A₂ were statistically high and in turn those of 6-keto PGF_1α were low, compared to the controls.
Patients with ischemic heart disease subjected to submaximal treadmill test showed a high response to thromboxane B₂ and a low response to 6-keto PGF_1α and CAMP, as compared with those of healthy age-matched controls.
All these data clearly demonstrate the important roles of a balance in the interrelationship of thromboxane and prostacyclin for physiological functions in the vessel wall and that any disturbance in this balance will lead to vascular injury, platelet aggregation and enhance the prospect of athero and thrombogenesis. Chemotherapy to right these unbalances is essential for prevention and treatment of atherosclerotic and thrombotic disorders.

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The results obtained in a number of clinical trials which have been conducted to evaluate thrombolytic therapy of pulmonary embolism, acute myocardial infarction and cerebrovascular disease in western countries were summarized. There is now good evidence that thrombolytic therapy with either urokinease or streptokinase accelerates the rate of lysis of acute major pulmonary emboli. More controversial is the usefulness of the activator preparations in the management of acute myocardial infarction. Many trials in patients with acure myocardial infarction admitted to CCU have not demonstrated a favorable results when patients' fatality is used as the end point. Currently recommended regimens in western countries are not indicated for the management of an acute cerebrovascular accident, although further studies with different dosage schedules and with more sensitive measurements seem to be needed.
According to the hypothesis of Sherry and his group on thrombolysis, a quantity of plasminogen is taken up by the thrombus during its formation, and plasminogen administered intravenously is either adsorbed upon thrombus or diffused into it and plasminogen activation is due to the thrombolysis. It is, however, noted in the present study that plasminogen content of plasma and serum was almost identical. This indicates low plasminogen content in the thrombus as already pointed out by many other investigators. Since α₂ plasmin inhibitor inhibits not only plasmin activity but also urokinase itself and binding of plasminogen to fibrin, in vivo thrombolysis must be greatly impaired in normal concentration of α₂ PI. In the present study, the result is obtained that clot lysis was much accelerated when α₂ PI in plasma was depressed by an addition of anti-α₂ PI serum. It has been reported by Thorsen et al that UK is only slightly bound to fibrin during clot formation while highly purified porcine tissue plasminogen activator is strongly bound. Taking into consideration of rapid plasma clearance of UK without any evidence for concentration into the thrombus, these results above mentioned are sufficiently promising to suggest that the regimen adopted Urokinase-Streptokinase Pulmonary Embolism Study Group seems to be the optimal regimen for complete resolution of thromboemboli in vivo. However, the prolonged fibrinolytic state produced by use of a large amount of systemic UK or SK interferes with normal hemostasis resulting in a high incidence of hemorrhagic complications.
Attempting to reduce these disadvantages, active investigation on altering dosage schedules or using different approaches, such as local therapy in which the UK or SK is introduced as close to the site of the thrombus as possible, intermittent administration of relatively small amounts of SK with plasminogen and so forth has been started. Thrombolytic agents have been investigated in numerous clinical situations in this country. However, many of the trials have been criticized because of the lack of adequate control studies. Further studies are required to establish proper dosage schedule and to evaluate the efficacy of the thrombolytic therapy in this country.

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The clinical effects of Urokinase (UK) were observed in 96 patients with peripheral arterial and venous occlusion comparing with those of heparin. UK was administered at a dose of 60, 000 I. U. once to several times daily for 7-8 days, total dosis being 932, 000 I. U. in an average. The results were as follows.
1) UK treatment was more effective than heparin treatment not only in clinical signs or symptoms, but also in angiographical findings.
2) In 12 cases FDP level was measured before and after UK administration. Three out of four cases with increased FDP level after UK treatment showed marked improvement angiographically, whereas, in eight cases with unchanged FDP level only one showed improvement.
3) Side effects were observed in two out of 66 cases (3.0%) treated with UK, while three out of 30 cases (10.0%) treated with heparin. There was no severe side effects in the groups of UK treatment. From our experiences, the dosis of UK we have used seems not to be enough for the treatment. The data suggest that UK treatment must be further monitored more carefully concerning dosage and time to obtain a good result and that one shot intrevenous injection of high dose UK following by intravenous drip infusion at initial stage of treatment should be considered to expect the accerelation of the fibrinolytic activity.

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A β-blockade, propranolol, was found to reduce lipogenesis from glucose stimulated by insulin in mice, suggesting that it inhibited an insulin action in vivo. When nicomol was administered with the blockade, its inhibitory action on insulin was remarkably relieved. It was demonstrated that propranolol also inhibited insulin-stimulated lipogenesis from glucose in isolated fat cells. In contrast to the in vivo experiments, nicomol did not relieve the inhibitory action on insulin in the fat cell system. On the other hand, nicotinic acid, one of metabolites of nicomol, was found to reduce the inhibitory action of propranolol on insulin. Then experiments were carried out to clarify the effect of micomol and its metabolites on PGI₂ and TXA₂ formation in the platelet rich plasma-arterial ring system. Nicomol was found to elevate PGI₂ level and reduce TXA₂ level in this system. On the other hand, nicotinic acid did not affect both PGI₂ and TXA₂ formation. Nicotinamide, one of metabolites of nicomol, increase PGI₂ formation, but did not affect TXA₂ content. Based on these experimental results, clinical effects of nicomol were discussed.

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Fatty acid oxidation activities in brain microvessels of spontaneously hypertensive rats (SHR) were decreased with persistent of hypertension, which is closely related to the formation of angionecrosis. Pantethine restored this decrease of fatty acid oxidation activities in vivo. Mechanism of this activating effect of pantethine was investigated in liver mitochondria and brain microvessels.
Pantethine, pantetheine and 4′-phosphopantetheine activated three steps of fatty acid oxidation, that is, acyl-CoA formation, acyl-CoA incorporation into mitochondria (carnitine acyltransferase) and intramitochondrial oxidation, 4′-phosphopantetheine being most effective.
For these incubations, pantethine was metabolized to pantetheine, 4′-phosphopantetheine, dephospho-CoA and coenzyme A, and the amount of 4′-phosphopantetheine was most and that of CoA was least, suggesting that the activating effect of pantethine on fatty acid oxidation may be mainly due to 4′-phosphopantetheine.
Pantethine increased phospholipid synthesis in brain microvessels of SHR with activation of fatty acid oxidation on both aerobic and anaerobic conditions. This effect might be a mechanism of activating effect of pantethine on fatty acid oxidation.

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The current studies of the growth altering effects of plasma lipoproteins on cultured cells, such as vascular endothelial and smooth muscle cells, have shown conflicting results in various experimentals models. No informations are available about the effects of plasma lipoproteins on the growth of cultured tumor cells. In the present study, the human tumor cells used were HeLa cells, which were cultured with human lipoproteins and the growth of tumor cells was investigated.
The human lipoproteins, low density lipoproteins (LDL: 1.019<d<1.050g/dl) or high density lipoproteins (HDL: 1.063<d<1.210g/dl), each was prepared by sequential ultracentrifugation and dialyzed exhausively against 0.154M NaCl at 4°C. Each of the lipoprotein fractions was concentrated by ultrafiltration and filtrated through a Millipore filter (pore size=0.22μm) for cell culture experiment.
HeLa cells were obtained from the department of virology in our institute, and they have been maintained by passage twice weekly in plastic flasks containing Dulbecco's modified Eagle's medium (DMEM) supplemented with 5% fetal calf serum under 5% CO₂/95% air at 37°C.
Approximately 1.5×10⁵ of the cells were harvested and cultured in DMEM with various concentrations of HDL (80, 200μg protein/ml) or LDL (35μg protein/ml). The cells were dissociated by trypsinization and the number of cells was counted every day for 4 days. Morphology of the cells was examined under a light microscope.
Phase contrast photographs of the cells cultured with HDL on day 3 showed almost all the cells were flattened in shape and formed a monolayer sheet partly. In the medium without lipoproteins, the cells were round in shape and small in size and the plating efficiency was very low. An addition of HDL resulted in significant proliferation in the number of cells per dish for 4 days. The number of cells on day 4 was inversely proportional to concentration of triglyceride (r=0.698) and of cholesterol (ester) (r=0.538) in the medium. While neither concentration of phospholipid nor of cholesterol (free) was correlated with cell proliferation. As the concentration of HDL increased in the medium, the incorporation of ³H-Tdr into DNA increased after the cells were cultured with HDL for 3 days. An increase of HDL concentration above 1, 500μg protein/ml, however, showed no change in the level of DNA synthesis compared with control value.
These results suggested that LDL and HDL, especially HDL, has growth promoting effect on HeLa cells at a low concentration in vitro.

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It is reported that cholesterol ester (CE) deposition was accerelated by acetylated low density lipoprotein (acetyl LDL). To clarify the mechanism of CE deposition by denatured LDL, metabolism of cholesterol ester in reconstituted LDL (rLDL-CE) or acetylated rLDL (acetyl rLDL-CE) was investigated using particulated fraction of rat arterial wall or homogenates of rat peritoneal macrophages.
Hydrolytic activity of rLDL-CE and acetyl rLDL-CE was mainly located in lysosomal fraction. Arterial wall and macrophages were shown to hydrolyse acetylated rLDL-CE in preference to rLDL-CE. So, it is unlikely that CE deposition in acetylated LDL is attributable to deficiency of CE hydrolytic activity in lysosomes.
Next, CE synthesis in microsomes was investigated as a cause of CE deposition. CE synthesis in microsomes from free cholesterol produced by CE hydrolysis in lysosomes was examined. ³H-cholesteryl oleate incorporated rLDL was incubated with macrophage homogenates at pH 4.5, and then [1-¹⁴C]oleoyl-CoA was added and further incubation was performed at pH7.5. Although CE hydrolysis was higher in acetyl rLDL-CE than in rLDL, CE synthesis in the case of acetyl rLDL was remarkably higher than that of rLDL. It might be concluded that CE synthesis in microsomes surpass CE hydrolysis in lysosomes because of unknown mechanisms in the case of acetylated LDL.

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An inherited deficiency of low density lipoprotein (LDL) receptors in familial hypercholesterolemia (FH) is assumed to be the cause of the decreased catabolism of LDL. In contrast, in hyperthyroidism there is an increased rate of LDL catabolism. A 58-year-old female patient with heterozygous FH revealed a normal level of serum cholesterol (193mg/dl) in coexistence of hyperthyroidism. After the therapy of hyperthyroidism with radioiodine the patient's lipid profile revealed an increased concentrations of cholesterol (Whole serum 338, VLDL 68, LDL 199, HDL 42mg/dl). There was a significant inverse correlation between her serum cholesterol levels and her serum thyroxine levels (r=-0.815, p<0.01).
Since LDL catabolism in vivo is thought to be mediated largely through the LDL receptor, the role of thyroid hormone in this receptor-mediated degradation of LDL was studied in human skin fibroblasts. Confluent cells were exposed to a medium containing 5% lipoprotein deficient serum prepared from a myxedematous patient by ultracentrifugation for 2 days with or without added triiodothyronine (T₃). After ¹²⁵I-LDL was added, the cells were incubated for 6 hours at 37°C for the determinations of ¹²⁵I-LDL uptake and degradation. In the cells from 2 normal subjects, LDL uptake and degradation was enhanced 29% by preincubation with T₃ (1.0μg/ml). In the cells from 2 heterozygous patients with FH, LDL uptake and degradation was enhanced 23% with T₃(1.0μg/ml). While, in the cells from 2 homozygous patients with FH, no effect of T₃ was observed. These results suggest that normal serum cholesterol levels in a heterozygous patient with FH result in part from an enhanced degradation of LDL by extrahepatic cells exposed to excess thyroid hormone.

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The experiments were carried out to clarify the role of phospholipid on triglyceride (TG) hydrolysis during the catabolism of triglyceride-rich lipoprotein. Maximal TG hydrolysis of TG-phospholipid liposome by purified bovine milk lipoprotein lipase (LPL) was obtained when liposome containing 0.1mM phosphatidylcholine and 1mM triolein was used as a substrate. When [¹⁴C] POPC-[³H] triolein double labelled liposome was incubated with phospholipaseA, TG hydrolysis by LPL was remarkably increased. Incubation with phospholipase C also increased the hydrolysis by LPL. These results suggest that PL hydrolysis may play an important role on TG hydrolysis.
Purified human hepatic triglyceride lipase (HTGL) from post heparin plasma had more phospholipase activity than that of LPL. The human post heparin LPL hydrolysed VLDL-TG for 3 hrs but not in further incubation. Addition of HTGL remarkably increased the hydrolysis of TG, while LPL could not hydrolyse TG in intermediate low density lipoprotein (IDL). These results suggest that LPL can hydrolyse the triglyceride in VLDL but not in IDL and that HTGL might be able to hydrolyse the triglyceride in IDL due to the decrease in PL by the action of phospholpase A in HTGL.

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A kindred was presented in which 4 members in 3 generations showed absent or reduced serum concentrations of thyroxine-binding globulin (TBG). TBG was undetectable by radioimmunoassay in males and decreased in females. The mode of transmission of the trait was consistent with X-chromosome linkage.
Hyperlipoproteinemia was also present in 3 members with TBG abnormalities. Hyperlipoproteinemia was combination the type II and the type IV.
The mechanisms responsible for hyperlipoproteinemia in TBG abnormality remained obscure.

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Serum Apo A-I and A-II levels in 42 cerebrovascular patients, 26 with cerebral thrombosis and 16 with cerebral hemorrhage, and 30 healthy subjects were determined by laser nephelometry. Apo A-I and A-II were purified from delipidated HDL by the combination of gel filtration and ion exchange column chromatography. Antisera against each apoprotein were prepared by injecting rabbits with purified Apo A-I or A-II.
Average serum Apo A-I and A-II concentrations in healthy subjects were 131±17.8mg/dl and 79±10.9mg/dl, respectively, and the calculated A-I/A-II ratio was 1.66±0.18. Apo A-I and A-II levels in thrombotic patients were 103±18.8mg/dl and 50±10.4mg/dl, and the values in hemorrhagic patients were 97±21.5mg/dl and 55±17.7mg/dl, respectively. All of these values were significantly lower than those in the age-matched healthy controls. No statistically significant difference was seen between two groups of patients. Average values of A-I/A-II ratio in thrombotic and hemorrhagic patients were 2.08±0.31 and 1.84±0.37, respectively. These values were significantly larger than the value in controls, and a statistically significant difference was observed between thrombotics and hemorrhagics.
From these results, it was noted that protein components, as well as lipids, of HDL were lower than normal in cerebrovascular patients. In addition to such quantitative changes, qualitative alteration in the composition of HDL, which is reflected in large A-I/A-II ratio, may be a characteristic of lipoprotein metabolism in cerebrovascular patients, and this alteration may be closely related with vascular lesions leading to the outbreak of cerebral thrombosis or hemorrhage.

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Behavior of apoA-I and apoC-II in aqueous solution was studied by using high performance liquid chromatography (HPLC), sedimentation equilibrium (SE) and circular dichroism (CD). ApoC-II is dominantly monomeric with rather randomized structure in solution. This has been indicated, by the monomeric molecular weight obtained by ultracentrifugal SE using Beckman Airfuge and Model E, by molecular seive type HPLC showing single peak at the position of larger molecular weight than its monomer, and by CD spectra with residual ellipticity at 222nm of 4600 deg. cm²/dmol that showed little change depending on the concentration of the protein.
ApoA-I, when its lyophilized specimen was solubilized in aqueous buffer, showed two peaks in HPLC with apparent molecular weights of tetra-and dimer. The first peak disappeared irreversibly with reciprocal increase of the second peak when the solution was heated. The kinetics of the disappearence of the first peak was the first order and the rate constant showed a straight line in Arrhenius plot, giving the activating energy=-120kcal/mol. The second peak could not be broken down any further by heating or by treating with chloroform: methanol, SDS, urea and DTT. SE resulted in compatible observation to the previous ones that apparent molecular weight increased from monomeric to origomeric as the concentration increased. CD study revealed its highly helical structure but the decrease in helicity as the first peak in HPLC disappeared and as the solution was diluted. Consequently, one must consider p rapid origomerization equilibrium of apoA-I in aqueous solution but there are another type of origomers which is the intermediate step in solubilizing apoA-I.

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Chick bioassay of angiotoxicity of 7-ketochoiesterol was performed in this study. Dietary cholesterol feeding induced more lipid deposition of connective tissue cells than neighboring smooth muscle cells of ascending and thoracic aorta. Abdominal aorta had little response to cholesterol. 7-ketocholesterol supplementation to diets with or without cholesterol resulted in frequent dead and dying smooth muscle cells especially in abdominal aorta. These data support the hypothesis that oxigenative sterol is more potent angiotoxin than cholesterol itself.

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The composition of lipids existing in hair, epidermis and dermis of rat was analyzed by thin-layer chromatography and the composition and amount of sterols were determined by gas-liquid chromatography.
The major constituent of the hair lipids was sterol esters and the sterol fraction (21.4mg/100g body wt.) consisted of 26.6% cholesterol and 44.9% lathosterol. On the other hand, in epidermis and dermis, the main component of the lipids was triglyceride. Cholesterol and lathosterol composed 49.7% and 21.6% of the sterol fraction, respectively, and the total amount of sterols was 43.9mg/100g body wt.
In the sterol fraction of the lipids of the other organs, there existed predominantly cholesterol (96.4%) and a small amount of plant sterols (campesterol and β-sitosterol) was observed.
The proportion of sterols existing in the skin including hair was 36.1% of the total sterols in whole body, which was about 28 times larger than that of serum cholesterol pool. Since the wet weight of the skin was 17.4% of whole body, it was concluded that the sterol concentration in the skin was higher than the mean concentration in the other organs.
These results indicated that the skin lipids might play an important role in the lipid metabolism of whole body. Although many investigators have studied the effects of hypolipidemic drugs in rat on the lipid levels in serum and liver and on the excretion of sterols and bile acids into bile or feces, the effect of drugs on the skin lipids must be examined as well.

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The contents of glycosaminoglycans (GAG) and lipids of normal and atherosclerotic intima and media of aorta were determined. The percentage of chondroitin-4-sulfate (CS-4-S), chondroitin-6-sulfate (CS-6-S) and dermatan sulfate (DS) were calculated on the basis of unsaturated disaccharide subunits obtained by paper chromatography after exhaustively digestion of chondroitinase ABC (Chase ABC) or chondroitinase AC (Chase AC).
Although, the existense of the disaccharide-OS in aortic GAG with the paper chromatogram after treatment of Chase ABC or AC was reported, there was no disaccharide-OS in all cases of this study.
The lipid contents, including total cholesterol, free cholesterol, esterified cholesterol, lipid phosphorus and triglyceride, was increased significantly with the advance of atherosclerosis in intima. Although, the differences of the content of total cholesterol and esterified cholesterol between nor mal and atherosclerotic lesion was stastically significant in media, those in atherosclerotic lesion of media was less than those of normal of intima. These findings were suggested that the intima was main lesion of atherosclerosis.
With the progression of atherosclerosis, there was a pronounced decrease in the uronic acid of both intima and media.
The percentage of DS of total GAG (DS%) in intima was increased with the advance of atherosclerosis, while the percentage of CS-4-S, CS-6-S, HS and HA were no apparent relationships with the advance of atherosclerosis. The differences of DS% between normal and atheroma, or fibrous plague and atheroma of aortic intima was significantly stastically significant (p<0.01). There was also statistically significant positive correlation ships between DS% and total cholesterol, free cholesterol, esterified cholesterol and lipid phosphorus in intima, while there was no such a correlationships between lipid contents and GAG in media. The percentage of CS-6-S (CS-6-S%) and heparan sulfate (HS%) had the statistically negative correlationships with the contents of total cholesterol, free cholesterol, esterified cholesterol and lipid phosphorus in intima.
With these results, it was suggested that the DS might have some role of the lipid deposition in arterial intima. The role of CS-6-S and/or HS, however, is still remained obscure, and the further study will be needed.

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Fatty acid oxidation activity in brain microvessels of spontaneously hypertensive rats (SHR) was decreased with persistence of hypertension compared with that of normotensive rats (W-K). The mechanism of this decrease was investigated.
Fatty acid oxidation pathway was divided to three steps, that is, fatty acid activating step (acyl-CoA synthetase), incorporation step of acyl-CoA (carnitine acyltransferase) and intramitochondrial oxidation step. No steps were significantly decreased in activities of brain microvessels of SHR compared with W-K, suggesting the decrease of fatty acid oxidation activity is not due to decrease in enzyme capacity but to disturbance of proper reaction of enzymes. Acyl-CoA synthetase and carnitine acyltransferase are membrane-bound enzymes. So the properties of mitochondrial membrane in brain microvessels were next investigated.
In the anoxic state, fatty acid oxidation activity of SHR was much more decreased compared with in the aerobic state than that of W-K, and fatty acid incorporation into phospholipid was decreased in SHR whereas that in W-K was in creased. This indicates that membranes of SHR respond to anoxic state differently from those of W-K. To investigate properties of membrane, release of enzymes such as acyl-CoA synthetase, carnitine acyltransferase and succinate cytochrome C reductase was estimated. The amount of release of these enzymes to supernatant fraction of brain microvessels treated with sonication or Triton X-100 was higher in SHR than in W-K.
From above results we conclude that mitochondrial membranes in brain microvessels of SHR were more unstable than those of W-K, which may result in disturbance of proper reaction of membrane-bround enzymes.

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The mechanisms of low density lipoprotein (LDL) binding to arterial elastin were studied.
Ratios of cholesterol/phospholipid bound to elastin from LDL at various incubation periods were constant, 1.4-1.6, when the ratio in the original LDL was set at 1.0. On the other hand, ratios of ¹²⁵I-LDL-protein/cholesterol or triglyceride increased with increasing amounts of LDL incubated with elastin. These results suggest that LDL does not bind to elastin as lipoprotein particle, but the binding activities of various components in LDL are different.
From the results on rosette formation around the elastin-LDL complexes after treatments with anti-LDL serum and protein A-coated sheep red cells, on the binding activities of various modified LDL, and on the trypsin digestion of elastin-LDL complex, it is demonstrated that the binding of elastin appears to occur dependently on the protein moiety of the lipoprotein, whereas the maintenance of the complexes appears to occur independently on the apoLDL.
The binding activities of LDL to elastin were compared with LDL preparations from normal and various kinds of hyperlipoproteinemic sera.
Those of LDL from types IIa and IIb hyperlipoproteinemia were significantly higher than that of LDL from normal serum.

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In 53 patients with essential hypertension during treatment with β-blocker (Alprenolol) and calcium antagonist (Diltiazem) the change on serum total cholesterol and triglyceride has been observed at same month or at least same season for 10-120 months (mean 48 months).
In the patients treated with β-blocker and calcium antagonist changes of total cholesterol and triglyceride levels in mean were only 2-5mg/dl and 6-9mg/dl respectively. These changes were not significant.
The variation over 20% of serum triglyceride was seen in several cases, and its variation was influenced not by drugs, but by the marked increase of r-GTP or body weight.

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Enhanced platelet aggregation (PA) is thought to play an important role in progression of arteriosclerosis with relation to fibrinolysis and we have previously reported that PA in arteriosclerosis is enhanced in chronic state. The aim of this report is to investigate the effects of a coronary vasodilator, trapidil, 5-methyl-7-diethylamino-s-triazolo- (1, 5-a)-pyrimidine, on PA, fibrinolysis, pulse rate and blood pressure in patients with arteriosclerosis. We measured PA, plasma euglobulin lysis time (ELT), pulse rate and blood pressure in 21 arteriosclerotic patents (mean age 65.2±11.7 years) including chronic state of ischemic heart disease, cerebral infarction and transient cerebral ischemic attack before treatment with oral trapidil 300mg/day, 4 weeks and 8 weeks after treatment. ADP-, collagen-and adrenaline-induced PA were studied by the turbidometric method. The aggregation agents employed were ADP in a final concentration of 2×10-6M, adrenaline in a final concentration of 1×10-6M and collagen solution using tendon of bovine and equine. ELT was also measured at the same time by using clot lysis recorder. Pulse rate and blood pressure were measured just before blood withdrawal. The following conclusions were drawn from this study.
1. All of these types of induced PA decreased gradually after administration of trapidil. The value before initial administration was significantly higher than the value of 8 weeks after (ADP: P<0.01, bovine collagen: P<0.01, equine collagen: P<0.01, adrenaline: P<0.05). The maximum optical density (Max. O. D.) showed the most significant difference in all of these types of induced PA.
2. ELT seemed to be shortened within the 8 weeks, but there was no significant difference.
3. Pulse rate prior to initial administration and that of 8 weeks after showed no significant difference, but within this period there was a significant increase 4 weeks after administration of trapidil. There was no significant difference in systolic blood pressure and diastolic blood pressure witin the 8 weeks.
4. Correlation of change between Max. O. D. of PA and blood pressure at the end 8 weeks was rather higher than the first 4 weeks after administration of trapidil. All correlations were not significant, but all correlation coefficients were positive. Among them correlation coefficient between Max. O. D. of adrenaline-induced PA and systolic blood pressure or diastolic blood pressure was higher than the others (systolic B. P.: r=0.42, diastolic B. P.: r=0.27).
These results show that PA decreases gradually after administration of trapidil and change of PA may be related to change of blood pressure and suggest that trapidil may have beneficial actions in the treatment of patients with arteriosclerosis who show enhanced PA.

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There are many substances effecting on the platelet aggregation. But the detail mechanism of platelet aggregation is unknown yet.
We observed many substances which have the effect on platelet aggregation. Consequently, we found two interesting substances which have similar chemical composition with Vitamin E. The effects of these substances on platelet aggregation (human platelets) were studied with Evans's aggregometer. 6-Hydroxy 2, 5, 7, 8, tetramethyl chroman-2-carboxylic acid (vitamin E analog) inhibited platelet aggregation induced by noradrenaline (55.5γ/ml), ADP (55.5γ/ml) and arachidonic acid (555.5γ/ml). But it didn't inhibit the aggregation induced by collagen (22.2γ/ml) or 0.5μ/ml of thrombin.
On the other hand, dl-α-Tocopherol diphosphoric acid ester induced the platelet aggregation by itself. The aggregating ability of 100γ/ml-150γ/ml of dl-α-Tocopherol diphosphoric acid ester is as same as ADP (555.5γ/ml). Its molecular weight is almost as same as ADP. So the ability of aggregation of this substance is one half of that of ADP. Vitamin Eis known as antioxidative agent. Though these substances have similar structure to Vitamin E, 6-Hydroxy, 2, 5, 7, 8, tetramethyl chroman-2-carboxylic acid inhibited platelet aggregation, while dl-α-Thcopherol diphoric acid ester initated the aggregation.
From these results obtained, it might be difficult to explain the difference between Vitamin E and substances examined.

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In rats fed a cholesterol-free, low-fat (1% cornoil) diet containing soybean protein isolate as a nitrogen source, serum cholesterol levels were consistently and significantly lower than those of the animals fed the corresponding casein diet. Soybean protein caused a decrease in apo A-I and an increase in apo B. Pantethine when added to soybean protein diets increased apo A-I and decreased apo B without influencing serum cholesterol levels. Tocopherol did not alter serum levels of cholesterol, but significantly raised apo B. The effects of dietary protein and/or these two vitamins on serum apo E were not unequivocal.
In rats fed soybean protein, the mesenteric lymph flow and apo A-I content decreased markedly. Addition of pantethine ameliorated these modulations to a level compatible with that of rats fed casein. However, when the dietary fat level was increased to 5%, effect of pantethine on these parameters became rather obscure. Soybean protein, compared with casein, stimulated the excretion of fecal steroids, but pantethine and tocopherol showed no definite effects on the steroid excretion. Pantethine increased cholesterol content of the adipose tissue irrespective of the source of dietary proteins.
These observations show that soybean protein manifests its effects on serum cholesterol and apolipoproteins partly through the regulation of intestinal contribution. Alternatively, the stimulation of intestinal function seems at least responsible for the HDL-cholesterol-rising potential of pantethine.

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In order to determine the Co Q₁₀ concentration and its distribution in plasma lipoproteins blood was obtained in the fasting state in 6 normal and 27 clinical subjects on Co Q₁₀ (30mg/day).
In normal subjects, mean concentration of Co Q₁₀ was 0.994μg/ml and its major part was fbund to be present in LDL.
After ingestion of 90mg Co Q₁₀ with 30gr of margarine rich in linoleic acid, the mean Co Q₁₀ concentration in plasma increased and reached the peak value of 1.553μg/ml at 4 hours.
Linoleic acid and Co Q₁₀ firstly appeared in chylomicron and later in VLDL or remnant, then in HDL and LDL. Absorption and transport of CO Q₁₀ seemed to be similar to those of fatty acid.
From the analysis of VLDL lipid composition, Co Q₁₀ appeared to exist in both core and surface part of the lipoprotein.
The concentration of Co Q₁₀ was related to the phenotypes of hyperlipoproteinemias. In type II, Co Q₁₀ was distributed more in LDL and in type IV, it was present more in VLDL.

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In order to get the ideal standard substance to evaluate serum apo AI in geriatric patients such as IHD and cerebral thrombosis, HDL₂ AI was semi synthesized with purely separated apo AI, lecithin and lysolecithin by urtrasonic synthesis method.
The properties of this semi-synthesized HDL₂ AI was strictly examined by CD spectrum, zone electrophoresis, lipo-dyed microultracentifugation method and immunoelectrophoresis and found to be identical to serum HDL₂ AI.
By using semi-synthesized HDL₂ AI as primary standard, apo AI value in QS serum (lyphilized pooled fresh Japanese serum by more than 3000 healthy people) was standardized as 108±6mg/dl, and the serum from 19 patients (9 cerebral thrombosis, 3 IHD and others) were evaluated by this standard value, which showed fairly lower value.
These results suggest that semi-synthesized HDL₂ AI can be used for the evaluation of the clinical meanings of apo A, apo AI in the patients of IHD and cerebral thrombosis.

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The effects of one month clofibrate administration on glucose tolerance, plasma immunoreactive insulin (IRI), total cholesterol, triglyceride and HDL-cholesterol levels were studied in 10 patients with non-insulin dependent diabetes mellitus (NIDDM). All patients were treated with ordered diet and/or exercise before and during the administration of clofibrate, 1, 500mg daily after every meals and the study was started after the establishment of the constant body weight and plasma glucose levels.
At the beginning and at 4 weeks of treatment, 75g glucose tolerance test and the measurement of total cholesterol, triglyceride and HDL-cholesterol were made. Plasma glucose, IRI and HDL-cholesterol were determined by the glucose oxidase method, the radioimmunoassay using single antibody method and the precipitation method using Heparin-Ca⁺⁺, respectively. All data were analyzed by the paired Student's t-test and were expressed as mean±S. E.
1) Fasting plasma glucose levels were 173.4±11.1mg/dl and 137.5±5.9mg/dl (p<0.01) before and at 4 weeks of clofibrate, respectively. While fasting IRI levels were decreased from 12.9±1.7μU/ml to 10.6±2.0μU/ml, but it was not significant statistically.
2) Plasma glucose values after 75g glucose loading showed significant fall in each point (P<0.01), especially at the latter points. On the other hand, IRI response was decreased, while it was not significant, statistically. However ∑IRI/∑glucose ratio before and at 4 weeks of clofibrate were constant and significant correlation was observed.
3) Total cholesterol and triglyceride levels were decreased, while HDL-cholesterol levels were increased statistically after 4 weeks administration of clofibrate. Atherogenic index (total cholesterol-HDL-cholesterol/HDL-cholesterol) was decreased from the value of 4.13±0.20 to 3.13±0.23 (p<0.01).
These observations suggest that clofibrate would be a useful drug to improve the disturbed glucose metabolism in NIDDM together with the enhancement of the HDL-cholesterol levels which are reported to be decreased in diabetics treated with oral hypoglycemic drugs.

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