Antiserum against cholesterol skeleton and free OH group at C₃ position of cholesterol and that against cholesterol skeleton and the side chain of cholesterol were obtained by immunizing rabbits with etiocholenic acid-poly-L-lysine complex and cholesterol succinate-poly-L-lysine complexes, respectively.
HDL inhibited strongly the PHA reaction between Etio-anti-Etio antiserum system, and LDL had the inhibitory activity on the PHA between CHS-anti-CHS antiserum system.
These results showed that the free OH group C₃ directed toward the surface of HDL and the side chain of cholesterol appeared on the surface of LDL molecule.

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Very low density lipoprotein (VLDL) is a major transport vehicle of triglycerides (TG) of endogenous organ in plasma. A major factor determining the amount of TG secreted in VLDL is an availability of free fatty acid (FFA) of either exogenous or endogenous origin. From a practical standpoint, a release of VLDL-TG from liver was associated with a level of plasma FFA concentration, and a hydrolysis of VLDL-TG might be dependent on an energic source in human metabolism. It is possible, therefore, that a VLDL-TG play a role on an initiative factor in lipoprotein metabolism. On the other hand, experimental studies using glucose, noradrenaline, and nicotinate in the rabbits showed as follows: 1) Plasma FFA concentration in the rabbits treated with glucose or nicotinate was significantly decreased, while that in the rabbits with noradrenaline was markedly increased as compared with that in the rabbits with saline. 2) Radioactive plasma FFA and VLDL-TG after the infusion of ¹⁴C-palmitate were the highest values in the noradrenaline group, but those were the lowest values in the nicotinate group as compared with those in each group. Under the condition loaded with glucose, a significant change in radioactive plasma FFA was not observed, but the radioactive VLDL-TG was increased as compared with that in the control. Additionally, half life of the both plasma FFA and VLDL-TG were more prolonged than those in the other 2 groups. 3) Radioactive TG in adipose tissue was markedly increased by the glucose, while those in the both groups treated with noradrenaline or nicotinate were decreased. These results suggests that a synthesis or a hydrolysis of VLDL-TG is regulated by an energy metabolism. Under the condition utilized FFA as an energic source, VLDL-TG synthesis is apparently accerelated, but it is result from the increased plasma FFA releasing from the adipose tissue. It is possible, therefore, that a release or a synthesis of VLDL-TG is controled by the homeostatic balance which is dependent on a certain level of plasma FFA concentration. Under the condition utilized glucose, plasma FFA is availabled for a VLDL-TG synthesis or an accumulation of TG in adipose tissue. However, VLDL-TG hydrolysis is reduced, and an abnormal lipoprotein metabolism of VLDL to low dencity lipoprotein can be observed at this condition.

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VLDL catabolism to LDL is accelerated by the intravenous injection of heparin due to the stimulation of the lipoprotein lipase (LPL) activity. But about 50% of cholesterol in VLDL could only be recovered in LDL during the constant infusion of heparin. This result suggested that the intravenous heparin injection removed the lipoprotein cholesterol from the blood circulation.
The aim of this study was to examine the effects of the intravenous heparin infusion on the distributions of the lipoprotein cholesterol amounts in order to estimate the action of heparin on the lipoprotein cholesterol removal from the blood circulation.
Two normal, three heterozygous type IIa hyperlipoproteinemic and four type IV hyperlipoproteinemic subjects were used for this study. VLDL (d<1.006), LDL₁ (1.006<d<1.019), LDL₂ (1.019<d<1.045) LDL₃ (1.045<d<1.063) and HDL (d>1.063) were harvested by the method of Havel et al. Cholesterol and triglyceride amounts were measured in each lipoprotein fraction.
The following results were obtained during the constant infusion of heparin.
Cholesterol amounts did not change in the normal subject but cholesterol/TG ratios increased markedly in LDL₂, LDL₃ and HDL. However cholesterol levels decreased specifically in LDL₂ of the heterozygous type IIa hyperlipoproteinemia. Cholesterol/TG ratios markedly increased in LDL₃ and HDL.
The reciprocal changes of cholesterol levels were seen in VLDL and LDL₂ in the type IV hyperlipoproteinemia. Cholesterol/TG ratios increased very much in LDL₂, LDL₃ and HDL.
LDL₂ was incubated with the postheparin plasma from a normal male at 28°C for one hour in order to check up the possible conversion of LDL₂ to LDL₃ in the LPL rich plasma.
The decrement of cholesterol level in LDL₂ could not be seen. Cholesterol/TG ratios did not change so much as in vivo phenomena in LDL₂, LDL₃ and HDL.
These results suggested that the heparin administration stumulated the removal of the plasma LDL₂ cholesterol in the heterozygous type IIa hyperlipoproteinemia not due to the stimulation of LDL₂ conversion to LDL₃. Intravenous administration of heparin seemed to work on LDL₃ and HDL because cholesterol/TG ratios increased very much during the constant infusion of heparin.

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In order to investigate the influence of humoral factors to the lipoprotein uptake into the arterial wall, following experiments have been performed. The isolated aortic wall obtained from male rabbit was incubated in Warburg's apparatus for measurement of two hour's intake of cholesterol-C¹⁴-labelled low and high density lipoproteins. During the incubation period, 95% oxygen with 5% CO₂ was administered and various concentrations of renin, angiotensin II, norepinephrine, epinephrine and insulin were added in the incubation medium. Radioactivity was counted by the liquid scintilation counter.
Result and discussion: 1) Addition of renin in various concentrations into the incubation medium did not show significant effects liprotein uptake into the arterial wall. 2) Intermittent administration of angiotensin II at intervals of 15 minutes significantly increased the uptake of both LDL and HDL into the arterial wall. However, one shot administration of this substance did not show any influences to the lipoprotein uptake. 3) Norepinephrine did not give any influences to the lipoprotein uptake. 4) Epinephrine also did not show any influences. 5) Addition of insulin to the medium showed significant increase of LDL uptake.
From the above mentioned results, it may be suggested that progress of vascular damege in high renin hypertension may not be affected the direct action of renin but be affected other factors which related to the high renin states, such as angiotensin II etc. It is difficult to explaine the reason why only angiotensin II increase the lipoprotein uptake. It maybe suggested that the temporal constriction of the artery may not be related to this phenomenon, because both angiotensin II and catecholamines constrict the artery. So angitensin II may be considered to cause the organic change of endothelium directly. On the other hand, catecholamine may cause the arterial damege by the result of long term continuation of arterial constriction. This may be the reason why catecholamine did not give any influences to lipoprotein uptake in this study.

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The objective of this study is to demonstrate the coronary risk factors with regard to nutriments, serum lipoproteins and lipids, especially in the subjects without hypercholesterolemia who are noticiable in the provinces of Japan.
Five year follow up has been undertaken in 107 subjects aged 45 to 55 years who has been in the similar circumstance and has the same occupation (relative metabolic rate 1.0-1.9), and the average intake of nutriments are: 2086 Cal. of total Calory, 72.1g of protein, 318g of carbohydrate, 15.2g of vegetable fat and 23.7 of animal fat per day. In the subjects, there were 4% of hypercholesterolemia (average conc. of Ch.>220mg/dl), the and 52% of hypertriglyceridemia (average-conc. of TG>120ng/dl), proportion of the six basic lipoprotein PAG disc patterns representing type b, bp, Pb, PB, BP and B were 37.4%, 33.6%, 21.5%, 3.7% 2.8% and 0.9%, respectively. Total number of cornoray heart disease are 8 in 107 subjects, 5 coronary heart diseases, including 4 effort angina pectoris and 1 myocardial infarction, occurred during 5 year follow up and the other three patients had angina pectoris already at the begining of this study.
The data obtained during 5 year follow up were examined by Principal Component Analysis and the first and the second principal components were computed. The normalized eigne vectors indicated that the first principal component was composed of vegetable fat, carbohydrate, serum triglyceride level and degree of obesity, the second principal component was composed by beta/alpha lipoprotein ratio, alcohol intake, proportion of animal fat to total dietary fat and serum triglyceride level. It was also indicated that the factors composing the first and the second principal component had influence on the lipoprotein PAG disc patterns, i. e. the above mentioned factores induces increase of pre-beta lipoprotein in plasma.
It was shown that the factors composing the second principal component had higher values in the patients with coronary heart disease than in the normal subjects. While such a characteristics was not presented from the first principal component.
In conclusion, only alpha lipoprotein deficiency which are detrimental to the transport of triglyceride rich lipoproteins but increase of the proportion of animal fat to total dietary fat and a lot of alcohol drinking should be contributory to coronary heart disease particularly in the subjects without hypercholesterolemia.

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Our previous studies showed that adult obese subjects, like patients with arteriosclerosis and cholelithiasis, has a decrease in α-lipoprotein and an incresae in β/α ratio of lipoprotein cholesterol, even if the total serum cholesterol level was in normal range.
In this study, analysis of serum lipoprotein patterns and the glucose tolerance test were performed in obese children. Obese children with body weight more than 20% above the ideal weight were approximately 6% of the school boys and girls examined.
There was no difference between obese children and the control (non-obese) in serum cholesterol and triglyceride concentration. However, in obese children, the concentration of α-lipoprotein cholesterol was significantly lower and β/α ratio was significantly higher than in the control. Electrophoresis of lipoproteins also showed a high frequency in obese children of a pattern similar to type IIa hyperlipoproteinemia.
Oral glucose tolerance test revealed that the glucose tolerance itself was in normal range in most of the obese children, but the half of them had marked hyperinsulinemia.
Data obtained in this study present an evidence that the obese children had abnormalities in glucose and lipid metabolism similar to obese adults. The importance of the prevention of obesity in young people must be stressed.

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Preventive effect of EG626 on atherosclerosis of cholesterol-fed rabbits has been shown in this experiment with 30 male adult rabbits. The placebo control group of 15 animals was given a capsule with potato-starch and the treated group of the same number of animals was given daily a capsule with EG626 (1mg/kg) once a day and all animals were kept on 1% cholesterol pellet for 16 weeks. At the end of 16 weeks of the treatment, all animals were sacrificed for the chemical and histological analysis.
The cholesterol content of aortic wall was significantly smaller in the treated group (24.0±7μg/mg D.W.) than that of the placebo control gorup (57.5±9μg/mg D.W.) (P<0.01) and the percent surface involved by fatty streaks was also signicantly smaller in the treated group (15.5±2%) than the control group (35.5±7%) (P<0.05), while there was no significant difference between both groups in the body weight, the weight of liver, heart, lung, brain, kidney, adrenal gland, and testis and in the serum cholesterol level.
The cAMP-level of atheromatous lesions and medial layers of the aortas of the placebo control animals exhibited an abnormally low level, while that of the healthy male adult rabbits was 3.43±0.21pmoles/mg D.W. and that of the treated group was 1.66±0.23pmoles/mg D.W. and was significangly higher than that of the placebo control group ((0.81±0.13)pmoles/mg D.W.) (p<0.05) and the cAMPPDE activity was abnormally elevated in atheromatous lesions and medial layers of the placebo control group and it was significantly lowered in the treated group (P<0.05) suggesting the possible significance of cAMPPDE inhibiting effect of EG626 in prevention of atherosclerosis.

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A coincidence between a location of myocardial ischemia by angiography and by an electrocardiography was studied on the 22 patients with myocardial infarction and 32 angina pectoris.
These patients having both ischemic ST-T change in the electrocardiogram and significant stenosis of the coronary arteries were selected from the cardiology department of the Center for Adult Diseases.
A mean value of the number of the affected vessels in the myocardial infarction was 2.5 and higher than that in angina pectoris in which the number was 1.7 (p less than 0.005).
In the cases with myocardial infarction, the Q wave localization coincided nicely with both coronary artery stenosis (68.8%) and myocardial asynergy (74.1%).
On the other hand, the location of ischemic ST-T change in the cases with angina pectoris coincided with coronary artery narrowing in 50.0%.
In the study of each coronary artery, 67.9% of the stenosis of left anterior descending artery represented abnormal Q wave in V₁-V₄ in myocardial infarction, 47.8% of that of right coronary artery represented Q wave in II, III and _aV_F and 30.4% of that of left circumflux artery represented Q wave in I, _aV_′L, V₅ and V₆.
On the other hand, 94.7% of significant stenosis of left coronary artery represented ischemic ST-T change in anterior chest leads or I, _aV_L after Master's double 2-step test in angina pectoris and 64.0% of that of right coronary artery represented ischemic ST-T change in II, III and _aV_F.
Although the accuracy of electrocardiogram as a reliable indicator of ischemic heart disease has remained a controversial subject, elctrocardiogram is often of value in indicating sites of coronary arterial stenosis and ventricular asynergy.

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The heart is suspended by the great vessels, especially by the aorta and its main branches and the surrounding pulmonary tissue, In this circumstance, the suspending points would be influenced continuously under cyclic cardiac motion, so that pathologic changes were induced in these points, such as the sinus of Valsalva and the orifices of the coronary arteries. In this report, the pathological changes seen in these portions of the aorta were studied in cases with ischemic ECG findings with control group without any ischemic changes.
Results obtained were as follows;
1. The diameter of the aorta at its origin and volume of the aortic valves were well related to the aging.
2. There was no difference between the cases with and without Ischemic changes of ECG with reference to stenotic grade of the coronary atreries.
3. There were some differences in the position of the orifice between the cases with and without ischemic ECG changes. However, its meaning was not clarified at the present time, so intensive study must be done in future.

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The influences of nitrites (pentaerythritol tetranitrate PETN) and dipyridamole on the myocardial tissue oxygen tension were determined in five anesthetized open chest dogs using polarographic method.
The results obtained were as follows;
1. Administration of PETN (2mg/Kg) caused evident elevation of tissue oxygen tension in the subendocardium, simultaneously the evident decrease in left ventricular endodiastolic pressure (LVEDP) was observed.
2. Administration of dipyridamole (0.3mg/Kg) caused consistent increase of mycoardial oxygen tension in subendocardium, while no constant response in subepicardium.
It was considered that the elevation in subendocardial tissue oxygen tension after administration of PETN was induced by the decrease in the preload (LVEDP).

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A total of 35 coronary arteries were obtained from 34 autopsy cases with acute or remote myocardial infarcts. Multiple step sections with approximately 4mm intervals were made from the arteries which were causative for the infarcts. Serial sections with 4 micron intervals were also made for the required cases. Intramural hematoma was found in 13 (37%) and microhemorrhage or hemosiderosis were demonstrated in 14 (40%). Among 13 arteries with intramural hematoma, four were considered to have followed fissured or break-down of the atheromas, as Friedman reported, whilst nine arteries suggested that the hematomas might be caused by rupture of the intramural capillaries. Among these nine arteries two had intramural hematoma only and no thrombus without intimal rupture. Seven had communication between the intramural vessels, hematomas and intraluminal thrombus. It seems reasonable to think that rupture of the intramural capillaries might be followed by hematoma formation, and subsequently rupture of the intima due to sudden pressure elevation in the atheromas, and thrombus formation. Frequent local edema, foam cell accumulation, concentric fibrosis and microhemorrhage around the intramural capillaries are suggestive of circulatory disturbances in the intramural locations of the coronary arteries, which may be one of local factors in progression of coronary sclerosis.

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It is well known taht non-thrombotic cases can be seen in myocardial infarction in wide variety of frequency. In these cases coronary dysfunction was believed a priori to be the cause of myocardial necrosis without offering some reliable morphological evidenses. This type of myocardial infarction itself becomes a contradictory fact to Virchow's classical theory of mechanical (physical) obstruction of an irrigating artery“ on one hand and may give a key to solve a few difficult problems concerning myocardial infarction on the other hand; namely (1) why a maine coronary thormbus is constantly found at a very proximal portion of a coronary trunk? and (2) why the severest atherosclerosis can without exception be found at that part?
Yajima et al. could observe the “mechanical destruction and bloody imbibition” at an attaching portion of a maine thrombus to a superficial layer of an atherosclerotic intima of a coronary trunk in cases of thrombotic cases of myocardial infarction and similar usually slighter lesions at a similar portion in non-thrombotic.
These lesions were searched for by Yajima and his collaborrators based on a hypothesis of a sudden elevation of inner pressure due to derangement of systemic coronary circulation.
In this report results of observatins on prethrombotic lesions are reported and discussed. The pre-thrombotic lesions are very clearly demonstrated by electron microscopy.

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Cardiocerebral apoplexy is nowadays applied to a syndrome of atypical acute myocardial infarction with cerebrovascular accident with a little vagueness in the definition. Ozawa and Fujii found by considerable reserches that this syndrome develops in the aged over 70 and that some cases reveal macroscopical findings of encephalomalacia in autopsy and others lack of those. But this syndrome is wanting pathological basis because of lack of detailed histopathological examination of its case.
For the purpose of investigating the pathogenesis of this syndrome, we studied microscopically cardiac lesions in cases of cerebrovascular diseases (cerebral hemorrhage, subarachnoid hemorrhage) and obtained histopathological findings of the heart which could explain the bringing about of this syndrome.
(1) There was no complication of myocardial infarction in all cases of cerebral hemorrhage and subarachnoid hemorrhage in our cases.
(2) Marked myocardial fibrosis was revealed in 10 (37%) out of 27 cases of cerebral hemorrhage and slight myocardial fibrosis was showed in all 9 cases of subarachnoid hemorrhge.
On the contrary, in cerebral infarction studied by Iida there was complication of myocardial infarction in 18 (29%) out of 62 cases and was marked myocardial fibrosis in 21 (60%) out of 35 cases.
Our reserches showed that there are differences in the degree of occurrence of coronary insufficiency between cerebral hemorrhage and subarachnoid hemorrhage and cerebral infarction. They explained that it should be understood that cardiocerebral apoplexy is formed on the basis of severe coronary insufficiency in cerebrovascular disease.

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Cardiocerebral apoplexy is difined as atypical myocardial infarction the onset of which is symptom of cerebrovascular disease which characterized by frequent occurence in older age and malacic lesion of brain, as discribed by Fujii. But, its detailed pathological process remains to be a matter of much controversy. It is therefore necessary to perform further studies of cardiac lesions of cerebrovascular diseases as well as cerebral lesions of myocardial infarction for elucidating the pathology of cardiocerebal apoplexy. Now, we examined cardiac lesions of 35 autopsy cases of cerebral infarction. A study of coronary lesions and myocardial fibrosis has been attempted. 11 cases were associated with myocrdial infarction and 4 cases of them were regarded as cases of typical cardiocerebral apoplexy. Moreover, in many cases over 60 years old involving cases of myocardial infarction we observed marked myocardial fibrosis which strongly indicated the presence of coronary insufficiency. Marked myocardial fibrosis and myocardial infarction were apperrently prominent in cases of cerebral hemorrhage and subarachnoid hemorrhage as discribed by Katoh. If we classify cerebral infarction to massive cortical encephalomalacia, multiple encephalomalacia and few encephalomalacia, myocardial infarction and marked myocardial fibrosis are usually outstanding in the 2nd group.
Based on these observations, it may be presumable that the hitheto described cardiocerebral apoplaxy is a pattern of extreme severity and various cases of slight to moderate clinical coursed should be existed.

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The purpose of the present study is to observe the vasa vasorum of human intracranial arteries and to discuss their role in the development of cerebral atherosclerosis and thrombosis.
Postmorten angiography was performed in 24 cases and 67 cases were examined histologically. Serial section technique was employed in 21 cases with cerebral thrombosis.
Adventitial vasa vasorum, which were derived from the small branches of the intracranial arteries or from the vasa vasorum of the extracranial arteries, were always seen in the proximal part of the vertebral artery, and were seen in 44.4% of its distal part. They were found in 16.0% of the distal part of the basilar artery, 46.8% of the intracranial internal carotid artery and less frequently in more peripheral arteries.
Two types of intimal vasculature were distinguished: one originated from the adventitial vasa vasorum and the other from the arterial lumen. The adventitial vasa vasorum occasionally penetrated into the media and even into the thickened intima through disrupted internal elastic lamina, where the arterial lumen was narrowed more than 40%.
Intimal vascularization from the arterial lumen was seen in the arteries over 40% of narrowing and was more frequently found in the arterial segments with thrombi. However, it was less frequent in the intracranial arteries than in the coronary artery. Intimal vasculature showed several pathologic changes such as marked dilatation, stenosis, obstruction and hemorrhage. Intimal vessels distributed in deeper layers of the laminated thickened intima. These intimal thickening was usually eccentric and atheromatous necrosis was also localized in each layer accompaning foam cell accumulation and hemosiderin deposition.
These findings suggested that the eccentrically thickened intima with intimal vascularization was a sequel of the organization of thrombus. In the vertebral, basilar and internal carotid arteries, the adventitial vasa vasorum penetrated into the thickened intima through disrupted internal elastic lamina and anastomosed with intimal capillaries from the arterial lumen. This fact also suggests an important role of the adventitial vasa vasorum in organization of thrombi in the cerebral arteries.

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The important role of hyperlipoproteinemia to the development of atherosclerosis have already been recognized. Most of studies on hyperlipoproteinemia have been made from the aspects of the lipid metabolism. However, studies on apolipoproteins should be very important to approach to the development of atherosclerosis. In this study immunological properties and amino acid compositions of apolipoproteins in the normal and hyperlipoproteinemic subjects (familial hyperlipoproteinemia, non-familial hyperlipoproteinemia, nephrotic syndrome, diabetes mellitus, hypothyroidism, obstructive jaundice and liver cirrhosis) were examined.
Antisera were prepared by injecting male rabbits with purified LDL, apo LDL, HDL, apo HDL and whole serum emulusified with complete Freund's adjuvant. The immunological studies were carried out by the techniques of double diffusion and immunoelectrophoresis. Protein hydrolysates (6N HCl, 110°C) of delipidated specimens of LDL and HDL were analyzed on a Hitachi KLA-5 amino acid analyzer.
Apo LDL in the normal subjects reacted with anti-whole serum, anti-LDL and anti-apo LDL, but did not react with anti-HDL and anti-apo HDL. Apo HDL in the normal subjects reacted with anti-whole serum, anti-HDL and anti-apo HDL, but did not react with anti-LDL and antiapo LDL. Apo LDL in the hyperlipoproteinemic subjects reacted with anti-apo LDL obtained from the normal subjects. These reactions were consistent with those between normal apo LDL and anti-apo LDL. The same results were observed in the apo HDL of hyperlipoproteinemic subjecta. No differences of immunological properties of apo LDL and apo HDL existed between normal and various hyperlipoproteinemic subjects. Above mentioned immunological results show that the lipoproteins increased in the various patients are same quality with those obtained from the normal subjects.
Gel-filtration of apo LDL through Sephadex G-200 produced two fractions, and fraction I was a major component. Apo HDL was seperated into six fractions, and fraction III was a major component.
Amino acid composition of apo LDL fraction I was almost consistent with that of apo LDL. Amino acid composition of apo HDL fraction III was also almost similar to that of apo HDL. Amino acid compositions of apolipoproteins obtained from the hyperlipoproteinemic subjects were almost consistent with those from the normal subjects. It is interested that there exist the patients among familial hyperlipoproteinemia whose amino acid compositions of apolipoproteins were significantly different from those of other subjects.
The precise mechanism is still unknown that, in spite of differences of amino acid compositions, no difference was observed in the immunological properties of these familial hyperlipoproteinemia.

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