Fifty-one patients with lacunar stroke admitted to our clinic during a period of 16 years were selected for the present study. There were 39 cases with pure motor hemiplegia, nine cases with pure sensory stroke and three cases with dysarthria clumsy hand syndrome. Hypertension was observed in 33 cases (64.7%), and diabetes mellitus in 16 cases (31.4%). Younger patients with lacunar stroke were more often hypertensive than older ones. Computerized tomography and autopsy upon cases with pure motor hemiplegia revealed responsible cerebral lesions in the basal ganglia. Majority of cases with lacunar stroke showed good recovery from neurological deficits.
Hypertension is considered to play a significant role in the pathogenesis of the lacunar stroke. The incidence of hypertension among patients with lacunar stroke in the present study, however, is much less frequent than that shown in the Fisher's series. Consequently, less often, lacunar stroke is conceivable to occur as a result of functional changes in circulation, or as a consequence of atherosclerotic involvement of penetrating arteries and larger arteries.

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The present report dealt with hypertrophy and acute vascular lesions of cerebral arterioles in hypertensive rabbits. Hypertension was produced by clipping of the renal artery with intact contralateral kidney (two-kidney hypertension) or with contralateral nephrectomy (one-kidney hypertention).
(1) Wall areas of cerebral arterioles were measured in 47 animals which had been killed one week after clipping of the renal artery (two-kidney hypertension) or sham-operation. The wall areas were calculated from the external and internal diameter of arterioles in hypocampus on coronal sections of the brain. The incidence of arterioles with greater wall areas was significantly increased in hypertensive animals as compared with normotensive ones. These results indicate that hypertrophy of cerebral arterioles develops already in the early stage of hypertension.
(2) Acute vascular lesions and increase in vascular permeability were studied in one-kidney hypertensive rabbits with or without cerebral hemorrhage. Colloidal carbon technique was used to detect increase in vascular permeability. Necrotic changes of cerebral arterioles were found in 14 of 18 animals with cerebral hemorrhage, but was found in none of 15 animals without it. Arteriolar necrotic changes of other organs were frequently found in both animals with and without cerebral hemorrhage. Increase in vascular permeability of arterioles was more widely distributed than the necrotic changes. Vascular permeability was increased not only in arterioles but also in larger arteries including aorta and carotid artery in some of animals with cerebral hemorrhage.

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In this symposium, I would like to show the actual clinical statistics of two major pathologies, intracerebral hemorrhage and cerebral infarction which are closely related to the arterio- and arteriolosclerotic change in the brain and bring forward two problems in the field of neurosurgical clinic; one regarding to the mechanism of hematoma formation in intracerebral hemorrhage and another concerning to the STA-MCA anastomosis for cerebral infarction.
Since 5 years ago, we admitted 600 cases of cerebro-vascular disease in our hospital which included 137 cases of the lateral type of intracerebral hematoma. As our principle, we admitted all these cases as early as possible after the attack and operated them in the peracute stage or within 7 hours after the attack if it is indicated. In order to elucidate the mechanism of hematoma formation, we took the biopsy specimen from the tip of ruptured lenticulo-striate artery at operation in these cases and examined microscopically and electron-microscopically under the cooperation of professor Takebayashi who is supposed to give an address after me. From the statistics of the lateral type of intracerebral hematoma, a quarter of them was the severe type which showed disturbance of consciousness of more than semicoma and marked hemiplegia on admission. A half of them was the moderate type namely they showed disturbance of conciousness of stupor and apparent hemiplegia on admission. The rest of a quarter was the mild type of mild disturbance of consciousness. By the examination of computerized tomography, the severe type was proved to have larger hematoma than the moderate type. And the mild type showed the smallest hematoma.
This difference of the size of hematoma seemed to be caused mainly by the size of ruptured lenticulo-striate artery. In the early craniotomy for these cases, the bleeding source could be clearly identified; Lenticulo-striate artery was usually ruptured not at its trunk but at its branch of the caliber of approximately 150-200μ. Bleeding source was mostly single and the hemorrhage could be controlled by its single hemostasis.
Pathological findings of these ruptured arteries will be stated by Prof. Takebayashi later. Anyhow, from the above findings, we concluded that intracerebral hematoma is as a rule, caused by the rupture of a single branch of the lenticulo-striate artery and is accomplished by the arterial bleeding mostly within 3 hours after the apoplectic attack.
At the last, I discussed shortly on the problem of STA-MCA anastomosis which is a bypass operation between the superficial temporal artery and the middle cerebral artery in the case of cerebral thrombosis.

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Light microscopic and ultrastructural changes and morphogenesis of intracerebral arteriosclerotic lesions were studied in autopsy cases. The lesions included were not only arteriosclerosis itself but also miscellaneous arterial changes which might stimulate cell proliferation and fibrosis in the intima resulting in arteriosclerosis.
Cellulofibrous intimal thickening occurred predominantly at bifurcations of the intracerebral arteries more than 150μ in diameter in aged and hypertensive patients. Elastin and collagen were formed in the basement membrane (BM)-like substance in the subendothelial tissue and also around intimal smooth muscle cells, particularly markedly in the latter in the deep intima. Collagen replaced the BM-like substance which seemed to be used as a material for collagen formation.
There are two kinds of atherosclerosis depending upon the atherogenesis, that is, fiber disintegration type and cell disintegration type. In the intracerebral arteries the latter was more common than the former. Various sized arteries down to 60μ in diameter were involved in the latter atherosclerosis.
The accumulation of foam cells or ceroid in the intima with neither intimal cell proliferation nor fibrosis was designated as intimal lipoidosis in this paper. The lesions were found in small arteries less than 150μ in diameter, sometimes encountered in ones as small as 50μ. Foam cells in the lesions were presumably derived from circulating monocytes. Ceroid occurred microscopically in the whole intima as rims of acid fast material surrounding large fat globules. The vascular ceroid in the brains retained the similar staining properties to the ceroid in liver cells. It was disclosed electron microscopically as a wavy laminated structure with a fingerprint pattern having an electron-dense osmiophilic layer of about 70Å thick alternating with an electron lucent layer about 100Å thick.
The arteries involved in intimal edema had several layered myointimal cells just beneath the endothelial lining. It was strongly suggested that the myointimal cells could be derived from the endothelial cells, because of having similar ultrastructures to them.
Pseudocalcification was observed in and about small blood vessels of the globus pallidus. Electron microscopic studies revealed that dense granular substances, consisting of phosphorus, calcium, iron, and sometimes magnesium were deposited in necrosed smooth muscle cells and in BM-like substance increased around the smooth muscle cells in the media.
Infrequent cases had meningeal and cortical arteries involved in amyloidosis. Intracerebral microaneurysms which were the direct cause of hypertensive intracerebral hemorrhage were formed on degenerated arterial segments with amyloidosis in a few cases.
In large microaneurysms over 350μ in diameter, thrombosis was frequently formed in their lumina. Organization of the thrombi was brought about by endothelial proliferation on the surface of the thrombi and by new capillary formation in the thrombi from the surface. Intimal cells, which were presumably derived from endothelial cells, and BM-like substance appeared beneath the endothelium. Fibrous nodular arterial lesions were completely organized microaneurysms with occluded thrombi.

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On the emergency surgery against hypertensive intracerebral hemorrhages of 18 patients, arteriolar biopsies were performed from each lenticulostriate arteries with ruptured hemorrhages. They were, then, serially sectioned and were examined by electron-microscopy as well as light microscopy.
The examination revealed six disrupted ostiums of the arterial walls causing massive hemorrhages, in 18 individuals. Four of them were ruptured at bifurcations of the arteries with arteriosclerotic changes which were characterized by smooth muscle cell degeneration in media as well as in intima, if thickened, and termed “exhausted degeneration” in this paper. That is, irregular or moth eaten-like atrophy of the smooth muscle cell with remarkable increase of its basement membranoid materials. The internal elastic lamella was sharply disrupted without any previous alterations at disrupted ostium. Ultrastructurally, there is, however, neither fibrinoid nor insudative changes at the disrupted portion, as a previous alteration. The disruption was not respected to lipid deposition in intima and/or media.
Microneurysm (“angionecrosis”) was another cause of the rupture in two remained patients. The wall of the microaneurysm, however, ultrastracturally comprised quit various components; only lamellar fibrin sheats or with intermingled inflammatory cells and/or lipophage, or with partially or completely regenerated endothelium and imigration of immatured smooth muscle cell (or indeterminated cell), intercellular deposition of calcium as well as ferrum as an electron dense granule, which were detected by electron microscopic x-ray analyzer. Those are quit different from that in hypertensive fibrinoid thickening of arterioles in other organs. Thus, the microaneurysm which thought causally relate to primary intracerebral hemorrhage, is rather easily understandable to be a resorbed state of a smaller or incomplete, subclinical hemorrhage.
On conclusions, hypertensive intracerebral hemorrhages occure more frequent from primary rupture of arteriosclerotic lesions than from microaneurysm which is more likely to be old recanalized, small hemorrhage.

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Serum lipoproteins were investigated in 151 patients undergoing chronic hemodialysis for renal failure. Hyperlipoproteinemia (over 700mg/100ml) was demonstrated in 34 cases of them. According to the criteria of WHO, those were classified as follows: Type III, 12 cases; Type IIb, 8 cases; Type IV, 7 cases; Type IIa, 5 cases; Type V, 2 cases. With polyacrylamide-gel disc-electrophoresis, abnormal lipoprotein bands such as extra pre-beta, extra beta and mid bands were recognized in many cases of 151 patients.
It is well known that the metabolism of the circulating lipoproteins is mediated by some enzymes. The activities of the following enzymes were determined: Total postheparin lipolytic activities were decreased in all subjects, and protamine-inactivated as well as protamine-resistant lipase activities reduced in almost all of them. Lecithin cholesterol acyltransferase activities were markedly decreased in about one half of the subjects, more and more reduced by heparin infusions associated with the hemodialysis procedure.
Basal serum fatty acid concentrations were supra-normal in only 6 cases of 49 patients. By contrast, there were considerable increases of free fatty acid in all subjects during the hemodialysis procedure. Hypertriglyceridemia was demonstrated in 21 cases of 71 patients, and hypercholesterolemia was observed in 11 cases of 71 patients. Concentrations of serum lipid peroxide were increased in 33 of 105 subjects.
It is said that accelerated atherosclerosis is a major risk to long-term survivors on maintenance hemodialysis. Although the nature of the causal chain linking plsma lipids and lipoproteins to atherosclerosis is as yet unclear, clinical and experimental studies have provided evidences that the concentration of plasma beta, pre-beta lipoproteins and remnants play an essential role in atherogenesis. Therefore, we must consider therapeutic measures for eliminating hyperlipidemia as well as hyperlipoproteinemia (abnormal lipoproteinemia) in order to protect the patients with chronic renal failure from atherosclerosis and subsequent arteriosclerotic diseases.

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Changes in the serum lipoprotein pattern following dietary treatment, the genetic relationship and the postheparin lipolytic activity (PHLA) were investigated in patients with hyperlipoproteinemia associated with the presence of an intermediate lipoprotein.
In these patients the serum lipoprotein pattern observed on polyacrylamide gel disc electrophoresis (PAGE), exhibited a “Mid-band” or the broad band ranging between pre-β- and β-lipoproteins. But the major peak of the mid-band was located at different positions for each subject, in some cases by the side of pre-β- and others by the side of β- lipoprotein. Most of the intermediate lipoproteins showing the mid-band on PAGE in these patients were detected as the intermediate lipoprotein of the density range 1.006-1.019 by means of the ultra-centrifugal separation and the estimation of the lipid composition.
In M family with hypertriglyceridemia, the propositus (G. M.) had an extremely high serum triglyceride level, above 2, 500mg/dl on a regular or high-fat diet. At this time the lipoprotein electrophoresis demonstrated a pattern of type V hyper-lipoproteinemia. While he received a low-fat diet (fat<15g per day), his serum triglyceride concentration was reduced to 500mg/dl and the serum lipoprotein pattern showed an accumulation of intermediate lipoprotein with a disappearance of chylomicron. G. M. and his wife were first cousins. One of their daughters (F. M.) suffered from abdominal pain and diarrhea with “fat-induced” hypertriglyceridemia and also showed an intermediate lipoprotein. Another daughter died of acute pancreatitis with severe hypertriglyceridemia more than 2, 000mg/dl. Both G. M. and F. M. had a deficiency of PHLA both for hepatic triglyceride lipase (H-TGL) and extrahepatic lipoprotein lipase (LPL) activities, so that the M family was considered to be affected with a subtype of familial lipoprotein lipase deficiency.
S family had three siblings with hyperlipoproteinemia and two of them had tendon xanthomas. The propositus (H. S.) was admitted with acute myocardial infarction and her lipoprotein pattern had intermediate lipoprotein by the side of β-lipoprotein. All of the family members with hyperlipoproteinemia had similar lipoprotein patterns. Following a calorie-restricted diet, H. S.' serum triglyceride concentration was reduced from 245mg/dl to 96mg/dl and her lipoprotein pattern turned to a typical type IIa phenotype with the disappearance of the intermediate lipoprotein. Thus, the S family was characterized essentially by familial hypercholesterolemia.
In most of the other subjects with hyperlipoproteinemia in this study, H-TGL was below normal but not severely deficient. Only one of these cases (K. O.) that was defined as typical “broad-β disease” by clinical symptoms and the chemical index showed a markedly reduced H-TGL. One of the other cases, R. F., with serum triglyceride level more than 300mg/dl had reduced H-TGL on a regular diet. However, when his triglyceride concentration returned to normal value on a fat-restricted diet, his H-TGL was also within the normal range. At both of the periods, the pattern of lipoproteins characterized by the presence of an intermediate lipoprotein was not significantly changed. Unknown mechanisms other than the deficiency in H-TGL must be involved in the impaired metabolism of the intermediate lipoprotein.

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An inverse relationship has been demonstrated between plasma high-density lipoprotein (HDL) cholesterol level and the incidence of coronary heart disease. Therefore, measurement of HDL cholesterol is of great importance.
In the present study we compared three different methods for the estimation of HDL cholesterol in plasma samples.
The methods were as follows.
(1) Ultracentrifugation.
Cholesterol was determined in the d<1.063g/ml fraction from 2ml of plasma after centrifugation at 100, 000 G for 20hr.
(2) Heparin-Mn²⁺ precipitation.
To 2ml of plasma we added 80μl of a heparin solution (5, 000units/ml) and 100μl of MnCl₂ solution (1 Mol/l), and immediately mixed. After the sample had stood for 30min. in an ice bath, the sample was centrifuged (1, 500G, 30min., 4°C) and cholesterol was determined in the supernate.
(3) Sodium phosphotungstate-Mg²⁺ precipitation.
To 2ml of plasma we added 200μl of a sodium phosphotungstate solution and 50μl of MgCl₂ solution (2 Mol/l). After mixing in a vortex mixer, the sample was centrifuged (1500G, 30min., 4°C) and cholesterol was determined in the supernate.
HDL cholesterol levels of 138 subjects were measured by two precipitation methods and for 32 of them the three methods were used simultaneously.
The correlation coefficients were 0.939 between ultracentrifugation and heparin-Mn²⁺ precipitation methods, 0.919 between ultracentrifugation and phosphotungstate-Mg²⁺ methods, and 0.966 between the two precipitation methods.
Our results showed similar results for the cholesterol content of HDL determined by the three methods.
Both precipitation methods are simple and appropriate for routine clinical laboratory use.

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Selective coronary cine-angiography is a standard method for the assessment of the degree of coronary artery stenosis. This study was undertaken to evaluate the association between serum lipid and lipoprotein levels, and the degree of coronary artery disease (CAD) in 110 patients studied by selective coronary cine-angiography. Coronary atherosclerosis were assessed by signs of narrowing of caliber exceeding 25% or more in at least one of the three main branches of coronary arteries. All the patients were devided into four groups depending on the number of coronary artery involved. There was no significant difference among these groups with regard to age, sex, obesity, glucose intolerance, hypertension and smoking. The results were as follows:
1) The mean serum cholesterol levels in the groups with CAD were significantly higher than that in the group without CAD, and the higher the serum cholesterol level was, the more the number of involved coronary vessels increased.
2) The mean serum triglyceride levels in the groups with two and three involved vessels were significantly higher than that in the group without CAD, and the higher the serum triglyceride level was, the more the number of involved coronary vessels increased.
3) In the group with involved three vessels, the concentrations of serum cholesterol and triglyceride were the highest of all the groups.
4) The mean VLDL-cholesterol levels were significantly higher in the groups with two and three involved vessels than that in the group without CAD and the mean VLDL-triglyceride levels were significantly higher in the group with involved three vessels than that in the group without CAD.
5) The mean IDL-cholesterol levels were significantly higher in the groups two and three involved vessels, but there were no significant differences in the mean IDL-triglyceride levels among these groups.
6) The mean LDL-cholesterol and LDL-triglyceride levels in the groups with CAD were significantly higher than that in the group without CAD.
7) In the group with three involved vessels, the concentrations of cholesterol in VLDL, IDL and LDL fractions were the highest of all the groups.
8) The ratio of HDL-cholesterol to serum cholesterol was significantly lower in the groups with two and three involved vessels than that in the group without CAD.
9) HDL-phospholipid levels were significantly lower in the groups with two and three involved vessels, and the ratio of HDL-phospholipid to serum phospholipid was significantly lower in the all groups with CAD than that in the group without CAD.

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Serum low density lipoprotein (LDL) and very low density lipoprotein (VLDL) levels of 254 patients with cerebrovascular disease and 215 cotrols were determined by the heparin-Ca⁺⁺ precipitation method. Serum high density lipoprotein-cholesterol (HDL-C) concentrations in the supernatant of heparin-Ca⁺⁺-precipitated serum were also measured.
Serum LDL concentrations of healthy controls, who had no abnormalities in physical findings, blood pressure, urinalysis, ECG findings, etc., in-creased with age in female subjects. In male controls, the value in fourties was highest. The average values of healthy men and women under 39 years old were 424±170.2mg/dl and 377±130.6mg/dl, respectively. Serum VLDL concentrations were unaffected by age, and average values of male and female controls under 39 were 113±114.1mg/dl and 52±35.5mg/dl, respectively. Serum HDL-C concentrations increased with advancing age in male controls but were unaffected by age in female controls. The average HDL-C values of healthy men and women under 39 were 57±16.9mg/dl and 63±16.5mg/dl, respectively. In the obese subjects aged from 40 to 59, serum VLDL value was higher and HDL-C was lower than the values in age-matched healthy controls.
Average serum LDL value of the patients passed more than one year from stroke was higher than that of those within one year. Therefore to eliminate the possible influence of time after stroke on serum lipid profile, patients passed more than one year from onset were excluded. Average serum LDL and VLDL values of 89 patients with cerebral infarction were 574±162.8mg/dl and 159±100.9mg/dl, respectively, and the values of 34 patients with cerebral hemorrhage were 460±124.3mg/dl and 123±79.1mg/dl, respectively. Serum LDL value of patients with infarction was significantly higher than those of patients with hemorrhage and healthy controls. Serum VLDL levels of both patients were significantly higher than control value. Average serum HDL-C value of 123 cerebrovascular patients was 57±16.8mg/dl and it was significantly lower than control value. There was no difference between the values of two types of diseases.
These results may suggest the close association of abnormal lipid metabolism with cerebrovascular disease. Further investigation considering many factors should be necessary and it may contribute to the elucidation of the role of lipid metabolism in cerebrovascular disease that is not evaluated simply by the measurements of serum cholesterol or triglyceride levels.

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Quantitative analysis of HDL, LDL, VLDL, cholesterol and triglyceride in sera were carried out in patients with ischemic heart disease. In addition, analysis of serum lipoprotein by polyacrylamide gel disc electrophoresis was performed in the same subjects. The concentration of serum HDL was measured by rocket immunoelectrophoresis using an anti-α₁-lipoprotein-serum (BEHRINGWERKE AG) and that of serum LDL, VLDL was determined by precipitating method.
Subjects consisted of 51 patients with ischemic heart disease and 42 healthy men and women. Cases with ischemic heart disease were classified into groups of old myocardial infarction, acute myocardial infarction, angina pectoris and arrhythmia, i.e., atrial fibrillation and/or A-V block.
Mean values of serum HDL were lower in each group of ischemic heart disease than in the healthy group. Mean levels of serum cholesterol were significantly higher in each group of ischemic heart disease than in the healthy group, but each mean level was under 200mg/dl. There were no significant differences of the mean levels of serum triglyceride, LDL and VLDL between both groups. In polyacrylamide gel disc electrophoresis, the midband appeared more frequently in each group of ischemic heart disease than in the healthy group.
Results of our study suggest that low level of serum HDL play an important role in the development of ischemic heart disease. Frequent occurrence of the midband in patients with ischemic heart disease suggests another abnormal lipid metabolisms in these patients.
In addition, the consistence of the result of lipid analysis in the group of arrhythmia with those in the groups of myocardial infarction and angina pectoris is notable.

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Livers from fasted pigs were perfused with Krebs-Ringer bicarbonate buffer containing glucose, bovine serum albumin, amino acids, and washed human erythrocytes.
Hepatic catabolism of very low density lipoprotein (VLDL) to intermediate density lipoprotein (IDL) or low density lipoprotein (LDL) was investigated by adding ¹²⁵I-VLDL to the perfusate, perfusing livers for 4 hours, and measuring the radioactivity in the IDL and LDL ranges after sequential and rate-zonal ultracentrifugations. It was proved that VLDL had been catabolized neither to IDL nor to LDL by the liver.
Since VLDL was shown not to be catabolized by the liver, it would be proper to consider that VLDL obtained from liver perfusion represent a nascent form of VLDL.
Very low density lipoproteins from 7 liver perfusions were analyzed by laser self-beat spectroscopy. The autocorrelation functions were accurately described by a single exponential which is considered to represent the uniformity of particles. The size of VLDL was calculated by measuring correlation time (τ_c), calculating translational diffusion constant, and putting it into the Stokes-Einstein's equation. The diameter of VLDL synthesized by the perfused liver from fasted pigs was estimated to be 647±39Å.
It was concluded: (1) The liver is not able to catabolize VLDL (2) The nascent VLDLs synthesized by the liver are uniform in size (3) Variable VLDLs which are seen in the circulation are produced during catabolism of these VLDLs by lipoprotein lipase. It was suggested that the size and composition of nascent VLDL, which is synthesized by the perfused liver, may depend on the perfusion and/or animal conditions.

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It is indicated that liver plays an important role in lipoproteins metabolism. Recently relationship between plasma concentrations of high density lipoprotein (HDL) and coronary heart disease has been reported. The present study was to investigate lipoproteins synthesis, especially HDL₃ or Apo A-I, using a culture system of rat liver parenchymal cells. The cell culture system has several advantages such as cell homogeneity and the possibility of longer incubation than liver perfusion system that has been used by many investigators.
The isolation of rat hepatocytes was performed accordint to the method of Berry and Friend with some modifications. The hepatocytes were cultured in the complete synthetic culture media HI/WO₅/BA₂₀₀₀ (International Scientific Ind.) under 5% CO₂/95% air at 37°C. Twenty-four and 48 hours after cell inoculation, light and electron microscopic study and the following metabolic function of the cells were examined: 1) gluconeogenesis from pyruvate and lactate (179.5 nmoles/mg cell protein/hour), 2) induction of tyrosine aminotransferase by 10^-5M dexamethasone (43.8mU/mg cell protein), 3) membrane transport of C¹⁴-α-aminoisobutyric acid (AIB)(9.5 AIB distribution ratio) and 4) albumin synthesis (156μg/mg cell protein/hour). The results of metabolic and morphological studies showed that integrity of cultured cells was well maintained at least for 48 hours after inoculation.
Rat HDL₃ (1.110<d<1.210g/ml) was isolated by sequential ultracentrifugation. Blood was obtained from male rats weighting 250 to 300g who had been fasted for 24 hours. The density of plasma was adjusted to d=1.110g/ml with solid KBr and centrifuged in a Type 40.3 rotor on the LS-50 preparative ultracentrofuge (Beckamn Instruments Inc.) for 22 hours at 105, 000×g. After 22 hours, the top 2ml was removed by tube slicing technique. Then, the density of the infranatant solution was adjusted with solid KBr to d=1.210g/ml and centrifuged at 105, 000×g for 44 hours. HDL₃ was washed at d=1.210g/ml to remove contaminating serum albumin and was dialyzed exhaustively against 0.154M NaCl, pH 7.5 or double distilled water at 4°C. The rationale for using HDL₃ is that this density fraction contains predominantly Apo A-I and a minimal amount of other apolipoproteins, as compared to other density classes. Anti-rat HDL₃ serum was prepared by several intraperitoneal injections of rat HDL₃ with equal volumes of Freund's complete adjuvant at intervals of 7 days. The anti-sera were examined by immunodiffusion and immunoelectrophoresis with various antigens such as VLDL (d<1.006g/ml), LDL₂ (1.019<d<1.050g/ml), HDL₃ and peak II (containing only Apo A-I) and peak III (containing mainly C peptides) fractions of apo-HDL₃ from Sephadex G-100 column chromatography. Anti-rat HDL₃ serum used for current study reacted only with HDL₃ or Apo A-I. The concentrated culture media in which hepatocytes were cultured for 20 hours, were examined immunologically. A single precipitin line was observed between culture media and anti-rat HDL₃ or anti-rat Apo A-I on immunodiffusion and immunoelectrophoresis, indicating the presence of Apo A-I in cultured media. HDL₃ fraction (1.110<d<1.210g/ml) isolated from pooled culture media was analyzed for identification of apolipoproteins. The electrophoretic patterns of totally delipidized HDL₃ fractionated from culture-media on 7% polyacrylamide gel with 8M urea were comparable with electrophoretic patterns of rat plasma Apo-HDL₃. Not only the Apo A-I band but also the Apo C bands were observed. These findings indicated that HDL₃ or Apo A-I synthesis and secretion by cultured hepatocytes.

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The concentrations of apolipoprotein AI, a major peptide of high density lipoprotein (HDL), have been measured by immunoelectrophoresis in samples of serum from twelve subjects who subsequently developed a coronary event during 2 years of follow-up and compared with those in serum from sixteen matched control subjects. The mean apolipoprotein AI concentration in the cases was significantly lower than that in the controls, independently of the serum total cholesterol and triglyceride concentrations. There was no significant difference between the cases and controls in mean HDL cholesterol: apolipoprotein AI ratio. Within several case-control pairs, however, the difference in apolipoprotein AI concentration was proportionately much less than that in HDL cholesterol. On discriminant function analysis, apolipoprotein AI concentration was a less powerful predictor of coronary heart disease than was HDL cholesterol.

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Acyl-CoA synthetase was estimated as an amount of acyl-CoA synthesis from radioactive palmitic acid in the presence of ATP, CoASH, and MgCl₂.
Characteristics of acyl-CoA synthetase in arterial wall extract were examined. Km values for palmitate or ATP or CoASH or Mg²⁺ were 0.29, 3.45, 0.03, 4.0mM, respectively. The optimum pH was around 8.0 and the highest specific activity was found in microsomal fraction. These characteristics were almost similar to those of liver acyl-CoA synthetase.
Acyl-CoA synthetase was inhibited by divalent cations such as Cu²⁺, Fe²⁺, Zn²⁺ and Ca²⁺ and was activated by serum, bovine serum albumin and lipoproteins. Changes of these enzyme activities were found in some pathological conditions. Increase in aged or hypercholesterolemic rat was found and in experimental hypertensive or diabetic or tocopherol deficient condition, the activities were found to be decreased. The relationship between regulatory factors and changes of this enzyme activities in pathological conditions was discussed.

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Relationship between ischemic heart disease (THD) and serum lipids, high density lipoprotein (HDL) lipids and lecithin: cholesterol acyl transferase (LCAT) activity was studied.
Serum levels of total cholesterol, free cholesterol and triglyceride in the patients with IHD were significantly higher than those in normal controls (P<0.01). The levels of total cholesterol, free cholesterol and triglyceride in HDL were lower in the patients with IHD than those in normal controls (statistically insignificant). However, ratios of HDL-total cholesterol/serum total cholesterol and HDL-free cholesterol/serum free cholesterol were significantly lower in the patients with IHD than those in normal controls (P<0.02, P<0.05, respectively). It has been proposed that reduction of HDL concentration or HDL/total lipoproteins ratio may accelerate the development of coronary atherosclerosis.
LCAT activity was positively correlated to serum total cholesterol, free cholesterol, triglyceride and phospholipid and free cholesterol/total cholesterol ratio. While, LCAT activity was negatively correlated to HDL-total cholesterol/serum total cholesterol and HDL-free cholesterol/serum free cholesterol ratio.
There was no difference in LCAT activity between the patients with angina pectoris and old myocardial infarction, and normal controls. The patients free of hyperlipidemia, hypertension, obesity, and heavy smoking had a significantly lower LCAT activity than normal controls (P<0.05). These results suggest that a low LCAT activity is a risk factor of coronary atherosclerosis. Thus, HDL and LCAT may be protective system of atherosclerosis that transfer cholesterol from the arterial wall to the liver.

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It has been reported by many investigators that phospholipids are markedly increased in atherosclerotic lesions as well as cholesterylesters. We studied the characteristics of, and the mechanisms of regulation of CDP choline: 1, 2-diacylglycerol cholinephosphotransferase (CPT) activity, one of the enzymes which synthesize phosphatidylcholine, in rat arterial wall comparing with those in rat liver.
CPT activity showed a linear relationship with incubation time and concentration of enzyme protein, upto 30 minutes and 0.1mg of protein per tube, respectively. Apparent Km value for CDP choline was 0.019mM in aorta and 0.015mM in liver. A linear activation of the enzyme was observed with upto 3.2mM of diolein in aorta and 1.6mM in liver. A maximum activity was observed around pH 8.5 in both in aorta and liver. In sub-cellular fractionaton of aorta, the highest specific activity per protein content was located in microsomal fraction.
When Ca²⁺, Zn²⁺, Cu²⁺ and Ba²⁺ were added, CPT activity was remarkably inhibited, but Mn²⁺ and Co²⁺ increased the activity in aorta in the presence of Mg²⁺.
CPT activity was increased by addition of a low concentration of serum. Very low density lipoprotein (VLDL) and low density lipoprotein (LDL) activated CPT activity, though high density lipoprotein (HDL) inhibited the activity. This may imply that LDL induced CPT activity in arterial wall, directly or indirectly.
CPT activity was also compared in various pathological states. Higher activity was found in hypercholesterolemic and vitamin E deficient rats than the controls. Fifty week-old rats showed significantly lower CPT activity in aorta than 4 weekolds. In spontaneous hypertansive rats, CPT activity was significantly lower in Wistar-King strain rats.
Further investigations are required in order to clarify the mechanism of regulation of CPT activity in arterial wall from the point of view of various pathological states of arterial wall.

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Acid lipase and acid cholesterol esterase (CEase) in rat arterial wall were largely associated with lysosomal fraction. Then lysosomal fraction of arterial wall was preincubated with 0.04% Triton-X 100 for 5min. at 0°C. Acid lipase and acid CEase were released from lysosomal fraction. The released lipase and CEase were more inactivated than those associated with lysosomal fraction, when they were preincubated for 10min. at 37°C.
When lysosomal fraction was treated with various concentration of Triton-X 100 for 5min at 0°C and centrifuged, acid lipase was released from lysosome in a small amount than acid CEase. From these results, the relationship between stability of lysosomal enzyme activities and lipid accumulation in lysosome was discussed.
Higher activity of lipase was observed using sonicated substrate than one homogenated with a Hiscotron (Nichion, Japan). High activity was observed when substrate mixed with phosphatidylcholine and arabic gum was sonicated. Lipase activity was elevated by an addition of 0.25mM phospholipid especially phosphatidylserine and phosphatidylcholine to reaction mixture, and was lowered by an addition of 2mM or more dosage of those phospholipids. It is suggested that phospholipids might be an important regulatory factor of lipase activity in the arterial wall.
We reported that serum nonspecific catboxyl esterase (tributyline hydrolase) might be converted to lipase (triolein hydrolase) in aorta. Further study has been done using lysosomal fraction of the arterial wall. Purified serum esterase was incubated with lysosomal fraction which was pretreated with heparin to remove lipase. Elevation of acid lipase activity was observed. But when boiled esterase was incubated with it, no elevation of lipase activity was observed. These results suggested that serum nonspecific carboxyl esterase might be converted to lipase in lysosome of aorta. Further studies were now in progress to certify this possibility.

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Serum lipid and lipoprotein levels were studied in 23 normal subjects, 50 heterozygous and 4 homozygous patients with familial hypercholeste-rolemia (FH).
1) The mean±SD of serum cholesterol (CHOL) levels in the normal subjects, the heterozygotes and the homozygotes were 165±22, 342±47 and 532±44mg/dl, respectively.
2) The hyperlipidemias observed in FH were selectively due to the raised LDL concentration in itself.
3) The LDL-CHOL levels in the normal subjects, the heterozygotes and the homozygotes were 102±21, 265±66, 463±63mg/dl, respectively.
4) The serum CHOL levels correlated highly with the LDL-CHOL levels (r: 0.980, p<0.01).
5) The serum phospholipid (PL) levels highly correlated with that of the LDL-PL (r: 0.925, p<0.01).
6) FH is a disease characterized by high concentrations of LDL with an increased proportion of CHOL and a decreased proportion of triglyceride and PL.

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Familial homozygous hypercholesterolemia is a fatal disease in consequence of coronary insufficiency or vascular disease. In addition, drug and diet therapy is often ineffective. Intravenous hyperalimentation is recognized as one of therapeutic procedures available for this disease. We treated one patient with familial homozygous hypercholesterolemia using intravenous hyperalimentation followed by good result in decreasing serum cholesterol level.
This 18 year-old female patient was suffered from anginal attack, xanthoma and her exercised-ECG showed ST-depressing in leads II, III, _aV_f and v_2-4. Coronary angiography showed the 90% stenosis in LAD with other stenotic lesions. Serum cholesterol level ranged from 758mg/dl to 1003mg/dl. On August 23, 1977 intravenous hyperalimentation was started with solution which contains 21% dextrose and 4% crystalline amino acids and electrolytes. Initially hyperalimentation provided 1, 800cal/day. However, serum cholesterol did not decrease so much while body weight increased. For that reason callory was reduced to 1200cal/day. After calloric reduction, serum cholesterol fluctuantly decreased into 363mg/dl on the 106th day after intravenous hyperlimentation therapy. Xanthoma, especially in the subcutane of Achilles tendon read, reduced as the decreasment of serum cholesterol level.
But anginal attacks showed no change in frequency. During this therapy, her general condition was kept in good status and serum total protein and albumin were maintained around 6.6-8.6g/dl and 3.3-4.2g/dl, respectively. On November 29, 1977, A-C bypass operation was done. After the 8th day of this surgery, the oral intake was started and intravenous hyperalimentation was ended and then serum cholesterol level again increased to 800mg/dl. As described above, intraveous hyperalimentation cause decline of serum cholesterol level, but the mechanism of this effect is not well known.

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Obesity, hyperlipidemia and some other metabolic abnormalities were investigated for school boys and girls in Shinjo City, Nara Prefecture. In 1968, obesity (body weight excess more than 20% of the ideal) was found in 4.5% of the children in primary school, and in 5.8% of the boys and girls in junior high school. Half of the obese children succeeded in weight reduction through dietary care and exercise and the body weight was in normal range when examined in 1972. Girls showed much better results than boys (rate of recovery: 62% for girls and 36% for boys). In spite of the effort against obesity, the frequency of obesity in general was not reduced at the last examination performed in 1972, because approximately the same number of children as those who succeeded in weight reduction newly developed obesity during the period between the first and the last examinations.
The mean value (±standard deviation) of serum cholesterol in boys and girls in junior high school was 166±28mg/dl and the value of serum triglycerides was 79±21. There were no sexual differences in serum lipids, but uric acid level was higher in boys than in girls (4.4±0.9 vs 3.8±0.6mg/dl, Folin method). Frequency of abnormalities which exceeded the normal range (mean+double s. d.) was 4.1% for cholesterol and 5.8% for triglycerides. Serum lipid levels of the obese were 177±33mg/dl for cholesterol and 97±35 for triglycerides.
Distribution ratio of cholesterol between β±pre β/α lipoproteins was 2.46±0.86 for males and 2.16±0.64 for females. Obese children showed a significantly higher value compared with nonobese children (3.27±1.17 vs 2.18±0.58, P<0.001). Compared with the non-obese and leans, obese children showed a significantly higher value of serum GPT and a lower value of serum alkaline phosphatase activity.

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During a period of 5 years, from 1973 to 1977, the total cholesterol values of 6, 352 males, and 10, 417 females, over 10 years old was measured in fasting state, using autoanalyzer. Blood pressure was measured in the spinal position. The electrocardiogram was classified by the Minnesota Cord. The results are as followed.
1) The mean total cholesterol value in Okinawa, according to age and sex was significantly higher than that of mainland Japan.
2) The incidence rate of hypercholesterolemia is also significantly higher in Okinawa compared to mainland Japan, (Kyushu).
3) The total cholesterol level increased in relation to blood pressure and the degree of obesity.
4) The major cause of the higher incidence rate of hypercholesterolemia in Okinawa, compared with mainland Japan (Kyushu), is the difference of fat intake between the two areas.
5) The mean total cholesterol value in the groups with ischemic change in the electrocardiogram was significant higher than that of normal group.

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Various column chromatographies were performed on the sera of patients with hypertension, diabetes mellitus or calcification of the aortic arch to study relationships between lipoproteins and binding hexoses in arteriosclerosis.
1) Serum binding hexose with lipid and serum binding hexose without lipid were fractioned from human sera by methods of sepadex G-100 column chromatography, bio-gel p-300 column chromatography, dialysis and aceton. chloroform absorption.
2) Though the concentration of serum binding hexose in patients with hypertension, diabetes mellitus or calcification of the aortic arch is obviously higher than that in healthy person, the cause of high values of serum binding hexose in these patients is mainly due to high values serum binding hexose without lipids.
3) Not only lipoprotein in serum but also binding hexose without lipid will have roles of importance as the causes of the mechanism in arterioscleosis.

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The mechanism of regulation of cholesterol esterase (E. C. 3. 1. 1. 13) in rat arterial wall was investigated. Cholesterol oleate [1-¹⁴C], sonicated with phosphatidylcholine, was used as a substrate. The enzyme has pH optima around 4.5 and 7.5. The acid and neutral cholesterol esterases were found to be mainly located in lysosomal and microsomal fraction, respectively, by differential ultracentrifugation method. Both enzyme activities were remarkably increased when the substrate which was prepared with phosphatidylcholine or phosphatidylserine was used. On the other hand, no increase in enzyme activity was observed with phosphatidylethanolamine or sphingomyelin.
Acid cholesterol esterase activity was increased when lower concentration of VLDL, (1.5μg/tube of protein concentration) was added to the reaction mixture, but it was decreased when a 12μg/tube of protein of VLDL or LDL or HDL was added. Neutral cholesterol esterase activity was increased by addition of each lipoprotein. When apoVLDL or apoLDL or apoHDL was added, both cholesterol esterase activities were decreased.
Further experiments are now in progress on the mechanism of regulation of cholesterol esterases.

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Wistar-King strain rats and spontaneous hypertensive rats were fed on a α-tocopherol deficient diet. A α-tocopherol supplemented diet (0.02% α-tocopherol added) was used as a control. Furthermore, α-tocopherol deficient diet group was divided into two groups. One was fed on a high cholesterol diet, and the other was fed on a normal cholesterol diet. A remarkable decrease in serum tocopherol concentration was observed in the α-tocopherol deficient diet groups, and in these groups, thiobarbituric acid reactive substance (TBA-RS) was considerably increased in the liver, brain, heart, kidney, and aorta. In these conditions, arterial enzyme activities concerning lipid metabolism, such as acid and neutral lipase activities, acid and neutral cholesterol esterase activities (CEase), cholesterol ester synthesizing activity (CEsyn.), triglyceride synthesizing activity (TGsyn.), choline-phosphotransferase activity (CPT) and acyl-CoA synthetase activity (aryl-CoAsyn.) were estimated. Acid and neutral lipase activities, acid CEase, CEsyn., acyl-CoAsyn. and TGsyn. decreased in α-tocopherol deficient Wistar-King strain rats fed on the normal cholesterol diet. On the other hand, CPT increased in that condition. In the high cholesterol diet condition, decrease in acid lipase activity, acid and neutral CEase, CEsyn. and acyl-CoAsyn., increase in CPT and no change in neutral lipase activity and TGsyn, were observed. In high cholesterol fed spontaneous hypertensive rats, increase in acid lipase activity and CEsyn., decrease in both CEases and CPT and no change in neutral lipase activity, TGsyn. and acyl-CoAsyn. were observed. Differences in lipid metabolism between tocopherol deficient and aged rat arterial wall were indicated, too. We discussed relationships between these changes in enzyme activities and lipid accumulation in arterial wall.

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It was previously reported that there was the decrease in linolate ratio and increase in oleate ratio of cholesteryl ester (CE) of serum lipoproteins (LPs) in atherosclerotic disease.
To clarify the significance of these changes in fatty-acid-compositions (FAC) in atherosclerosis, an attempt was made in this paper to investigate the changes in FAC of CE of serum LPs, liver and aortic wall in the process of development and regression of experimental atherosclerosis, which was induced with 0.5% cholesterol diet in female albino rabbits weighting around 2.5kg.
Atheromatous lesions were remarkably pronounced 10 weeks after the initiation of this experiment. Experimental animals were divided into 6 groups and all of them were sacrificed at the end of the experiment.
Part I; observation of the process of development of atherosclerosis.
Subgroup A: fed a normal diet for 20 weeks.
Subgroup B: fed a cholesterol diet for 24 weeks. Part II; observation of the process of regression for 8 weeks.
All of the subgroups C, D, E and F were fed a cholesterol diet for 12 weeks, and subgroup C was sacrificed at 12 weeks.
After this 12 weeks, subgroup D fed a cholesterol-free diet up to 20 weeks, subgroup E fed a cholesterol-free diet with 200mg of PPC a day up to 20 weeks, subgroup F fed a cholesterol-free diet with 200mg of BCPMP a day up to 20 weeks.
1) In subgroup B, the FAC of CE of serum LPs (VLDL, LDL) gradually increased in oleate ratio and gradually decreased in linolate ratio from 4 weeks.
2) The changes of FAC of CE in liver and aortic wall were similar to those of serum LPs in subgroup B.
3) Both subgroups B and C showed the similar change in FAC of serum LP, liver and aortic wall. In both subgroups D and E were recognized the increase of linolate ratio and the decrease of oleate ratio in serum LPs at 20 weeks. The changes of subgroup F in FAC of serum LPs were similar to those of subgroup C. The changes of subgroup D, E and F in FAC of aortic wall were similar to those of subgroup B.
4) The proportion of atheromatous lesions in thoracic and abdominal aorta were ranged from 30 to 60% in subgroup D, E and F. This might imply that there were no significant differences in three subgroups.
5) On histological findings, the intimal foam cells were markedly noted, but the intimal smooth-muscle cells were less remarkable in subgroup D. While, in subgroup E the intimal smooth-muscle cells were markedly ovserved, but the intimal foam cells were less prominent. Both the intimal smooth-muscle cells and foam cells were moderately observed in subgroup F.
6) Consequently, these results lead us to believe that the continuous increase in oleate ratio and decrease in linolate ratio in CE of serum LPs were one of the pathogenic factors and the potent factors of development in experimental atherosclerosis in rabbits.
We also observed the decrease in oleate ratio and increase in linolate ratio of CE of serum LPs and liver, in subgroups D and E, these changes would be indicative of regression in atherosclerosis.

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A variety of drugs is currently in use for treatment of the various forms of hyperlipoproteinemia (HLP) with the ultimate purpose of preventing and regressing atherosclerosis. In man, clofibrate, nicotinic acid, metformin, colestipol resin and propranolol have been reported to affect the lipoprotein patterns and compositions. The purpose of the present study was to investigate the effects of oxandrolone (OX) on plasma lipids and lipoprotein compositions in patients with type IIa, IIb and IV HLP. Subjects were three males and nineteen females. Their mean age was 63 years, ranging from 30 to 78 years old. They had been admitted in the hospital and had constant diet. Types of HLP were defined according to the WHO recommendation and criteria of Lipid Research Clinics Program. The patients had been selected on the basis of elevation of low-density-lipoprotedin cholesterol (LDL-Ch) above 150mg/100ml and/or plasma triglyceride (plasma-TG) above 160mg/100ml which were decided according to our data of lipid values in healthy men and women. (i) A patient with type IIa HLP was characterized by predominant increase in LDL-Ch and normal plasma-TG. (ii) A patient with type IIb HLP was characterized by increase both in LDL-Ch and plasma-TG. (iii) A patient with type IV HLP was characterized by predominant increase in plasma-TG and normal LDL-Ch. (iv) A patient with type III HLP was excluded by patterns of agarose gel electrophoresis and chemical indices which were calculated by cholesterol and triglyceride values in lipoprotein fractions. Oxandrolone, 4mg/day was given for 28 days. Blood was obtained from patients who had been fasted for more than 14 hours before and after treatment with oxandrolone. Plasma lipoproteins were fractionated by sequential ultracentrifugation at densities 1.006 and 1.063g/ml in a Type 40.3 rotor on the Beckman L5-50 ultracentrifuge. Ch and TG concentrations of very low density lipoprotein (VLDL), LDL and high density lipoprotein (HDL) fractions were determined. Ch and TG were measured according to the methods of Zlatkis A. and Zak B. and Fletcher M. J., respectively. Apolipoprotein B (Apo B) concentrations in plasma and three major lipoprotein classes were determined according to the method of Curry M. D. et al based on the electroimmunoassay of Laurell.
In patients with type ha HLP, plasma-Ch decreased significantly. LDL-Ch, HDL-Ch and VLDL-Ch showed a tendency to decrease, while LDL-Ch/HDL-Ch ratio increased. In patients with type IIb HLP, plasma TG, VLDL-TG and VLDL-Ch decreased significantly. There were no significant changes in plasma-Ch, LDL-Ch, HDL-Ch and LDL-Ch/HDL-Ch ratio, whereas LDL-TG and HDL-TG tended to decrease. In patients with type IV HLP, reduction of plasma-TG, VLDL-TG and HDL-TG were significant. VLDL-Ch was significantly reduced and reciprocal elevation of LDL-Ch was observed. HDL-CH was significantly lowered. Consequently, LDL-CH/HDL-Ch ratio was significantly elevated.
There was a positive correlation between the changes in LDL-Ch and the pretreatment VLDL-TG so that higher pretreatment VLDL-TG levels responded with an increase of LDL-Ch and lower pretreatment VLDL-TG responded with a decrease of LDL-Ch. This regression predicts that, when pretreatment VLDL-TG is greater than 138mg/100ml, treatment with oxandrolone will cause an increase in LDL. There was the inverse correlation between the changes in LDL-Ch and the pretreatment LDL-Ch. Subjects with higher pretreatment LDL-Ch concentrations manifested a decrease of LDL-Ch and vice versa. This regression predicts that when LDL-Ch is above 154mg/100ml, a decrease in LDL will be induced by treatment with OX. When LDL starts at lower levels, an increase of LDL will occur. In patients with type IV HLP, significant reduction of plasma-Apo B and VLDL-Apo B and inverse LDL-Apo B increment was observed after treatment.
Ehnholm C. et al reported that OX increases the activities of postheparin plasma hepatic lipase. It is sugg

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The effect on plasma total cholesterol and triglyceride has been observed for 3-19 years (mean 9.5 years) with treatment of antilipidemic substances. These treatments were effective for primary prevention of incidence of arteriosclerotic vascular diseases.
In group treated with CPIB with anabolic steroid, total cholesterol and triglyceride levels decreased respectively from 275.5 to 226.1 and from 200.7 to 105.6mg/dl. This combined method was more effective than another single treatment.

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Cholesterol concentration of eight kinds of cephalopoda or crustacea, eight kinds of eggs, ten kinds of meat, nine kinds of processing meat, 23 kinds of fishes, five kinds of processing fish meant, three kinds of processing fats and three kinds of treated milk, that is 69 kinds as a whole, is determined using colorimetric, enzymatic, thin layer chromatographic and gas-liquid chromatographic analysis.
Values obtained are as follows, in cephalopoda or crustacea, 139mg/100g for madako, 180 for surume-ika, 123 for mongo-ika, 156 for ashiaka-ebi, 132 for taisho-ebi, 164 for kuruma-ebi, 53 for taraba-gani, and 72 for matuba-bani. In bird eggs, 1030mg/100g is obtained for yellow of chicken egg, 950 for uzura (yellow), 740 for duch (yellow), in fish eggs, 446mg/100g for tarako, 485 for suzuko, 225 for kazunoko, 345 for tainoko, and 338 for tainoshirako. In kinds of meat, 54mg/100g for sirloin of caw, 69 for average beef, 66 for loin of pork, 69 for average pork, 114 for thigh of chicken, 63 for teba (chicken), 391 for liver of chicken, 175 for sunazuri (gizzard), 58 for sheep meat, and 62 for meat of whale. In treated meat, 59mg/100g for ham (loin), 54 for superior ham, 48 for average ham, 41 for low grade ham, 47 for roast pork, 53 for bacon, 40 for corned beef, and 47 for sausage. In kinds of fishes, 53mg/100g for hamo, 50 for maguro, 52 for tai, 56 for hamachi, 64 for katuo, 60 for hirame, 85 for iwashi, 68 for tachiuo, 38 for azi, 41 for karei, 72 for mebaru, 67 for saba, 150 for kisu (seed), 79 for isaki, 61 for bera, 76 for itoyori, 79 for kamasu, 99 for sannma, 62 for sake, 262 for shisyamo (seed), 122 for unagi, 137 for ayu (seed), 131 fir shirasagi (seed), 65 for koi, for funa, and 212 for moroko (seed).
In treated fish meat, 41mg/100g for kamaboko, 19 for chikuwa, 49 for tenpura, 17 for anpei, 136 for atuyki, and 30 for sausage. In kinds of fats, 100mg/100g for cheese, 220 for butter and 60 for margarine. In kinds of treated milk, 65mg/100g for cream, 28 for yogurt, and 10 for milk.
These values may be helpful for patient with atherosclerosis in dietary cure to prevent a development of the disease.

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Lipid metabolism in the arterial wall plays an important role in the formation of atheroma (1). Glavind et al. reported that lipoperoxide increased in atheromatous lesion (2). Therefore the experiments were carried out to clarify the relationship between lipoperoxide and the changes of enzyme activities concerning lipid metabolism in the arterial wall of tocopherol deficient rats fed for 16 weeks. Lipoperoxide, which was estimated as thiobarbituric acid reactive substance (TBA-RS), increased in the brain, the lung, the liver and the arterial wall of tocopherol deficient rats compared with these of tocopherol supplement rats. Both acid cholesterol esterase and acid lipase which were present in lysosome as previously reported (3), were decreased significantly in the arterial wall extracts of tocopherol deficient rats compared with those of tocopherol supplement rats. These results suggest that lipoperoxide might cause a decrease in these lysosomal enzyme activities. These facts suggest that decreased cholesterol esterase in the arterial wall of tocopherol deficient rats might cause the accumulation of cholesterol ester in lysosome, because cholesterol ester was less hydrolysed.
Acyl-CoA synthetase, triglyceride synthesizing activity and cholesterol ester synthesizing activity, which were considered to be present in microsome (3), decreased and only cytidine diphosphatecholine 1, 2-diacylglycerol choline phosphotransferase activity increased in the arterial wall extracts of tocopherol deficient rats. The mechanism of decrease or increase in these enzyme activities in this experiment must be further investigated.
Ratio of synthesis to hydrolysis of cholesterol ester in microsome was decreased in tocopherol deficient rats compared with that of tocopherol supplement rats. This ratio decreased in high cholesterol diet rats (4). Therefore, these results suggest that the changes in these enzyme activities might not cause cholesterol ester deposition in microsome of the arterial wall in tocopherol deficient rats.
1. Chobanian, A. V.: J. Lipid, Res. 13: 201, 1972.
2. Glavind, J. et al.: Acta Path. Microb. Scand. 30: 1, 1952.
3. Shirai, K. et al.: Proc. J. C. B. L. 20: 121, 1978.
4. Matsuoka, N. et al.: J. Jap. Atheroscler. Soc. 6: 121, 1978.

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Influences of humoral factors such as renin, norepinephrin, epinephrin, angiotensin-II and insulin to serum lipoprotein entry into the arterial wall have been performed in in vitro. Rabbit thoracic aorta was incubated in Warburg's apparatus for measurement of two hours' intake of cholesterol-¹⁴C-labelled low and high density lipoproteins. During the incubation period, 95% oxygen with 5% CO² was administered. Two procedures in which the humoral factors were added into the incubation medium in various concentrations (in vitro administration) and were injected into the rabitt vein with a continuous infusion pump (in vivo administration).
Results and discussion: 1) Renin, norepinephrin and epinephrin did not show any influence to the lipoprotein entry into the arterial wall in the both study. 2) Addition of angiotensin-II into the medium at intervals of 15min. and continuous infusion in vivo significantly increased both LDL and HDL entries. 3) In the both procedure to administrate the insulin, this substance increased only LDL uptake significantly. 4) From the above mentioned results, it may be concidered that progress of vascular damage in high renin hypertention is not affected from the direct action of renin but is affected from the other factors which relate the high renin state such as angiotensin-II. Even though both catecholamine and angiotensin-II constrict the arterial wall, only angiotensin-II increased the lipoprotein entry. this may be suggests that mechanism which increased the lipoprotein entry is not the constriction of artery but the direct arterial damage caused by angiotensin-II. As compaire with the mechanism of angiotensin-II, mechanism increased the lipoprotein entry in the insulin administration may be considered as functionally since only the LDL uptake was affected by the insulin.

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In order to investigate the entry of serum lipoprotein into the arterial wall and influence of various humoral factors such as renin, norepinephrine, epinephrine and angiotensin-II to the lipoprotein entry, immuno-fluorescient antibody method has been utilized to identify tissue antigens of serum lipoprotein in the rabbit thoracic aorta. Two procedures have been employed.
In the in vitro study, the humoral factors were added into incubation medium directly in which the arterial wall incubated with LDL. In the in vivo study, the humoral factors were injected into the rabbit vein with a continious infusion pump for 6 hours. Then, the thoracic aorta was obtained as a material.
Results and conclusions were follows.
1) In both in vitro and in vivo studies, no immuno-fluorescience of the LDL has been observed at the additions of renin, norepinephrine and epinephrine.
2) Immuno-fluorescience of LDL has been observed when angiotensin-II was administered intermittently at intervals of 15min. in vitro, and injected cotiniously in vivo.
3) In the in vitro study, immuno-fluorescience of the incubated LDL was observed inside of endothelium and not identified in the other dimensions which exope to the LDL directly.
4) From the above mentioned results, it may be concluded that endotherial cells play important role to uptake the LDL into the arterial wall and angiotensin-II is one of the most important factors which affect the lipoprotein uptake.

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