There are many reports which regard thrombogenic theory (Duguid) or other mechanical obstructive theories as pathogenesis of myocard infarction. These theories are true in some cases but questionable in others. Because nonthrombogenic infarction can be seen at necropsy. Moreover those areas are not wedge form but patchy, even if thrombi are found out. Yajima et al offered “Peripheral circuratory disturbance theory”; disorders of peripheral circuration rebound centripettaly and result in thickening and inhibition of blood at a point of coronary artery trunk. Constanitides has the same view, too. Kohmoto decleared thrombi and prethrombotic lesion take place at about 2.5cm. from left coronary ostium. If these are true, we must be able to find out marked changes in central side of extramular coronary artery and in contrast slight changes in periphery.
Author has examined histologically the left coronary arteries of 62 autopsy hearts (Infarction 16 cases, Hypertension 20 cases and Control 26 cases). It is detected most cases of myocard infarction and myocard fibrosis have thickened intima and/or severe medial dameges (loss of muscle fibers, fibrosis and thinning) in their central side. But in periphery these are more slight. When we discuss them, we must investigate both changes of intima and media.
According to these, author can recognize the changes as results of high internal pressure due to “rebound”. Because these have the same predilection place to thrombi and prethrombotic lesion of acute myocard infarction. And because some cases of infarction have no severe stenosis.
In regard to coronary stenosis of infarct hearts we have been able to find out not only thrombogenic ones but also atherosclerotic. Duguid's thrombogenic theory cannot be true in all cases.

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The agewise progression of severe atherosclerosis and the relationship between the coronary stenosis and the ischemic myocardial lesion were investigated in 583 consective autopsies over the age of 60 at the Tokyo Metropolitan Geriatric Hospital.
The prevalences of severe coronary atherosclerosis were 42.8% in male and 33.3% in female at the seventh decade, and increased to 66.7% and 75.0% respectively at the tenth decade. The similar agewise tendency was observed in the iliac and femoral arteries. Severe atherosclerosis in the aorta as well as in the cerebral artery revealed the equal prevalence over the age of 65 in male, while in female the prevalence increased up to the nineth decade.
Fifty percent or greater stenosis of major coronary arteries was most frequently observed in the anterior descending branch, followed by the right coronary artery. The agewise increase in the stenotic lesion was seen in these branches, but no such increase was demonstrated in 75 percent or greater stenosis.
The complete coronary occlusion was observed in 68 cases in 583 consecutive autopsies (11.7%), in which myocardial infarction of 2cm or greater was found in 48 cases (70.6%), myocardial scar being found in 10 cases (14.7%). The obstructions of anterior descending and right coronary arteries were equally appeared in myocardial infarction group, while right coronary obstruction was found predominantly in cases without myocardial infarction. The coronary stenoss in the remaining two branches were mostly less than 75% in cases of coronary occlusion without myocardial infarction.
The coronary stenoss of 75% or more in one, two and three vessels were observed in 13.2%, 8.2% and 2.7% respectively, in which myocardial infarctions were found 17%, 29% and 56%. The myocardial infarction was usually small and found in the posterior wall, when the corresponding coronary stenosis was less than 75%.

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Although the etiology of atherosclerosis remains obscure, acid mucopolysaccharides (AMPS, glycosaminoglycans) have been assumed to play an important role to precipitate the plasma lipoprotein in the arterial wall. Furthermore, in many tissues besides artery, focal deposits of cholesterol have been demonstrated to be associated with AMPS.
Since AMPS is rich in the renal papilla and administration of papain has been reported to liberate AMPS from various connective tissues, the effect of papain on the renal papillary AMPS and the associated change in total cholesterol content have been evaluated.
Male Wister rats weighing 200-250g were injected intraperitoneally with 1-3mg/0.1kg body weight of a papain solution. After 24hrs, the kidneys were obtained and analysed for AMPS and total cholesterol of the papillary region. Insoluble collagen was employed as reference base.
Significant decrease in AMPS and the rather large loss of cholesterol have been observed by papain treatment. However, there has been no dose dependency in the effect of papain concentrations used in this study.
Analysis of AMPS composition revealed a marked decrease in the hyaluronic acid fraction, but all other fractions, i. e., heparan sulfate, chondroitin sulfate A/C and dermatan sulfate were also decreased.
These results suggest that changes in AMPS content and composition of renal papilla may affect the binding capacity of lipoprotein, resulting decreased concentration of cholesterol in this region.

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45 cases of myocardial infarction and 44 cases of cerebral infarction have been observed for past 20 years (mean 6 years). Oral glucose tolerance test (G. T. T.) in them was studied.
Abnormal oral glucose tolerance was found 44% in M. I. and 52% in C. I.
It was found the recovery of curve of G. T. T. was needed three months after attack of C. I.
For prevention of arteriosclerotic vascular disease, it must be important to control not only hyperlipidemia, hypertension but also abnormal oral glucose tolerance.

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Plasma lipids such as cholesterol, triglycerides and free fatty acid were observed in a correlation with atherosclerosis or ischemic heart disease (IHD). However, we described that the abnormal lipoprotein metabolism regardless of plasma lipids concentration was more important problem in the patients with IHD. On the other hand, atherosclerosis and IHD occur more frequently in the patients with diabetes mellitus. Therefore, it take an interest in a characteristics of lipoprotein metabolism with abnormal GTT in a patients with IHD.
In this study, we attempted to investigate the relationship between abnormal glucose tolerance test (abnormal GTT) and lipoprotein metabolism in the 64 patients with IHD. Particularly, lipoprotein metabolism was the metabolic pathway of VLDL to LDL.
Frequency of the abnormal GTT was observed in the 42 patients (65.6%), but the dietary treatment for four weeks reduced the incidence of the patients with abnormal GTT from 65.6% to 32.8%. Similar changes of the frequency were observed in each of three groups according to the analysis of plasma lipids concentration: normolipoproteinemias (N=21) or hyperlipoproteineimas (N=23) before and after the dietary treatment were from 66.7% to 33.3% or from 65.0% to 35.0%, and twenty patients whose plasma lipids concentration were changed from hyper to normolipoproteinemia by our dietary treatment were from 65.2% to 30.4%. According to these results, relationship between the frequency of abnormal GTT and the concentration of plasma lipids was not observed in the patient with IHD. On the other hand, lipoprotein metabolism such as the metabolic pathway of VLDL to LDL was reduced in the patients who were still hyperlipoproteinemia after the dietary treatment: proportion of VLDL-triglycerides decreased with an increase in VLDL-protein, and a decrease in LDL-protein was observed simultaneously. Additionally, 66.7% of the patients with abnormal GTT had the Beta-K band using polyacrylamide gel-disc electrophoresis (the patients without abnormal GTT were 14.8%). The abnormal metabolic pathway of VLDL to LDL and the Beta-K band were not correlation with plasma lipids concentration. According to these results, lipoprotein metabolism with abnormal GTT was abnormal lipoprotein metabolism regardless of plasma lipids concentration in the patients with IHD. This abnormal lipoprotein metabolism is produced by incomplete hydrolysis of VLDLtriglycerides and another metabolic pathway as a remnant lipoprotein, Beta-K band.

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Lipoprotein farction was made visible by polyanion precipitation in the 1% agarose gels after electrophoresis, according to the method descreibed by Wieland and Seidel (Clin. Chem. 19, 1139 (1973)). The electrophoresed samples were placed for 30min. into a bath of 0.1mol/liter MgCl₂ containing 1g/liter sodium heparin and 10g/liter NaCl (solution 1), to selectively precipitate the lipoprotein band corresponding to VLDL fraction, or into a bath of 0.2mol/liter CaCl₂ containing 6g/liter sodium dextran sulphate to recipitate all plasma lipoproteins (solution 2).
It was possible by this method to differenciate type III and IV. The hypoalphalipoproteinemic serum showed the almost same intensity of precipitin band in the alpha position as control.
From the results of immunoelectrophoresis, it was found that the protein precipitated by solution 2 contained not only lipoproteins but also albumin, IgG and other serum proteins.

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Authors investigated the recovery rate of cholesterol (TC), triglyceride (TG) and phospholipid (PL) from each gel-block contained lipoprotein after electrophoresis. Method: Authors separated in several lipoprotein fraction from the 4ml to 5ml of serum by Wada's method (Clinical Chemistry 19: 235-239, 1973). Electric current was 8mA in 130-140 volts. The period of phoresis was 6-8hrs. Each fraction was cutted and each gel-block was weighed and it was crushed to pieces (2-3mm²) through pushout-type-crusher acompaning wire netting. The lipoprotein fraction was extracted by stirring saline with crushed piecies at 4sC for 20-24hrs. After extraction, the saline contained the crushed pieces was centrifuged in 27500g for 30min. at 4sC, then the supernatant was concentrated by riofilyzing in polyethylenglycol 4000. The concentrated solution was diluted to the original serum volume again. TC, TG, PL, β-lipoprotein (β-Lp) and α₁-lipoprotein (α₁-Lp) in each extract-solution were estimated. The β-Lp was measured by turbidity method with dextran sulphate (β-Lp I) and immunodiffusion method (β-Lp II). The α₁-Lp was measured by immunodiffusion method. Results: I) Recovery rate of β-Lp by β-Lp II in Sudan black B stained block was better than in unstained block. It is difficult to explain the cause of this result for the present.2) Recovery rate of β-Lp by β-Lp I in each block was poor regardless of staining in all cases. It was concluded that electric charge on surface of β-Lp-molecule was changed by phoresis with POLYACRYLAMIDE-GEL. 3) Recovery rate of alubumin was almost perfect. It was supposed that protein in serum was not degenerated by phoresis. Therefore, the decrease of basic charge on β-Lp by electrophoresis was resulted from degeneration of lipid part in β-Lp or caused by degeneration of lipid part with Sudan Black B staining. 4) A patient suffered from primary biliary livercirrhosis had a few α₁-Lp but her serum contained much more PL, TC, TG, and β-Lp than in normal subjects, especially the concentration of TG in pre-β-Lp was higher than in normal subjects. It was concluded from above results that there were apoprotein-C-deficiency in serum of patients (P. B. C.) which was not able to convert from pre-β-Lp to β-Lp by lipoprotein lipase. 5) The percentage of recovery of each lipid fractions were as follows;

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All the major classes of the serum lipoproteins may be precipitated with sulfated polysaccharides and divalent cations.
Burstein, et al. (1972) have shown that at neutral pH and in the presence of Ca⁺⁺, Mg⁺⁺ and heparin, lipoproteins can be precipitated from human serum. Methods have been developed for the isolation of the different classes of lipoproteins, i. e., low density lipoprotein (LDL), very low density lipoprotein (VLDL), and chylomicron by the stepwise addition of the reagents and increasing the ionic strength by the addition of sodium chloride.
We developed quantitative precipitation method using heparin and CaCl₂ solution by establishment of the standard solution, based on comparison between precipitation and ultracentrifugation methods. Quantitative estimation of the individual lipoprotein classes provides major advantages over other simplified techniques for various hyperlipoproteinemias.
1) Heparin-Ca⁺⁺ precipitation method
i) Reagents: (I) 0.005% heparin in 0.05M CaCl₂ solution, (II) 0.005% heparin and 0.6% NaCl in 0.05M CaCl₂ solution, and (III) 0.005% heparin and 0.9% NaCl in 0.05M CaCl₂ solution.
ii) Methods: 4ml of each solution is added to each of three tubes. To each added is 0.1ml of serum, respectively. The tubes are mixed and after 15min. at room temperature, turbidity is measured at 650nm. Turbidity in tube No. 1 represents the sum of chylomicrons, VLDL and LDL. Tube No. 2 turbidity corresponds to chylomicrons plus VLDL and tube No. 3 turbidity to chylomicron alone.
2) Quantitative ultracentrifugation
VLDL fraction is prepared by ultracentrifugation at 105, 400×G at 20°C for 25hrs. The LDL plus VLDL fraction is prepared form the serum-NaCl mixture with the density of 1.1315+0.0005, by ultracentrifugation at 105, 400×G at 20°C for 16hrs. Each lipoprotein fraction weighed after desiccation at 60°C and also after dry asking at 600°C, respectively. From the difference of both weights, the amount of each lipoprotein fraction is calculated by the established formula.
3) Correlation between heparin-Ca⁺⁺, and ultracentrifugation method
The correlation coefficient of the LDL plus VLDL fraction for the two methods was 0.75 and the regression line was Y=0.38X-9.8, and VLDL fraction for both methods giving a correlation coefficient of 0.82 and the regression line of Y=0.33X-19.4.
4) Preparation of standard solution
Because the standard lipoprotein solutions can not be obtained, the adequate latex emulsion has been prepared by the correlation between heparin-CaCl₂ and ultracentrifugation method.

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The antiskeletalmuscle- and antimyocardial antibodies in the sera of the cerebrovascular patients and controls were detected in order to inspect the patients from the immunological standpoint.
Pieces of M. gastrocnemius, and heart muscle were obtained at autopsy within 6 hours after death homogenized and suspended in 9 volumes of the isotonic saline solution.
After 10 minutes centrifugation at 2, 500rpm, the sediments were used as antigens. Steffen's antiglobulin were used consumption test using frseh skeletal muscle and lyophilized myocardial antigens was adopted.
The antiskeletalmuscle antibodies were detected significantly frequently in the cerebrovascular patients compared with controls: 32 of 74 patients (43.2%), 5 of 70 controls (7.1%), and significantly frequently in the patients elapsed 7 months or more after attacks compared with those within 6 months: 2 of 22 patients within 6 months (9.1%), 30 of 52 patients after 7 months or more (57.7%).
The ADL-T values of the lower limbs were inferior in the patients with strongly positive antiskeletalmuscle antibody to those with moderately positive or negative ones.
The antimyocardial antibodies were detected significantly frequently in the controls over 70 years old in comparison with those under 69 years: 12 of 36 over 70 years old (33.3%), 6 or 69 under 69 years old (8.7%), and significantly frequently in the group with abnormal electrocardiograms compared with those with normal ones in both controls and patients.
The ADL-T values of the lower limbs were inferior in the patients with positive antimyocardial antibody to those with negative one.
These results indicate that the antiskeletalmuscle- and antimyocardial antibodies are present in the sera of the cerebrovascular patients, and it is conceivable that the antiskeletalmuscle antibody is closely related to atrophy of patient's extremities and the antimyocardial antibody to myocardial involvement in patients and controls.
The report of immunological study on the cerebrovascular disorder has not been published except an observation on the anticerebral antibody by Murakami et al. The anthor's study is regarded as the first trial to the observation of the antiskeletalmuscle- and antimyocardial antibodies in the cerebrovascular disorders. The data obtained suppose the existence of the autoimmune response in the pathophysiologic status of the disorder. However, no relation was found between the antibody titer and γ-globulin concentration in this study.
Therefore, the conclusion of the discussion about the existence of the autoimmune response in the pathophysiologic status of stroke patients could be drawn from the further detailed and versatile investigations.

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Recently cyclic nucleotides have been highlighted as a key substance in metabolic and pathophysiological condition of tissues and clinical approach has been pursued in every field. Plasma cyclic AMP phosphodiesterase activity, a disintegrating enzyme of cAMP, has been studied to be deviced a new assay with the combination of Lowry's mcroassay method and Scott & Solomon's thin layer chromatographic analysis. 10μl of plasma taken from 2ml of haparinized venous blood was mixed with 20μl of reagent mixture, consisted of 0.36μM 8-³H cyclic AMP, 150μg of snake venom, 10mM MgCl₂ and 960μl of 50mM Tris-HCl buffer (PH 7.4). After incubated for 30min, under the temperature of 37°C, 1μl of incubated mixture was spotted on Polygram Cell DEAE thin layer chromatography and developed with the solution of 1M CH₃COONH₄ and 95% ethyl alcohol (30:75v/v) for 2 hours. Each fraction of adenosine, cAMP, 5′AMP was collected in vials and radioactivity of each was counted by liquid scintillation counter. Concerning this method, optimal conditions of concentration of substrate, incubation time, comparison between serum and plasma, volume of sample, and Michaelis constants were studied. Under these optimal conditions, plasma cAMP-PDE was measured in 33 healthy men (44.5±3.4yrs) and in 47 healthy women 38.8±2.5yrs). Plasma cAMP-PDE activity was 2.23±0.19pmole/ml/min, in man and 1.37±0.13pmole/ml/min. in woman, both of which activity exhibiting statistically significant difference. Moreover, man did not show changes of PDE activity in spite of his advanced age. On the contrary, women exhibited lowest value of 1.00±0.11pmole/ml/min. in the twenties and a gradual increase of values with advancing years. These data suggest the different activity in sex and they will be more studied in the comparison of the pathophysiological condition in women.

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