Bilirubin load test was performed to differenciate hyperbilirubinemia in various liver diseases. For this purpose, bilirubin excretion pattern, serum direct and indirect bilirubins, and the molar ratio of glucuronic acid to bilirubin in the ester-form bilirubin fraction separated by Kosaka and Hera's method were investigated. Phenobarbital was effective to decrease serum bilirubin in Gilbert's syndrome, so this phenomena was investigated from the point of bilirubin metabolism. The following results were obtained:
1) Serum total bilirubin markedly elevated after bilirubin loading in Gilbert's syndrome, and direct bilirubin was increased significantly, and the molar ratio of glucuronic acid to bilirubin was lower than normal, which was coincided with the low activity of UDPGA bilirubin glucuronyl transferase in liver tissue.
2) Serum total bilirubin after bilirubin loading in Dubin-Johnson syndrome showed high retention, and many peaks were observed in direct bilirubin, which showed high value. Molar ratio flucutiated in normal range.
These findings support the prolonged excretion of serum bilirubin in this disease due to the bilirubin circulation from liver cell to microbili.
3) In Rotor syndrome, retention of serum total bilirubin after bilirubin loading was not so high, and serum direct bilirubin increased and the molar ratio was in normal range.
4) Excretion of serum bilirubin slightly delayed in posthepatitic hyperbilirubinemia after bilirubin loading, and serum direct bilirubin and molar raio showed normal pattern.
5) Phenobarbital administration in Gilbert's syndrome was effective to decrease serum bilirubin, and serum bilirubin increased after stopping the drug administration.
This phenomen was not only elucidately by the induction of UDPGA bilirubin glucuronyl transfesase activity, but the activation of oxydative and redox state of microsomal enzyme system and the normalzation of liver cell membrane would be participated in the nomalization of serum bilirubin.

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As almost no study has been made on the mass spectroscopic analysis of bilirubin obtained from the bile collected from the patients with liver diseases and Gunn rat, it was attempted to determine the isomeric composition of bilirubins by the use of mass spectrometry. Bilirubin was crystallized from the bile by Ostrow's method. Bilirubin IXα dimethylester was yielded by Küster's method from the commercial bilirubin (Daiichi pure chemical Co. Ltd.), and was purified by thin layer chromatography. Photochemical natures of the fifth of six bands was coincided with that of bilirubin IXα dimethylester. The following results were obtained;
1. Mass spectra of bilirubin IXα dimethylester from bilirubin IX: No molecular ions were obtained by our method. The base peak at m/e 314, 300, or 227 was found and these peaks were derived from the cleavage of central methylene bridge of bilirubin IXα and IXγ dimethylester. No peaks, showing the exsistence of bilirubin IXβ and IXδ dimethylester, appeared on the mass spectrum at m/e 360 and 239.
2. Mass spectra of bilirubin IX dimethylester from various liver diseases and from Gunn rat: Mass spectra of the bile bilirubin from various liver diseases and Gunn rat were observed at m/e 314 or 300 as the base peak and there was no significant peaks showing other isomers such as IXβ or IXδ dimethylester. These findings coincided with that of bilirubin IXα dimethylester standard.
From these results, the exsistence of other isomeric compositions except bilirubin IXα (IXγ) was not able to find in the bile of human being in health and various liver diseases.

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Hemodynamic effects caused by ethanol infusion systemically in dose of 75mg per kilogram per minute for 20 minutes have been studied in anesthetized open-cest dogs, and following results were obtained. Cardiac output, stroke volume and heart rate did not change and total peripheral resistance decreased with a fall in systemic blood pressure. Reduction in coronary vascular resistance, as same as a fall in total peripheral resistance, was observed with no change in coronary blood flow. Diminished left ventricular function, manifested as rise in end-diastolic pressure and a decline in deflection of segmental contraction curve of left ventricle, was observed in most cases. Isoproterenol (0.1μg/kg) injected intravenously after the in fusion of ethanol, produced a rise in systolic blood pressure and an elevation in left ventricular end-diastolic pressure (LVEDP), while a fall in blood pressure and a decline in LVEDP were caused with the intravenous injection of isoproterenol before the infusion of ethanol. Effects of isoproterenol on heart rate and cardiac output after the infusion of ethanol gave no essential change when compaired with those obtained without ethanol infusion. It is concluded that ethanol dilates both of coronary and systemic arteries with depression of left ventricular function, and makes a remarkable rise in left ventricular work when a positive inotropic stimulant such as isoproterenol was given, though an increased cardiac output caused by the stimulant was not influenced with ethanol essentially.

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It is usually considered that isolated left ventricular hypertrophy is not responsible for marked left axis deviation, and it is associated with left anterior fascicular block. From this point of view, morphological features were studied on the vectorcardiograms recorded with the Frank's system in 63 cases. They had diseases producing systolic overload on left ventricle (mainly essential hypertension) revealed out left ventricular hypertrophy in Ecg findings. They were divided into three groups based on electrical QRS axis as follows: (i) NAD group consisted with+90°-0°. (ii) LAD' group consisted with 0°--30°. (iii) LAD group consisted with -30°--90°. The patterns of QRS Loop classified into 5 types in horizontal plane (N_{I, II}, L_I-III) and 4 types in frontal plane (A-D). In horizontal plane, 75% of NAD group showed type N_I, or N_II, and type L_I, or L_II occupied in 75% of LAD group. In frontal plane, a half of LAD belonged to type D and type A was less than 10% in LAD, compared that type A was observed in 85% of NAD. Left ventriular hypertrophy with marked left axis deviation was classified into three types on the base of QRS loop as follows: the terminal portion of QRS loop directed superiorly to the left (type B) and superiorly to the right (type C), and the majority of QRS loop located to leftward and superiorly (type D). It was considered that the electrical force caused by hypertrophied left ventricle directed more posteriorly and to the left in type B, and type D was the pattern added to type B with the delayed excitation in antero-lateral wall of left ventricle caused by left anterior fascicular block. On the other hand, it was considered that the majority of type C, showing type N_II, was the resultant of the delayed excitation in postern-basal portion of ventricles and was not closely related with left ventricular hypertrophy or left anterior fascicular block.

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The effects of variable antero-posterior distance of the trunk upon standard 12 lead systems and three different corrected orthogonal VCG lead systems were investigated with the reference to the lead vector. Human torso model of an average Japanese adult male and four elliptical cylinders, which had fixed height and left to right distance and variable antero-posterior distance were employed as a homogeneous volume conductor. Lead vectors were measured when the artificial current dipole was placed at each of 15 points in the heart area. The following results were made in the present study.
1) The presence of proximity effect in V1, V2, V3 and V4 leads were demonstrated. These proximity effects were greater value in the models with short antero-posterior distance of the trunk.
2) The normality and orthogonality of the Frank lead system were better than those of the modified Frank lead system in the models with long antero-posterior distance of the trunk.
3) The normality and orthogonality of the modified Frank lead system were better than those of the Frank lead system in the models with relatively short antero-posterior distance of the trunk.
4) The Mc Fee lead system had better orthogonality and worse normality.
5) The effects of variable dipole locations upon the lead vectors measured with each lead systems became considerably greater, according as the shortening of the antero-posterior distance of the trunk.

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The experiment was undertaken to study the effect of total parenteral nutrition (TPN) on nitrogen balance and organs in the postoperative period, 73 of six-week old rats were devided into four groups as follows.
Group I Simple laparotomy and oral feeding
Group II Simple laparotomy and TPN
Group III Intestinal bypass operation and oral feeding
Group IV Intestinal bypass operation and TPN
TPN groups received 338 cal/kg/day and 1, 96 Ng/kg/day administered continuously throughout 13 days.
The body weight increase in group I, II, III, and IV were 66, 6g, 55, 6g, 4, 6g and 42, 5g, respectively.
The weight gain was observed in TPN groups during postoperative period.
The nitrogen balance study in group II and IV showed that positive nitrogen balance was able to achieve in both group and cumulative nitrogen balance in group II and IV was 1, 65g and 1, 31g, respectively.
This study shows that positive nitrogen balance can be obtained in the immediate postoperative period by means of TPN.
Small intestine during TPN showed significantly lesser in weight and shorter in villous height than that of oral feeding.
The lack of absorption of nutrients from intestinal mucosa seems to be responsible.
Histological studies of TPN group revealed that liver and spleen had fatty infiltration and there was swelling of reticuloendothelial cells which is supposed to be due to infusion of fat emulsion.
Swollen cells of tubules in the kidney were observed.

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A survey on anemia and parasite infection was conducted at Khonkaen district, Thailand. The number totaling about 1000 was examined. Physical examination revealed that hypertension and obesity were extremely rare and also that anemia and symptoms of the gastrointestinal tract were common. Hookworm infection was prevalent (67.5%). Anemia was seen in 31%. Judging from hemoglobin levels, 36.4% of children below 6 years, 35.1% of children 6-14 years, 14.2% of adult males, 36.5% of adult nonpregnant females and 36.4% of pregnant females had anemia respectively. Hypochromia and microcytosis were frequently seen in the blood smears. Eosinophilia was also marked in all age and sex groups. Serum iron levels and % saturation of transferrin were low in high percentage except in adult males. In regard to serum vitamin B₁₂ and folic acid, low values were scarecely observed in all groups. Megaloblastic anemia was not detected at all. Total serum protein was relatively low (though within normal limits) in children below 6 years.
It can be said that the anemia is due to iron deficiency in almost every cases in this district and vitamin B₁₂ and folic acid do not contribute to the anemia. Protein deficiency may play some role for the anemia of children below 6 years.

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Dietary survey on 42 families in 7 villages in Khonkaen district was carried out. Though mean calorie intake was sufficient (2296 Cal), protein intake from animal sources was extremely low. This was due to the fact that farmers ate large amount of rice and other dietary items were used only to make the rice more flavorful. Eightynine percent of the total calorie was taken from rice. Mean intake of iron was 7.9mg/person/day. Standard man ratio made it possible to calculate individual iron intakes from the data of family survey. Iron intakes of each group were 5.6mg in 1-3 years, 8.6mg in 4-5 years, 10.3mg in 6-9 years, 11.2mg in 10-12 years, 16.1mg in males over 13 years, 12.5mg in nonpregnant females, 11.2mg in females over 50 years and 17mg in pregnant and lactating women respectively.
Folic acid contents of 52 Thai foods were assayed microbiologically using Lactobacillus casei. Calculation of folic acid intake was also performed using the results of folate assay of foods and other data from literatures. Intake of free folate was 217.4μg/person/day and total folate 346.1μg/person/day.
In conclusion, iron intake of the farmers in this district was insufficient except in adult males to meet the increased requirements of children and women. Folic acid intake was rather abundant. The results can explain well the results of the blood examination showing prevalence of iron deficiency anemia and rare occurrence of folic acid deficiency in this district.

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Electrocardiographic, vectorcardiographic and phonocardiographic changes were studied before and after surgical correction of isolated pulmonic stenosis.
1. Before operation, the maximal right ventricular systolic pressure were correlated well with the T wave changes, such as mean electric T axis, direction of maximal T vector in each plane, spatial maximal QRS-T angle, ratio of spatial magnitude of T to QRS vector, and the QRS wave changes, such as height of Rv₁, sum of Rv₁ and Sv₅, direction of maximal QRS vector in frontal plane, rotation of QRS loop in horizontal plane, configuration of QRSv₁. After operation, in addition to those above, R/Sv₁, Rv₁+Sv₅/Sv₁+Rv₅ and mean electric QRS axis tended to improve much more within the first one year after operation than thereafter.
2. The appearance of RBBB type were found more frequent after operation than before, and thought to be due to the decreasing right ventricular systolic overloading, or its combination with operative invasion to peripheral network of right bundle branch.
3. The more or less abnormal findings were revealed in twenty one cases, the remaining one was appreciated to be normal one hundred and twenty five months after operation.

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Catecholamine in tissue, serum and urine has been determined by the fluorometric method. It still remains some problems, especially sensitivity in the measurement and purification of catecholamines according to this method. Therefore, I determined by a gas chromatographic method (using an Electron Capture Detector: ECD) modified from the method of Imai (1971). Catecholamines in cerebrospinal fluid (CSF) have not been determined by gas chromatographic method so far. A new technique for the determination of catecholamine in CSF of human will be presented in this paper.
The results are as follows,
1) According to the gas chromatography, 20mg of tissue, 2.5ml of serum and 3.0ml of CSF are enough to determine with the grade of one nanogram.
2) Dopamine, norepinephrine, epinephrine, 3, 4-dihydroxyphenylalanine (DOPA), tyramine, octopamine, metanephrine, normetanephrine and synephrine were determined. Retention time of gas chromatography for these substances was decided. Heptachlor epoxide or dieldrin were available to the internal standard substances.
3) Dopamine was determined in serum of normal human subjects and in experimental animals (rat and cat).
4) Epinephrine was detected in brain tissue of rats and cats. This evidence will show that the central nervous system contains enough epinephrine to evaluate.
5) Micro-quantitative analysis of catecholamine and its derivatives in various samples will contribute to the neurochemical researches and to the clinical works in the central nervous system.

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Catecholamine and its metabolites level in cerebrospinal fluid (CSF), serum and urine of the 62 patients with epilepsy (38 males and 24 females, 12 cases of Lennox's syndrome, 48 cases of centrencephalic epilepsy, one case of focal cortical epilepsy and one case of temporal lobe epilepsy) were determined by gas chromatographic method (using an Electron Capture Detector) and fluorometric method. Studies on the epilepsy was performed with a special reference to catecholamine metabolism. As the results,
1) 3, 4-dihydroxyphenylalanine (DOPA) and dopamine in CSF of the epileptics were significantly lower than in the control group.
2) Homovanillic acid (HVA: end-product of dopamine) in CSF of the epileptics was also significantly lower than in the control group.
3) In epileptics tyramine, octopamine and synephrine were detected in CSF.
4) It is speculated in epileptics that the suppression of the pathway from tyrosine to dopamine may act to enhance the side pathway from tyrosine to tyramine followed by octopamine, synephrine and epinephrine.
5) Serum DOPA levels of epileptics being treated with anticonvulsants were abnormal in about half of the cases.
6) Serum MAO activity of epileptics was significantly lower than in the control group.
7) HVA in urine of epileptics tended to increase, but its level was changeable.
Based on these clinical studies of epileptics, a penicillin focus was made on the cortex of cats. And effects of the catecholamines to penicillin focus were electroencephalographically examined.
8) Penicillin spikes were strongly inhibited by topical application of dopamine.
9) When octopamine was injected into cisterna magna after the appearance of penicillin spikes, spike frequency increased and abnormal spike discharges were observed.

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Combined effects by L-DOPA and peripheral decarboxylase inhibitor (L-alpha-hydrazinomethyldopa: MK 486) were studied in cats and eighteen parkinsonian patients, with special reference to catecholamines and amino acids metabolism.
Catecholamine levels in brain tissues and plasma were measured in cats following the intraperitoneal injection of L-DOPA alone (100mg/kg b. w.) or combined with MK 486 (10mg/kg b. w.) by gas chromatographic method (using an Electron Capture Detector).
1) DOPA level in cat plasma in the combination with MK 486 was three-fold as high as in L-DOPA alone. Dopamine levels in caudate nucleus of cats were increased up to about six-fold in the combination. Dopamine level in plasma was markedly decreased in the combination with MK 486. Norepinephrine and epinephrine levels in cat plasma were no significant change in both L-DOPA alone and combination with MK 486. Norepinephrine level in the brain was no significant change except slight increase in pallidum and caudate nucleus.
2) After oral administration of L-DOPA in combination with MK 486, DOPA level in plasma of the patients with parkinsonism was maintained at a higher level and for longer than after the oral administration of L-DOPA alone. Plasma dopamine level with the combination was markedly lower than with L-DOPA alone. Dopamine level in plasma at three hours after the administration tended to be decreased at a 1:5 ratio of MK 486 to L-DOPA in dosage. Plasma epinephrine level was not significant change at all. Dopamine level in cerebrospinal fluid was maintained at a higher level and for longer than the oral administration of L-DOPA alone.
3) Urinary homovanillic acid level in parkinsonism was lower in the combination of MK 486.
4) Plasma amino acids which might affect catecholamines metabolism was not significant change following the administration of L-DOPA alone as well as L-DOPA plus MK 486.
5) In clinical studies of 18 patients with parkinsonism, the effectiveness of the combination therapy with L-DOPA and MK 486 (mean dosage of L-DOPA: 750mg/day and MK 486: 75mg/day) was observed in all cases. Marked improvement was noted in 10 cases out of 18 (56%) with akinesia, in 12 cases out of 18 (67%) with rigidity and in 6 cases out of 18 (33%) with tremor. Maximum plasma DOPA levels were higher in cases with marked improvement than the other, and were highest in patients with dyskinesias of a side effect.
6) Nausea and vomiting of the side effect, which were less severe than those experienced with L-DOPA alone, were noted in 5 cases (28%). Dyskinesias in extremities, face, mouth and tongue were observed in 6 cases (33%).

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I have investigated islets of chronic pancreatitis patients histochemically and electron microscopically, and classified chronic pancreatic lesions into six grades according to the degree of fibrosis (Grade O to Grade V).
1) In Grade II, some acinar cells showed irregularly dilated rough endoplasmic reticula, swollen mitochondria in their cytoplasm, and reduction in the number of zymogen granules. In Grade III, these alterations were more prominent.
2) As far as islet cells were concerned, before Grade II most of the islet cells remained almost normal in structure but some B cells in Grade II are characterized by cellular structures that are irregular because of their dilated endoplasmic reticulum. In Grade III cells, these alterations were more prominent and mitochondria became rounded in shape and endocrine granules showed irregular formations and a reduction in number.
A and D cells, in comparison with B cells, usually exhibited relatively mild alterations.
3) In grade IV, most of the islet cells were almost wholly occupied by the cisternae of dilated endoplasmic reticulum and cytoplasmic vacuoles; and those few granules that remained had lost almost all their density. Almost all of the A, B and D cells presented these marked alterations, but alterations in the B cells were especially prominent. Even in Grade IV, since normal nerve endings still existed around the islets and at least one third of the islets still had almost normal granules, the endocrine function of the pancreas seems to be preserved.
4) In Grade IV, it is of prime interest that in the basal region of ducts islet cells containing almost normal granules were frequently demonstrated. These cells were observed as isolated single cells in the duct at first, most of which were A cells. On the other hand, most of the B cells were located in the clusters. The presence of small islets close to a duct epithelium was also observed frequently. I am convinced that islet cells develop from ductules in human adult pancreas because all these findings are as same as that in human fetus.
5) I observed the other cells than A, B and D cells in human pancreas. They have many granules elongated and irregular in shape. I believe them to be F cells, same as enterochromaffin cells (EC cells) in digestive mucosa.

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Studies were attempted to know biochemically on bacteriophage T₄ head proteins, especially those which have been known to participate in its head morphogenesis. Both IPI and IPII were proved to be cleaved and the cleavage of all the following proteins, P23, IPI, IPII and IPIII, was proved to occur near to their N-terminals during this morphogenesis. In vitro examination revealed that this cleavage is caused by some enzymatic reaction. Amino acid compositions of P23^c, P24, Wac and the phage-λE protein were rather similar to each others as the structural proteins, while P22, IPI^c, IPII^c and IPIII^c were quite rich in lysine and were basic as the nuclear protein histon. The preliminary examination for the sequencing protein structure was attempted on the major protein, P23 and P23^c.

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When the crystalline beef liver catalase was treated with SDS (sodium n-dodecyl sulfate) or LIS (3, 5-diiodosalicylic acid lithium salt) which is a kind of the anionic surfactant, its catalase activity dwindled and disappeared at last, and its peroxidatic activity was exhibited.
It was found by the ultracentrifugal analysis that the molecule of beef liver catalase treated with SDS splited into two subunits with sedimentation constants of about 10 S and 2 S.
The same phenomena as mentioned above were compared in the crude catalase extracts of livers from normal mice and acatalasemia mice. In the crude catalase extracts from both mice, catalase activities dwindled and disappeared in the same way and peroxidatic activities appeared if treated with SDS. But peroxidatic activity of acatalasemia liver extract was higher than that of normal liver extract. It seems that the liver catalase in acatalasemia mouse is more labile by SDS treatment than that in normal mouse, resulting in showing the conformational changes of catalase molecule in acatalasemia mouse liver.

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An EB virus-carrying lymphoblastoid cell line established from peripheral blood of a patient with SMON was super-infected in vitro with murine Rauscher leukemia virus. Since then, the cell line has been dually infected with both viruses for more than 1.5 years, as repeatedly demonstrated by the presence of herpes type virus particles and C type virus particles. The C type virus particles liberated from the human cultured leukocytes were no longer leukemogenic when inoculated into otherwise susceptible BALB/c mice, but this procedure rendered them considerably resistant to challenge infection with leukemogenic Rauscher leukemia virus that had been maintained by passage through BALB/c mice. The infectivity of the human cell-derived C type virus to other lymphoblastoid cell lines established from different patients and to primary BALB/c mouse embryo cells could not be demonstrated by the method employed. The implication of these findings is discussed in the light of possible viral etiology of human malignancy.

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