As a part of the study to elucidate the influence of rhythmical sound on the central nervous system activity, the effect on the recruiting response of rabbits was investigated and the following observed.
1) The recruiting response in the cerebral cortex EEG induced by 8 Hz stimulation of the nucleus ventralis anterior (VA) or the nucleus reticularis (RET) was facilitated by the rhyt hmical sound of 60/min or 120/min beat frequency and 500 Hz pitch.
2) The recruiting response induced by 8 Hz stimulation of V A or RET was inhibited by the rhythmical sound of 450/rnin and 1000 Hz.
3) The rate of change in the amplitu d e of this recruiting response by sonic stimulation was analysed quantitatively by determination of the wave pattern, using the digital computer (ATA C-201)
With a sonic stimulation of 60/min or 120/min and 500 Hz, there was an increase in amplitude of the recruiting response of an average of +61% while with 450/min and 1000 H z an average decrease of -30% was noted.
These results suggest that the pr e viously reported decrease in excitability of the brain stem reticular formation (RF) by rhythmical sound of a low frequency and low pitch and the incr ease in excitability of RF by the rhythmical sound of a high frequency and high pitch can be attributed to a rise in indirect inhibition and a decrease in indirect inhibition, respectively, via the diffuse thalamocortical projection system.

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Previous methods for measuring plasma free tryptophan (f-Trp) are not completely satisfactory, and therefore we developed an improved method for this purpose. Free tryptophan was separated from bound tryptophan b y centrifugation through Amicon ultrafiltration membrane cones in a quite short period (within 2 minutes). The first fraction of filtrate obtained in 30 seconds by centrifugation at 3,000g at controlled pH was assayed with a high performance liquid chromatography system.
The first filtrate showed a rath er higher tryptophan concentration than that obtained by prolonged centrifugation. We propose that tryptophan concentration in the first filtrate is nearest value to f-Trp concentration in plasma, since a change of equilibrium between bound and free tryptophan during separation procedure is quite small.
Furthermore we compared our improved met h od with previous methods and got following results.
1. The fresh human plasma after replacing the air in the test tube with CO₂-N₂ gas mixture kept pH and pCO₂ almost constant but without replacing the air, pCO₂ decreased a n d pH increased.
2. The linear correlation between Trp concentration and the intensity of fluorescence was observed upto 1000 μmol/1 of Trp, but with the Denckla and Dewey method modified by Bloxam and warren it was observed upto 20 μmol/l.
3. In case of ultrafiltration under pressure to obtain free tryptophan fraction, the pCO₂gradually increased and pH decreased from 7.4 to 7.0 with the progress of ultrafiltratio n. The f-Trp concentration determined by this method showed lower value t han that by our method.
4. Our improved method proved to keep pH and pCO₂ of plasma almost constant for more than 30 minutes and the necessary sample to determine the f-Trp concentration was obtained in a short time.
5. The f-Trp concentration increased as the temperature of plasma rose, and it changed after freezing without showing definite tendency.
The dialysis method to determi n e f-Trp concentration was not proper, because as the volume of outer solution increased, the f-Trp in plasma increased until all bound tryptopha n got free.

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Summary: Free tryptophan in plasma was separated by centrifugation through an Amicon ultrafiltration membrane cone. The value obtained without control of pH was found to be lower than that obtained with control of pH by an improved method (Hijikata et al. (1981) Anal. Biochem.118,10-16).
For determina tion of the total tryptophan concentration in the plasma, high performance liquid chromatography (HPLC) was better than the method of Denckla and Dewey as modified by Bloxam and Warren ( (1974) Anal. Biochem.60,621-625), as judged on the basis of sensitivity, recovery rate and coefficient of variance.
The total tryptophan concentration in the plasma determined by HPLC was lower than that determined by the Bloxam and Warren method. The total tryptophan concentration (t-Trp), free tryptophan concentration (f-Trp) and f-Trp/t-Trp ratio were 55.8±10.2μmol/1,11.6 ± 1.5μmol/1 and 0.211 ± 0.03 (mean ± 1 SD) respectively, in healthy subjects (controls). No significant difference was observed between the values of controls and those of patients with liver cirrhosis, hepatocellular carcinoma, liver cirrhosis with hepatocellular carcinoma and hepatic encephalopathy of liver cirrhosis without bleeding. But in liver cirrhosis with bleeding, free tryptophan concentration (48.0 ± 23.3 prno1/1, p<0.001) and f-Trp/t-Trp ratio (0.645 ± 0.289, p<0.001) were significantly highe r than those of controls. In fulminant hepatitis free tryptophan concentration (106.6 ± 79.9 pmo1/1, p<0.02), and f-Trp/t-Trp ratio (0.973 ± 0.017, p<0.001) were significantly higher than those of controls, and free tryptophan concentration (p<0.005) and f-Trp/t-Trp ratio (p<0.005)were significantly higher than in liver cirrhosis with bleeding. In other words the increase of free tryptophan concentration in plasma may be related to the bleeding, and in fulminant hepatitis the binding ability of free tryptophan to albumin may decrease.
In the examined cases of liver cirrhosis, the correlatio n of the concentration of plasma tryptophan with those of other amino acids and indicators of liver function was changed when hepatocellular carcinoma was also present.

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Cancer immunotherapy and immuno-chemotherapy have made a remarkable progress in recent years and are frequently used in clinical cases. However, the mechanisms of antitumor effect in these therapies have not been well established. Particularly, there have been few reports on the tumor cell kinetics in the therapies. The purpose of this study is to examine the influences of the therapies on the tumor cell kinetics and to elucidate the mechanisms of antitumor effect.
Immunotherapy: Female Donryu rats (130-140g, body weight) inoculated with Yoshida Sarcoma (Y. S. ) cells (2.5 x 10⁶ i. p. ) on day +0 were treated with OK-432 (JOKE/kg x 5 i. m. ), a streptococcal immunopotentiator, or with PSK (125mg/kg x 5 i. p. ), a proteinpolysaccharide immunopotentiator obtained from mycelia of the Coriolus vesicol, on day -2-+2.
Immuno-chemotherapy: Some animals were given cyclophosphamide (CPM) (40,80 or 160rng/kg x 1 i. p. ) on day +3 in combination with immunotherapy.
Application of Coenzyme Q₁₀ to immuno -chemotherapy: Coenzyme Q₁₀, a physiological activator of the electron transfer system in mitohondrias, was also used (5mg/rat x 5 i. p. ) in combination with immuno-chemotherapy, on day ? 2-- +2. The cell kinetics was represented by the DNA histog ram of Y. S. obtained using flow-cytometry on day +4 or day +6.
The results obta ined were as follows.
Immunotherapy: Y. S. cells in S and G2M phase were reduced by OK-432 and PSK, but this effect varied with stage of tumor growth.
Immuno-chemotherapy: Accordi ng to results of the survival rate and the inhibition of tumor cell growth anti-tumor effect was more remarkable in the PSK-treated group than in the OK-432-treated group. The DNA histogram showed a decrease in components of G₂M phase and arrest of the cell cycle. This change of the histogram was not simply explained, indicating that in part of the mechanisms of the immuno-chemotherapy, drugs for immunotherapy and chemotherapy interact with each other.
Application of Coenzyme Q₁₀ to immuno-chemotherapy: Coenzyme Q₁₀ showed remarkable combinatory effect in survival rate and inhibition of tumor cell growth, but not represented the change of the DNA histogram.
These findings sugg est that this new method using DNA histogram obtaind by flowcytometry is one useful approach to elucidating the mechanisms of immunotherapy and immunochemotherapy, for cancer.

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To investigate the effect of vasodilator therapy on the cardiac function, the radionuclide angiocardiography by the first pass method was performed in patients with old myocardial infarction (OMI), using the computerized multicrystal gamma camera system 77. The ointment containning 12 mg of nitroglycerin (NTG-O) was applied on chest wall and abdominal skin of 23 patients and 10 mg of Nifedipine (NFP) was orally given to 20 patients.
The changes of ejection fraction (EF), left ventricular end syst olic volume (LVESV), end diastolic volume (LVEDV) and cardiac output (CO) were assessed before and after the therapy. Moreover, the response to the drugs in patients with normal ventricular function (EF>45% group I) was compared with that in those with EF of 45% or less (group II ).
Before the therapy, the patients in group II had larger LVEDV, as compared with that in group I, suggestive the maintenace of the left ventricular pump function in patients with decreased EF. LVEF in both groups was decreased after administration of NTG-O, because of preload reduction. NFP, which is known to reduce afterload, was decreased blood pressure and LVESV and increased EF and CO in all patients. In patients in group I, NTG-O reduced the preload, while NFP reduced the afterload.
In contrast, in patients with group II, both NTG-O and NFP were decreased not only afterload but also preload.
The fin ding suggest that vasodilators improved left ventricular performance by reduction of both preload and afterload of left ventricle.

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The present study was designed to assess the left ventricular (LV) wall motion abnormality using regional ejection fraction (EF) image and the moving EF image. The changes in LV wall motion after administration of Nifedipine (10mg) were also evaluated in the present study. The study group consisted of 31 patients with old myocardial infarction and 2 pa tients with angina pectoris as a control.
Radionuclide angioc ardiography was performed by computerized multicrystal gamma camera system 77 by first pass method using ^99mTcO4. Moving EF images made it possible to diagnose small anterior infarction which was impossible identify by conventional LV regional EF image. Infarction site could not be diagnosed neither by LV regional EF image nor by moving EF image in patients with subendocardial infarction. Entire LV contraction was decreased including non-infarcted zone in low EF patients. The infarcted area with greater wall motion abnormality could be identifid using enhanced moving EF image.
Improvement of LV wall motion including infarcted area was seen in patients with normal left ventricular EF, slight improvement of the motion in infarcted area by Nifedipine administration was also seen in depressed left ventricular EF patients with small infarction, while no improvement was seen in patients with large myocardial infarction.
These finding suggest that after load reduction not only improves LV pump function but also improves wall motion by decreasing wall tension.

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Nowadays, corticosteroids are indispensable for treatment of various disorders, especially for allergic and immunologic ones. Immunological suppression resulted from corticosteroid administration can improve such disorders, but, on the other hand, this suppressive state may induce opportunistic infections.
In this stud y, five cases, i. e. two cases of systemic lupus erythematosus (SLE) and three cases of bronchial asthma, having been treated with corticosteroids, were examined on the correlation between corticosteroid administration and immunologic alterations, as well as on their relationship to the incidence of infections, especially of tuberculosis which was found in three of the cases.
Case 1. M. K.,21 y. o., F., SLE. She had been administered with corticosteroids for her original disease. During this therapy, she suffered once from herpes zoster, and about one year thereafter died of complicating systemic tuberculous infection which had grown worse rapidly, in spite of potent anti-tuberculous chemotherapy. In this case, the term of corticosteroid therapy was about 30 months, the total dosage being 12g, and the daily dosage was equivalent to 5mg prednisolone (PSL) when the tuberculous infection was found. In this case, suppression of cellmediated immunity (CMI) was suggested on the basis of false positive of tuberculin reaction and decreased peripheral lymphocyte count (820/cmm)
Case 2. M. O.,33 y. o., F., SLE. During corticosteroid therapy, she suffered from cholecystitis, ostitis of mandible and pulmonary tuberculosis. All of these infectious complications were completely cured by conventional anti-microbial chemotherapy. In this case, corticosteroids had been administered for about 24 months,16.5g in total, and its daily dosage was 17.5 mg PSL when the, tuberculosis was found. No findings suggesting CMI suppression were obtained.
Case 3. Y. N.,57 y. o., M., bronchial asthma and diabetes mellitus. His b ronchial asthma had become steroid-dependent and his diabetes mellitus had grown worse due to the steroid therapy. The seral IgG level was 1,564mg/d1 under PSL 5mg/day administration, which went down to 749mg/d1 under 10mg/day of it. On the other hand, seral IgA level rose from 445mg/dl to 836mg/d1 coincidently. The peripheral lymphocyte count was low (680-1,000/cmm). About one year later, his old pulmonary tuberculosis relapsed, which well responded to conventional anti-tuberculous chemotherapy.
Case 4. M. T.,66 y. o., M., bronchial asthma. His bronchial asthma, too, had been steroiddependent. Seral IgE level rose as PSL dosage was reduced: Namely, the IgE level was 81.2--143 u/m1 under PSL 10 mg/day,851 u/ml under intermittent administration of 10-7.5 mg/day and 1473.5 u/ml under 5 mg/day administration. Throughout the corticosteroid therapy, his peripheral lymphocyte count remained within normal limit, but in regard to its subsets examin ed using monoclonal antibody of OK-series, the percentage of helper T cell tended to be high, while that of suppressor T cell tended to be low.
Case 5. H. T.,20 y. o., F., bronc h ial asthma. Her bronchial asthma had been steroiddependent for a long time, being treated with PSL 25mg/day combined with ACTH 20u/week. Later, she died of anaphylactic shock caused by intramuscular injection of ACTH. In this case, seral IgE level tended to increase (65→641u/ml) in concomitance with the decrement of PSL dosage down to 5mg/day.

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Here reported a case of esophageal cancer showing high level of anti-erythrocyte autoantibody of IgG type possibly caused by his professional long-term exposure to chemicals such as hydrogen fluoride, chloride and sulfuric acid.
Case: Y. Y.,54 y. o., M.. He had been working as a house washer over twenty years using hydrogen fluoride, chloride and sulfuric acid as cleansers and bleaching agents, used to taste the mixtures of the chemicals in order to obtain the best ratio and condition of them. As he had been quite healthy since his birth, he had never used any medicines habitually, and had never received blood transfusion. On Apr.1983, he consulted our hospital, mainly complaining of dysphagia, and was diagnosed to be suffering from esophageal cancer (squamous cell carcinoma) by means of radiological and endoscopic examinations. Immunological tests detected high level of anti-erythrocyte autoantibody (IgG), i. e. positive direct and indirect anti-globulin tests using polyspecific and anti- IgG antiserum without addition of the chemicals, both his serum and eluate from his red cells showing panaggulutination against several panel cells of blood type O, and these antibodies were absorbed completely by his own red cells. None of other autoantibodies such as LE factor, anti-nuclear antibody and anti-DNA antibody were detected.
For the operation to rem ove his cancer, concentrated human red blood corpuscles of 960ml in total were transfused to him, although pretransfusion cross match tests between these CRCs and his blood yielded incompatible results. The transfusion seemed not to be sufficiently effective, and caused a slight elevation of seral GOT, GPT and indirect bilirubin, suggesting extravascular hemolysis, for about two weeks thereafter, although no remarkable clinical side effects such as fever or renal dysfunction were recognized.
About ten mon ths after the operation, while pulmonary metastases of the cancer occurred, the anti-erythrocyte autoantibody titers in his eluate and serum went down. During these ten months, he had never been exposed to these chemicals.
The observation of this case sugges ts that a long-term oral exposure to the chemicals such as hydrogen fluoride, chloride and sulfuric acid might cause esophageal cancer, and on the other hand, it might cause production of anti-erythrocyte autoantibody. Hitherto, no papers reporting such cases as ours could be found in the literature. It should be necessary to pile up such cases, in order to provoke hygienical and experimental researches which may result the establishment of the concept of an occupational disease newly found.

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Energy dispersive X-ray microanalysis of the silicotic lung tissue of a 57-year-old founder revealed that high electron-dense material deposited in numerous macrophages found in silicotic nodules consist of silicon-containing substances such as qualz, pyrophyllite, muscotive, biovite and aluminum silicate.
This seems to be the first report which provided calculation of X-ray energy intensity ratios of elements in human silicotic lungs using analytical electron microscope in our country.

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A case of granular cell tumor of the esophagus is presented. The patient is a 51-year-old man, who developed dysphagia and loss of appetite with symptoms of the common cold. Rentgenographic and endoscopic examination of upper gastro-intestinal tract revealed findings consistent with carcinoma of the esophago-cardiac region, and esophago-cardiectomy was performed. Pathological examination of resected specimen revealed tubular adenocarci noma of the esophagocardiac region. In collision with adenocarcinoma, a small submucosal tumor of the esophagus, measuring 10 x 5 x 5 mm, was detected incidentally.
This small tumor showed typical histopa thological features of granular cell tumor. Tumor cells had small, round nuclei and abundant, eosinophilic cytoplasm which contained numerous PAS positive granules, and were arranged in clusters or cords. A portion of the stroma showed myxomatous change. S-100 protein was demonstrated in cytoplasm of tumor cells immunocytochemically. So-called “angulate body cells” were scattered in the stroma of this tumor. These cells had neither S-100 protein nor lysozyme in their cytoplasm.
Ultrastructurally, cytoplasm of tumor cells (“granu lar cells”) were filled up with numerous autophagolysosomes which contained laminated fragments of membranous structure, electrondense granules and electron-dense bodies in various proportion. “Angulate body cells” contained saccular structures in their cytoplasm which were packed with microtubules-like, tubular structures 13-16 nm in diameter. No transitional or intermediate cells between tumor cells (“granular cells”) and “angulate body cells” were detected.
Granular cell tumor arising in the eso phagus is rare and heretofore only about 20 cases are reported in Japan. Histogenesis of this tumor is still obscure, but the theory of Schwann cell origin is widely accepted in recent years. Demonstration of S-100 protein in cytoplasm of the tumor cell in our case provides further evidence supporting this theory. So-called “angulate body cells”, which appear characteristically in the stroma of this tumor, contain glycolipid and do not show the characteristics of histiocyte or tumor cell. Origin and significance of this peculiar cell still remains to be elucidated.

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