A histochemical study on alkaline phosphatase, acid phosphatase, β-esterase, aminopeptidase and β-glucuronidase has been caried out to clarify the relationship between the function and morphology of the human breast tumors; mastopathia: 63, fibroadenoma: 8, gynecomastia: 7 and cancer: 34. The surgically removed tumors were cut at 20μ in -20°C cryostat, then the sections were incuvated in each histochemical reaction mixture. For the histochemical demonstration of hydrolytic enzymes, azo-coupling methods were employed, i.e., alkaline phosphatase, acid phosphatase and β-esterase were demonstrated by the methods described by Pearce, 60) β-glucuronidase by Seligman et al, 61) and aminopeptidase by Nachlas et al.62) Alkaline phosphatase activity was the strongest in capillary vessels and myoepithelium and moderate in the normal lobulus and ducts. This enzyme activity increased in proliferating area, but in precancerous area and in cancer it decreased markedly. Acid phosphtase activity was usually low in normal glands and not prominently decreased in cancer than that of alkaline-phosphatase. β-Esterase activity was moderate in breast tumors but increased in periluminal region and necrotic tissue. Aminopeptidase activity was moderate in normal glands but strong in proliferating area. This enzyme activity was elevated in precancerosis, but decreased in cancer cells. β-Glucuronidase activity was increased in proliferating area and precancerous area, but slightly decreased in cancer. This enzyme activity was the highest in duct papillomatosis.
Histochemical observation of human breast tumors were caried out on lactic dehydrogenase, glucose-6-phosphate dehydrogenase, isocitric dehydrogenase, succinic dehydrogenase, malic dehydrogenase, glutamic dehydrogenase, α-glycerophosphate dehydrogenase and β-hydroxybutyric dehydrogenase. The surgically removed breast tumors were examined; mastopathia:, fibroadenoma:, gynecomastia:, and cancer:. The tumors were cut at 20μ in-20°C cryostat, then the sections were stained by the methods described by Pearce60) with Nitro-BT as the electron acceptor. The following results were obtained. Lactic dehydrogenase activity was the strongest of all dehydrogenases studied in this report. It increased in proliferation and slightly decreased in cancer. The stromal reaction was moderate. Glucose-6-phosphate dehydrogenase activity did not decreased in cancer tissues so much as that of other dehydrogenases. These results showed that anaerobic glycolysis and hexose monophosphate shunt played an important role in breast cancer metabolism. For investigation of tricarboxylic acid cycle, succinic dehydrogenase, isocitric dehydrogenase and malic dehydrogenase were studied. These three enzymes showed similar distribution to each other. Their activity were increased in proliferation, but decreased in cancer. Their activities were especially decreased in poorly differentiated cancer cells except for scirrhous carcinoma. Glutamic dehydrogenase activity revealed a simillar pattern to that of TCA cycle enzymes. As enzymes of fatty acid metabolisum, α-glycerohosphate dehydrogenase and β-hydroxy-butyric dehydrogenase were examined. The activity was relatively weak in benign tumors and very weak in cancer.
Immunological studies were performed to determine the cancer-specific fractions in the sera taken from the gastric cancer patients. The antigen, which used in this studies, was the pooled sera from the advanced incurable gastric cancer patients and the antisera were from the rabbits sensitized with the intramuscular injection by Freund's complete adjuvant. 1. With immunoelectrophoretic examination, the increased density of the precipitate lines was demonstrated in α-and β-globulin portions in the majority of the patients. however, it had no definite proportional relation to the development of cancer. 2. Density of precipitate lines maintaned its level up to the period of 4 to 5 weeks after extended radical operation for gastric cancer. It showed remarkable increase immediately after the operation, unexpectedly. 3. There was little difference in the pattern of the cancer serum and the non-cancer by means of the plate method of Ouchterlony.
Modified Boyden's hemagglutination test was performed with the gastric cancer sera and the tannic acid treated human O-type red cells conjugated with the mitochondria fraction from the pooled gastric cancer tissues for the demonstration of cancer-specific humoral antibody. 1. The increase of humoral antibody was recognized in the serum from the gastric cancer patient sensitized with his own cancer tissue. 2. These titers of the humoral antibody were seemed not to be proportional to the development of cancer, the extent of infiltration in stomach serosa and lymphatic metastasis. 3. No significant relatiohship could be noted between these titers of humoral antibody and the immunoelectrophoretic patterns in gastric cancer sera. 4. There was some relationship of inverse proportion between the titers of humoral antibody and the immunoelectrophoretic patterns in the cancer sera before and after extended radical operation for the gastric cancer.
Microscopical studies were performed on the infiltration of mononuclear cells in the interstitum adjacent to cancer. It was examined whether the extent of the cellular infiltration had some correlation to the titer of humoral antibody and immunoelectrophoretic pattern in sera from each patient. 1. No significanct correlation was obtained in the extent of the cellular ifiltration and the development of gastric cancer. 2. The extents of the infiltration were proportional to the titers of humoral anti body by Boyden's hemagglutination test. 3. There was no relationship between the extent of the infiltration and the immunoelectrophoretic pattern. These data would seem to suggest that human gastric cancer has some homograftic aspects in the immunological point of view.
The function of megakaryocytes in femur bone marrow of dog after left side lumbar sympathectomy or left side parasympathectomy, sectioning the posterior roots of fifth, sixth and seventh lumbar spinal cord as well as the first sacral segment, was observed by means of clinical bone marrow tissue culture. And also observed the change of platelet counts in nutrient vessel of tibia after these operations, comparing with those on the another right side as control. The results are as follows. 1) In the cases of sympathectomy, megakaryocyte functions (number of megakaryocytes in growth zone, percentages of moving type of megakaryocyte and tentacle formation type for platelet formation) on bone marrow tissue culture was accelerated. And the platelet counts in nutrient vessel of tibia was increased on the sympatheticomized side. The maxima of these changes were showed on around 10th day after the operation, then these changes had been disapeared until 60th day. 2) Megakaryocyte function on the parasympatheticomized cases was diminished. And also the platelet counts in nutrient vessel of tibia was decreased on the parasympatheticomized side. The maxima of these changes were on around 10th day. But the change of number of megakaryocytes in growth zone was maximum on 5th day after the operation. These results should be presumed that sympathetic denervation introduce increase of blood flow in the bone marrow and parasympathetic denervation introduce decrease of blood flow in the bone marrow, then secondarily lead up to increase or decrease the function of megakaryocytes. Futhermore, it may be thought that parasympatics influences some direct effect for the function of megakaryocytes in bone marrow.
By means of clinical bone marrow tissue culture influences of sympathetic stimulating or blocking drugs and parasympathetic stimulating or blocking drugs on the function of megakaryocytes were observed. The solution of these drugs at various concentrations were added directly to the culture media and obtained the following results: 1) The function of megakaryoctes was diminished when epirenamine or nor-epirenamine being sympathetic stimulating drugs added to culture media. 2) 2-Benzyl-imidazoline-hydrochloride being sympathetic blocking drugs accelerated the function of megakaryocytes. 3) In the cases administered acetylcholine hydrochloride or bethanechol chloride which are parasympathetic stimulating drugs the function of megakaryocytes were accelerated. 4) No effects on the function of megakaryocytes were observed when atropine sulfate or dl-tropyltropate-n-methylbromide were added to culture media directly. These results may conclude that the function of megakaryocytes are diminished when adrenaline that is produced at the end of sympathetic nerve affects on bone marrow directly and also the function of it is accelerated when acetylcholine which is produced at the end of parasympathetic nerve affects on bone marrow directly.
The diagnosis of cancers by meas of polarography is of a common practice in clinics and among others the Protein Index Method devised by Muller is considered to be quite useful for the identification of cancer bearing persons from normal person. With this Protein Index Method, the authors comparee the protein indices of patients beating various cancers with those of normal persons, and also studied the changes in the protein index before and after the operation performed on stomach cancer, gastro-duodenal ulcers, and stomach polyps in order to find out the relationship between the changes in symptoms and the Protein Index. As the results the following points were clarified. Although the serum reaction of polarography is non-specific and hence is not sufficiently applicable to all kinde of cancer, it does show significant differences in gastro-duodenal ulcer group and stomach cancer group. It is also valuable method for the clinical diagnosis of lung cancer. It seems that this is one of the methods also useful for knowing a possible recurrence of stomach cancer by meanse of the changes in the Protein Index befor and after the surgical operation.
The experimental studies were carried out to examine the relation between the hemorrhagic tendency after massive transfusion of preserved blood and serotonin. The results were as follows: 1) After massive transfusion of preserved blood, whole blood serotonin and platelet serotonin levels were decreased, while plasma serotonin levels were slightly increased. Tissue serotonin levels were increased in the lungs and slightly elevated in the liver, but there was no change in the small intestines. 2) After massive transfusion of preserved blood, the 5HIAA levels in the urine were decreased. The same tendency was also observed in surgical cases without the transfusion. This change was correlated with post-operative oliguria. 3) The effect of the reduction of bleeding time by administration of synthetic serotonin was not constant, but passing. 4) In coagulation studies on serotonin added to blood in vitro, no effect was found concerning coagulation time, prothrombin time, recalcification time or clot retraction test. On the thrombelastogram, it was observed that coagulation was accelerated in lower serotonin concentration, while suppressed in higher serotonin concentration. 5) In preserved blood, whole blood serotonin and platelet serotonin levels were decreased. On the contrary, plasma and red cell serotonin levels were increased as time passed. 6) From the experimental results above, it is concluded that serotonin contained in platelets should not be recognized as one of the factors causing hemorrhagic tendency after massive transfusion of preserved blood.
The authors reported the results of screening tests for hypocatalasemia at several places in Japan during the year of 1963. Among 10, 358 individuals submitted to the test, 24 proved to be hypocatalasemia, i.e., the state of acatalasemic gene carrier. The examinee consisted of five different populations; 1, 050 students of Okayama University, 2, 363 school-children at Akasaka, Gifu Pref., 598 Korean school-children around Hiroshima City, 2, 109 people including school-children at Kozagawa, Wakayama Pref. and 4, 238 school-children at Tsushima Is., Nagasaki Pref. The rate of hypocatalasemic cases varied from zero to 0.67% in each place. Concerning acatalasemia, the reported cases amounted to 34 families and 76 individuals by the end of 1963.
The application of ultrasonics to medical diagnostic method has not been so much successful, because the complexity of the sound reflecting structures echos which are so numerous and variable that controlled experiments and consistent results are difficult to obtain. In this point, fundamental electrical and ultrasonic study in medicine has been very important; electrical circuit of the pulse generater, receiver, amplifier and indicator, pulse length and intensity of the transmitter, the directibility of the ultrasonic beam and scanning mehtod, they would be concerned with upper problem. A new type of apparatus for this study has been designed and produced by the author; Sensitivity of the recciver is 90100 dB, measuring distance is 2mm1.8m and the freguency of the pulse generator is 0.5MC10MC. A 500pF capactior is charged to about 1, 800 V through a high resistance and is then discharged by a thyratron tube (3C45) through the primary winding of a pulse transformer. The amplitude of the transmited ultrasonic pulse rises from 1μs to its maximum value in 20μs. Fifty or sixty such pulses are transmited each secound to human body. This apparatus has been used to diagnostic medicine and fundamental study in the author's clinic.
New application method of ultrasonic probe to diagnosis of digestive organs was deviced by the author. The special ultrasonic probes for this study have been designed and producel by the author. Transdnsducers with barium titanate ceramic vibrator (2.25MC) and 10mm or 7mm in diameter have been used at the tip of the flexible probes. Probes are 23m in length and 5mm in diameter and can be rotate in all directions in the oesophagus, stomach, rectum and discending colon. Clinically, digestive organs would be examined by transoral or transanal application of these probes. In the experimental and clinical studies, the echos from the vagus, heart and mediastinum were obtained from the probe which was located in the oesophagus: the echos from the liver, gall bladder, diaphragm, pancreas, spleen and retroperitoneum were obtained from the probe which was located in the stomach; the echo from the urinary bladder, small intestine and kidney were obtained from the probe which was located in the descending colon. In the clinical studies, the echos from the heart, aorta and inferier vena cava showed pulsating, fine and multiple wave form; the echos from the cancer tissues, scharp and multiple wave form; the echo from the diaphragm, sharp wave form and movable by respiration.
Transorbital ultrasonic examination was deviced by the anthor. The echo from the IVth ventricle were obtained by using this method. In order to record normal echo-gram, the probe (1MC or 2.25MC) was placed directly on the upper eyelid and the echos from the orbit, pyramid, IVth ventricle and occipital bone were recorded by this method. Clinically, this method would be applied to diagnosis of the infratentorial tumors. Abnormal echos would be obtained between the echos from the pyramid and the occipital bone on the side of tumor or the echo of the displaced IVth ventricle would be obtained.
Experimental tuberculosis in guinea pigs infected with cultures of H37Rv strain exhibing high degrees of resistance to SM, PAS and INH and subsequently treated with usual dose and large dose of INH, combined administration of INH and sulfonamide compounds, SM and KM was carried out. Also a clinical observation of reversal of infectiousness of the tubercle bacilli in the sputum of the patients with chronic pulmonary tuberculosis was performed by this treatment regime. The results lead to the following conclusions. In the experimental tuberculosis, administration of the usual dose and large dose of INH and combined administration of INH and sulfonamides revealed remarkable effectiveness to the SM- and PAS-resistant strains and considerable to the INH-resistant strain. Administration of the large dose and combined treatment of INH and sulfonamides were more effective than the usual dose of INH. In clinical observation, these kinds of INH treatment showed same tendency as in the animal experiment. So it is thought that these treatment regime of INH should be tried in the treatment of the patients with INH resistant tuberculosis. However, in this experiment, administration of the further larger dose of INH did not show the more remarkable effectiveness and it may be considered that there is a limit in the effectiveness in the treatment of the large dose of INH. Administration of SM to the animals with even low degrees of SM resistant strain scarcely showed its effectiveness and it should be warned to administer SM to the patients with SM-resistant tubercle bacilli. Although KM revealed somewhat lesser effectiveness than SM of the same dose, KM brought marked effectiveness to each resistant strain and showed no cross resistance to SM-, PAS- and INH-resistant respectively. The sulfonamides revealed low antituberculovs effectiveness in the experimental tuberculosis in guinea pigs and it is thought that single administration of the sulfonamides to tuberculosis is unable to be expected thier effectiveness. But it is a point of interest that the blood level of sulfadimethoxine was showed to keep a high and long-lasting.
Experimental tuberculosis in mice infected with cultures of the H37Rv strain exhibiting high degrees of resistance to SM, PAS and INH and subsequently treated with single administration of KM, CS and TH respectively was carried out, Also a clinical observation of reversal of infectiousness of the tubercle bacilli of the sputum of patients with chronic pulmonary tuberculosis by this treatment regime was performed. Kanamycin treatment of 1 mg per mouce scarcely showed the effectiveness to all of the resistant strains. Cycloserine treatment of 2.5mg per mouce did not show any improvement of the tuberculous lesions. It is assumed that many inactive factors to CS in mouse body take part in this uneffectiveness of CS, but especially the activity of D-alanine does, While, ethionamide treatment of 1 mg per mouce revealed marked effectiveness to each resistant strain and it is noted that TH treatment of 1 mg per mouce was as effective as INH treatment of 0.1mg per mouce. In the clinical observation, single administration of KM, CS or TH revealed no remarkable effectiveness to the patients with triple resistant tubercle bacilli to SM, PAS anp INH. Especially this treatment regime to the patients with chronic far advanced, active pulmonary tuberculosis scarcely brought reversal of infectiousness. It may be concluded that the single administration of TH should not be done because of easy appearence of the resistance in short time.
Experimental tuberculosis in mice infected with cultures of H37 Rv strain exhibiting high degrees of resistance to SM, PAS and INH and subsequently performed following combined treatment; 1) TH 1 mg and KM 1 mg per mouce, 2) TH 1 mg and CS 2.5 mg per mouce, 3) TH 1 mg, KM 1 mg and CS 2.5 mg per mouce, 4) TH 1 mg, KM 1 mg and SF (Sulfisomezole) 2.5 mg per mouce, 5) TH 1 mg, CS 2.5mg and SF 2.5mg per mouce was carried out. The results showed remarkable effectiveness to each resistant strain. But the effectiveness of these combined treatment was chiefly based on the effectiveness of TH and the synergism of the combined drugs scarcely noted in this experiment and the combined treatment including CS or SF revealed rather antagonistic. In clinical observation of reversal of infectiousness of the sputum of the patients with chronic active pulmonary tuberculosis resistant to SM, PAS and INH, each mentioned above treatment regime showed marked effectiveness in the moderately advanced pulmonary tuberculosis but scarce in the far advanced cases. Also the combined treatment with three drugs were more effective than that with two drugs.