岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 線維芽細胞抑制剤による悪性腫瘍の治療に関する研究 クロロキン剤の基礎と臨床（第一報） 297 316 EN Kiyoshi Hiraki Ikuro Kimura Zensuke Ota Kenwo Asano Hiroshi Kageyama Koichi Shibuya Hidenari Kotani Ryozo Matsuura Junichiro Tsuchida Tatsuo Sezaki Toshinobu Hiraoka Hajime Himei Yoshiaki Moritani Masatoshi Yamana Followings are the results derived from the treatment of animal and human cancers with a fibroblasts inhibiting agent such as chloroquine, based on the unique idea of ours. 1). In implanted tumors of animals, the effect was noted in Bashford cancer and Brown-Pearce cancer relatively rich in connective tissue in terms of life prolongation, an inhibition of tumor growth and of decrease of liver catalase activity, an improvement of iron metabolism, an enlargement of necrotic area in histology, an inhibition of connective tissue conponents, and a decrease of acid mucopolysaccharides. A tendency for a decrease of amount and cell numbers of ascites was almost the only effect noted in Ehrlich cancer, Yoshida sarcoma, and MH 134 poor in connective tissue. 2). 75 cases of humau cancer were treated with this agent and among these 75 the effect was evaluated in 40 cases with definite histological diagnosis in respects with subjective and objective improvement. It was effective in 28 cases, resulted failure in 11 cases, and gave an obscure result in 1 case. Cases which were treated for more than 2m onths never resulted in therapeutic failure, indicating that the agent was somehow effective for all cases but those in the last stadium. In respect with organs involved, the effect was most prominent in lung cancer and urinary bladder cancer, somewhat less in gastric cancer, and often seen in carcinomatous peritonitis and many other advanced cancers. This effect was noted in a comparatively short period in terms of subjective improvement, regression in size of tumors in a number of cases, a decrease of serum lactic acid dehydrogenase, and improvement of general condition, a tendency for necrosis of tumor and an inhibition of interstitium. 3). The above results indicate that the agent is more effective for tumors rich in connective tissue and that it's secondary effect on tumors through the inhibition of interstitial connective tissue is considered as the operative mechanism of chloroquine against malignant tumors, but it's anti-inflammatory effect and generalized influence on hosts are to be taken into consideration. The agent is indicated in inoperable cases, postoperative relapse, or prior and following an operation. A search for stronger fibroblasts inhibiting agents and an evaluation of combined therapy with so called anti-cancer agents are being made and that with mitomycin C is giving a fairly promising result. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 サポニン投与による実験的類白血病反応に関する研究 第3編 骨髄血管構造所見 283 295 EN Tadashi Sato The detailed vascular structure of the bone marrow of saponin-treated mice was studied with clarified bone specimens (Spalteholz's method), because the technic appeared encouraging in elucidating the problems of the mechanism of both granulocytic liberation into the peripheral circulation and development of the extramedullary hematopoiesis. The results obtained were as follows. 1. In the normal control, unit sinuses were connected with each other, and a unit sinus net was primarily composed of 6 unit sinuses in a hexagon form, and many of these sinus nets were mutually connected to make up the complex network. The sinus wall was composed of a single layer of endothelial cells. The vascular system of the bone marrow was a completely closed system, no fenestrated vessel-walls being demonstrable in normal mice. 2. In saponin-treated mice the sinus network was disordered. There was enlargement or narrowing of the sinuses with destruction of their wall. These alterations were seen already at 24 hours after an injection of saponin to mice. At this stage, no extramedullary hematopoiesis was yet seen in spleen tissue culture or in histologic specimens of the spleen. It was considered, therefore, that saponin acted upon sinuses causing destruction of their thin walls. This was followed by the liberation of blood cells into the peripheral cirulation from the destructed sinuses. Finally, these blood cells were transported and multiplied in the organs such as the spleen and liver. The author believe that this is the mechanism of the extramedullary hematopoiesis (colonization theory). The simultaneous finding that megakaryocytes increased in number in the spleen, while their number decreased in the bone marrow gave support to this view. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 X線及び20-Methylcholanthreneにより惹起せしめたRF系マウス白血病の無細胞濾液移植に関する研究 213 231 EN Shozo Irino Zensuke Ota Koichi Okada Isao Miyoshi Tatsuo Sezaki Norio Kawamura Mineo Kunisada Koji Ikejiri Masanobu Sato Susumu Sota Masanori Suzaki Incidence of leukemia in RF mice exposed to a single whole body irradiation of 350 r X-rays was 77.4%, 41 out of 53 animals. Of them, 20 developed the thymic and other 20 the non-thymic lymphocytic leukemia, while only one developed myelogenous leukemia. Incidence of leukemia by 20-methylcholanthrene painting was 88%, 22 out of 25 animals. Of them, 12 developed lymphocytic leukemia and other 10 myelogenous leukemia. Experiments of inoculating cell-free filtrate prepared from these X-ray and 20-methylcholanthrene-induced leukemic RF mice revealed the following facts. 1) Cell-free filtrates from the leukemic mice which were induced by a single whole body X-irradiation were inoculated to the newborn mice of the same strain less than 16 hours old. Two (16.7%) of the twelve incculated animals developed leukemia 357 th and 374 th days, respectively, after the inoculation. Inoculation of the cell-free filtrates from normal mice to the newborn mice of the same strain did not produce leukemia. 2) Attempts to inoculate cell-free filtrate from 20-methylcholanthrene-indeced leukemic RF mice to newborn mice of the same strain were successful. 3) Cell-free filtrate from myelogenous leukemic mouse caused lymphocytic leukemia in one of three inoculated mice. This is suggestive of the possibility of transformation of the type of leukemia during cell-free transmission. 4) Intracerebral inoculation of cell-free filtrate from the brain of a mouse with lymphocytic leukemia also produced lymphocytic leukemia in one of two inoculated mice. 5) Inoculation of cell-free filtrate from the leukemic mice developed mammary carcinoma in one of three inoculated female mice. 6) Serial cell-free passage in two generations of 20-methylcholanthrene-induced leukemic mouse was possible. One of two inoculated mice developed leukemia in the first transmission and one of eight inoculated mice in the second transmission. 7) Only two cases of spontaneous lymphocytic leukemia were observed in RF mice during the past four years and the cell-free transmission of one leukemic mouse was also successful. 8) Virus-like particles were demonstrated by electron microscope in the lymph nodes of 20-methylcholanthrene-induced leukemic mouse. These facts suggest that the X-ray and chemical carcinogens may act to induce leukemia or mammary carcinoma by the "activation" of a latent virus naturally resident in the RF mouse. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 腹水腫瘍の細胞化学的研究 第�V編 脱水素酵素系に対する制癌物質皮下投与の影響 201 211 EN Takao Kawashima The four kinds of anti-tumor agents i. e. endoxan (0.5 mg per mouse, three times), nicotinamide (15 mg per mouse, three times), DPN (40 mg per mouse, one time) and colchicine (4 mg per mouse, one time) were subcutaneously administered to dds-mice which had been transplanted with Ehrlich ascites tumor seven days previously. And the changes of the activity of the DPN diaphorase, TPN diaphorase, succinic degydrogenase (SD) and α-glycerophosphate degydrogenase (α-GD) of the ascites tumor cells were cytochemically examined in the smear method and suspension method. 1) In general, their was no definite morphological change of the cells except for inhibition of mitosis in the above dosages, nor destruction of the enzymes. But a few variations of the enzymatic activity were noted. 2) After the administration of endoxan, stainabilities of the DPN diaphorase, SD and α-GD were slightly increased, while that of the TPN diaphorase showed no change. 3) After the administration of nicotinamide, the number of tumor cells with the high activity of the DPN diaphorase and TPN diaphorase was increased about five per-cent. NO change of the activity of the SD and α-GD was observed. 4) After the administration of DPN, the number of tumor cells with the high activity of the DPN diaphorase was increased about ten per-cent. But no change of the TPN diaphorase, SD and α-GD was observed. 5) After the administration of colchicine, there was no change of the activity of the DPN diaphorase, TPN diaphorase, α-GD, while the SD activity was increased in lesser degree. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 腹水腫瘍の細胞化学的研究 第�U編 脱水素酵素系に対する制癌物質腹腔内投与の影響 189 199 EN Takao Kawashima The four kinds of anti-tumor agents i.e. nitromin (0.5 mg per mouse, four times), colchicine (0.05 mg per mouse, one time), sakomycin (5 mg per mouse, one time) and mitomycin C (0.04 mg per mouse, one time) were intraperitoneally administered to dds-mice which had been transplanted with Ehrlich ascites tumor seven days previously. And the changes of the DPN diaphorase, TPN diaphorase, succinic dehydrogenase (SD) and α-glycero-phosphate dehydrogenase (αGD) of the ascites tumor cells were cytochemically examined in the smear method and suspension method. 1) In general, moderate morphological changes of the tumor cells by these anti-tumor agents were observed, and the changes of the stainability of the oxidative enzymes were often observed. 2) After nitromin administration. no change of the stainability of the DPN diaphorase and TPN diaphorase in the smear method and slight increase of their stainability in the suspension method were observed. The stainabilities of the SD and α-GD were decreased in lesser degree in both methods. 3) After colchicine administration, no change of the stainability of the DPN diaphorase and TPN diaphorase in the smear method was observed, but their stainability of all tumor cells showed a tendency down to keep same intensity. A slight decrease of the stainability of the SD and α-GD was observed in the smear method, while their stainabilities were slightly increased in the suspension method. 4) After sarkomycin administration, stainabilities of all four enzymes were decreased in certain degree in the smear method, but the DPN diaphorase and TPN diaphorase staining intensity were remarkably decreased in the suspention method and their stainabilities of all cells showed a tendency down to keep same intensity. The SD and α-GD staining intensity were also decreased slightly in the suspension method. 5) After the administration of mitomycin C, changes of the staining intensity of the DPN diaphorase and TPN diaphorase were scarcely observed in both methods. The staining intensity of the cells in the end stage of the interphase showed to be decreased in lesser degree. Stainabilities of the SD and α-GD were slightly decreased in the smear method, while their stainabilities were slightly increased in the suspension method. 6) It might be assumed that the decrease of the stainability reacted with the anti-tumor agents in the smear method specimen showed decrease or destruction of the oxidative enzymes and that the change of stainability reacted in the suspension method specimen showed change of the activity of the enzymes. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 eP系マウスの病態生理に関する実験的研究 第3編 ep系マウスの尿成分について 179 188 EN Ryosuke Sugiu 1. Routine urinalysis revealed that nothing characteristic was found in the ep-mouse in comparison with the control, the CF-1 group. 2. Specific urine study for electrolytic substances revealed that specific difference was not found in Na, K, Cl and Mg but in Ca of which concentration in 100 ml urine volume was measured by about 160 % of the control group. Such data were obtained in case of young ep-mouse which had not manifested the convulsion yet, and it appears, therefore, to show that it is of the characteristic for the ep-mouse. 3. No difference was found of 17-OHCS in urine between the ep-mouse and the control group. 4. No significant difference was either found in amino acids in urine between the two group. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 ep系マウスの病態生理に関する実験的研究 第2編 ep系マウスの脳波ならびに各種抗痙攣剤のこれにおよぼす影響について 161 178 EN Ryosuke Sugiu An attempt was made to find out the effect of anticonvulsive agents such as diphenylhydantoin, phenobarbital and homocarnosine (γ-aminobutyryl-L-histidine) on the abnormal resting potentials of the ep-mouse. 1. Bilateral and sporadic spikes were noted to be characteristic abnormal potentials in the resting E. E. G. of the ep-mouse comparing with the one of the control, the CF-1 group. It was observed in 95% of the ep-mouse. 2. The spikes were found to be inhibited by the administration of phenobarbital: the absolute abolishment was obtained with 12 day successive administration of the 50〜60mg/kg/day. 3. Such inhibition was also obtained by the administration of diphenylhydantoin: the absolute abolishment was obtained with 15 day successive administration of the 120〜130mg/kg/day. 4. So was found in case of homocarnosine intracerebrospinal administration. The absolute abolishment of the spikes was found after the 7th post-administration day includingly. 5. The abolishment of the abnormal spike potential by these anticonvulsive agents was preceded by inhibition of convulsion. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 ep系マウスの病態生理に関する実験的研究 第1編 Homocarnosineのep系マウスにおよぼす影響について 145 159 EN Ryosuke Sugiu Homocarnosine, one of the GABA associates was administered on the ep-mouse into cerebrospinal fluid through frontal region of intact scalp and skull. 1. Increased respiration and heart-beat and behavioral inhibition were observed in all cases immediately after the administration. Slight loss of body weight was followed for the first week in a half of the cases. 2. Anticonvulsive effect was obtained with the administration of homocarnosine. It is to be noted that the minimum effective dosage was 4-5 mg/kg which appears to be almost the same dosage contained in the cerebrum. 3. The anticonvulsive effect tended to be gradually accelerated by the time passing and to show the peak at the 3rd post-administration day, which was hardly varied during the following 2 month observation period. 4. Several associates of homocarnosine were examined in standpoint of anticonvulsive effect. Carnosine has also anticonvulsive effect, but less than homocarnosine. GABA, GABOB, histidine and alanine do not have such effect. 5. Homocarnosine does not cause any effect on acetylcholine and cholinesterase activity in cerebrum. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 Bilirubinoid-Azo色素に関する研究 第3編 黄疸尿より分離した直接bilirubinに於けるdiazo反応について 139 144 EN Kotaro Kaneda According to the method described by Komuta, jaundiced urine was salted out with ammonium sulfate, and the bilirubin, collected on a filter paper, was extracted in methanol. After the addition of the double dose of chloroform to the methanol solution, the dark-reddish precipitation was obtained, that was the ester-form bilirubin. On the other hand, its supernatant was dried in vacuo and then dissolved it in water, followed by the addition of chloroform to that solution. The dark-brown flocculate appeared at borderline of the both solution, that was considered to be an unknown bilirubinoid forming the intermediate zone. A spectrophotometrical examination was performed on the diazo reaction's process of the ester-form bilirubin and the unknown bilirubinoid. The following results were obtained; After the addition of the diazo reagent to the methanol solution of ester-form bilirubin, a color with reddish-violet ton was appeared rapidly, while a maximum on the absorption curve of the azopigments appeared after a little while, and its absorption maximum was shown at 530 to 540 mμ. A blue-violet color appeared macroscopically after the addition of HCl to the ester-form bilirubin-azopigment, its absorption maximum moved to 560 mμ, while no change in its absorption curve was seen after leaving this acid azopigment in the room temperature for 48 hours. In adding the diazo reagent to the methanol solution of the bilirubinoid forming the intermediate zone, its absorption curve also moved as like as that in the ester-form bilirubin. On the other hand, the absorption maximum of its azopigment showed at 550 mμ, and there was no appearance of a color with the blue ton by the addition of HCl to this azopigment. Moreover, its absorption maximum moved only a little toward 560 mμ, namely to the side of the long wavelength, and this acid azopigment showed no particular change on its absorption curve after leaving it in the room temperature for 72 hours. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 Bilirubinoid-Azo色素に関する研究 第2編 数種bilirubinoidのdiazo反応の分光化学的研究 125 137 EN Kotaro Kaneda The process on the indirect diazo reaction after adding methanol to the bilirubinoids of bilirubin, mesobilirubin and bilirubindimethylester have been studied spectrophotometrically, the diazotised sulfanilic acid was used on the diazo reaction. The following results were obtained; Bilirubin-azopigment showed an absorption　maximum at 560 mμ, and also at 420 mμ before the reaction was completed. The time to reach the absorption maximum at 560 mμ prolonged in parallel with a decrease of the amount of the diazo reagent, and also the absorption maximum at 420 mμ remained higher. The same results to the above were found in mesbilirubin-azopigment, except showing the absorption maximum at 550 mμ and at 390 mμ before the reaction was completed. Again the same results were found in bilirubindimethylester-azopigment, except showing the absorption maximum at 540 mμ and at 410 mμ before the reaction was completed. The absorption maximum of the bilirubin-azopigment at 560 mμ was flattened by leaving it in room, showing the more remarkable change along with a increase of amount of the diazo reagent. The same results were also found in the azopigments of the mesobilirubin and bilirubin dimethylester, but having slight differences in positions of the maximum absorption. After leaving the bilirubinoid in room, a maximum absorption was appeared at 400 to 420 mμ in the case having a plenty amount of diazo reagent, which was found to result from the excess amount of the reagent remained in the reacting solution. All of the acid azopigments, yielded after the addition of HCl to the each azopigments of bilirubin, mesobilirubin and bilirubindimethylester, showed their maximum absorption at 580 mμ, a very stable character and no significant change on the absorption curve by leaving them in room for a week. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 Bilirubinoid-Azo色素に関する研究 第1編 数種Bilirubinoid-azo色素のPaperchromatographyによる検討 115 124 EN Kotaro Kaneda Azopigments, yielded after the addition of diazotised sulphanilic acid to the each of bilirubin, mesobilirubin and bilirubindimetbylester, have been studied by paper partition chromatography if there were some differences in Rf on the chromatograms among these azopigments. The following results were obtained; Regarding developing solvents, methylethylketone-n-propanol-water (15:5:6), n-propanol-water (7:3), n-propanol-acetic acid-water (4:1:5) and saturated n-butanol with 10% citric acid were found as giving the most adequate partition of these samples from the chromatographies done with uses of more than forty kinds of solvents. Using these developing solvent systems, the distinct diferences in the partition were found among these three bilirubinoid-azopigments. In developing process on the chromatography of these azopigments, a reddish-violet spot changed its color partially to blue, and was finally divided into two spots of red and blue. This indicated the coexistence of two kinds of the azopigments in the crude samples. By leaving or heating the paper after developing the chromatography, a yellow spot appeared with the Rf common to these three azopigments, and it was possibly throught that this spot resulted from excess amount of the diazonium salt remained in the reacting solution. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 移植動物癌の抗腫瘍性抗体産生に関する研究 第3編 制癌剤投与担癌マウスおよびX線照射癌細胞感作マウスの抗体産生機序について 103 114 EN Kiyoshi Takeda Tumor isoimmunity has been questionable; thus, two experiments were carried out using mice of isogenic lines (C(3)H×dd: F(1)). Experiments were with tumor-bearing mice treated with anticancer drugs, and with mice immunized with heavily irradiated tumor cells (MH 134). 1. By treating the tumor-bearing mice with anticancer drugs, the γ-Globulin titre increased in the subject serum, followed by a marked decrease as the tumor increased. With anaerobic glycolysis and with neutralization test of tumor cells, the inhibitory action of sera were ineffective. 2. The result of x-ray irradiation on MH 134 tumor cells in vitro, produced total stain using Eosin when irradiated with more than 8000r. With the same dose of irradiation, tumor cells lost their transplantability in sensitive F(1) mice. 3. F(1) mice, immunized with heavily irradiated MH 134 tumor cells which lost their transplantabilty, became resistant to the transplantion of relatively few, fresh MH 134 tumor cells. 4. F(1) mice immunized with heavily irradiated MH 134 tumor cells, were bled and their organs excised. These sera, organ homogenates and organ suspensions on being tested for anticancer activity, revealed greatest antitumor activity in the spleen followed by serum and liver. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 移植動物癌の抗腫瘍性抗体産生に関する研究 第2編 癌細胞感作家兎吸着血清中の抗腫瘍性抗体の消長について 91 102 EN Kiyoshi Takeda Rabbits were immunized with Ehrlich ascitic tumor cells and then bled periodically. Following adsorption of the antisera with normal mice liver powder or packed Ehrlich tumor cells, the antisera was examined by electrophoresis, by anaerobic glycolysis by Warburg's manometry and for life prolongation study by neutralization. 1. In case of intravenously immunized rabbits sera, adsorbed γ-Globulin titre was comparably greater in the earlier stage with normal mice liver powder. Furthermore, the titre of γ-Globulin was greater and prolonged than the titre of preimmunization. 2. After adsorption with liver powder, the inhibitory action of antisera on anaerobic glycolysis of tumor cells was weak in the earlier stage, but still retained its former effectiveness in the later stage. 3. In the neutralization tests of adsorbed antisera, life prolongation was noted equally in the initial and later stages, with a decrease in the early period following adsorption. 4. Following adsorption of the antisera with the packed Ehrlich tumor cells, an ineffectiveness was revealed in both anaerobic glycolysis and neutralization. 5. It was deduced through points 2-4, that, the antisera produced in Part-1, immunized with Ehrlich tumor cells, contained antiumor antibody. 6. a) In IV immunized rabbits sera, adsorbed γ-Globulin titre was greater in the early stage. b) In anaerobic glycolysis and neutralization, the inhibitory action was weaker in the earlier stage. In supposition, the unadsorbed antisera produced from heterogenically and intravenously immunized animals contains species specific antibody chiefly in the earlier stage with tumor specific antibody chiefly in the later stage. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 移植動物癌の抗腫瘍性抗体産生に関する研究 第1編 癌細胞感作家兎非吸着血清中の抗腫瘍性抗体の消長について 77 90 EN Kiyoshi Takeda Rabbits were immunized with Ehrlich ascitic tumor cells by intravenous injections or intramuscular injections of Freund's adjuvant. Rabbits were bled periodically, with the antisera being separated from the blood. Using these sera, the γ-Globulin titre was examined by paper-electrophoresis, anaerobic glycolysis with Warburg's manometry, and for life prolongation studies by neutralization. I wanted to know whether the antisera contained the antitumor antibody or not. 1. After immunization, the γ-Globulin titre in rabbits sera increased, with the Albumin titre decreasing. In the once intravenously injected group, such change was insignificant, but remarkable in the groups injected 3 or 6 times. In the adjuvant group, almost identical change occurred as in the latter two groups. 2. The inhibitory action of antisera on anaerobic glycolysis of tumor cells was weak in the groups injected 3 or 6 times. The antisera of the adjuvant group also had strong inhibitory action for a long duration following slow initiation. 3. Antisera of the once intravenously injected group were ineffective in the neutralization test. But in the groups injected 3 or 6 times and in the adjuvant group, it was very effective, resulting in life prolongation of the mice. 4. The peak of the γ-Globulin titre preceded the peak of glycolysis and neutralization in the group injected 6 times, whereas in the adjuvant group, peaks of the three coincided. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 動物腫瘍の蛋白基，各種酵素およびこれに及ぼす制癌剤の影響に関する組織化学的研究 第3編 Ehrlich固型癌，NF肉腫の各種水解酵素，脱水素酵素およびこれに及ぼす制癌剤の影響に関する組織化学的研究 55 75 EN Kiyoshi Ishihara Activities of various kinds of hydrolytic and oxydative enzymes in the Ehrlich solid carcinoma and NF sarcoma were histochemically examined. And the influences of the antitumor agents, i.e. Colchicine, Nitromin, Mitomicin C, Endoxan and Merphyrin on these tumor tissues were also observed, and the following results were obtained. 1. The hydrolytic enzymes: The activities of alkaline-phosphatase and acid-phosphatase in both tumors were negative. The activities of β-Esterase and β-glucuronidase were slightly positive in the proliferating area of tissue, while their activities increased in the degenerated area. Aminopeptidase showed stronger activity in the NF sarcoma than in the Ehrlich solid carcinoma, but its activity decreased promptly if the tumor cells degenerated. 2. The oxydative enzymes: Succinic, lactic and α-glycero-phosphate dehydrogenase showed moderate activities in the proliferating area of tissue, while their activities decreased, gradually in the area of necrobiosis or necrosis. The activities of malic, glutamic and β-hydroxybutyric dehydrogenase were slightly positive in the proliferating area of tissue, and they increased at the transitional part between the proliferating and necrobiosis area. The activity of glucose-6-phosphate dehydrogenase was negative in the NF sarcoma, but moderately positive in the proliferating area of Ehrlich solid carcinoma. When the tumor cells degenerated, its activity was promptly decreased. 3. The degeneration and necrosis of the tumor cell were observed in various degree by the administration of the antitumor agents. The enzyme activities did not show the special changes by the administration of the antitumor agents. The giant cells were observed in the administration of Endoxan, Nitromin and Merphyrin. The giant cells showed the highly increased activities in β-glucuronidase and all oxydative enzymes. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 動物腫瘍の蛋白基,各種酵素およびこれに及ぼす制癌剤の影響に関する組織化学的研究 第2編 固型腫瘍の蛋白基およびこれに及ぼす制癌剤の影響に関する組織化学研究 41 54 EN Kiyoshi Ishihara The protein and protein-bound amino-bases in the Ehrlich solid carcinoma and NF sarcoma were histochemically examined. And the influences of the antitumor agents, i.e. Colchicine, Nitromin, Sarcomycin, Mitomycin C, Endoxan and Merphyrin on these tissues were also observed, and the following results were obtained. 1. The stainability of the protein-bound SH and SH-SS was slightly increased by the administration of the antitumor agent. Both tumors showed moderate reaction in the proliferating area of tissue, while they showed always moderate or high reactions in the area of necrobiosis or degeneration, and especially remarkable by the administration of Mitomycin C and Sarcomycin. 2. The protein-bound SS reaction did not reveal so remarkable changes by antitumor agents in both tumors, but was moderately increased by Colchicine administration. 3. The tyrosine reaction did not show the definite regularities in both tumors by the administration of the antitumor agent. 4. The protein-bound NH(2) reaction did not show the definite regularities in both tumors by the administration of the antitumor agent. Especially, the reaction in the proliferating area of the NF sarcoma was moderately increased by the administration of Sarcomycin and Mitomycin C. 5. The phenol and its compounds reaction did not show the remarkable changes in both tumors by the administration of the antitumor agents. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 動物腫瘍の蛋白基，各種酵素およびこれに及ぼす制癌剤の影響に関する組織化学的研究 第1編 腹水腫瘍細胞の蛋白基およびこれに及ぼす制癌剤の影響に関する細胞化学的研究 25 39 EN Kiyoshi Ishihara The protein and protein-bound amino-bases in the Ehrlich and Yoshida tumor cells were cytochemically examined. And the influences of the antitumor agents, i. e. Colchicine, Nitromin, Sarcomycin and Mitomycin C on these tumor cells were also observed, and the following results were obtained. 1. The protein-bound SH showed the moderate reaction in cytoplasma and nucleus in both tumors. The nucleolus of the Ehrlich tumor cell showed a weak reaction, while that of the Yoshida tumor cell a remarkably positive reaction. By Colchicine administration, the stainability of SH was increased. 2. The protein-bound SH-SS reactions revealed almost the same localisation in both tumors as the SH reaction, but the intensity of the reaction was stronger than that of SH. And the cytochemical pattern of SH-SS caused by the antitumor agents was almost the same as SH. 3. The protein-bound SS reaction was weak in both tumor cells. Generally, SS reaction diminished by the administration of the antitumor agents, especially by MitomycinC administration to the Yoshida tumor, Nitromin and large amount of Colchicine to the Ehrlich tumor cells. Only by MitomycinC administration to the Ehrlich ascites tumor, the reaction of SH was increased. 4. In the Yoshida tumor, by the administration of all antitumor agents, the stainability of SH, SH-SS and SS were diminished, while in the Ehrlich tumor the changes of the stainability were variable. 5. Tyrosine reaction was moderate in both tumor cells. The nucleus of the Ehrlich tumor showed the negative, and that of Yoshida tumor the slightly positive reaction. 6. The NH(2) reaction was moderate in the cytoplasma of both tumors, and very strong in the nucleus. In the Yoshida tumor, the stainability was diminished by the administration of all antitumor agents, while in the Ehrlich tumor the reaction was diminished by the large amount of Colchicine, and the reaction was elevated by Nitromin and small dosis of Colchicine. 7. The phenol and its compound reaction were moderate in the Ehrlich tumor and weak in the Yoshida tumor. By the administration of the antitumor agent, these reactions were generally diminished in both tumor cells, but Mitomycin C administration to the Ehrlich tumor these reactions were elevated. No potential conflict of interest relevant to this article was reported.

岡山医学会 Acta Medica Okayama 0030-1558 75 1-3 1963 口腔剥離細胞の電子顕微鏡的研究 1 24 EN Toshiro Kobayashi Electron microscopic observation was applied to both twenty cases of normal mucosa and eighteen cases of squamous cell carcinoma of the oral cavity and their exfoliative cytologic findings were investigated in comparison with the histologic findings. �T. TISSUE CELLS Desmosome, which are regarded as a universal structure of cohesion of epithelial cells, was much poorer in carcinoma cells than in normal ones and intercellular spaces displayed a marked variation in width in carcinoma tissue cells. Thus intercellular cohesion was morphologically very poor and loose in carcinoma. In both cells an intercellular border was distinct and no syncytium was found to be formed. Tonofilaments, which can be distinctly found in normal cells, could not be noticed and a higher development of endoplasmic reticulum and mitochondria was shown in carcinoma cells. However, the mitochondria of carcinoma cells was remarkable in degenerative findings and was larger in size than that of normal cells. There was a strong resemblance between basal cells of the normal tissue and carcinoma cells in an indefinite nuclear shap and a decrease of cytoplasmic: nuclear ratio. In carcinoma cells a fine structure of their nuclear membrane was well-preserved and their nucleoli could be remarkably noticed. �U. EXFOLATED CELLS In normal cells spinous processes of almost definite size and shape were extending regularly around them, while in carcinoma cells poorer processes of various shape and size were extending irregularly around them or sometimes they were missing. Mechanical destruction was particularly marked in the area of peripheral cytoplasm of normal cells of the deeper layer under the superficial spinous cell layer probably because of a tight intercellular cohesion. Generally speaking, findings of tonofilament, endoplasmic reticulum and mitochondria corresponded to those of tissue cells in both normal and carcinoma cells, while a slightly severer degeneration of intra-cytoplasmic organelles in exfoliated carcinoma cells could be noticed in comparison with carcinoma tissue cells. A fine structure was well-preserved in nuclei of carcinoma cells, but disintegration and disappearance of the nuclear membrane could be noticed in severely degenerated carcinoma cells and no thickening of the nuclear membrane could be found. From these results obtained the cytologic findings of the smear in Papanicolaou staining and electron microscopy were compared and discussed. No potential conflict of interest relevant to this article was reported.