This experiment suggest that Bile and Bile acid has an antidotal effect for every strain of Rickettsia tsutsugamushi (orientalis), and that immunological hetegenity among strains of Rickettsia orientalis can be differentiated by the serological cross test with the immune state of mice surviving infection as a result of treatment with Bile and Bile acid. Observation was made in many mice which were injected with the “Umayado”, Ozeki and Shichito strain of Rickettsia tsutsugamushi (orientalis). Material of inoculation as follows; 1, 3, 5% Solution of Bile, and 10-4-10-5.5 Natrium salt of Bile acid mixed with 10-2, 10-3 brain emulsion of infected mice by these strains, in same quantity, for 10, 20, 30 minutes at 4 C. 1. 0.2ml of this material (Umayado), in each dencity and time, was inoculated subcutaneously for primary immunization of mice and these mice were challenged intraperitoneally after 65 days inoculated by 0.3ml, 10-2 Emulsion of three strains as a cross immunization test. Antigenic relationship among three strains were investigated from mortalities of mice after primary infection for 65 days and after challenge for 20 days. 2. One paralled series of observation was made in mice. Employing in subcutaneous infecting dose of 10-2-10-3 dilution of these strain mixed with 1-5% Bile for 20-30 minutes, greater part of 150 mice survived and in every case antidotal effect of Bile was demonstrated distinctly. The same result was noticed in Ozeki and Shichito strain. 3. Cross immunization test was observed in mice survived after primary inoculation of Umayado and Ozeki strain treated with Bile same qualified. Greater part of immunized mice remained resistent to reinfection, by the challenge of the strain of Umayado and Ozeki, but in case of Shichito strain, mortality of survived mice after primary infection was higher than other homologous strains. The greater part of mice by primary inoculation with Shichito strain treated with Bile succumbed to reinfection with Umayado and Ozeki strain. These result of cross immunization test suggest that Umayado strain is similar to Ozeki strain in its antigenic structure. 4. It was difficult to examin about the antidotal effect of Natrium cholate and Natrium desoxycholate for Rickettsia orientalis, because these bile acids were toxic for maus, but immunogenicity of R. orientalis treated with Bile acid seemed to be maintained. In short, it is recognized that Bile has not influence on the immunogenicity but a antidotal effect for R. orientalis. This fact suggest that Comparsion of heterogenity among strains of tsutsugamushi disease are investigated with surer confidence than the immunological test hitherto in common use.
One of the most important factors for the study of endemic Tsutsugamushi Disease is to isolate surely Rickettsia tsutsugamushi from morbid materials. Judging from the fact that the results of the ordinary isolation by the intraperitoneal blind passage in mice were not necessarily satisfactory, investigations were in the first generation of mice carried on comparing intraperitoneal with intracutaneous inoculation of Rickettsia strain. Material of inoculation is 0.2ml, 10-2-10-8 dilution of hepatolienal emulsion of mice which were infected and diseased by IV-strain of “Umayado”. 2 groups of mice, the one inoculated with definite quantity of material intraperitoneally and the other intracutaneously, were observed for 30 days in the 1st generation. From 2nd to 4th generation, intraperitoneal passage was tried with 10-1, 0.4ml hepatolienal emulsion of conditional mice: in the 1st group of the 1st generation that employed intraperitoneal inoculation, the survived during the observation or the suceumbed to infection and yet Rickettsia in their peritoneal fluid was not proved on the microscope, while then in the 2nd group of the 1st generation that employed intracutaneous inoculation, for all mice, living or dead. By these blind passage from 1st to 4th generation, Rickett is in the peritoneal fluid was observed on the microscope, and disseminating limit was investigated as follows: in case of 1st group was 10-4 dilution, and in 2nd group 10-6 dilution of the emulsion by the inoculation at the 1st generation. In short, it is recognized that Rickettsia of the same density and the same quantity, is more easily and surely preserved, when inoculated in the 1st generation intracutaneously than intraperitoneally. This fact suggests that the adoption of the intracutaneous inoculation should be recommended for the isolation of Rickettsia from diseased person and mouse.
I have made researches on the kernel of an eruptive febrile disease which is usually seen, exceptionally in summer, at Aioi-mura, Okawa-gun, the eastern part of Kagawaken. The nature of this disease was unknown but I have succeeded in the isolation of Rickettsia from a patient, and made it clear that bacteriological natures and the Rickettsia isolated are identical with Rickettsia tsutsugamushi (orientalis). A similar disease in Tsuda-cho, Okawa-gun has been proved also to be Tsutsugamushi disease. while it can be seen in winter. This proves that there is a serious kind of Tsutsugamushi disease that develops in summer and a comparatively mild kind that develops in winter. The following shows the research I have made through clinical findings on each of the different types of tsutsugamushi disease. 1. Both Aioi-mura and Tsuda-cho are topographically located near the sea. It is usually the case that the patients houses are situated in lower land districts. 2. It develops only in summer in Aioi-mura with the duration of months from June to September. 3. It develops irrespective of age. In the block of Umayado (Aza Umayado) and Sakamoto (Aza Sakamoto) of Aioi-mura, I have found all the people, regardless of their ages, exposed in their home to the attack of the disease. 4. People of the contaminated areas are engaged in farming with fishing as their side job. Almost none of them raise pigs. This shows that pigs are not the cause of it. It is not as yet clear whether it is from the migratory birds. 5. No relation of cause and effect can be found between the occupation of the villagers snd the outbreak of the disease of the patients. 6. Clinical findings of the patients Mr. Mitani who had the disease in summer in Aioimura, and Mr. Tanizawa, in winter in Tsuda-cho are both found to be the same with those of Tsutsugamushi disease, and antibiotics especially the Terramycin is effective for it. Serums of the patients agglutinated the OXK remarkably, and assurance of the rising of the agglutination titers shows that it does not differ from the Tsutsugamushi disease. 7. The antibiotics especially the Chloromycetin and Terramycin are very effective. With these antibiotics the fever goes down. After the fever drops, it is difficult to isolate the Rickettsia from the blood. In brief, I have analyzed the clinical findings of de-velopment of “Umayado Disease”, a local eruptive febrile disease, and at the same time, classified the peculiar local type of Tsutsugamushi disease according to the clinical findings into the summer type and the winter one. This leads to the fact that in a single prefecture and moreover at the places quite close to each other there can be seen a serious and a mild type of Tsutsugamushi disease in summer and winter respectively, which I think is sure to contribute to the study on the distribution and epidemic researches of Tsutsugamushi disease.
In Aioi, a village in the eastern part of Kagawaken near the border of Tokushima, an eruptive febrile disease with glandular swelling developed in summer. The death rate is found to be comparatively high. 1 made the study on the so called “Umayado Disease” broken out in Aioi-mura in summer to Mr. Mitani and Mr. Tanizawa of Tsuda-cho, a town about 15km northwest of Aioi in winter. From each of the patients, a Rickettsial strain is isolated by intraperitoneal transmission through the mice. I have performed serological tests about the Rickettsia isolated. I have discovered that these Rickettsia strains are nothing but Rickettsia tsutsugamushi. The data examined are as follows. 1. By intraperitoneal transmission, Rickettsial strains isolated have high virulence against mice. I can have succesive transmission easily and LD50 of brains, liver and spleen are 6.1, 6.9, and 7.1 respectively in Tanizawa strain, 6.3, 7.1, 7.3 respectively in Mitani strain. Again, the transmission of strains isolated by intranasal infection is possible. 2. The inoculation test into rats shows that the exudation of the abdominal fluid is scare, the Rickettsia is rarely proved by smearing and Maxcy's phenomenon is not observed. Transmission by intranasal inoculation is possible and we can see consolidated lung about 10 days after inoculation, and impression smears stained with Giemsa's stain revealed Rickettsia. 3. The inoculation test into guinea pig shows that Rickettsia appears in its peritoneal cavity by intraperitoneal inoculation but the Neill-Mooser's Reaction is not seen usually. 4. The intratestinal inoculation into rabbits causes its testinal swelling frequently. and can have successive transmission of strain by the testinal route. The intraocular inoculation test give rise to the acute iritis and the typical Rickettsia appears in the cells of Descement membrane. 5. The guinea pig, mouse. and rabbit inoculated with the isolatod Rickettsia strains produce antibodies which agglutinate OXK only. 6. The complement fixation test between the antisera of the isolated strains and R. prowazeki or R. mooseri is negative usually. So we can recognize that Rickettsia isolated by me is different from R. prowazeki or R. mooseri. 7. The intra or subcutaneously inoculated monkey with the Mitani strain produce ulcer on the inoculated place, and glandular swelling, with critical condition. Serum from the monkey 21 days after inoculation shows the positive agglutination titer of 1:1280 against OXK. By reinoculation with Tanizawa strains, the monkey develops some slight symptoms 14days after reinoculation. 8. The strains isolated by me did not increase at thioglycolate-broth, blood-agar, and glucose-broth during 7 days after inoculation. 9. Concerning the neutralization test between the strains isolatd and the other standard strains, I can say as follows. The Mitani strain showed that it belongs nearer to the Ozeki strain than to the Shichito strain or Karp strain. On the contrary the Tanizawa strain to Shichito strain rather than to the Ozeki strain or Mitani strain. Consequently, it became clear that the “Umayado Disease” of which the one type breaks out in summer in Aioi-mura and the other in winter in Tsuda-cho is nothing but the Tsutsugamushi disease. These two types differ only in antigenic structure of Rickettsia isolated from patients. It means that in Kagawaken we have two kinds of Rickettsia tsutsugamushi strains that differ in developing places and time and etiological nature.
Patients with gastric cancer today coming under the care of surgeon often considerably malnourished. Because of a high incidence of postoperative complications and deaths in these patients, a study was instituted to investigate the state of their plasma proteins. This paper deals with the plasma protein concentrations in patients with gastic cancer, the effect of operation on them and the significance of plasma protein determination in therapy and as a prognostic aid. The concentrations of plasma protein in 43 patients with gastric cancer were pre- and post-operatively measured by copper sulfate specific gravity method, and 10 normal persons and 26 patients with gastroduodenal ulcer were studied as the control. The results obtained were as follows: 1. A significant decrease in the preoperative concentrations of plasma protein was observed in patients with gastric cancer. 2. Whether radical operation could be performed or not, and whether the gastric passageway was patent or not, made no difference in the plasma protein levels of patients with gastric cancer. 3. The determination of plasma protein was of little significance in the differential diagnosis between gastric cancer and gastroduodenal ulcer except in special cases. In the absence of massive hemorrhage or severe obstruction, plasma protein concentrations below 6.5 g/dl had some significance, those below 6.0 g/dl having a important significance. 4. The lowest level of plasma protein concentration in the patients who overcame a partial gastrectomy for gastric cancer was 4.72 g/dl. 5. Following resection of the stomach for gastric cancer, it was in most cases found that a decrease in plasma protein concentration occurred in the patients having the preoperative level abov 6.5 g/dl, whereas an increase occurred in those showing a level lower than 6.5 g/dl before operation. 6. The most pronounced drop in plasma protein concentration occurred about 7 to 12 days after gastric resection for cancer of the stomach. 7. Most of the patients with gastric cancer who underwent merely exploratory laparotomy or gastrojejunostomy because radical operation was impossible to be performed, showed a increased concentration of plasma protein after operation. 8. Death following operation for gastric cancer was found to occur in the patients who had low levels of plasma protein concentration.
The data presented in the first paper demonstrated that patients with gastric cancer suffered from a significant decrease in plasma proteins concentration on admission to the hospital, but that the postoperative course was frequently associated with a increased plasma protein content. It is the purpose of this paper to discuss these findings by the determination of plasma protein fractions. The pre- and post-operative concentrations of plasma protein protein fractions in 47 patients with gastric cancer were measured by refractometry and electrophoresis. As control material 15 normal individuals including mild patients near normal, 13 cases of rectal cancer, 13 cases of various types of cancer other than that of the alimentary tract, 9 cases of sarcoma and 29 patients with gastroduodenal ulcer-a total of 90 cases-were selected. The following results were obtained: 1. Patients with gastric cancer showed a marked decrease of albumin fraction. This is the cause of the hypoproteinemia in these patients. 2. The decreased albumin concentration in patients with gastric cancr was noted to become more profound with the progress of the disease. 3. The reduction of albumin content in patients with gastric cancer was more pronounced in the cases where the passageway of the stomach was not patent than in those where it was patent. 4. In patients with gastric cancer an increase of both globulin and fibrinogen fraction was demonstrated. 5. With the occurrence of metastasis of gastric cancer in the liver, there was found an increase in β- and γ-globulins. 6. Albumin-globulin ratio in patients with gastric cancer was lowered and became lower with the progress of the disease. 7. The albumin level and the albumin-globulin ratio in patients with gastric cancer proved to be the index of the prognosis, and albumin levels below 2.70g/dl and albumin-globnlin ratio under 1.0 were associated with an increased incidence of the cases where radical operation was not indicated and of postoperative complications. 8. The determination of plasma protein fractions was of little significance in the differential diagnosis between gastric cancer and gastroduodenal ulcer. 9. The albumin concentrations in patients with gastric cancer decreased after gastric resection. The degree of the decrease was more profound in the cases where preoperative levels of albumin were higher. On discharge from the hospital the albumin contents were, in most cases, found to increase over those before operation. 10. An increase in both fibrinogen and globulin fractions occurred after gastric resection for cancer. On discharge from the hospital these levels, although lowered, yet remained slightly higher than those before operation.
The data presented in the second paper indicated that in Patients with gastric cancer the preoperative determinations of albumin content and albuminglobulin ratio were of value for judging the patient's condition of nutrition and operative prognosis, but that the alternation occurring in the concentrations of plasma protein fractions following operation was not always proportional to the preoperative degree of protein deficiency and the clinical picture. Therefore this paper is concerned with the study on hypoproteinemia in patients with gastric cancer on the base of the examination of both the concentration and the total circulating mass of plasma protein. Blood volume, plasma volume, cell volume, total circulating plasma protein mass, total circulating hemoglobin mass, plasma protein contration and hemoglobin concentration in 14 patients with gastric cancer were measured before and after operation. As control material similar studies were made in 11 normal persons, in 13 patients with gastroduodenal ulcer, in 5 cases of cancer of the brest and in 4 cases of Graves' disease. Additional studies were done to determine a standard of blood requirement to be transfused by investigating the effect of transion in the cases in which blnod had been administered before and after operation. The resnlts of these studies were as follows: 1. In patients with gastric cancer there was a decrease in blood volume and in cell volume. 2. Plasma volume was above normal in many patients with gastric cancer and decreased in only a few cases. 3. The decreased blood volume in patients with gastric cancer chiefly was due to a reduction of cell volume, and there were a few cases where plasma volume was concerned in it. 4. The prognosis of the patients with gastric cancer who had a decreased blood volume was poor. 5. In patients with gastric cancer a decrease of both total circulating hemoglobin mass was noted, the loss of the later being more pronounced. 6. In patients with gastric cancer the transfusion of at least 1000 cc each of whole blood and plasma was considered necessary before operation. 7. In patients with gastric cancer blood volume, plasma volune and cell volume were reduced 4 to 8 days after the resection of the stomach. After the 12th postoperative day, plasma volume increased, whereas cell volume did little. 8. Total circulating plasma protein mass in patients with gastric cancer decreased 4 to 8 days after operation, but increased after the 12th postoperative day. 9. Total circulating hemoglobin mass in patientients with gastric cancer.was markedly decreased following resection of the stomach. The value showed a especially sharp decrease on the 4th postoperative day and still further decreased gradually afterwards and there were a few cases in which it shifted to the increase on discharge from the hospital. 10. There were the cases of gastric cancer where a discrepancy between the concentration and the total circulating mass of blasma protein was noted and this tendency was more striking in both of hemoglobin. It was often difficult, therefore, to estimate total circulating mass of plasma protein and hemoglobin by the determination of only concentration of plasma protein and hemoglobin. 11. Total plasma protein mass rather than total total hemoglbin mass was suitable as an index of tissue protein deficiency in the postoperative course of patients with gastric cancer. 12. The variations in values of blood determined after operation in patiens with gastric cancer depended on the preoperative state of body nutrition and on the amount of blood postoperatively transfused. In well nourished patients and in those transfused over 800cc of blood, the degree of reduction in blood volume aud plasma volume was low, and on discharge from the hospital, there was a tendency for total circulating plasma protein mass to increase above the preoperative value.