Using the typhus bacilli taken from the departmental stock, the author obtained crude nucleoprotein fraction, Julianelle substance and Boivin substance by respective fractionating procedure and investigated the antigenic properties of these fractions with the use of hemolysis test, precipitation test and the inhibition test for hemagglutination in the presence of influenza virus. The following results were obtained.
1) As the result of hemolysis test, crude nucleoprotein fraction showed most favorable antigenic capacity through the fractions obtained from Sal. typhi and Sal. paratyphi A and B, then Boivin substance followed that, but the other fraction had no significance for antigenic capacity. In addition to this, chick's erythrocytes was most favorable for the hemolysis test to carry the sensitivity.
2) By the precipitation test, it was noticed that Julianelle substance, Boivin substance and crude nucleosprotein fraction were avilable respectively as the precipitinogen. Especially, on the study of precipitation test using Julianelle substance and Boivin substance both obtained from various type of Sal. typhi, and antiserum obtained by injection of these substances, it could be supposed the existence of type specificity on antiserum. From old days, the members of Salmonellae were classified depending upon the antigenic properties of agglutination reaction. The author studied the type specificity from the point of precipitation reaction, and noticed fairly evident specificity.
3) As the cellular fractions of bacteria used for the inhibition test of hemagglutination, the author noticed the superiority on crude nucleoprotein fraction or Boivin substance than Julianelle substance, which was reported to be favorable for inhibition. However, these action for inhibition was seemed to be really unspecific, therefore it could be said no type specificity was present at all.

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Using the hepatitis virus isolated from hepatitis patient, the author cultured the virus by serial passage in chorio-allantoic cavity of chick embryo, collected it in purified form by means of ultracentrifuge and observed the resistance to heat that was supposed to be a important feature of the virus. For the infection of the virus it does not render the lethal effect on mouse, only pathological findings have the significance for identification of the infection. On account of this, the author adopted pathological findings on the liver of the mouse, that was inoculated the heat-treated purified virus, as to determin the resistance to heat. The following results were obtained.
1) From the results of repeated test that the heat resistance of purified virus was examined, it could be concluded that the heating at 55°C for 30 min. did not affect its activity at all; but the heating at 60°-65°C for 30 min. showed serious effect on the activity, however, whole virus could not be inactivated by this treatment; and the heating at 70°-75°C for 30 min. inactivated the virus completely.
2) The result of the inactivation test described above, in that the complete inactivation of the virus was achieved by heating at 70°-75°C, was confirmed further by the serial passage of the virus. However, the resistance of the virus was supposedly varied to some extent by the heating method and the pH of medium at heating.

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Using the hepatitis virus isolated from hepatitis patient. the author purified the virus by HASHIMOTO'S method, and studied some properties that were supposed to be important for the virus; the resistance to disinfectant and ether, the inactivation by ultraviolet rays and the filtrability. The following results were obtained.
1) For the study of the properties of the virus the author adopted the pathological findings as to determine the causable change by the infection. This method was assumed to be reasonable in spite of the dispute for the good agreement of both results obtained by tests on the purified and unpurified virus.
2) The virus was resistant fairly good for disinfectant, hence it needed for over 3 weeks to inactivate the virus completely by the action of disinfectant, and the virus gradually lost its activity as the duration of disinfectant treament.
3) It was confirmed that the virus was filtrable well through the Seitz EK filter and resistant to the action of ether. Also the virus could stand against the irradiation of ultraviolet rays for less than 40 min.
4) By the intraperitoneal injection of inactivated virus for 3 times, the animal could be immunized for the challenge of the same or the other type virus. But in sometime slight pathological changes were observed, and this implied the immunization thus obtained was not so stiff as capable for immunising the all challenge.

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In order to study the enzymatic properties of human type tubercle bacilli, H37 Rv, H37 Ra and BCG, and avian type bacilli, Takeo's strain, Myc. ATCC 607 and Myc. phlei, the auther prepared the soluble fraction and particulate fraction of these cells by means of ultracentrifuge frnm the ground cells that were cultured in Sauton's media. Using the fractions thus obtained, the author observede nzyme activities of these fractions. And then, the author picked out the soluble fraction of H37 Ra; dialyzed it and studied the dehydrogenase activity of the dialyzed soluble fraction for lactate and malate with the addition of Cofactor by means of Thunberg's methylen blue reduction method. The following results were obtained.
1) It was observed that the enzyme activities of two fractions, soluble and particulate, were distinctly different in any organisms.
2) It could be comfirmed that the specilic enzyme activity of each fraction for ehe substrates and the intensities of that activities were varied as the strains.
3) The dehydrogenase activity of the dialyzed soluble fraction of H37 Ra for lactate was raised by the addition of CoI. And riboflavin could substitute for CoI.
4) The more increased dehydrogenase activity for malate was observed by the simultaneous addition of GoI and riboflavin to the dialyzed soluble fraction of H37 Ra compaired with the single addition of CoI to that fraction. However riboflavin failed to replace the role of CoI.

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Using the same procedure and organisms as in the Part I, the author prepared the soluble fraction and the particulate fraction, and carried out further enzymatic study of these fractions by means of Warburg's manometer techinipue. The following results were obtained.
1) The soluble fraction of H37 Rv and H37 Ra could oxidize lactate and other substrates fairly successfnlly; but on the other hand, the particulate fractions of that coule oxidize lactate and succinate specifically and failed to oxidize the other substrates. And this fact was also noticed in the case of the particulate fraction of BCG.
2) In the case of Takeo's strain one of the avian type bacilli, the enzyme system that oxidise malate was found merely in the particulate fraction.
3) It could be observed that the soluble fraction of Myc. ATCC 607 showed intense endogeneous respiration, and the particulate fraction of the organism could oxidize fumarate and malate specifically.
4) The soluble fraction of Myc. phlei could oxidize lactate successfully, but oxideze the other substrates not so satisfactorily. And in the case of the particulate fraction, it could oxidize any of lactate, succinate or malate, but, in a small extent.

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MURAKAMI and his collaborator had reported already the properties of the hepatitis virus isolated from hepatitis patient in detail, and they pointed out some similarity of the virus with the IZUMI fever virus. The author obtained both viruses in purified form by means of ultracentrifuge, and carried out further investigation on the virological properties of these viruses. The following results were obtained.
1) It was confirmed that both viruses could be obtainable easily and efficiently in purified form by means of ultracentrifuge, and showed powerful pathogenecity to mouse capable of inducing the typical pathological changes by infection; however, the infection of either virus did not render the lethal effect and was detectable as latent infection in that only the pathological findings were informative. And by the electronmicroscopical study, both viruses were observed in minute round or oval form whose diameter was 50-70 mμ.
2) From the viewpoint of the virological properties, both viruses were much different in the resistance to heat and disinfectant: the purified hepatitis virus could stand aginst heating at 70°-75°C and against the action of disinfectant, 0.2% formalin or 0.01% marzonin, for less than 3 weeks; on the other hand the IZUMI fever virus was inactivated by heating 55°-60°C for 30 min. or by action of the disinfectant for 2 weeks.
3) By the irradiation of ultraviolet rays, it is needed to inactivate the purified hepatitis virus for 40 min. irradiation, but in the case of IZUMI fever virus it was completely inactivated by only 5-20 min. irradiation.

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As has been reported in the part I, the author obtained the hepatitis virus, and IZUMI fever virus in purified form by means of ultracentrifuge method proposed by HASHIMOTO, and studied comparatively on some serological properties of these purified viruses. The results obtained were following.
1) The antiserum obtained by inoculation of the purified IZUMI fever virus (YASHIRO'S strain) had high titer and showed the titer of 1:64 on the complement fixation test to the YASHIRO'S strain, but did not give the positive reaction to the purified hepatitis virus when it was used as antigen. Vis-a-vis, the antiserum obtained by inoculation of the purified hepatitis virus showed the titer of 1:64-1:128 to the hepatitis virus and negative reaction to the IZUMI fever virus. Hence, in the view of complement fixation test, the serological properties of the viruses were evidently different.
2) Conerning to the results of neutralization reaction and “Absätigungsversuch” after WILDFÜHR, it could be postulated by the pathological findings the occurence of effective neutralization reaction when the test conducted between a virus and the antiserum obtained by inoculation of the same virus; contrarily to this the neutralization reaction between a virus and the antiserum obtained by the inoculation of other virus did not occur. Since the reaction thus observed was not so evident as the complement fixation reaction described above, sometimes it was hardly possible to identify the definite reaction. However, this reaction was still informative, especially in referential use of the reaction to complement fixation test, because of the occurence of certain inhibition or mitigantion of pathological changes as the result of the neutalizing effect.
3) From the facts described above, it could be concluded that the evident difference of properties was noticeable in both viruses tested despite of the presence of some similarity on serological properties.

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When high hydrostatic pressure was applied to isolated frog heart, following changes were observed on the cardiac action:
1) At relatively low pressure, heart rate increased, but at moderate high pressure, it decreased.
2) There occurred no change of conduction time, namely P-Q interval.
3) Upon appling pressure, the amplitude of action potential of ventricle muscle, i.e. the height of R wave augumented.

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Experiments were performed to examine the action of high hydrostatic pressure on the isolated frog heart ventricle.
The refractory period of the heart muscle prolonged by about 20 per cent. It seems also that cardiac ventricle under high pressure responds either all or none to electrical stimulus.

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When high hydrostatic pressure was applied to isolated ventricle muscle, the results were obtained as follows:
(1) The excitability of cardiac muscle augments by the pressure from 50 to 200 atm.
(2) The response to electric stimuli disappears at high pressure (400 atm. or more).
(3) It is interesting that the excitability of ventricle muscle rises transiently upon release from high pressure.

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As objects for study, the following three types of work were taken:
a) Work in a railway hospital---light labor.
b) Work of baggage-transfer men at the station---medium labor.
c) Maintenance work---heavy labor.
Regarding the above three, I examined the worker's fatigue mainly by the Ogata's method and observed the development of the density of urine-protein and the quantity of it excreted per hour and also compared the influences on the urine-protein by giving water to the worker. The following results were obtined.
1) In the light labor, the density of urine-protein and the quantity of it excreted per hour did not increase.
2) In the medium and heavy labor groups, the density of urin-protein and the quantity of it excreted per hour did increase.
3) In all three, light, medium and heavy labor, as food and water given, the urine quantity increased and the density of urine-protein decreased. So, in putting the Ogata's method into practice, it is advisable to restrict the taking in of water and to observe both the density of urine-protein and the quantity of it excreted per hour.
4) In the light labor group it is presumed that the urine-protein is excreted comparatively constantly.
5) Testing by the Ogata's method, the fatigue could not be measured in the railway hospital worker, but in the work of the baggage-transfer men at the station and the maintenance worker a slight fatigue was noticeable.

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As an example of the heavy labor for a short while, I chose a model stocking and measured the fatigue of the stocker in spring, summer and winter mainly by the Ogata's urinary protein measuring method and observed the development of the density of urine-protein and the quantity of it excreted per hour before and after the work and also studied the influences affected on the urine-protein by giving much water to the stocker just before the work.
In addition, in summer I measured the quantities of Cl in the urine and the perspiration excreted while at work. In winter I measured how the work affected the pulse frequency and the physical strength (the breathing capacity, the strength of the back and the grasping power).
The results were as follows:
1) The density of urine-protein and the quantity of it excreted per hour markedly increased with work.
2) The density of urine-protein and the quantity of it excreted per hour shour showed the highest figures just after the work.
3) As compared with the cases in summer, in winter the density of urine-protein and the quantity of it excreted per hour showed a tendency to rapidly recover.
4) Comparing the cases where water was not given, to the cases where water was given just before the work, the urine quantity increased and the density of urine-protein decreased after the work, but the density of urine-protein and the quantity of it excreted per hour increased with work.
5) As water was given, the density of urine-protein and the quantity of it excreted per hour showed a tendency to recover more rapidly than the cases where it was not given.
6) In summer the quantity of urine-Cl excreted per hour decreased after the work and when water was given it showed a tendency to recover more rapidly than the cases where water was not given.
7) In summer loss of Cl was more remakable in the perspiration than in the urine.
8) In summer the Cl quantity of the perspiration while at work conspicuously increased when water was given.
9) After the work, the pulse frequency increased, while the breathing capacity decreased and the grasping power declined in the right hand and in the left hand it did not decline. In the back, however, strength was increased.

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In order to ascertain the distribution of the bacteria introduced into an animal body by tracing radioactivity of radioisotope labeled organism, the author studied the properties of P³² labeled Sal. 57 S and the distribution of radioactivity in mouse that imply the presence of the organism. The following results were obtained.
1) To obtain an appropriate organism for the investigation, the author observed the stability of bound radioactivity on the cells that were obtained by three different labeling methods. i.e. on resting cells, on 3 hrs cultured cells and on 18 hrs cultured cells respectively. Concerning to dissociation of radioctivity from labeled cells by repeated washing or incubation of the cells, it was noticed in vitro that the labeling on resting cells was most labile, then came next the labeling on 3 hrs cultured cells, but that on 18 hrs cultured cells was most stable. Moreover, the study of. P³² incorporation into phosphorus fructions of the cells by Schneider's method showed the marked incorporation of P³² into nucleic acid and protein fractions by the labeling on 18 hrs cultured cells compaired with other labelings.
2) The radioactivity of labeled cells was observed to be stable in vitro compaired with living cells, when the labeled cell was killed by application of heat.
3) From the observation of interrelation between the presence of living cells and the radioactivity in the body of mouse, it could be said that the presence of bacteria could be traced out by detection of radioactivity within 6 hrs after injection and that the situation became to be disturbed beyond that time possibly owing to the dissociation of P³² from the organisms caused by metabolism and divisions of the cells. While in the case of killed organism the situation was remained undisturbed for a fairly long time than in the case of living cells for absence of metabolism and division of the cells.
4) When the labeled organism was injected intraperitoneally into mouse, it was observed the marked accumulation of radioactivity on spleen and liver of the animal 2-3 hrs after the injection. In the case of intravenous injection, a conspecious accumulation was occured even 1 hr after the injection. However, by subcutaneous injection the radioactivity did not spread out into the body and ramained the site of injection for a fairly long time.
5) In the study on a group of previously immunized mouse by a intraperitoneal in jection of killed organisms, a wide distribution of radioactivity into the body of animals was inhibited by the intraperitoneal challenge of the labeled cells. While, the distribution was almost the same as the control group excepting slightly increased accumulation of radioactivity into the spleen by the intravenous challenge of the labeled cells.

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Exfoliative cytologic findings of the oral mucosa in three cases of Acatalasemia (Takahara's disease) were investigated by means of Papanicolaou's method. Three cases were sisters of the same family, 19, 22, and 15 years of age. Oroantral fistula was noticed in the left molar area in Case 2 and Case 3 complained of loosening of anterior lower teeth with marginal gingivitis in the left lower molar ares.
Smears were prepared by the following technics:
1) Scraping four different areas of the oral mucosa---hard palate, gingiva, radix linguae and buccal mucosa without causing bleeding.
2) Sediment taken from centrifuged mouth washings by physiologic saline solution.
3) Cells caught by membrane filter or both of them.
All the smears were stained by Papanicolaou's technic and histologic examination of the gingiva was made in comparison with cytologic smears in Case 3. Thus keratinization of the epithelial cells were studied.
1) Keratinizing tendency was the same as those of normal mucosa in Cases 1 and 3. In the ulcerating area of Case 3 keratinized cells showed an increase as healing was going on, while they showed a marked decrease in Case 2, which might be due to the decrease of local resitance associated with ulcerative changes and the influence of partial denture.
2) Exfoliated cells of the whole oral mucosa in case of mouth washings Cases 1 and 3 showed the same findings as normal cases and Case 2 showed a marked decrease of keratinized cells. These findings were same as those of four different areas of the oral mucosa.
3) Gingiva of Case 3 showed little difference from normal mucosa in histopathologic findings.

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The author studied on the changes of the lipid and mitochondria in experimental degeneration of the retina of the rabbit caused by intravenous injection of 5% solution of sodium iodate.
Lipids in the pigment epithelinm slightly decrease with the exception of those in the lipochrin.
No remakable change was observed in the lipids of the outer segments and ellipsoid of vesual cells.
Most of the mitochondria in pigment epithelial cells disappeared as the cells degenerate, but some of them remained unchanged even in rather advanced stages of degeneration.

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MURAKAMI and his collaborator had isolated the hepatitis virus from hepatitis patient and cultured in the chorio-allantoic cavity of chick embryo. The author newly isolated the hepatitis virus from the patient and studied on virological properties of the virus having obtained the purified virus. The following results were obtained.
1) In the treatment by heating or by disinfectant, the purified virus showed high resistance to these treatment, and these resistances were thought to be the featuring properties of the virus.
2) Also it was observed that the virus was highly resistant to irradiation of ultraviolet rays, and to action of ether.
3) The purified virus was highly infections to mouse by peroral administration of it. The pathological changes were observed conspicuously, and moreover the changes induced were just the identical in any case.
4) By the serological study, it was observed that the purified virus had an eminent antigenic capacity and reacted with the immune serum obtained by inoculation of the virus to rabbit giving the antigenic titier of 1:64-1:128. Also the virus showed distinctive reaction in the complement fixation test using the convalescent patient's serum from hepatitis; hence, this test might be highly valuable for the serological diagnosis of hepatitis despite of rather low titer than the above.
5) The most difficult problem was the identification of neutralizing effect in the neutralization test and “Absättigungsversuch”. For this purpose the author adopted the mitigation of pathological changes caused by the neutralizing effect and observed the marked mitigation of the changes on the test using the virus and corrsponding immune serum; but this evidence was not observed in the test of the virus and uncorresponding serum. The mitigation of the changes was not distinct in the “Absättigungsversuch” possibly by the occurence of combination of the virus and its anti-body in the body of animal instead of in the test tube. It is to be expected the further study on the test procedure and the method of identifying the neutralization effect.

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Rabbits were used as experimental animals. Total calcium contents in serum and weights of rabbits were determined. When 1 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 30 mg/kg and 40 mg/kg of NaF were administered for 10 weeks.
(1) Total Calcium contents in serum
(a) When small amounts of fluorine (1 mg/kg and 10 mg/kg of NaF) was administred, total calcium contents in serum increased.
(b) When 20 mg/kg, 30 mg/kg and 40 mg/kg of NaF were administered, amounts of calcium in serum decreased remakably.
(2) Weights of rabbits
It was found that rabbits administered with 1 mg/kg, 5 mg/kgand 10 mg/kg of NaF tend to gain weight and to loss when 20 mg/kg, 30 mg/kg and 40 mg/kg of NaF was given.

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By performing tissue culture of the bone-marrow aspirated from the sternum of normal persons, ribs removed at the thoracoplastic surgery, and from the femur of normal rabbit, the author observed the course of the bone-marrow tissue culture, and obtained the following results.
1. In the bone-marrow tissue culture of the human sternum the growth area keeps on increasing up to 18 hours of the culture and its average relative growth rate is 10.64; in the case of the rib removed at thoracoplastic surgery it lasts for 36 hours and the average of its relative growth rate is 16.90; and in the case of the rabbit femur it lasts for 48 hours and the average of its relative growth rate is 45.37. The fluctuation curve of the average relative growth rate in each case is somewhat like a parabola.
2. The average of the cell density index in the case of human sternum is 52 at 18 hours after the start of calture; in the case of the rib removed at the thoracoplastic surgery it is 54 after the 24-hour culture; and in the case of the rabbit femur it is 67 after the 24-hour culture.
3. In the case of the sternum of normal persons the average value of wandering velocity of neutrophils reaches the maximum of 15.24μ/m three hours after the start of the culture and neutrophils cease wandering on the fourth-sixth day of the culture; in the case of the rib removed at the thoracoplastic surgery the average of the wandering velocity reaches the maximum of 10.51μ/m, and the wandering of neutrophils ceases in the sixth to seventh day of the culture; and in the case of normal rabbit femur the average wandering velocity of pseudoeosinophils reaches the maximum of 14.17μ/m immediately after the start of the culture and the wandering of pseudoeosinophils stops on the 5th to 6th day of the culture.
4. The cell growth area is divided into the central, intermediate and peripheral zones. The course of cell activity in the growth area is divided into the initial, middle, and terminal stages, and obsewations have been carried on from stage to stage.

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In human and rabbit bone-marrow tissue culture the author carried out vital observations on the movement cycle, pseudopodia and movement pattern of mature and immature leucocytes along with moving pictures of them, and Obtained the following results.
1. The movement cycle of leucocytes in culture is divided into six stages, namely, the preparatory stage, pseudopodia forming stage, migratory stage of the main body, stage of caudal retraction, stage of distribution of granules, and resting stage.
2. In the case of immature leucocytes the movement cycle is not complete, and in most of them different stages can not be clearly distinguished.
3. The pseudopodia of leucocytes are classified into five types, namely, tongue shaped, saw-tooth shaped, bubble-shaped, flag-shaped, and tentacle-shaped types, starting from the most active wandering velocity down.
4. The movement pattern of leucocytes is classified from the higher wandering velocity down into A to I types; and A type is still further divided into three sub-types and B type and B type into two sub-types.
5. Observations have been carried on the relationship between the pseudopodia type and cell movement type, and various leucocytes in bone marrow.

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1. By performing the culture of human and rabbit bone marrow the author carried out vital obseruvations under the light microscope and the phase-contrast microscope on the leucocyte series as well as studies on the moving pictures of these cells in culture.
2. Those cells presenting transiently the nucleus in the string shape constriction in the middle or the nucleus in the shape of a dumb bell should be considered as undergoing a temporary transformation and they should not be taken as the cells with string shaped nucleus or with dumb bell shaped nucleus.
3. Since the caudal string-like projections have no motive function, they shoule be called as the tail strings rather than the cilia or pseudocilia.
4. The centriol of the cell does not actively determine the direction of the cell movement.
5. The power of the cell to push other cells or foreign particls aside is in the descending order of eosinophils, pseudoeosinophils, neutrophils, basophils, monocytes and lymphocytes.

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There were known that the much production of ascites would be observed by the inoculation of the tsutsugamushi disease rickettsia to the chlorpromazine injected guinea pig. But there were still suspended the problem on the factors to result in much production of ascites.
The author studies the relationships between the production of ascites and body weight of animal and between the former and breeding environment of animal, especially the temperature of environment. As the results of this study, it could be confirmed on animals having body weight 300-500 gms. that the body weight of animal had no relation with the production of ascites and the proliferation of the rickettsia in the animal so far as the animal was injected previously chlorpromazine; however, the temperature of environment affected powerfully to those. Namely, the production of ascites, the proliferation of the rickettsia and the presence of the specific cells for infection were found to be much on the infection in spring, fall and even in winter if the temperature kept at 15°C, but these evidence were not found on the infection in summer.
These informations were thought to be valuable in order to obtain the tsutsugamushi disease rikettsia in large amount.

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It has been already shown that the tsutsugamushi disease rickettsia obtained from the infected mouse of this disease, that mouse was repeatedly injected small dose of chloramphenicol, is highly capable of inducing the disease to the chlorpromazine treated guinea pig leading to the much production of ascites and the antigenic capacity of this ascites was inferior to that obtained from the infected animal by the original strain of the organism owing to the variation on some properties of the organism by chloramphenicol.
The author studied the complement fixation reaction using the original strains, OSEKI's, MITANI's, and HICHITO strains and the strains obtained by variation of these and found some specificity on the antigen. In any case of the test, the ascites was obtained from the organism inoculated guinea pig and used as antigen for complement fixation reaction. From the view of the results, all ascites obtained by inoculation of the original strains showed difinite strain specificity to the corresponding antiserum, but those by the varied strains showed only slight inhibition on hemolysis in any case just like having no strain specificity. Hence, it was confirmed that the occurence of variation on the properties of the tsutsugamushi disease rickettsia by action of chloramphenicol.

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While the use of chloramphenicol showed marked curative effect on tsutsugamushi disease, especially in the case of a large dose at the same time. W_OODWARD and K_ATSURA expressed a doubt on this effect, above all K_ATSURA reported the possible occurence of re-infection of this disease following after the therapy by such method of dose. On the other hand, the repeated use of small dose of the drug might allow the pathogenic rickettsia to survive the action of the drug, also allow to cause the variation of the rickettsia.
From the viewpoint of this, the author investigated the variation of pathogenicity of the tsutsugamushi disease rickettsia, O_SEKI's and M_ITANI's strains, by action of chloramphenicol: the variation was determined by the use of LD₅₀ for a intraperaperitoneal inoculation of the organism on the previously treated mice, that were given chloramphenicol perorally for 10 days so as each mouse to have the different drose daily, namely 5 mg., 2 mg., 1 mg. and 0.5 mg. per 10 g. of body weight respedtively. Contrarily to the control, it was noticed the very low value for the LD₅₀ of the test animals. Further the author conducted the study on the varied organism thus obtained, namely varied O_SEKI's and varied M_ITANI's strains, in that the rickettsia was inoculated intraperitoneally to the guinea pigs that were previously injected 20 mg. of chlorpromazine per Kg. of body weight daily for 4 days; and the result of the test was determined on the basis of the amount of ascites produced, the amount of free cells and the presence of the rickettsia in the ascites.
In this study, it was demonstrated that the infection of the original strain did not reveal the illness during summer and winter, but the varied strain could reveal the marked sign of illness at any time of seasons and the much production of ascites and marked proliferation of the rickettsia were observed in abdominal cavity.
These facts showed supposedly that the marked variation was occured on the pathogenicity of the tsutsugamushi disease rickettria treated by chloramphenicol compared with the untreated organism.

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The author reported already in the Part I that the pathogenicity of the tsutsugamushi disease rickettsia was varied by the continual action of small amount of chloramphenicol in the body of mouse. Further to study the protective effect of ascitic fluid vaccine prepared by the varied tsutsugamushi disease rickettsia, the author inoculated the original microorganism, , OSEKI'S and MITANI'S strains, and the varied organisms, varied OSEKI's and varied MITANI's strains, respectively into abdominal cavity of the guinea pigs that were previously given chlorpromazine and prepared the ascitic fluid vaccine from these animals. The vaccines thus prepared were injected into mice and then the protective capacity against the infection was studies on these animals.
From the results of the study, it could be confirmed that the protective capacity of the vaccine obtained from the original strains were very satisfactory, while the capacity of the vaccine from the varied strains were weak and could not be capable to prevent the infection, hence, being concluded that the variation on the organism affected the properties of the organism and let loose the antigenic capacity.

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Direct smear technic has been adopted in almost all the study on exfoliative cytology of the oral cavity, but cytology of the mouth washings and garglings has not been made so far. Therefore the author determined the most satisfactory centrifuging factors of the mouth washings and garglings to collect cells in the previous study.
Now in this study cells were collected by means of centrifugation, membrane filter method and cell block method in normal healthy controls-25 newborns and sucklings, 30 adults. and 28 edentulous old persons with normal healthy oral cavity and in various clinical cases-24 malignancies, 6 benign tumors, 5 various other oral lesions, and 41 smokers. All the specimens were stained by Papanicolaou technic.
I. NORMAL CYTOLOGY
1) Both nuclei and cytoplasms were larger in red and blue precornified epithelial cells, while somewhat smaller in orange keratinized cells showing pycnosis or disappearance of nuclei.
2) Fewer white blood cells were noticed in newborns, sucklings and old persons, while much more in the adults. These results may be due to the presence of gingival pockets, in other words the presence of teeth.
3) With the progress of age keratinized cells or those with keratinizing tendencies showed an increase, and those with regressive changes revealed the same findings. These results could be statistically proven significant and there was an aggreement between these cytologic and histologic findings.
II. PATHOLOGIC CYTOLOGY
Centrifugation
1) Most of the exfoliated malignant cells showed blue cytoplasm, which could be considered they were low-keratinized immature cells.
2) Of 21 cases confirmed of carcinoma by biopsy cytology-positive cases were 81 per cent in direct smear method and 52 per cent in centrifugation befor operation and 35 per cent in the former and 50 per cent in the latter after operation.
3) Sarcoma and other various lesions were cytology-negative, but plasmocytoma associated with intraoral ulceration revealed cytology-positive.
4) More keratinized cells or those with keratinizing tendency could be noticed in both edentulous and dentulous persons in smoker's group than in non-smoker's group. And it could be found that 15 cases, i.e. 62.5 per cent of 24 malignant cases were smokers.
Cell Block Method
Epithelial cells of both normal and pathologic oral cavity showed so marked morphologic changes that this method was not suitable for examination of cell elements. However, cells showed crowdings so that keratinizing tendency could be easily noticed at first sight by the difference of staining and the tumor cells could be distinctly observed.
Membrane Filter Method
No morphologic change could be seen and it was considred to be a useful method of diagnosis of malignancy, although cells overlapped each other.
As a conclusion examination by centrifugation of the mouth washings and graglings was easy to collect cells, gave little irritation to the tissue and was a simple technic. It also could be repeated many times so that it was a very satisfactory method in cases of the following subjects: observation of postoperative progress, for example when the remnant of malignant tissue was suspected, examination of the cells after radiation therapy, early discovery of recurrence and examination of extensive lesions of the oral mucosa.

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This study was made for the purpose of establishing the criteria of normal exfoliative cytology of the oral cavity. Clinically healthy twenty male adults with normal well-cared-for mouth were used and their smears were prepared from four areas of the oral mucosa-hard palate, gingiva, radix linguae and buccal mucosa, and stained by Papanicolaou and Giemsa stainings. And the distribution of keratinized cells was examined statistically. Concurrently histclogic sections were taken by gingivectomy and the specimens were staind by hematoxylineosin, azan, and Papanicolaou technics whose findings were compared with each other.
The results obtained are as follows:
1) By means of Papanicolaou technic cytoplasm of the epithelial cells showed different colors---non-keratinized blue, mixed with red or red mixed with blue, red and highly keratinized orange.
2) Sizes of nucleus and cytoplasm showed somewhat difference according to the areas where smears were taken.
3) Keratinization of the epithelial cells are highest at the hard palate, then gingiva and radix linguae are followed in the order. Almost no keratinized cells are noticed at the buccal mucosa.
4) Neither parabasal cells nor atypical cells could be found in normal cases.
5) White blood cell count showed a gradual decrease as epithelial cells showed the progress of keratinization.
6) Nuclear-cytoplasmic ratio varied to some extent according to the localization. Nuclei of the epithelial cells of hard palate and gnigiva showed a higher percentage in their disappearance.
7) Both keratinization grade and structural density were higher in the superficial layer and lower in the deeper layer in normal gingival epithelium.
8) Minute strucutures of the cytoplasm and nuclei could be noticed more remarkably in Papanicolaou staining than in Giemsa staining.
9) Almost same agreement could be found between exfoliative cytologic and histologic findings.

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In this comparative study on the toxicity of various alkylphoshosphates, commercial substances in general use, the author obtained the following results.
1. LD₅₀ of alkylphosphates subcutaneously injected in the form of emulsion into mice is as follows (in mg/kg): in the case of TEPP it is 0.39 (0.33-0.45); Pestox-3, 8.2 (5.5-12.1); Ethylparathion, 10.0 (8.3-12.1); EPN, 42.8 (24.8-73.6); Methylparathion, 48.5 (23.6-99.5); Diazinon, 57.8 (37.8-88.5); Malathon, 221 (180-272): and Dipterex 252 (165-386). The toxicity of TEPP is markedly higher than others.
The duration of survival after injection is within 45 minutes in the case of TEPP; 3 hours with Ethyl- and Methyl- parathion as well as with pestox-3; 6 hours with Malathon; 12 hours with EPN; 24 hours with Dipterex; and within 48 hours with Diazinon.
2. In the oral administration to mice LD₅₀ of Ethylparathion is 11.5 (8.1-16.4). showing hardly any great difference from that in the case of subcutaneous injection. Furthermore, in testing LD₅₀ of Methylparathion emulsions manufactured by different makers, LD₅₀ ranges from 30.5 (24.8 -27.5) to 19.4 (14.4-26.0), showing a great difference though statistically not significant. From this it is obvious that the toxicity differs according to the emulsifying materials used by different makers. In addition, the toxicity of Methyl-parathion is definitely lower than that of Ethylparathion. Therfore, as the agricultural insecticide it is advisable to use the former.
3. After subcutaneous injection of alkylphosphates mentioned above and DFP into rabbits and examining the degree of inhibiton of blood cholinesterase, the order of such inhibition coincides exactly with that of LD₅₀ in the case of the mice mentioned above. This suggests that intoxication is mainly brought about by the inhibition of acetylcholinesterase and it is not much affected by other factor. Moreover, the period of time necessary for the inhibition is appoximately in proportion to the duration of time elapsed before death in the case of mice.
4. In measuring the amounts of parathion and p-nitrophenol excreted in the urine of the rabbits previously administered with Ethyl- and Methylparathion, the concentration and the duration of excretion are greater when the doses given are greater, but there is no other prominent characteristic.

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The author studied the effect of pyridine-2-aldoxime methiodide (PAM) on various alkylphosphate poisoning from the standpoints of (1) its effect on LD₅₀ in mice; (2) on the recovery of blood cholinesterase (ChE) in rabbits; and (3) on human cases.
1. The effectiveness of PAM against Ethyl- and Methyl- parathion was already established in human by Hiraki et al. The auther carried on animal experiments and ascertained the effectiveness.
2. In the cases of EPN and Diazinon poisoning PAM proves to be effective both in human and animals.
3. As for TEPP, Pestox-3, Malathon, Dipterex, and DFP, it was impossible to encounter human cases, but in animal experiments with these alkylphospnates PAM seems to be also effective. Therefore, PAM is to be applied for these alkylphosphate poisoning in human.

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With the use of two new oximes that have just recently made debut, namely, diacetylmonoxime (DAM) and monoisonitroscacetone (MINA), the author made comparative study of their effectiveness on alkylphosphate poisoning with that of pyridine-2-aldoxime methiodide (PAM). Alkylphosphates used in the present experiment were commercially available ones such as Ethyl- and Methylparathion EPN, TEPP, Pestox-3, Malathon, Daizinon, Dipterex and DFP.
1. LD₅₀ of DAM given intravenously to mice is 356 (337-376) mg/kg, and that of MINA is 159 (133-191) mg/kg.
2. In determinig LD₅₀ of various alkylphosphates excluding DFP by injecting intravenously 80 mg/kg DAM or 40 mg/kg MINA, there can be found no difference from the control group of mice given only alkylphosphates.
3. After injecting intravenously 100 mg/kg DAM or 30 mg/kg MINA into rabbits and observing the effects of these drugs on the serum cholinesterase activity that has been inhibited by various alkylphosphates, DAM is slightly effective in the case of Malathon and DFP poisoning while MINA on EPN, TEPP, Diazinon, and DFP, but the effect of either one is quite inferior to that of PAM.
From these results the effects of DAM and MINA on the alkylphosphates mentioned above are quite disappointing, and their effects at leaet can never be said to exceed that of PAM.

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With the purpose to elucidate the influence of parathion exposure on the human body while spraying it in the rice field, the author studied 37 farmers in the Bizen rice-producing farmland as the subjects; and obtained the following results.
1. There is no difference between the physical findings before sparying and those after the spraying.
2. As for subjective symptoms 4 cases (10.8%) complained of the symptoms that can be thought relatively charateristic to parathion poisoning, and 12 cases (32.4%) complained of symptoms that are not characteristic. However., judging them in conjunction with the values of serum cholinesterase, these symptoms are not due to poisoning but they are due to fatigue or incidental ones. In any event, the total of 16 cases (43.2%) had complained about some symptoms on account of spraying parathion.
3. Twenty-four cases (14.8) showed urine p-nitrophenol which is the evidence of absorbing parathion into the body. As those who absorbed parathion at early stage of the spray may no longer show p-nitorphenol in the urine, if these possible cases are added, the actual number of absorbing parathion will be greater.
4. What is a convincing evidence of parathion invasion into the body, namely, inhibiton of serum cholinesterase, can be recognized in 15 cases (40.5%). The ones that showed the greatest reduction of serum cholinesterase as low as minus 64.8 per cent are thought to be on the verge of onset of poisoning.
5. Those who showed either p-nitrophenol in the urine or inhibition of serum cholinesterase, in cther words, those who were really exposed to danger during parathion spraying amounted actually as much as 30 cases, 81 per cent of the total studied. Therefore, at the present method of spraying those actually attacked dy parathion poisoning are rare, but as they are really exposed to that danger, it is desirable to take all possible precautionary measures during the work.

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Hematological researches in peripheral blood, especially the changes of eosinophils were made from various points in order to solve the nature and etiology of alveolar pyorrhea.
The results obtained are summarized as follows:
1) Little difference was recognized on red cell count, hemoglobin quantity and index in comparison with healthy control cases. No remarkable quantitative and qualitative changes of white cells except eosinophils were observed. In addition, any apparent difference between peripheral blood taken from the ear lobe and local gingiva could not be found in these above-mentioned points,
2) Increase of eosinophils was recognized in about 45 per cent of 78 cases, and, moreover, its remarkable relationship to clinical type, especially that of inflammatory findings characterstic of the disease, was suggested. Many cases showing morphological changes, especially degenerative changes of the cells, were observed, and, furthermore, relationlhip between changes of the cells and those of the disease conld be suggested as the experimental results
3) Peroxidizing reaction of the peripheral blood cells by the vitamin C method was investigated in 20 cases showing an increase of eosinophils. Consequently, quite difference could be observed in staining ability of granules between clinical cases and control cases, and thus it could be noticed that granules within the cells in case of alveolar pyorrhea were quantitatively different from those in normal ones.
4) As a result of vital observations on movement of eosinophils by the peripheral blood culture method in 10 cases of the disease, it was recognized that the migration velocity of the cells showed a rapid decrease as time went by and was markedly inferior to that of normal cases and that in the mobility form the cells suggestive findings of declining mobility increased much more than those of normal cases, especially showing a increasing tendency with the progress of culture.
All of these various findings of the cells can not necessarily be considered to be the phenomena characteristic of this disease, but pathologic findings and characterics of the cells of this disease may suggest both correlation of the cells to chronic inflammatory changes in case of alveolar pyorrhea and possibility of allergic process as an etiologic factor of the disease.

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0.25M sucrose homogenate of dog's liver was fractionated by differential centrifugation as described by Schneider. About half of histamine of the whole homogenate was found in the mitochondrial fraction. Smears were made from each fractions, fixed in 1 per cent basic lead acetate in 50 per cent alcohol and stained with 0.1 per cent aqueous solution of toluidine blue. Microscopic observation showed that the mitochondrial fraction contains a large amount of very small round metachromatic particles, which are indistinguishable from extruded granules outside of the mast cells similarly stained in the stamp preparation from the liver slice. These observations confirm the finding of Mota and his co-workers and this fact in all probability indicates that the higher histamine concentration of the mitochondrial fraction of dog's liver might be due to contamination with mast cell granules. Intravenous administration of 3 mg/kg sinomenine or 150 to 500 mg/kg peptone into dogs resulted in a significant release of histamine from the mitochondrial fraction with some variable loss from other subcellular fraction.

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In vitro histamine release from intracellular large granules of the dog liver was determined and compared with that from the chopped tissue. The histamine release from the large granules depended on the concentration, duration of action, temperature and pH of histamine liberators. Histamine was released by all the liberators tested even at 0°C less than at 37°C. The action of basic liberators such as sinomenine increased with increasing alkalinity of medium.
Dibucaine hydrochloride, decylamine hydrochloride, HgCl₂, quinine hydrochloride and tutocaine hydrochloride at pH 7-8 and 6mM concentration released over 50 per cent of histamine from the large granules. Under the same conditions ethylmorphine hydrochloride, toluidine blue, procaine hydrochloride, saponin (0.1 per cent), tropacocaine hydrochloride, sodium cholate, Compound 48/80 (0.1 per cent), sinomenine hydrochloride, Tween 20 (0.2 per cent), cocaine hydrochloride and xylocaine hydrochloride revealed the histamine liberating action in the descending order named. Histamine release from the granules by these substances was larger in variable degrees as compared with that from the chopped tissue.
Sodium salicylate inhibited the release from the granules and chopped liver by other substances, while diphenhydamine and guaiazulen did not reveal such an inhibitory action but rather accelerated the release though slightly. The histamine release from the chopped tissue by sinomenine and decylamine was inhibited by uranil nitrate, but it was not the same from the large granules. The lack of oxygen accelerated the histamine release by decylamine from both the granules and chopped tissues, but did not reveal any significant effect on the action of other liberators. In in vitro anaphylaxis histamine release occurred in the chopped tissue but not in the granules. As far as basic liberators are concerned, there seemed to be some correlation between the histamine release ability and the heparin combining power. However, the surface activity or the hemolytic power of liberators and the histamine release ability was not to be necessarily in parallel with each other in degree.
Interpreting the intracellular large granules as the granules of mast cells and in the light of the above findings the author discussed the respective mode of action of the liberators on mast cells and on their granules.

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Clinico-Statistical findings were summarized in 252 cases of clinically diagnosed pericoronitis of the lower third molar.
1) Sex and Age Incidence:
Generally speaking it could be seen a little more frequently in male, while more often in female before about 30 years of age, but a contrary tendency could be noticed with the progress of age. Eight per cent of all the cases were between the ages of 20 and 29 years.
2) Seasonal Incidence:
It showed a decaease in the order of fall, spring, summer and winter.
3) Side Incidence:
Almcst no difference of incidence could be seen between right and left sides. As for inclination of the eruption normal position was most frequent in acute cases, while mesial and horizontal inclincations more frequent in chronic cases. Twenty-eight per cent of all the cases showed the findings of bone resorption of the alveolar process.
4) Clinical Findings:
They were almost agreed with those of previous description except lower incidence of disturbed mouth opening and fever which had been considered to be the most frequent symptoms so far. According to the clinical findings all the cases could be classified into acute and chronic ones at the ratio of 15 to 85.
As a conclusion the fact that a cusp or a part of the crown had been exposed in all the cases examined may suggest that pericoronitis was due to infection of the dental follicle after eruption and had hardly connection with insufficiency of the space, direction of eruption, and difference of localiztion.

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Smear specimens were prepared from the inner surface of lower wisdom teeth of 20 normal healthy control cases and 252 cases of pericoronitis and stained by Papanicolaou and Giemsa.
1) Cytoplasm showed yellow, red mixed with blue or blue mixed with red, and blue according to the progress of keratinization by Papanicolaou staining. In chronic cases cells were more poorly stained than in normal control and acute cases.
2) Exfoliated epitheilail cells were classified accoding to keratinization or cytoplamic colors by staining and a definite relationship between progress of keratinization of acute and chronic cases could be statistically noticed as to rise and fall of the disease.
The severer symptoms shows the inflammation. the lower tends the keratinization, and parabasal cells and atypical cells increased in their number as a specific finding.
3) Regressive changes were most remarkable in chronic cases, which were followed by acute and normal cases in the order.
4) The largest number of white blood cells could be seen in acute cases and as the disease proceeds to chronic type, they showed a decrease, which could be helpful to observation of the clinical progress.

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Biopsy was taken from the gingivae in 42 cases of pericoronitis and 8 normal healthy control cases, which were the same materials used in previous cytologic study. Sections were stained by Hematoxylin-Eosin, Azan and Papanicolaou methods, investigated histopathologically and compared with the findings of exfoliative cytology.
1) In normal gingivae keratinous layer could be seen in various degrees, while in acute inflammatory ones typical keratinous layer could not be observed and keratinization showed a decrease.
2) In Azan and Papanicolaou stainings yellow and red stainings decreased in the superficial layer and a definite correlation could be seen between keratinization and staining effect according to structural density of the tissue. Findings of exfoliative cytology and histo-pathology were completely agreed in their comaprison.
3) The SH groups of the surrounding tissue in case of pericoronitis were investigated by Chévremout-Erédéricq's reaction but a decisive conclusion could not be drawn from these results.
It could be concluded that pericoronitis was induced by infection of the dental follicle after tooth eruption and could be divided into acute and chronic types clinically.
As the results obtained from exfoliated epithelial cells classified according to their staining effect of the cytoplasm by Papanicolaou staining, a considerable decrease of keratinization could be noticed in cases of pericoronitis in comparison with normal controls. This fact could be proven by histo-pathological examination.
Consequently clinical data, exfoliative cytogy and histo-pathology in case of pericoronitis coincided in their findings.

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