The minimum requirment and the assay of typhus vaccine in Japan were, even previous to other biological products, announced early during the month of February, 1947 (Kitaoka (1) ) . Among various tests to be done, the potency test most essential part, requires many guinea-pigs and the result is to be observed comparatively long some 45 days. Therefore simplifying the method and shortening the observation period have been tried by investigators as follows.
(1) By estimation of the antigenicity of the vaccine through the complement fixation test with the standard serum of known units.
(2) By estimation of the antibody produced in the serum of guinea-pigs to which the vaccine to be tested is inoculated through the complement fixation test against the rickettsial antigen of known units.
Those are, however, not yet in practical use for there exists sometimes discrepancy between the two results obtained by the complement fixation test and by the direct challenge on the animals.
The Federal Security Agency, National Institute of Health has announced of a new method of neutralization test on mice against toxin of Rickettsia prowazekii and R.typhi cultured on the yolk-sac of developing chick-embryo, with which estimation on mice of the neutralizing antibody developed by injection of the vaccine in guinea-pig serum has made certain that the result observed in such test resembling in most part to the findings in the direct challenge test upon guinea-pigs.
This method has shortened the two week period for observing the result whether or not infection developed in the guinea-pigs following the challenge with rickettsia 2 weeks after the last vaccination. The process requires, however, some 4 weeks.
Authors have, therefore, undertaken to simplify all the process in assaying the vaccine and invented a method to estimate the potency in mice by direct challenge with rickettsial toxin after injection of the vaccine to be tested, without using guinea-pigs.
Most essential part in our new process is preparation of the test toxin. It is well known that the toxin is contained in the emulsion of the yolk-sac in which the strains not R. prowazekii or typhi is cultivated, and is neutralized by the convalescent serum in a guinea-pig or a typhus fever patient. Special care should be paid to keep the toxin highest virulent possible because the influence upon its potency of pH, centrifugal effect, filtration, disinfectant (phenol and formalin) and temperature has been described by Gildemeister (3), Bengtson (4, 6) Henderson (5, 7) and Kitaoka (8) .

抄録全体を表示

In 1939 Rao (1) reported that Past. pestis could grow on media composed merely of hydrolysed gelatin, cystine and inorganic salts. Further, he specified, using chemically defined amino-acids media, proline, phenylalanine and cystine as indispensable substances and glycine as groth-promoting substance. In 1940 Rao (2) reported that hematin promoted the growth of Past. pestis sown in a chemically defined amino-acids media. Cozymase, nicotinic acid, thiamin also possessed similar but less activity.
As Rao's observation has been depended on a subculture only, to which an amount of carefully washed bacterial cells were sown, we attempted to find out the possible variations in the growth during the repeated subcultures and obtained some newer informations, which will be described in the following lines.

抄録全体を表示

Since Koch's discovery of tuberculin, the active principle of tuberculin has long been the subject of many researchers, and a significant advance was made in the study of the nature of tuberculin' when Seibert and Long and their associates (1, 2, 3, 4) wsucceeded in the isolation of the tuberculin active principle apparently in pure form, and as a result of which, as is well known, it has now become possible to compare accurately the tuberculin sensitiveness of human beings in the field testing.
Quite recently, working preliminarily with Maschmann's purified tuberculin (5, 6), and, following this, with Sauton's synthetic-medium Old Tuberculin, Ito and Koshimura of our laboratory have succeeded in establishing a simple procedure for preparing ο-aminophenol azo-tuberculin derivative quite equivalent in skin-reacting potency to the P. P. D. (purified tuberculin derivative) of Seibert. The new process is especially significant, in that it provides us with ο-aminophenol azo-tuberculin preparations possessed of practically uniform skin-reacting potency in good quantities.
Although much work has been done in the research concerning the effect of chemical modification on the immunological reactivity of protein antigens (7, 8), there has, insofar ns the writer is aware, been practically no work done towards the isolation of the active principle of tuberculin in the form of its azo-derivative.
In this paper I have tried to review the principal results of the research work which has been done in our Institute for these past two years.

抄録全体を表示

The exprimental as well as clinical studies in the therapy of typhoid fever have far advanced since the appearance of sulfonamide preparations and antibiotics. Nagabayashi (1945) reported that 80-90% of mice, infected with E. typhosa, survived by the treatment with self athiazole, the results of which could not be confirmed by the authors, perhaps because the virulence and the amount of bacilli used by us were different from that of Nagabayashi.
Next Bigger (1946) investigated the additive action of penicillin and sulf athiazole upon the growth of E. typhosa in vitro and in the same year Mc Sweeney reported that they had obtained good results in clinical cases of typhoid fever by the combined medication of penicillin and sulfathiazole.
Mikamo (1948) used patulin in combination with sulfathiazole for the patients of typhoid fever and concluded as follows : Bacillemia disappears in course of the use of drugs or directly after the interruption of drug administration ; the temperature does not rise temporarily and it seems that the general conditions turn for the better. The stop of medication invites, however, the relapse of the disease. Patulin itself is toxic and its great amount can not be used.
Prior to this a great success was expected about the effect of streptomycin upon typhoid fever but the report of Keefer and his collaborators (1946) in 51 clinical cases was contrary to their expectations.
And in recent times chloromycetin is the only hopeful substance to heal typhoid fever according to the report of Smadel, Joseph, Léon, Ley and Varela, while the details thereabout are not known to us as yet.
Now our report concerns also with the therapeutic study of typhoid fever and the discovery of complete protection of mice against E. typhosa infection by our procedure is believed to promote the study in this field.

抄録全体を表示

A study on the chemotherapeutic agent against C. diphtheriae was carried out already by Behring (1, 2) side by side with his study in the serotherapy. Boer (3), Eeiler (4-6), Burkard and Dorn (7), Langer (8), N euf eld, Schieman and Baumgarten (9), Reinhardt (10), Ritter and Schenkel (11), Brunner and Gonzenbach (12), Browning and Gulbransen (13), Goldschmidt (14), Braun and Goldschmidt (15), Shibanuma and Hata (16, 17) followed after him in the study of the same field.
In the early stages they investigated about inorganic substances but later chiefly about organic substances and especially about acridine derivatives. The strong bacteriostatic action of acridine compounds in vitro attracted their interest and they have succeeded also in saving animals from experimental diphtheria infection. But no clinician has intended as yet to treat the infection of human beings with acridine compounds, because of the uncertainty of effect. Injection of antitoxin serum is the most trustworthy-treatment; the serum is capable not only to neutralize the toxin, but to inhibit the growth of bacilli and cures the disease in its earlier stage with promptitude. But on the other hand the serum is somewhat inconvenient to treat with and troublesome in its production. The apparition of such a compound, as not only capable of inhibiting the growth of bacilli, but also to destruct or to neutralize toxin in vivo, if possible, might be desirable. Of course we have had comprehensive studies on the detoxication. of diphtheria toxin, while those works were to gain an effective anatoxin for the use in preventing the disease. Formol, ketene, vanilline, adrenaline, sodium salicylate, urea, quinine salt, gluthathion, soap, ricinoleic sodium salt, colloidal manganese, colloidal iron, various anorganic salts, ascorbic acid, hormones, enzymes etc, were namely the detoxicating agents in this sense, but none of these seems to be practically useful as therapeuticum, because they have no strong detoxifying ability in vivo.
Meanwhile penicillin appeared and a ray of hope has been thrown for a new treatment of diphtheria, yet so far we do not know about its final success.
On this occasion a number of compounds were investigated by us (18-21), in the bacteriostatic and bactericidal action against C. diphtheriae and among 120 chemicals, inclusive of 15 phenothiazine, 5 phenoxazine, 13 quinoline, 4 acridine, 31 diphenylsulf one, . 17 diphenylsulfide, 5 diphenyldisulfide, 15 diphenylether, 9 tetronic acid derivatives, citrinin, kojic acid, usnic acid and 3 others, usnic acid was found to be the only substance, that can not only save animals from experimantal diphtheria infection (22), but also detoxify the toxin.
The usnic acid, used in these studies, was extracted from Usnea longissima. Fp 203°C [a] ¹³_D= +491.8° (in chloroform) .
The acid has recently been reported to have strong bacteriostatic effect on the growth of the tubercle bacillus and the Staphylococcus aureus (23-25), but no one has ever reported in details about its effectiveness in diphtheria infection. The experiment here reported was designed in order to observe the effect of usnic acid in experimental diphtheria infection and its action on the toxin.

抄録全体を表示

Miyamoto and Inagaki (1) discovered that a special fold figure is formed in a thin layer of serum by a special drying process, by placing a cover glass on the serum on the slide glass and heating it for 20 to 30 minutes in a desiccator at 120°-130°C. Later, . Ohbuchi (2) made a detailed study on this special fold figure on tuberculosis and classified these figures as forms A, B, C, D, and E, according to the condition of the disease. Isome (3, 4) compared the variation of these fold figures and the condition of the disease with the blood sedimentation rate, the viscosity, the globulin-albumin ratio of the serum, and other clinical observations, on malnutrition, pneumonia, and other diseases, and. pointed out that this variation of the fold figures is the barometer of the general condition of the patients. He added, moreover, a new form F, to Ohbuchi's 5 forms. However, no decisive conclusions have been reached regarding the mechanism of the fold. figure formation.
Miyamoto and Inagaki (5), by observing the structure of the fold figure, pointed out that the formation of the fold figure was due to the fact that the foams caused by the vapor and other gases, when the serum is heated in the capillary field between the slide and the cover-glass, took various forms according to the serum; and also pointed out that the viscosity and the surface activity of the serum were the two chief causes determining the form of the figure. Meguro (6) placed the serum under decreased pressure and after removing the various gases dissolved in it, tried this experiment. He thought that the formation of the vapor by heating the serum was due to the heating of air and other gases dissolved in the serum and pointed out that the number of the fold figures that are formed was controlled by the quantity of gases dissolved therein.
This fold figure is characteristic of the serum, as shown by Miyamoto and Inagaki. When they used other albumin solution or other viscid materials, the figures formed were different from those of the serum. But they succeeded in forming figures similar to that of the serum, by using an albumin solution to which other materials were added to give it the proper viscosity and surface activity. Miyamoto and Inagaki (5) observed a figure similar to that of the serum by mixing ovalbumin with sodium glycocholate, and explained that these fold figures might be controlled by a proper viscosity and surface activity.
In order to investigate the mechanism of the fold figure formation, I tried to see by expanding the experiments by Miyamoto and Inagaki, how the fold figure changes according to the viscosity and the surface activity of the gel.

抄録全体を表示

When a small amount of human blood serum is placed between two slides and heated to 110-120°C, the serum boils and foams appear, and when the serum dries a certain kind of figure remains.
It was noticed that this foam appearance of blood serum, a phenomenon discovered by Miyamoto and Inagaki (1), differs between the human and the animal blood sera, and between the healthy and the sick persons. The writer and his colleagues experimented on this foam appearance of blood serum mostly on the tuberculous patients with the healthy as controls. The figure may not be definitely characteristic of tuberculosis, but, that there can be figures characteristic for each disease, can be expected when we observe the figure from the serum of a small number of jaundice and cancer patients.

抄録全体を表示