It is difficult or almost impossible to give a scientific definition of fatigue. In certain text books of physiology, fatigue is defined as the lowering of activity as the result of preceding activity of a whole organism or an organ. But such a definition is merely formal so long as activity is difficult to express in purely scientific terms. A great number of methods or tests have hitherto been proposed for the measurement of fatigue, and they may be useful so long as they express certain features of fatigue, for we must be acquainted with all functional changes of the body in order to get a complete information of fatigue. It may be right so long as academic interest is concerned, but in practice, it is demanded to resort to a few methods which represent the most fundamental feature of fatigue. The procedure should be as simple as possible, and the result should be clear cut and in good harmony with physiological and psychological common sense. Moreover, the procedure should neither inflict any pain on the subject nor lower his working efficiency. These demands seem, to some extent, to be satisfied by our method which will be reported in the following.

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Certain inorganic substances are endowed with the actions resembling those of enzymes, but these “inorganic enzymes” (Bredig) are, for the most part, known only for their action of decomposing hydrogen peroxide, id est, catalase activity. As to other types of enzyme action, palladium black in the classical experiments of Wieland may be regarded as an “inorganic dehydrogenase”. It is also known that peroxidase-like action is shown to some extent by, Fe, Co, Cu, Ni, and other metallic compounds.
We recently found that manganese peroxide possesses an exceptionally strong power to produce purpurogallin from pyrogallic acid in the presence of hydrogen peroxide (peroxidase action), far surpassing the similar activity of other metallic compounds.

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Now it is an important problem for the Japanese penicillin production to produce stable and crystalline penicillin sodium salt. While we failed to crystallize penicillin as various amine salt, Kusaka and Kawashima (1) isolated crystalline penicillin G Nethylpiperidine salt, according to the method described by Seehan, Mader and Cram (2) .
Afterwards we also confirmed that penicillin G N-ethylpiperidine salt could be easily obtained by this method, and the penicillin sodium powder extracted from the modified stone and Farrell's synthetic medium (surface culture) implantedd with Q-176 strainn contained penicillin G at about 60%. Penicillin G sodium salt was also easily obtained from its N-ethylpiperidine salt, and besides it was found that after the sodium salt obtained by freeze-drying had once dissolved in dry acetone, immediately it crystallizedd to the very stable salt. Present paper discusses the difference between crystalline penicillin G sodium obtained by freeze-drying and by the addition of dry acetone.

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The hemolysin produced by Streptococcus pyogenes has long been the subject of many researchers. (For early literature see M'Leod, 1912¹⁾, Hellens, 1913²⁾, and Kruif and Ireland, 1920³⁾.) Recent work by Todd, and by Weld are particularly noteworthy: Weld (1934⁴⁾, 1935⁵⁾) found that a powerful hemotoxin might be extracted from living hemolytic streptococci by agitating the organisms in normal serum and subsequently filtering the serum. In a satisfactory experiment the filtrate has a very high hemolytic titre and is lethal to mice in small doses intravenously. On the other hand, it has been demonstrated by Todd (1934⁶⁾, 1938⁷⁾, 1939⁸⁾) that group A hemolytic streptococci produce two distinct varieties of streptolysin:
(a) Streptolysin O which is oxygen sensitive and antigenic.
(b) Streptolysin S which is serum-extractable and apparently non-antigenic.
Although much work has been done in the research concerning streptolysin, all these experiments were carried out with the filtrate hemolysin or the supernatant fluid of the culture; thus making more difficult the most essential points in the study of streptolysin, that is its isolation and purification.
In 1939, the phenomenon of “the high promotion by nucleic acid of the production of hemolytic toxin of Streptococcus pyogenes, streptolysin, ” was discovered by Okamoto.⁹⁾ For the first time, it then became possible to carry out studies on streptolysin uncomplicated by the presence of serum.
Within a short time, the study on this phenomenon lifted our knowledge of the streptolysin (S') (the term “streptolysin S'”was designed by Bernheimer to indicate its similarity in properties to Todd's streptolysin S. See below.) from the status of “bacterial toxin of chemically unknown nature” to a point, where the essential quality of this toxin could be almost fully understood.
Thus, we are able to establish the isolation method^10)11)15)19) of the streptolysin in relatively pure form from the culture of hemolytic streptococci grown in 1% Nucleic-Acid-Broth.
It was found that the streptolysin (S') is not protein in nature, and it therefore probably follows that this toxin possesses no antigenic property. Further chemical investigations of the purified streptolysin-fraction (minimum hemolytic concentration=1: 200, 000, 000), finally led to the discovery that this hemotoxin is a nucleic acid derivative of polynucleotidic form.^12)13)16)17)
On the other hand, it was found that the dyestuff of trypan blue series possesses a distinct specific antidotal activity against this streptolysin.
The excised intestine (or uterus) suspended in Ringer's solution is aroused to contracture by streptolysin.¹⁸⁾
Our observations have been quickly confirmed and extented by Alan W. Bernheimer of the Department of Microbiology, College of Medicine, New York University^**.²⁰⁾²⁴⁾
Bernheimer reported in his paper²⁰⁾ as follows: “Streptolysin S”.-The sodium salt of yeast nucleic acid has been shown by Okamoto to induce the formation of a potent hemolysin in infusion broth cultures of hemolytic streptococci. The observations of Okamoto have been confirmed in this laboratory using the C203S strain of Streptococcus pyogenes. The hemolysin obtained by Okamoto's procedure was found to differ from streptolysin O in several respects among which were failure to be activated by cysteine and failure to be inhibited by cholesterol or by antistreptolysin O. Its properties, so far, as they have been studied agree in all respects with those of streptolysin S²¹⁾ but since complete proof of its identity with streptolysin S is lacking, it may provisionally be designated streptolysin S'.

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In the previous work, we ascertained the fact that the washed hemolytic streptococci, when suspended in nucleic acid solution, are able to produce streptolysin very luxuriantly.
As the continuation of our research work, we recently were able to demonstrate a remarkable inhibitory effect of O: O'-dihydroxyazobenzene and its tetrabromo derivative on the streptolysin production of hemolytic streptococci.
The present communication records the results of such experiments in brief.

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