Among the methods of the paper chromatography of digitalis glycosides, that of the Svendsen and Jensen's is widely used. It uses the mixture of chloroform, methyl alcohol and water for development and trichloroacetic acid for producing fluorescence. Our experiment has shown that there is a kind of digitoxin which does not produce fluorescence with trichloroacetic acid.
Digitoxin made by Merck Co. of Germany (Dm), when developed on filter paper strip with the mixture of chloroform, methyl alcohol and water and heated with trichloroacetic acid, evidently produced two fluorescent spots with Rf values characteristic of digitoxin and gitoxin, in ultra-violet light. But digitoxin made by Rosenthal-Bercow Co. of the U.S.A. (Dr), when treated by the same method, produced only one fluorescent spot of gitoxin and none of digitoxin even if a large quantity of material was used. If, in that case, other reagent, for example Raymond's, was used, digitoxin contained in Dr was found on the spot of filter paper that had the same Rf value as that of Dm.
As both Dm and Dr contained some quantity of gitoxin, they were purified by the chromatography. After materials dissolved in ethyl acetate containing mehtyl alcohol by 0.5 volume per cent, were passed through the column plugged with silica gel, the portions positive in the Keller-Kiliani's reaction were gathered-from filtrates. In these substances could no gitoxin spot be found by the paper chromatography. Therefore these substances were considered to be almost purified.
These two kinds of purified digitoxin were alike in several chemical reactions and had no appreciable difference in biological action on the frog heart. However, purified Dr did not produce fluorescence while purified Dm did.
As a result of these studies, it has been shown that there are two kinds of digitoxin and that it is not desirable to depend only upon the method of Svendsen and Jensen's in the paper chromatography of digitalis glycosides.

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Our previous studies (1-4) of the experimental chemotherapy of malaria have been done by the use of Plasmodium cathemerium in canary birds. Not only is the Plasmodium cathemerium unsatisfactory as a test subject, but it is necessary to study plasmodia other than the human malaria plasmodium in various animals in investigations of the action of cinchona alkaloids. It is a well known fact that Plasmodium cathemerium, relictum, gallinaceum and lophurae of birds, Plasmodium cynomolgi of the monkey and Plasmodium berghei of the rat and mouse, which are .test subjects for experimental malaria, have different sensitivities for plasmodicidal agents. The effects of plasmodicidal agents on the parasites of the cold-blooded vertebrates have not been studied.
Two different types of parasites exist in the peripheral blood of the frog (Rana esculenta). Their. development is not completely known. One is Lankestella minima (Chaussat). This was found by Labbe (1899) in the red blood cell of the frog, which was first seen by Chaussat (1850). This parasite has been studied chiefly by Noeller (5). It is a small vermicle, which may attain half the length of the corpuscle. Various observers have described the vermicle as becoming spherical and reproducing in the red cells by sehizogony, but Noeller believes the schizonts belong to Dactylosoma ranarum, which is another parasite in the frog's red cells. Infection of the frog is brought about by the leech, which introduces the sporozoites. They, then apparently enter the endothelial cells. Schizogony results in numerous merozoites, which bring about infection of further host cells. Ultimately macro- and micro-gametocytes are formed. The zygotes transform into oocysts in which a number of sporozoites develop. These sporozoites are set free upon disintegration of the oocyst wall in the plasma and enter the red cells. In the red cell of the frog another type of small parasite which is devoid of pigment and which multiplies by producing 4-16 merozoites was found by Kruse (1890). This is Dactylosoma ranarum (Kruse). Two years later Grassi and Feletti (6) described it under the name of Hoemogregarina ranarum and recognized it as having a close resemblance to human malaria parasites. According to Noeller (7) the asexual stage of the parasite. may be seen within the living red cell as hyaline masses of cytoplasm, either elongate or rounded. The schizonts have a diameter of 4 to 9 micra, while the number of merozoites produced is four to sixteen. When nuclear multiplication is completed, merozoite formation commences and this takes place frequently by the development, of buds on one side of the parasite only, producing eventually a fan-like appearance. The other type of merozoite, after entering another red cell, grows into a gametocyte instead of a schizont. They usually lie at one end of the red cell and the narrower end is often bent into a loop. In some of the gametocytes the nucleus, which is a spherical body, contains a small karyosome while in others there is a much larger one. The former may represent the male gametocytes and the latter the female.
Since the specificity of drugs in the treatment of malaria depends upon the type of infecting parasite, it is of interest to know the difference in sensitivity to the cinchona alkaloids between the avian parasite and .frog's parasite.

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There have been some investigations as to the influence of atropine on the activity of cholinesterase (ChE). Roepke (1) demonstrated that atropine inhibited horse serum ChE competitively with an affinity for the enzyme much higher than that of acetylcholine, and Schaller (2) also pointed out that atropine inhibited human blood ChE. On the other hand, Scoz and Michole (3) found that very small quantity of atropine increased blood ChE in vitro, and Yoshida et al. (4) reported that atropine intensified the destruction of acetylcholine by rabbit serum in vivo and in vitro. I have also undertaken this work using rabbit blood to investigate this problem.
The activity of abbit blood ChE was determined by the Warburg's manometer. Into the main compartment of each flask were placed definite quantities of enzyme material diluted twenty times with bicarbonate-Ringer solution saturated with C02. One cc of substrate (acetylcholine, to make a 0.0025 molar solution) saturated with CO₂ was added to a side arm of each series of flasks. Atropine sulfate was added in varying concentrations to the enzyme solution. The total fluid content of each flask was 5.0 cc. After equilibrating at 37.5°C, the content of the side arm was tipped into the reaction chamber. Controls and blanks were run simultaneously. Gas evolution resulting from the liberation of acetic acid and its action on bicarbonate was measured in the conventional manner. The results of the experiment are presented in the accompanying tables. Activity of enzyme was indicated in the vo'ume (cmm) of CO₂ gas liberated by 0.1 cc blood or serum in one hour.
In vivo: The activity of rabbit serum ChE after intravenous injection of atropine was compared with that before injection. There was no change between them (Table 1).
In vitro: Firstly, when atropine was added to rabbit serum directly, the activity of its ChE was inhibited (Table 2). Secondly, after defibrinated rabbit blood was incubated with atropine at 37.5°C for ten minutes, the activity of ChE in serum from the blood was determined. In that case the activity was higher than control in high concentration of atropine. But when whole blood under the same treatment was used, the activity of its ChE was inhibited in high concentration of atropine (Table 3). It was probably because atropine liberated the ChE contained in blood cell into serum that the activity of ChE in serum from the blood with atropine was higher than that without atropine.
There is an interpretation that the influence of atropine on the parasympathomimetic effect or acetylcholine action can be attributed to the activating action.of atropine on the ChE (4). However, this interpretation has been found inappropriate from my observation, because atropine did not activate rabbit blood ChE both in vivo and in vitro.
Acknowledgement: The author wishes to express his gratitude to Professor Z. Kanda for suggesting this investigation and for his continued. guidance and help throughout the course of this work.

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The discovery of choline acetylase (1) has revealed the position of acetylcholine (ACh) in general metabolism. In view of the fact that certain drugs including narcotics and convulsants have some effects on ACh metabolism of brain, it is reasonable to suppose that some of these drugs might associate with the activity of choline acetylase.
In this paper, we have studied several, problems regarding choline acetylase, especially the effects of certain drugs on this enzyme.

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Choline acetylase is in close connection with general metabolism ; this may be characteristic of this enzyme, in contrast to that of cholinesterase.
Nowadays, much attention is focused on the problem of acetyl donor in enzymatic synthesis of ACh. Nachmansohn has long been of opinion that acetate was the unique acetyl donor, while such metabolite, as citrate or glutamate was nothing more than accelerator (1), though recently he has withdrawn this opinion (2). On the other hand, there is valid evidence that citrate is the main acetyl donor (3, 4).
In this paper, the author has examined enzymatic synthesis of ACh in various tissues of animals, paying special attention to the problem of acetyl donor. The enlightenment of the nature of acetyl donor is concerned not only with this enzyme system, but also with the function of these tissues of organs themselves, in which ACh is formed.

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In the previous report (1), a small dose of morphine, dolantin and ohton (dithienyldimethylbutenylamine) blocked splanchnic afferents in the spinal cord, and vagal afferents in the medulla oblongata, while they did not block afferents of the sciatic nerve. In the present paper, the effect of analgesics on afferent pathways of other nerves was studied, and attention was concentrated mainly upon pain afferents. Furthermore the effect of these drugs on their relay nuclei, reverberating system and centrencephalic integrating mechanism was investigated.

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Lamson et al. (1-8) carried out extensive studies on the anthelmintic activity of a large number of alkylhydroxybenzenes and found that substances which apparently act through the cuticle of ascaris were ineffective when their melting points were too high. Harwood (9) demonstrated that if such chemicals were kept in the liquid phase by adulteration with certain organic liquids, their in vitro ascaricidal properties were heightened. Considering these reports which suggest a close relationship between the anthelmintic effect and melting points of these substances, the present authors attempted to increase the anthelmintic efficacy by combining certain alkylresorcinols with comparatively slight irritant action.
The present paper gives evidence for the potentiation of in vitro and clinical efficacy by the combination of such chemicals, and describes observations on the toxicity and local irritant properties. Certain physical properties arising from such a combination, were studied in order to elucidate the mechanism of their synergistic effect.

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Despite its severe toxicity, the globe fish—Japanese “Fugu”—has been eaten as a special delicacy for a long time among the Japanese because of its unusual taste. Previous experimental studies on the fish demonstrated that the toxic substance of this fish was found substantially in the viscera, especially in the ovary and liver. According to the earliest biological experiments performed by Ôsawa (1), the toxic substance has a curare-like action on the motor nerve endings. This finding received support from the work of Miura and Takesaki (2). Furthermore, Takahashi and Inoko (3) showed that the toxic substance exhibited a central depressant action on the medullary level in addition to the peripheral paralysis. Since then, several detailed reports regarding toxicologic and pharmacologic actions of the Fugu toxin have been presented [Hayashi and Muto (4), Iwakawa and Kimura (5), Yano (6), Nomiyama (7), Kawakami (8), Ueki (9), and Iwamoto (10)] . Some of them dealt with cocaine-like local anesthetic effects of the toxin.
On the other hand, the process of neuromuscular transmission was recently clarified following advanced studies on the end-plate potential (e.p.p.) ?? Eccles (11), Hunt and Kuffler (12), and Rosenblueth (13) ?? . Consequently, some light has been thrown on the mechanism of the action of drugs at the motor nerve ending. The object of the present paper is to show the features of the blocking effects of the Fugu toxin on neuromuscular as well as ganglionic transmission.

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