The investigation on the plant growth-promoting substances in the bamboo shoot (Phyllostachys edulis) were carried out, applying paper chromatography^{(2, 3, 4, 5, 6, 7)} as bioassay method. In this method the squares cut from the chromatograms of the ether extracts of bamboo shoot, were extracted with water, and each of them was tested on the extension of Avena coleoptile sections.
Each of the acid, non-acid, and hydrolysed acid fractions were extracted with ether, and the free amino acid fraction with warm water, by the modified extraction methods of YAMAKI⁽⁸⁾ and BONDE⁽⁹⁾ (cf. Table 1).
It may be concluded from the application of bioassay method of paper chromatography that the growth-promoting substances in the bamboo shoot were localized in the zones of R_F 0.0_??_0.1, 0.35_??_0.5 and 0.7_??_0.85, of which the last R_F value was found to coincide almost perfectely with that of synthetic indole-3-acetic acid (cf. Figure 1). As to the acid fractions in the top, middle and bottom of the shoot, the coleoptile extension was most conspicuous in the top (cf. Figure 2).
Tryptophan, supposed to be a precursor of indole-3-acetic acid, could not be identified in the free amino acid fraction, but sixteen amino acids-aspartic acid, glutamic acid, serine, asparagine, glycine, threonine, glutamine, lysine, alanine, tyrosine, arginine, γ-amino butyric acid, methionine, valine, leucine, phenylalanine were found by means of paper chromatography (cf. Figure 3).

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(1) The constituent change of Shoyu caused by heating was studied; total titratable acidic substances, nitrogen containing acidic substances soluble in ether, in both the free and conjugated form, respectively, and ether soluble acids of the free form, phenolic substances of the conjugated form and the extinction value of 470mμ, 530mμ and 610mμ, increased in accordance with heating. The phenolic substances of the free form showed a maximum content in the course of heating. The ether soluble acids of the conjugated form and the total nitrogen did not change, but amino nitrogen and reducing sugar decreased.
(2) The changes of ultraviolet absorption spectrum of weak acidic (phenolic), acidic and neutral fractions of the ether extract of Shoyu, were investigated. The maximum absorption of the weak acidic fractions shifted from 265mμ to about 280mμ by heating, and its content exceeded the maximum value in the course of heating. This phenomenon was believed to indicate the change of the content of 1-hydroxy-2-methoxy-4-ethyl benzene (which was identified as an important flavorous ingredient of Shoyu, by the author).
All of the acid fractions had the same maximum absorption wave length at about 265mμ, but their contents showed the maximum later than those that of the phenolic fractions; this fact was believed to suggest the content change of vanillic acid.
The ultraviolet absorption of the neutral fraction increased by gradually heating, always showing the maximum value at about 290mμ and this fact seemed to suggest the change of furan components, but the sample did not give its color reactions.

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The influence of sulfur containing drugs on E. coli phosphorylase and yeast hexokinase activities was studied to ascertain the mechanism of the respiratory inhibition which was investigated in the proceeding paper.
E. coli phosphorylase was inhibited to an extent of 13_??_30% by both B. S. A. and SCN-compounds, and yeast hexokinase was inhibted to an extent of 20% by B. S. A., while the SCN-compound not at all.
The inhibition of B. S. A. on the yeast hexokinase was competitive and reversible, and completely antagonized by cysteine. Tne inhibiting results of B. S. A. on microbe respiration were seemed to arise from the inhibition on many enzymes containing essential SH-radicals.

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The carbohydrates in the pistillate petal of Humulum Lupulum L., hop, were investigated (see Table 2). The following results were obtained.
Glucose, fructose, sucrose, maltose, isomaltose, raffinose (?) and one unknown spot were detected as the free sugars.
Galactose, fructose, glucose, rhamnose, xylose and one unknown spot were detected as the components of glycosides.
Galactose, mannose, glucose, arabinose, rhamnose, fructose, ribose, desoxyribose and galacturonic acid (?) were found as the components of the hot 50% ethanol-soluble polysaccharides.
Mannuronic acid, galactose, mannose, arabinose, rhamnose, ribose and desoxyribose were detected as the components of cold water-soluble polysaccharides, and in addition, galactose, mannose, arabinose, mannuronic acid (?), xylose, rhamnose, ribose and desoxyribose as those of hot water-soluble polysaccharides.
Galactose, arabinose, galacturonic acid and glucose were contained as the components of hot 0.5% ammonium oxalate aq.-soluble polysaccharides (protopectin fraction).
Glucose, xylose and arabinose were detected as the components of hemicellulose.
Cellulose and a small amount of lignin were also detectable.

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The carbohydrates in the staminate petal of Cucurbita moschata DUCH, pumpkin, were investigated (see Table 2). The following results were obtained.
Fructose, glucose, sucrose, raffinose (?), stachyose (?) and one unknown spot were detected as free sugars.
Glucose, fructose, galactose, arabinose, rhamnose and xylose were detected as the components of glycosides.
Galactose, arabinose, galacturonic acid, glucose, xylose, rhamnose, fructose, ribose (?) and desoxyribose were those of the hot 50% ethanol-soluble polysaccharides.
Glucose, galactose, arabinose and galacturonic acid were the components of cold water-soluble polysaccharides, and in addition, glucose, arabinose, galacturonic, acid, ribose and desoxyribose of hot water-soluble polysaccharides.
Galactose, arabinose, galacturonic acid and glucose were found as the component sugars of hot 0.5% ammonium oxalate aq.-soluble polysaccharides (protopectin fraction).
Glucose, mannose, arabinose, rhamnose, fructose, ribose and desoxyribose were as those of hemicellulose.
Cellulose and a small amount of lignin were also found.

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The carbohydrates in the petal of Hydrangea macrophylla SERINGL var. Otakusa MAKINO hydrangea, were investigated (see Table 2).
The following results were obtained.
Fructose, glucose, sucrose, maltose, raffinose and stachyose were detected as free sugars, and their contents are shown in Table 4.
Glucose, galactose, xylose, arabinose, rhamnose, fructose, galacturonic acid, ribose and desoxyribose were the component sugars of glycosides.
Glucose, galactose, xylose, arabinose, galacturonic acid, rhamnose, ribose, desoxyribose and fructose were detected as the component sugars of the hot 50% ethanol-soluble polysaccharides.
Glucose, xylose, arabinose, galacturonic acid, mannuronic acid (?) and ribose were found as the components of the cold water-soluble polysaccharides, and, in addition, glucose, arabinose and galacturonic acid as those of hot water-soluble polysaccharides.
Galactose, arabinose galacturonic acid and small amounts of glucose and mannose were found as the components of the protopectin fraction.
As the components of hemicellulose, glucose, mannose, xylose, rhamnose, fructose and ribose were detectable.
Cellulose and a small amount of lignin were detected also.

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The fundamental studies on the metabolism of organic acid by Corticium centrifugum have been made, and the following results were obtained.
1) It was identified with mixed melting point and paper chromatography that the organic acid produced by this fungus was only oxalic acid.
2) Thiamine was the growth promoting substance of this fungus, although the relation-ship between added thiamine and accumulated oxalic acid was not proportional.
3) The maximal accumulation of oxalic acid was obtained after one week's incubation, thereafter, oxalic acid was decomposed by the autolysis of this fungus.
4) The initial pH was optimum at either the neutral or the weakly acidic side; this fungus was resistant to acid and fragile to alkali. Optimal sugar concentration to the formation of oxalic acid by the fungus was apporoximately 5 per cent (sucrose), and relation between growth of the fungus and accumulation of oxalic acid was not proportional, in the case of a variety of sugars being used as the source of C.
5) Peptone as nitrogen source was more effective to maximal accumulation of oxalic acid by this fungus, and potassium nitrate, ammonium phosphate and ammonium sulfate yielded a lower maximal accumulation, in the decreasing order.
6) The effects of inhibitors to growth and oxalic acid formation were examined using fungus matter: The growth of this fungus was completely inhibited by 10^-3M of sodium azide, and considerable accumulation of oxalic acid in the culture media was found by the addition of 10^-3M monoiodo acetate, whereas thiourea and urethane were not effective.

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The phosphorylase inhibitor in the root of mangold is fractionated, in the same procedure as reported in Part 3 of this series. The precipitate fraction from 75% acetone solution is most inhibitory, showing the same reddish-violet coloration of saponin as other inhibitory fractions by the LIEBERMANN-BURCHARD's reaction. The saponin fractions are refined progressively, the result indicates that one of the phosphorylase inhibitors is a saponin.

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Although the literature contains several references concerning to the browning reaction, that is to say, the amino-carbonyl reaction of heated milk, investigation on the main factors of this reaction has not been extended beyond the milk protein-lactose system, as source of the reaction. In view of the facts described herein, the most reasonable conclusion to be drawn from available data is that the urea contained in milk is the other main factor taking place in the amino-carbonyl reaction, besides the protein-lactose system. Urea, however, has been dealt with in previous reports as an inactive reactant in the amino-carbonyl reaction.
(1) The urea commonly contained in milk is completely decomposed by the addition of 1cc of 10 per cent urease solution, prepared from soya bean powder by the method of VAN SLYKE and CULLEN, when 10cc of milk has been incubated at 20° for 3hours. (Fig. 1)
(2) The browning of milk in which urea has been decomposed by the urease action before heating at 120° for 2 or 3hours is not more remarkable than the control milk to which (NH₄)₂CO₃ was added, equivalent to the urea contained in milk, in order to cancel the reaction of (NH₄)₂ CO₃ produced in the samples by urease action. (Table 1).
(3) The loss of urea in milk is 67 per cent during heating at 100° for 10hours. It appears that these changes obtained are maximum in amino compounds, the amino-carbonyl reaction taking place as shown by HODSON and KRUEGER. (Fig. 2).
(4) The browning of heated milk is increased by increasing the addition of urea to milk before heating. (Table 2)

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The artificial culture of goat's rumen Infusoria which belong to oligotrich cliata have been studied.
Infusoria can survive for 5_??_7 days at various concentrations of the rumen liquor, and for 10_??_14 days by the addition of the fresh white clover leaves into the culture medium.
The life of Infusoria which include species of Entodinium caudatum, E. longinucleatum, E. simplex can be extended for more than 30 days in vitro by the addition of the fresh white clover leaves at the amount of 5% to the hay-salt solution extract culture medium which is incubated with rumen bacteria for 24_??_48hrs. at 38° with vitamin B₁₂ added.
It is observed that when the culture medium is renewed at intervals of 48hrs., Infusoria are in an actively motile state and have a frequent fission which, multiplies to numbers of 50, 000 organisms per ml medium.
Vitamin B₁₂ is not effective for these protozoa when it is supplemented without fresh clover leaves.

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(1) The determination of cobalt has been carried out on about thirty samples of indigenous feeds by colorimetric and polarographic methods, several factors affecting the determination being examined.
(2) Results obtained by the two methods coincided very closely.
(3) The cobalt content of indigenous feeds were in the same order with those reported in foreign countries, no sample with an extremely low cobalt content being found among the feeds examined (Table 5).

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1. A more satisfactory paper-chromatographic separation than that previously reported⁽³⁾ of unsaturated fatty acid esters as their mercuric acetate addition compounds has been achieved by employing the ascending technique.
2. The reversed-phase paper chromatography of the hydroxamic acids of higher fatty acids has been examined and found to be inferior to that of the 2, 4-dinitrophenylhydrazides of fatty acids⁽⁴⁾ in respect of their sensitivity and separability on the paper.
3. The component fatty acids of cow milk fat have been well separated and identified on paper by combining the preliminary fractional distillation of their methyl esters with various paper-chromatographic techniques, such as the mercuration, 2, 4-dinitrophenylhydrazide, and hydroxamic acid methods.
4. The fatty acid composition of cow milk fat has been estimated from the paper-chromatographic, spectrophotometric, and distillation data by a simplified procedure, using a relatively small amount of the original fat.

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Lecithin (I), cephalin (II), and acetone-soluble (III) fractions were separated from the heart of a rorqual as presented in Figure 1.
The following characteristics were found, I: P 4.13%; N 1.76%; choline 14.57%; iodine value 91.6; N:P=1:1.06 and N:choline=1:0.96; II: P 2.66%; N 1.37%; iodine value 122.3; N:P=1:0.88, ethanolamine 4.90%; and serine 2.52%; and III: acid value 1.0; sapon. value 127.8; iodine value 130.5 and unsaponifiable material 43.8%.
The composition of component fatty acids of I and II were estimated spectrophotometrically after they had been converted to conjugated acids by KOH-glycol reagent. The contents of saturated, monoethylenic, dienoic, trienoic, tetraenoic, pentaenoic and hexaenoic acids were: 20.6, 52.9, 4.1, 6.1, 7.6, 6.2 and 2.5% respectively, in I; and 26.4, 35.0, 0.8, 16.5, 10.0, 8.2 and 3.1% respectively, in II.
The composition of component fatty acids of III was as follows: myristic 2%; palmitic 12%; stearic 12%; zoomaric, 4%; oleic 24%; eicosenoic 8%; linoeic+linolenic 6%; arachidonic 16%; and clupanodonic 16%.
From the unsaponifiable material of III, cholesterol was identfied while the presence of cetyl, octadecyl, zoomaryl, oleyl, gadoleyl, and C₂₀-highly unsaturated alcohols was presumable.

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Phosphatidic acid obtained from the heart of a rorqual was found to possess the following characteristies: P 3.96%; N 0.07%; and the iodine value 112.1.
Glyceryltriglycero phosphoric acid was isolated as barium salt from its hydrophoslyzate, so the skeletal structure of the phosphatidic acid was supposed to be similar to cardiolipin.
Sodium salt of the phospatidic acid showed a strong activity as haptene for the serodiagnosis of syphilis.
The percentage composition of its component, fatty acids, estimated spectrophotometrically after they had been converted to conjugated acids by KOH-glycol reagent, was as follows: saturated 34.0; monoethylenic 13.8; dienoic 9.0; trienoic 23.6; tetraenoic 12.1; pentaenoic 6.2; and hexaenoic 1.3.

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Both α, α- and α, β-trehalose were isolated from Koji extract as their crystalline octaacetates, and the latter was identified by the mixed melting point with the synthetic specimen prepared by the method described by MICHEEL et al. Separation of these sugars from isomaltose was performed by removing of the latter as its water insoluble phenylosazone.
Kojibiose also gives no phenylosazone and is presumed to be 1, 2-α linked disaccharide; neither the free sugar nor its octaacetate could be isolated in the crystalline form.

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The antibiotic components were isolated from the broths of chlortetracycline-producing Streptomyces, fermented in natural mediums containing sodium bromide in place of sodium chloride. The components were shown to be the mixture of chlortetracycline and a new antibiotic separated by fractional crystallization. From the ultraviolet and infrared absoroption spectra, melting point, optical rotation, color test, elementary analysis and biological potency, this new antibiotic was determined to be identical with tetracycline.

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In order to establish a direct colorimetric method to estimate urea in milk, the α-isonitrosopropiophenone method was studied as follows:
(1) The color of urea with α-isonitrosopropiophenone was stable in the dark, slightly stable under scattered light, and unstable under the direct rays of the sun (Tbale 1).
(2) The regression coefficient in urea solution and milk were obtained with a 5% significance level (Tables 2 and 3). The relation between the reading in a Beckmann Spectro-phtometer (X) and content of urea in milk (Y), was indicated according to the following equation with a 5% significance level:
Y=104.35X
(3) The coefficient variation of this colorimetric and the standard urease method were 3.68 and 2.90, respectively, with a 5% significant level (Table 4).
Therefore, by means of the colorimetric method described above, the content of urea in milk was determined as follows:
(4) It appeared with a 0.1% significance level that the content of urea in milk is different for each cow (Table 5).
(5) The percentage of urea in market milk was 14.02_??_18.37mg% with a 5% significance level obtained from 56 samples.
The correlation coefficient between the color of milk heated to 120° for 1.5 hours and the content of urea in milk before heating was 0.0678_??_1.0278 with a 5% significance level. It is found that the content of urea in milk is probably a factor in the browning of heated milk which is different in each milk as shown by WEBB⁽²⁶⁾ (Fig. 1 and Table 7).

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In the present paper, the author has reported the fluctuation of amylase, water-soluble phosphorus compounds, nitrogen substances and ketonic acids during autolysis of green malt of American two-rowed barley, subjected to 2 days' germination, placed in a closed vessel containing a small amount of toluene as a disinfectant, and kept to autolysis.
It was found that activity of several enzymes except amylase decreased, while the increase of amylase activity (both saocharogenic and dextrinogenic powers) was observed by the anaerobical treatment mentioned above.

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