Studies were performed on the phenomenon of blood coagulation of American crawfish, Cambarus clarkii, and the fibrinogen contained in its blood plasma was demonstrated electrophoretically. The results are as follows:
1) In the case of crawfish, the course of blood coagulation can be observed separately in two stages. First, the blood cells contacted with and joined each other by the cytoplasmic processeses long dendritic or horn-like form grown out rapidly from the cells, involving a few of fine fibrilles, and yet the blood plasma maintained its liquidity.
The explosive breake down of so-called explosive cells might take an important part of this phenomenon.
The decarcifying anticoagulants, such as Na. citrate, could not inhibit the coagumechanism completely, and the fibrinclot appeared promptly yet maintaining the liquidity of blood plasma, by the addition of Na. citrate to freshly aspirated blood. Moreover the microcopic examination of extended preparation of this clot revealed the numerous blood cells involved in the fibrin-nets. So that it is reasonably considered that the Na. citrate may have some promoting effect on the first stage of coagulation.
2) In the second stage, the major part of fibrinogen in the blood plasma changed into fibrin-fibre and the complete gelation of blood was reached. The facts found in this stage are resemble to that of the higher janimals. Oxalate and citrate,, though having a promoting effect on the first stage, showed the inhibition of the second stage of coagulation. But heparin showed no inhibition.
3) The blood plasma obtained by inhibiting the second stage of coagulation using Na. citrate was gelated completely when Ca. chloride or homogenous thrombin solution was added to it.
The temperature, neither low (3-4°C) nor high (46°C) inhibits the first stage of coagulation, resulting the formation of a soft clot.
4) The electrophoretic pattern of crawfish serum on cellulose acetate membrane revealed a prominent protein fraction, hemocyanin which corresponds to the albumin in higher animals. and other several minor fractions than hemocyanin corresponding to the globulin. In the case of citrated blood plasma, a major fraction appeared in the globulin portion, which was never seen by serumelectrophoresis.
The electrophoretic pattern of the liquid obtained after the gelation of citrated blood plasma by adding Ca. chloride, seemed quite similar to that of the serum and no major fraction in the globulin portion was demonstrated.
5) The globulin fraction salted out from the citrated blood plasma by 30 per cent saturation of Ammonium sulfate showed gelation by the addition of homogenous thrombin solution.
From the facts described above, ethe major protein fraction in globulin portion of citrated blood plasma of crawfish is considered to be the fiblinogen of this animal.
6) It is a phylogenical interests to notice that in the case of crawfish, next of hemocyanin, fibrinogen is the most prominent fraction of serum protein while others than these are found in very small quantities.

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Eighteen strains of Ps. aerugincsa were cultured on the glutamic acid liquid medium and the production of the fluorescent substances were examined by the gel filtration of culture fluid filtrate and the paper electrophoresis of fluorescent fractions.
The results are as follows:
1. The common characteristic of all the 18 strains tested was the production of Fluorescence II fraction.
2. In all the instances of pigment producing strains, the yellow or yellowishgreen portion of fluorescent fraction, observed at the early stage of cultivation, may correspond to the Fluorescence I substance.
3. The paper electrophoretic patterns of Fluorescence II fraction of colorless strains (No.58,39 & 65) originated from a pigment producing strain, were quite the same as that of the ordinary colored strains and showed a clear C-spot.
4. In other colorless strains than these, a wide and undistinguishable zone of fluorescence was demonstrated without showing the C-spot distinctly, and the site of C-spot was involved in the wide zone.
5. The aceton soluble fuorescent substances were extracted from the washed cells of both the pigment strains and colorless variants, and the liquid soluble portion which was contained therein was similar to the Fluorescence II substance presented in the culture fluid filtrate.
6. The aceton extracts of bacterial cells of the other colorless strains than these showed only a faint fluorescence.

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The formation of red pigments by Ps. aeruginosa. strain NO.145, was studied.
1. _DL-glutamic acid was found to be a good substrate for the synthesis of the red pigments in culture media by Ps. aeruginosa.and it could be replaced by several related amino acids (_DL-alanine and _L-leucine).
2. The red pigments formation on the medium containing one of these amino acids varied according to its concentration and the incubation time of the bacteria.
The maximum yield of the red pigments was obtained at 1% concentration of _DL-glutamic acid or _DL-alanine in 80 hrs. after incubation, although the production of the red pigments was rather lower by leucine medium.
3. The yellow pigments produced by Ps. aeruginosa. strain No 145, in the up-right medium converted its color gradually into red by age, but on the slant medium they changed the color rapidly in red.
4. The fleshly isolated strains of Ps. aeruginosa.obtained from human speciemens have commonly the ability of the red pigments formation on the glutamic acid medium, although the ability was decreased gradually successive subcultures.
5. The synthetic medium containing _DL-glutamic acid at 1.0% (for 80 hrs. incubation)has been found to be the most suitable for production of the red pigments by Ps. aeruginosa.

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In the previous paper, it was reported that the synthetic medium containing 1 %glutamic acid is most suitable for production of the red color pigment of Pseudomonas aeruginosa (strain No.145).
The highest yield of the pigment producing can be obtained at pH 7.5, in 80 hours after incubation 37°C.
The present report deals with the isolation and some physiochemical properties of the red colored pigment.
Since the red pigment can not be extracted by common organic solvents, the culture filtrate was fractionated by chromatography, Sephadex G-25 M, using distilled water as a eluent, and the red colored portion was obtained.
By paper electrophoresis (2 sheets of Toyo filter-paper and Mcllvaine's buffer pH 6.0), the concentrated portion was fractionated in three red colored fractions, as a pink color fraction, a red color fraction and a purple fraction. The fractions were arbitrarily named P-I, P-II and P-III from the negative pole side.
The solution of each fraction was stable on a visible light, in air oxygen and considerably stable in aq. solution at pH 3-10, although unstable in alkaline solution in which it was converted to yellow color. In acidic solution P-I and P-II were converted to reddish violet color, and P-III solution to deep violet.
The spectra of the reduced and oxidized pigments did not show the band at 515,525 and 550 mμ. Reduction of each red pigment with Na₂S₂O₄ gave yellow compounds, which could be converted to the original color by hydrogen peroxide or free oxygen in air. Oxidation of these solution by hydrogen peroxide resulted in complete bleaching.
The maxima wavelengths of the pigments were shown as follows; in distilled water, λ max (mμ): P-I 234,283,396,515: P-II 258,393,525: P-III 241,297,404,550; in 0.1 N-HCI, λ max (mμ): P-I 236,287,390,546: P-II 258,388,546: P-III 242,298,394,560; in 0.1 N-NaOH, λ max (mμ): P-I 233,271,374,460: P-II 261,372,480: P-III 221,283,372,480.
The pattern of ultraviolet and visible absorption spectrum of P-I and P-III were very similer, However, the absorption curve of P-II showed only a single peak.
On the basis of these evidences, the three red pigments isolated from the culture broth of Ps. aeruginosa, strain No.145, were considered to be derivatives having the same skeleton, which were substituted for different auxiliary chromophores.

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Neonatal respiratory distress often leads to pulmonary hyaline membrane disease. Etiologically primary atelectasis or oxygen intoxication is suggested. It is also reported that cases of respiratory distress syndrom reveal patent ductus arteriosus which elevates pulmonary artery pressure, resulting in the exudation of blood plasma from pulmonary capillaries with a resultant formation of pulmonary hyaline membrane.
A loose ligation of the radix of venae pulmonalis at auricula sinistra resulted two days later in pulmonary hypertension with the exudation of blood plasma in alveoli, leading to a marked development of pulmonary hyaline membrane. The right ventricular pressure which used to be 4.5∼9.0mmHg. was elevated to 11.0∼17.0mmHg. following ligation. Such elevation suggested the development of pulmonary hypertension.

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In an attempt to confirm a possible decrease in pulmonary compliance in case of pulmonary hyaline disease, a study was made of young guinea pigs in which hyaline membrane disease was induced by intratracheal instillation of the guinea pig serum or by inhalation of CO₂.
The air was introduced into the animal's excised lungs. The pulmonary compliance of these animals obtained on measurements of intrapulmonary pressure was generally proved take less than that of normal lungs.
This finding had been obtained with acceptance in human neonates with respiratory distress and in normal neonates.
It was concluded that experimental pulmonary hyaline membrane disease was not only morphologically but also functionally similar to that found in premature infants.

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The fat metabolism has been studied in premature infants fed with several formula. In this study the auther has analized fatty acid components of several kinds of dried milk powder, the trade marks of which were M., L., E., P., S. and N., and also examined the relationship between their fat absorption and the growth of the premature infants.
The results of this study are summarized as follows:
1) The fat content was 18.0% in M.,17.0% in L.,14.0% in E.,6.6% in P.,27.6% in S. and 24.0% in N. milk powder.
2) The linoleic acid content in the fat of each milk powder was 11.5% in M.,12.7% in L.,1.6% in E.,2.4% in P.,19.4% in S. and 12.7% in N., respectively.
These linoleic acid content occupy 3.0%,3.3%, O.3%, O.2%,6.6% and 3.8% of the total calory of each milk powder.
3) The components of the fatty acid in the feces were noted to be influenced by the components of the fatty acid in the milk powder taken by the infants. The fatty acid components in the feces were different in some extent among each individual, even though they were fed with the same milk powder. However, any significant difference was not found in a single case when the fatty acid components of the feces were assayed according to the course of days after birth.
4) Some different results were observed between the weight method and the chrom green method to measure the rate of fat absorption. From the results obtained, it is concluded that the chrom green method should be performed to obtain an exact result.
5) The rate of fat absorption was not influenced by both age and body weight of the premature infants, but it decreased markedly when more fat was given to them. However, as for the absolute quantity of fat absorbed, it increased when more fat was taken by the infants.
6) The rate of fatty acid absorption was not connected with both age and body weight of the premature infants, and the rate of fat absorption was pararell,, neniethitehrer with the rate of absorption of oleic acid nor with that of linoleic acid..
7) As the fat contents of feces did not show any marked change even after the intestinal flora was suppressed by administration of antibiotics, the intestinal bacilli were not considered to give a significant influence upon the fat absorption.
8) The rate of absorption of poly-unsaturated fatty acids including linoleic acid was not so high, comparing with the rate of fat absorption.
9) Both the rate and absolute quantity of absorption were very low as far as the linoleic acid of E. milk was concerned, while the rate of fat absorption of E. milk was the highest among the examined milk. On the other hand, those of the linoleic acid of L. milk were most excellent.
10) The increase of body weight was most excellent in the premature infants fed with M. milk or L. milk, while it was not so good in the premature infants fed with E. milk. The latter fed with E. milk, however, did not show any significant clinical signs which might be caused by essential fatty acid deficiency such as anorexia, abnormal changes of the feces, or appearance of the dermal symptomes did not occur.
11) Many attempts are being tried to improve the dried milk powder at present, the ideal of which is the breast milk, but the fatty acid components of the breast milk are not always the same; they are found to be varied according to the differences of race, climate, food and the course of days after the delivery. It is one of important problems, in which percentage the fatty acid of the milk should be contained.
The auther has made it certain that milk powder which contained linoleic acid in 3 to 4 per cent of whole milk calory braught the most excellent growth in the premature infants whose birth weight were over 1500g.

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