In order to improve the method for obtaining the hypocalcemic proteinous substance precipitated from thymus gland of an ox with calcium chloride, which was found to be unsuitable for purification of the objective protein, fractionation with ammonium sulfate was attempted. The fractions precipitated at the concentrations of 15%, 25%, or 41.7% ammonium sulfate showed a significant effect, showing average decrease of serum calcium level by 10-17% of the initial value upon administrations of 1.0mg/kg body weight in rabbits. The fraction of 15% saturation was obtained in a crystalline-like form. It was also found that the leucocyte activity did not necessarily parallel the calcium activity.
The present investigation was undertaken to determine the effect of mechanical agitation on liquid-liquid dispersion. A turbine impeller (150 mm in diameter) with fixed guide vanes was employed at 200-350 rpm and 30-90° blade angle. A mixture of n-C7H16-CCl4 (D4=1.000) and distilled water were used as the dispersion phase and continuous phase, respectively. The particle diameter of dispersion was determined by a microscopic method. (1) Particle size distribution showed a logarithmic normal distribution after considerable agitation. (2) Mean diameter to agitation time decreased linearly on log-log coordinate. (3) The energy efficiency of the dispersion was in the range between 10-3 and 10-6. (4) With the increase of revolution number and blade angle, energy efficiency had a larger value on liquid-liquid dispersion.
Protonation equilibrium of a series of 1, 2, 3, 4-tetrasubstituted 3-pyrazoline-5-thiones (I) was examined by spectrophotometry and potentiometric titration. In aqueous solution of sulfuric acid, protonation of the sulfur atom in the thiopyrazolone ring of I occurs and the increase in their ionization rate is greater than the increase of Hammet's acidity function, h0, by increasing concentration of sulfuric acid. This phenomenon can be explained by the highly polarized structure of I resulting in the different degree of hydration between I and the Hammet indicator. Basicity of I agrees with that presumed from the I-effect of the substituent, and the basicity of 4-substituted thiopyrine derivatives fall in the order of CH3, H, C6H5, I, Br, Cl, NO2, NH3+, and NH(CH3)2+ as a substituent. Ionization constant of thiopyrine is -0.35 and it is a much weaker base than antipyrine.
Solvent effect on the ultraviolet absorption spectra of 1, 2, 3, 4-substituted 3-pyrazoline-5-thiones (I), and their intermolecule hydrogen bonding with phenols and alcohols were examined. The ultraviolet spectra of I in proton-donating solvents, such as water and alcohols, are much more hypsochromic than those in nonpolar solvents like cyclohexane and benzene. This phenomenon is due to the formation of hydrogen bonding of I with hydrogen of the solvent. This hydrogen bonding power of I is greater than that of antipyrine derivatives and is reverse of the strength of Brφnsted bases.
Reaction of 1-phenyl-2, 3-dimethyl-3-pyrazoline-5-thione (I) with oxidation agents and chemical properties of its oxidation products were examined. I undergoes reaction with iodine, nitrous acid, and selenous acid to form 5, 5'-dithio-bis (1-phenyl-2, 3-dimethyl-pyrazolium) salt, 1-phenyl-2, 3-dimethyl-5-sulfinopyrazolium hydroxide, or its anhydride inner salt (II), and also reacts with bromine and hydrogen peroxide to form II and 1-phenyl-2, 3-dimethyl-5-sulfopyrazolium hydroxide, respectively. II undergoes electrophilic reaction with proton and iodine cations in acid solution, and nucleophilic reaction with hydroxide and methoxide anions in alkaline solution.
Tautomeric equilibrium of 1-phenyl-3-methyl-3-pyrazoline-5-thiones (I) was examined from ionization constant and from ultraviolet, infrared, and nuclear magnetic resonance spectra. It was thereby presumed that I are present in SH form in nonpolar solvents and in NH form containing a few percent of SH form in aqueous solution, and are associated in crystalline state by forming an intermolecular hydrogen bonding by NH or SH form.
Lactic acid bacilli used for microbiological determinations all receive growth inhibition by 3.25-30.0 μg/ml of Lincomycin so that the parent strain cannot be used in the determination of vitamins in the presence of Lincomycin. If these lactobacilli are serially cultured in the medium containing increasing amount of Lincomycin, they can tolerate up to 2000 μg/ml of Lincomycin. The vitamin requirements of this resistant strain is almost the same as that in the parent strain. It was thereby found that the calibration curve will be entirely unaffected by the use of this resistant strain, even in the presence of 300-400 μg/ml of Lincomycin. Satisfactory results were obtained in the determination of urinary vitamins in patients administered Lincomycin, which had been impossible using the parent strain.
The metabolites of Streptomyces luteoreticuli KATOH et ARAI were investigated. Four new compounds, luteoreticulin, (V), C19H19O5N, luteothin, (VI), C22H25O5N, metabolite X, (X), C15H12O3N2, and metabolite XI, (XI), C15H14O3N2, were isolated from the acetone extract of the mycelial cake. 1-Methoxyphenazine (VII) and methyl phenazine-1-carboxylate (IX), which were first isolated from natural origin, were obtained in addition to aureothricin (I), thiolutin (II), aureothin (IV), and 1, 6-dimethoxyphenazine (VIII). The other two metabolites (III and XII) were also isolated but not identified.
To determine the configuration of A, B-ring juncture in lyoniol-A, examinations were made on the stereomodels and octant diagrams of 3- and 10-keto compounds derived from lyoniol-A, grayanotoxin-II, and analogous compounds, and it was found that only the 10-keto compounds (Fig.2) and not 3-keto compounds (Fig.1) are suitable for this purpose. The optical rotatory dispersion (ORD) curves of 10-keto compounds derived from lyoniol-A (Fig.3) and grayanotoxin-II (Fig.4) have typical negative Cotton effect. Thus, the A, B-ring juncture of these two compounds must be trans.
3-Oxo-1-thia-4, 8-diazaspiro [4, 5] decane derivatives (IV, VI) were prepared from 1-substituted 4-piperidones (I), primary amines (II-1, R2=H ; II-2, R2=alkyl ; II-3, R2=arylalkyl ; II-4, R2=aryl), and mercapto acids (III-1, A=-CH2- ; III-3, A=-(CH3)CH- ; III-4, A=-(C6H5)CH- ; III-5, A=-(CH3)2C- ; III-6, A=-(C6H5)2C-). Similarly, 4-oxo-1-thia-5, 9-diazaspiro [5, 5] undecane derivatives (VII) were prepared, though in low yields, from I, II-1, and 3-mercaptopropionic acid (III-2), but corresponding products were not obtained from I, II-2-II-4, and III-2. Sulfoxides (XI) and sulfones (XII) of IV, VI, and VII were also prepared. Reaction condition, byproducts and reaction mechanism were briefly examined.
Reaction of diketene with ethylenimine (Ia), 2, 2-dimethylaziridine (Ib), 2, 2-diphenyl-3-methylaziridine (Ic), ethylaziridine-2-carboxylate (Id), aziridine-2-carboxamide (Ie), and 2-benzoyl-3-phenylaziridine (If) afford the corresponding 1-acetoacetylaziridines (II) in a good yield. Treatment of II with sodium iodide or conc. sulfuric acid results in their isomerization to 2-acetonyl-2-oxazoline derivatives (III or IV). Isomerization of IIb-e preferentially occurs only in one direction but treatment of IIf with sodium iodide produces IIIf and IVf at the same time. Warming of IIf in benzene results in its isomerization to 6-methyl-2-phenacyl-3, 4-dihydro-2H-1, 3-oxazin-4-one (XVII), while treatment of IIf with sodium ethoxide gives 3-acetyl-5-benzyl-4-phenylpyrrolenin-2-one (XIX).
1-Phenyl-3-methyl-3-pyrazolin-5-thione, 1-phenyl-3, 4-dimethyl-3-pyrazolin-5-thione, and 1, 3-dimethyl-3-pyrazolin-5-thione were synthesized and it was found that their chemical reactivity indicated a thiol structure.
Difference in the potentiative action of amphetamine and cocaine in increasing the analgesic effect of morphine was examined by the hot-plate method and pressure-stimulation method in mice. Amphetamine and cocaine themselves did not show any analgesic effect but both potentiated that of morphine. Administration of reserpine or Tetrabenazine, followed by dl-DOPA, and then by morphine indicated that the analgesic effect of morphine returned to the value of that with single administration of morphine. Administration of tetrabenazine, followed by amphetamine and morphine resulted in potentiation of the analgesic effect of morphine but not when morphine was administered with cocaine. However, when administration of tetrabenazine and then dl-DOPA was followed by simultaneous administration of cocaine and morphine, potentiation of analgesic effect of morphine was observed, as in the case of cocaine and morphine alone.
In Acetobacter suboxydans ATCC 621, which requires pantoic acid for growth, mevalonic acid, which is its isomer having a similar structure and which is a growth factor for some kind of Lactobacillus, showed 1/400 activity of pantoic acid. N-Mevalonyl-β-alanine, which has a structure similar to pantothenic acid, had only a weak substitutive activity. This relationship parallels the relationship between pantoic acid and pantothenic acid for the growth of Acetobacter suboxydans ATCC 621.
Thirty-one kinds of N-(α-naphthylacetyl) amino acid were prepared by the Schotten-Baumann method. These derivatives were considered to be useful for the growth of plants. These derivatives had smaller specific rotation than phenacetyl-amino acids. N-(α-Naphthylacetyl)-L-homoserine was found to undergo cyclization to its lactone by treatment with hydrochloric acid.
Distribution of phosphotransacetylase in 48 strains of lactic acid bacteria, including Lactobacillus, Streptococcus, Leuconostoc, and Pediococcus, was investigated. Lactobacillus fermenti IFO No.3071 (ATCC No.9338) was found to contain the highest level of phosphotransacetylase among the lactic acid bacteria tested. Not any correlationship was found between fermentation patterns, such as homo and hetero fermentation, and levels of phosphotransacetylase.
The colorimetric determination of methanol with chromotropic acid was applied for assaying the activity of kallikreins to hydrolyze p-toluenesulfonyl-L-arginine methyl ester (TAME). This method was found to be useful for a fine determination of the specific activity in terms of μmoles of TAME hydrolyzed per minute for a certain amount of sample. The assay results from this method were in good agreement with those from the usual hydroxamate method. This method was more simple in manipulation, and was found to be 10 times more sensitive than the hydroxamate method.
A packed bed reactor was made of stainless steel tube which contained alumina catalyst and was heated by nichrome wire. Using this reactor, reactions between ethylenediamine (DA) and ethyleneglycol (GL) were studied under several conditions. This reaction yielded very interesting products such as pyrazine, methylpyrazine, ethylpyrazine, and 4-methylpyridine. From experiments performed under various conditions, it was concluded that the higher the molar ratio of GL to DA was, the higher the conversion of DA to pyrazines resulted, and also that the lower the reaction temperature was, the higher was the formation of ethylpyrazine than pyrazine.