A strain of hyphomycetes that produces a toxic substance was isolated from a fungus that was parasitically growing on the malt feed responsible for mass death of milking cows. The fungus penetrates into malt grains from an injured section, causes morbid change in the starch and aleuron layers, to brown the tissues, and produces a metabolic products that causes marked toxicity in animals. The fungus was found to grow on a medium with a wide variety of carbon source. The morphology of the fungus on the Czapek culture medium is as follows: Vegetative mycelium, 3-5×18μ; aerial mycelium, 2μ diam.; conidiophore, 3.0-4.5×400-500μ; metulae, 3.0-5.2×6.0-10.4μ; sterigmata, 2.3-2.7×6.0-6.7μ; conidia, 2.5×3.2μ. Since the morphology and other properties of this fungus are identical with the description for Penicillium urticae Bainier in the classification by Thom (“A Manual of the Penicillia” (1949)), the fungus is now determined as that strain.
1) The range of optimal temperatures for the growth of the hyphomycetes which produce toxic substance is 8-40°, the best range being 22-25°. 2) The allowable pH for growth is in a range of 3.5-8.5, the best being pH 4.5-6.5. 3) Essential nutritional sources are sugars, nitrogen, phosphates, and magnesium salts, the growth being dependent on the ratio of sugar to nitrogen content. 4) The fungus grows in aerobic conditions, absorbing oxygen from the air in contact and producing carbon dioxide. 5) The sporodium of this fungus dies in 1 minute in 0.1% AgNO3 solution, in 24 hours in 0.001% AgNO3 solution, and in 5 minutes in 70% ethanol, but is comparatively resistant to other chemicals. The sporodium was also made inert by the ultraviolet rays (2537 Å) of 1350 radiation energy and by moist steam of 58° in 10 minutes. 6) The enzyme of this fungus possessed weak amylase, lipase, and protease actions, and a markedly strong disaccharide decomposing action. The presence of phosphatase, reductose, and catalase was marked.
The production of toxic substance by the fungus on culture medium was found to be possible over a wide range of sugars as the basal medium, the strength of the toxicity being in proportion to the power of enzymatic decomposition of various sugars. The toxic substance is produced comparatively rapidly in a medium containing glucose or sugars which are easily decomposed to glucose. In various grains incubated for 14 days after inoculation of this fungus, only the malt grains showed toxicity, the maltose having been consumed to a large extent. The toxic substance produced in this case was the same as that obtained from the culture medium containing glucose. The toxic substance corresponded to the composition of C7H6O4 and the toxicity, LD50, was 15.6mg./kg. in mouse.
The fungus grew parasitically on sugar-containing medium or malt grains and produces a toxic substance (C7H6O4) as a metabolic product in early stages. The domestic animals which take the fungus with their feed falls prey to poisoning or succumbs by poison. Pathological dissection of these animals revealed the cause of death to be due to brain hemorrhage.
In the previous paper, isolation of lumicaeruleic acid from the ether extract of the bark of Phellodendron amurense Rupr. was described. In the present series of experiments, Phellodendron bark was extracted with alcohol, berberine and the bitter principles were removed from the extract, and an acid substance soluble in alkali was obtained. This acid substance gave lumicaeruleic acid, m.p. 167-168°, in a better yield than by the previous method. This acid substance was found to be present in an equimolar amount as berberine. Low-pressure sublimation (150-170° at 3mm. Hg) of lumicaeruleic acid from Phellodendron bark and that, m.p. 164-165°, from the rhizome of Coptis japonica Makino, yielded the same sample melting at 173° which was found by mixed fusion to be identical with ferulic acid, m.p. 173°, obtained from Asafetida. It was thereby shown that the structural formula of the capillary images of Phellodendron bark and Coptis are indicated by the following: α-(Br)⋅non-(B)L=Ferulic acid⋅β-(Y)=Berberine⋅γ-(Br)
It was found that the respective application of thionyl chloride to quinaldine ethiodide, lepidine ethiodide, and 2-methylbenzothiazole ethiodide yielded 2-trichloromethylquinoline ethochloride (I), 4-trichloromethylquinoline ethochloride (II), and 2-trichloromethylbenzothiazole ethochloride (III). Reaction of (I) with 2 moles of quinaldine ethiodide in acetic anhydride, in the presence of potassium acetate, yielded 1, 1′, 1″-triethyl-10-(2″-quinolyl)-2, 2′-trimethinequinocyanine diiodide (IV) with λmax 5850 Å. Similar reactions of (II) and lepidine ethiodide and of (III) and 2-methylbenzothiazole ethiodide respectively yielded 1, 1′, 1″-triethyl-10-(4″-quinolyl)-4, 4′-trimethinequinocyanine diiodide (VI) with λmax 6700 and 5950 Å and 1, 1′, 1″-triethyl-meso-(2″-benzothiazolyl)-2, 2′-trimethinethiocyanine diiodide (VIII) with λmax 5450 Å. The reaction of (I) and quinaldine ethiodide in xylene, with cadmium carbonate as the addition agent, gave hydriodide or iodide of (IV) with the same absorption maximum as that of (IV), and the same reaction of (II) and lepidine ethiodide gave the hydriodide or iodide of (VI) with λmax 7100 and 5950 Å, and of (III) and 2-methylbenzothiazole ethiodide, dihydriodide or diiodide of (VIII) with λmax 5750 Å, all in comparatively good yield. The dehalogenation of these with ammonia respectively gave (IV), (VI), and (VIII).
1) Antimicrobial activity of nonylpyrazole was tested. Their maximum dilutions were found to be 1:200, 000 and 1:2, 000, 000 against Trichophyton interdigitale and Staphylococcus aureus, respectively. 2) Nonylpyrazole was found to have the strongest antimicrobial activity among the various alkylpyrazoles tested, possesing alkyl group other than nonyl.
Thioformylphenylhydrazine was obtained by the application of potassium dithioformate to phenylhydrazine. Application of α-haloketone, α-haloacid, and their esters to the sodium salt of thioformylphenylhydrazine yielded the respective condensation products (V). Attempt to effect cyclization of these condensates to obtain hereto unknown 2-unsubstituted thiadiazine derivatives was found to be difficult, the majority undergoing decomposition. After trying various reactions, (Vc) and (Vd) finally yielded the corresponding objective compounds (VIa) and (VIc). Of the many decomposition products formed during cyclization, one obtained as yellow plate crystals of m.p. 190-192° was also obtained by the fusion of thioformylphenylhydrazine. Its molecular formula approximates C14H12N4 and its structure was assumed to be as shown in (VIII), although no confirmatory evidence had been obtained as yet.
As the compounds possessing structure similar to the procaine antagonist, sulfonamide, N1, N1-diethyl-, m.p. 105°, and N1-ethyl-p-aminobenzenesulfonamide, m.p. 107°, were prepared. The lethal dose, LD50, of the diethyl derivative against mouse was 5.5mg./10g. (subcutaneous injection, Behrens-Kärber method), 31.3±3.29mg./10g. (oral administration. V. d. Waerden method), and that of the monoethyl derivative, 5.1mg./10g. (subcutaneous injection of the hydrochloride, Behrens-Kärber method). The general action of these amides against mouse was ceasure of automatic motion, trembling, and respiratory paralysis. Neither showed any antagonism against procaine in either mouse or rabbit.
In order to clarify the mechanism of high frequency titration, a bridge suitable for high frequency wave was constructed and was so devised that the dielectric constant of a solution possessing conductivity could also be measured. This made it possible not only to measure dielectric constants and loss of dielectrics with ease and rapidity but also to carry out titration with higher accuracy and without any limit to the concentration. It has also effected the combination of the so-called high frequency titration and conductivity titration. Cells insulated by glass was constructed, equation to calculate its cell constant was obtained, and actual measurement was carried out.
1) Hydrogen bond energy of systems of pyridine 1-oxide (I) and quinoline 1-oxide (II), added with phenol (III) and methanol (IV) as the proton donor, was examined by the measurement of ultraviolet absorption. Such energy was 6 kcal./mole in the system (II)-(III), and 5 kcal./mole in (I)-(IV) and (II)-(IV) systems. The decrease of entropy, Δ, S, in these cases was -9 E. U. in (II)-(III) system, -13 in (I)-(IV) system, and -14 E. U. in (II)-(IV) system. The energy of hydrogen bonding was considered from the electrostatic and charge transfer forces. 2) In the above cases, absorption in the ultraviolet region shows a marked blue shift according to the formation of a hydrogen bonding. This fact has repeatedly been pointed out by the writer and it is assumed that the stabilization of the polar structure in the ground state in accordance with hydrogen bonding with the solvent is making the chief contribution. It is difficult to assume that the dielectric constant of the solvent acts primarily in such an effect. Some considerations were also made on this point.
Nuclear acylated compounds were prepared by heating catechol, resorcinol, resacylophenones, and their methyl ethers with acetic or propionic acid, in the presence of polyphosphoric acid, for 10-20 minutes in a boiling water bath. Catechol, guaiacol, and veratrole yielded acylocatechols, acylovanillones, and acyloveratrones, while resorcinol and its mono- and dimethyl ethers easily gave resacylophenones, their 4- and 2-methyl ethers, and dimethyl ethers in a good yield. Resacylophenones gave 4, 6-diacylresorcinols and resacylophenone 4-methyl ethers gave 4, 6-diacylresorcinol monomethyl ethers. However, nuclear acylation of resacylophenone 2-methyl ethers was difficult and resacylophenone dimethyl ethers failed to undergo nuclear acylation.
For the determination of organic thiocyanate compounds, the procedure of Pantschenko and Smirnow was reëxamined and the results obtained were unsatisfactory. The cause of such insufficiency of their determination method was concluded as being due to the complication of the reaction by the hydrolysis or solvolysis of alkali sulfide. Further, the reaction of benzyl thiocyanate and sodium trisulfide was studied in detail and it was clarified that thiocyanate ion was formed in equivalent amount during this reaction and that the disulfide was isolated and identified in almost equivalent amount. By the argentometry of the thiocyanate ion thereby formed, determination of benzyl thiocyanate was successfully concluded. The accuracy was ± 0.5%.
The determination method for benzyl thiocyanate using alkali polysulfide, described in the previous paper, was extended to other thiocyanate compounds and found that the method can be applied generally. A microdetection method for thiocyanate group was also proposed (in which a drop of Na2S3 or Na2S4 is added to one drop of the alcoholic solution of the sample, warmed for 1 minute in a water bath, and 1 drop of 10% H2SO4 is added. After warming the mixture for some time, 1 drop of 1% HCl solution of FeCl3 is added after the mixture has cooled and a red coloration appears in the presence of a thiocyanate compound. Limit of detection is 1 γ, and the limit of concentration is 1:10, 000 to 1:15, 000). The corresponding disulfides were isolated in 80-90% yield from the reaction of phenyl-, 2-carboxyphenyl-, 2, 4-dinitrophenyl-, and 4-methoxynaphthyl thiocyanate and thiocyanoacetanilide with sodium trisulfide. All the disulfides were isolated and identified. For the reaction of a thiocyanate compound and alkali polysulfide, a general formula (1) was forwarded and radical reactions (2-4) were assumed for it.
1) A brown color appeared when the solution of hydroquinone and hydrochloric acid was added to the alcoholic solution of ascaridol, and the light absorbancy (e) of the colored solution of different concentrations (γ) was measured by the Beckman spectrophotometer (λ=400 or 425mμ). From these experimental data (n=45), the following equations were calculated by the method of least squares: at 400mμ x=308.7y+15.7, at 425mμ x=473.5y+16.1 2) The standard deviation, σ, is ±0.0217 and the concentration limit corresponding to 2 σ limit is ±17.25mg. when measured at 400mμ. 3) The influence of a low boiling fraction of the chenopodium oil on the method was found to be negligible. 4) The data obtained by iodometry (N.F.), the Nelson's method (J.P.VI), and the present colorimetric method were compared.
In order to test the antibacterial action of thiazole derivatives, 6 kinds with diphenyl ether as the basic nucleus and 2 with diphenyl sulfide as the basic nucleus, shown in Table I, were synthesized. Growth inhibition tests in vitro with human type tubercle bacilli (H37 Rv) showed that the compound (V) alone possessed a strong antibacterial action while 7 other allied compounds showed only a weak action. All the 8 compounds tested also showed only a weak antibacterial action against other bacteria, such as Bacillus dysentereae and Salmonella typhosa.
Heating of anise alcohol and 2-aminopyrimidine, in the presence of alkali, results in dehydration-condensation to form p-methoxybenzylaminopyrimidine. By the examination of this condensation reaction, the condensation product was obtained in a comparatively good yield. The condensation products were also obtained by this condensation with the use of 2-aminopyridine and aniline in place of 2-aminopyrimidine, and benzyl alcohol in place of anise alcohol.
Solubility of vitamin A palmitate in the aqueous solution of various surface active agents of polyhydroxyethylene series was examined. It was found that a different rate of solubility occurred when the sample was added to the aqueous solution of surfactants and the mixture shaken and when the sample and surfactant was mixed and diluted with water. According to the second method of testing, the solubility of vitamin A palmitate increases, the larger the lypophylic radical and smaller the hydrophilic radical of the surfactants.
Datura Tatula, D. Metel, and D. meteloides were grafted on each other obtain Tat./Met., Met./Tat., Tat./met., and met./Tat. These plants were cultivated with a definite amount of fertilizer and were harvested when fully grown to examine the degree of growth and the yield. Variation in content of the componental alkaloids, i.e. hyoscyamine, scopolamine, and meteloidine, was also examined. It was thereby found that the rate of the growth was comparatively near the yield of D. Tatula while the varition in the ratio of alkaloidal content in leaves was approximately those in D. Metel and D. meteloides. It was concluded that not only the root plant but also the specie of the grafted plant had largely affected the alkaloidal content of the plants. There was little variation of alkaloidal content in the roots, showing about the same ratio as the roots in the control area and not so marked an influence of the grafted plant. In other words, it may be concluded that the variation of the alkaloidal content in Datura spp. was affected not only by the root but also by the function of the leaves.
It has been shown that the composition of the basic aluminum chloride solution prepared by different methods could conveniently be indicated by Al2+n(OH)3nCl6. It was found that even when this n is the same, the neutralization velocity by acid was different according to the manner of preparation. Such a fact indicates that there is a difference in the degree of formation of ol-bridge by the complex ions present in equilibrium. Therefore, the properties of the substance formed from the solution of the same basicity by SO4-- would probably be different. Basic aluminum solutions were titrated with acid and alkali and their pH curves were found to show some specificity. Especially, titration with alkali was found to give the second breaking point at pH 5-6, besides the first breaking point at pH on the formation of aluminum hydroxide. This second point appears the sooner, the larger the value of n, and it was assumed that this second point was caused by the formation of [Al3(OH2)x(OH)8]+.
Alkyl ethers of m- and p-aminophenols were prepared and antibacterial action of their hydrochlorides against tubercle bacilli was tested in Kirchner medium. In the alkylation of acetylaminophenol, the best result was obtained by the use of alkyl p-toluenesulfonate as the alkylating agent. The reactivity of the three isomers of acetylaminophenol was in the order of para, meta, and ortho. The compounds synthesized were, ethyl, butyl, hexyl, octyl, decyl, dodecyl, tetradecyl, and hexadecyl ethers of m-aminophenols, and butyl, octyl, dodecyl, and tetradecyl ethers of p-aminophenols. The antibacterial action of the hydrochlorides of these amines against tubercle bacilli was generally low but there was found a correlationship between the size of the alkyl group and the antibacterial action, such action increasing with the number or carbon atoms in the alkyl, reaching the maximum at around C12, and gradually decreasing over that number.
Alkyl and dialkyl ethers of resorcinol and hydroquinone were prepared in which the alkyl was methyl, ethyl, butyl, hexyl, octyl, decyl, dodecyl, tetradecyl, or hexadecyl. Antibacterial action of monoalkyl ethers against tubercle bacilli was tested in the Kirchner medium. All the ethers showed a small action but the action increased with the increase in the number of carbon in the alkyl, reaching the maximum at around C12, and falling thereafter. Summarizing the results obtained to date, the influence of the alkyl group appeared in the antibacterial action, irrespective of the difference in NH2 and OH, or the position of such functional groups and it was concluded that such influence was due solely to the alkyl group. In order to find the difference in antibacterial action according to the difference of substituent position in the three isomers of dihydric phenol monoethers and aminophenol ethers, the action of C8 and C12 ethers was compared by concurrent incubation. It was found that the activity of dihydric phenols was in the order or ortho ≥ meta > para, and that of aminophenol ethers was ortho > meta > para; in the case of difference of hydroxy and amino groups, the order in ortho and meta was NH2 > OH, while that in para compound was NH2 ≥ OH.
Using the Kofler-Adams method of measuring hemolytic index, which can judge the potency of saponin-bearing crude drugs to some extent, the saponin content of Campanulaceae plants was examined, especially the hemolytic action of the plants (chiefly the roots) in various species of this family. As a result of the comparison of the difference in the saponin content of Platycodon, with and without cork, by the hemolytic index, it was found that only Platycodon grandiflorum showed hemolytic action, and there was no significant difference in the saponin content in the two kinds of Platycodon, with and without cork.
The so-called Fleitmann test of reducing arsenic to hydrogen arsenide by aluminum in caustic alkaline medium is advantageous in that it avoids formation of hydrogen sulfide, hydrogen antimonide, or hydrogen phosphide that is inevitable by reduction in acid medium. The use of zinc as the reducing agent in alkaline medium, report on which has not been found in the past 20 years although such use in acid medium has been described often, was examined. The detection of hydrogen arsenide was made by paper chromatography by spraying 5% silver nitrate solution and the recognizable limit of arsenic was found to be 0.1 γ in 0.2cc., and its limiting concentration to be 1:2, 000, 000. These values indicated good sensitivity compared with other methods of detecting arsenic.
By the hydrogenolysis of hexamethylenetetramine over a Raney nickel catalyst, in the presence of formamide, at 130-140° and at a high pressure, formylmethylamine was obtained in 85% yield, by the following equation: (CH2)6N4+6HCONH2+6H2=6CH3NHCHO+4NH3 This process is valuable as an industrial method of preparing methylamine.
Antibacterial action of 14 kinds of 3-halo-4-hydroxybenzoic acids and their esters, 30 kinds of hydrazides, 8 kinds of phenol derivatives, 4 kinds of quinones, and 3 kinds of other compounds against Staphylococcus aureus 209P, Escherichia coli communior, and Bacillus subtilis was tested. It was thereby found that the growth of Staph. aureus 209P was completely arrested by hexyl 3-chloro-4-hydroxybenzoate in 1:160, 000 dilution and by isobutyl 3-iodo-, amyl 3-bromo-, and hexyl 3-bromo-4-hydroxybenzoate and by 4-bromo- and 4-iodo-thymol in 1:80, 000; and the growth of Bacillus subtilis was completely arrested by 1:80, 000 dilution of amyl 3-bromo-4-hydroxybenzoate, 4-bromothymol, and 4-bromothymol, and 4-iodothymol. Furacin used as the control also inhibited the the growth of these three kinds of bacteria in 1:80, 000 dilution.
Of the 62 kinds of compounds tested, 2, 4-dihydroxy-5-amylbenzaldehyde 4-p-tolyl-thiosemicarbazone and 2-cyclohexylphenol completely inhibited the growth of Staphylococcus aureus 209P in 1:80, 000 dilution, while 2-chloro-4-cyclohexylphenol and 2, 4-dihydroxy-5-chlorodiphenylethane arrested the growth of Bacillus subtilis completely in 1:80, 000 dilution. None of the compounds tested were effective in preventing the growth of Escherichia coli commuuior. Furacin used as the control was able to completely inhibit the growth of the three bacteria in 1:80, 000 dilution.
The method of reduction by the utilization of hydrogen generated at the time of developing nickel in aqueous alkaline solution was applied to the reduction of nitriles with successful results. The reduction vessel, in this case, a flask containing the alkali solution, was provided with a separatory funnel and a distillation tube so that amine formed immediately on the addition of Raney nickel can distill out with steam. The starting material is placed with the alkali solution in the flask and water in the separa-tory funnel is dropped gradually into the flask to supplement the water lost by steam distillation. In this manner, benzylamine, phenethylamine, homoveratrylamine, o-me-thoxybenzylamine, p-methoxybenzylamine, and p-ethoxybenzylamine were obtained from the corresponding nitriles.