To study the uptake and discharge of Cadmium by chlorella, Chlorella ellipsoidea (C-27) was cultured in Myers-4N medium by the autotrophic culture method, and following facts were observed. 1) When 1μg/ml (as cadmium) of CdCl₂ was added to the medium, in which chlorella has been grown for 3 days, cadmium was taken up rapidly, but it was taken up slowly when CdCl₂ was added to the medium after chlorella was grown for 5 days. 2) When 1μg/ml (as cadmium) of CdCl₂ was added to the medium, in which chlorella was grown for 3 days, cadmium content in the chlorella cells reached a maximum of 1.25×10^-8 μg after 12 hr. 3) When chlorella with cells containing cadmium was cultured for 20 days in a cadmium-free medium, slight excretion of cadmium from the cells was observed.

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On the way of the study on the metabolism of α-bromoisovalerylurea (BV), a series of new mass fragment ions (m/e 103, 115, 130, 144, 175, and 190) was observed in the mass spectrum of crude extracts from BV-administered rabbit urine. After purification of the extracts by silica gel column chromatography and subsequent thin-layer chromatography (TLC), small pieces of crystals were obtained and characterized to be 2-(methylthio)-3-methylbutyrylurea (BM) by comparison of melting point, Rf value in TLC, mass spectrum, and infrared spectrum with those of the standard BM, synthesized from BV and methylmercaptan. In order to know the amount of BM excreted by the isotope dilution method using a mass spectrometer, 2-(methylthio-d₃)-3-methylbutyrylurea (BMd₃) was synthesized from BV, thiourea, and methyld₃ iodide. After the administration of 1 g of BV to a male rabbit (ca. 3.5kg), approximately 4 mg of BM was found to be excreted in the urine within 30 hr.

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An analytical method for aromatic nitro compounds, such as nitrobenzene, o-, m-, and p-nitrotoluene, o-, m-, and p-nitroanisole, o-, and m-dinitrobenzene, and 2, 4, 2, 6-, and 3, 4-dinitrotoluene, in river and sea water is described. One liter of sample water was extracted with benzene in the presence of hydrochrolic acid. The extract was dehydrated and concentrated to 10 ml by a Kuderna-Danish (K. D.) evaporative concentrator. The extract was cleaned up by a Florisil column chromatography, and the components were eluted with benzene-ether (98 : 2, v/v). The eluate was concentrated to 10 ml and then quantified by a gas chromatograph (GC) with an electron capture detector (³H). A 5 ml portion of the solution was concentrated to 0.1 ml by the micro-K. D. evaporative concentrator and then qualified by a gas chromatograph-mass spectrometer (GC-MS) with a multiple ion detector. The recovery by this procedure was 86-99%, and as little as 0.025 to 0.25 ppb of the aromatic nitro compounds was able to be determined. The samples were obtained from the mouths of six rivers in Kanagawa Prefecture and the Tokyo Bay. Nitrobenzene was found in all of the samples and their content was 0.16-0.99 ppb. o-Nitroanisole (0.68 ppb) was detected in the water of the Tsurumi River in Yokohama City.

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Chemical composition of the bone and the activity of the enzyme responsible for bone resorption in rats were examined after oral administration of cadmium chloride for various lengths of time. The amount of sialic acid and hexose in the bone of rats treated with cadmium increased over that of control rats, but the amount of hexosamine and uronic acid did not change. The amounts of calcium and phosphorus in the bone decreased markedly after exposure of the animals to cadmium chloride for 6 months. In the rats which treated with cadmium for 6 months, the activities of acid phosphatase, β-glucuronidase, β-N-acetylglucosaminidase, and β-galactosidase in the bone all increased over those of the control rats. These results suggested that resorption of diaphysial bone of rats were accelerated by the continuous oral ingestion of cadmium.

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The Schiff bases of p-dimethylaminobenzaldehyde and aromatic primary amines exhibit a characteristic absorption maximum and emit fluorescence in ethanolic solution containing hydrochloric acid. Procaine in dental 2% procaine hydrochloride injection was determined by both colorimetric and fluorescence methods.

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Complex formation of L-tryptophan (Trp) or adenosine (Ado) with mercuric chloride (MC) or methylmercuric chloride (MMC) was examined in an artificial intracellular fluid (pH 8.0) and artificial extracellular fluid (pH 8.0). 1) A strong complex formation of L-tryptophan or adenosine with mercuric chloride was demonstrated in the artificial intracellular fluid containing oxianions (HPO^2-₄, HCO^-₃), while the complex formation in the artificial extracellular fluid was considerably weak, because of the high concentiation (110 mM) of chloride ion in this fluid. 2) Dissociation constants (K) of complexes of [MC·Trp], [MC·trptamine], [MC·indole], and [MC·Ado] were found to be 1.83×10^-3M, 7.60×10^-4M, 1.22×10^-5M, and 2.30×10^-2M, respectively, in phosphate buffer (32 mM, pH 8.0). 3) The complex formation of L-tryptophan or adenosine with methylmercuric chloride in the artificial intra or extra cellular fluid was much weaker than that with mercuric chloride.

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To investigate the direct effect of cadmium on the bone metabolism, tibia from 9-day chick embryo was cultivated for six days by Roller-tube method and CdCl₂ was added into the culture medium in 5 to 30 ppm concentration range. Cartilage ends and bone shafts were analyzed for bone matrix formation and calcification by measuring the chondroitin sulfate, hydroxyproline, inorganic phosphate and cadmium content and alkaline phosphatase activity. Both bone matrix content and alkaline phosphatase activity in two parts of the bone decreased with increasing the concentration of CdCl₂, although the rate of inhibition of cadmium was stronger to alkaline phosphatase than to the increase of matrix content. Cartilage ends and bone shafts were found to differ in mineral content of cadmium-treated bone unlike matrix content and alkaline phosphatase activity, as inorganic phosphate content of bone shafts in the presence of CdCl₂ were similar to control though those of cartilage ends were less than control. Cadmium content of cartilage ends per dry weight linearly increased with the increase of cadmium concentration while that of bone shafts exponentially increased. Histopathological examination of the tibia cultured with medium containing CdCl₂ showed that in bone shafts the formation of bone matrix was inferior to control and the ossification remained nearly in the state before culture. These results suggest that cadmium inhibits the bone matrix formation and brings about a damage to the ossification although cadmium unaffects the bone mineral deposition.

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The effect of cadmium on chondroitin sulfate synthesizing and releasing activities in embryonic chicken limb bone was studied. At the concentration more than 20 ppm CdCl₂, cadmium enhanced the release of chondroitin sulfate into the medium. However, at the concentration of 5 to 10 ppm CdCl₂, cadmium inhibited the incorporation of ³⁵S into chondroitin sulfate. These findings suggest that the primary action of cadmium to the chondroitin sulfate metabolism is the inhibition of the synthesis. p-Nitrophenyl-β-D-xyloside did not reverse the inhibition of chondroitin sulfate synthesis brought about by cadmium. Furthermore, cadmium inhibited the incorporation of [³H]-acetate into UDP-N-acetyl hexosamine. From these results, we suppose that cadmium inhibits mainly the synthesis of nucleotides, even though the inhibition of cadmium to core protein synthesis and chain elongation may be extensively excluded.

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^<14>C-Isocyanuric acid given orally to rat distributed rapidly, showing higher radioactive concentration in blood, liver and kidney, and the peaks of concentration in all of organs and tissues were observed at 30 min. after dosing. Thereafter, the radioactivities decreased rapidly. After repeated oral administration, a similar result was obtained, so any accumulating effect was not noted. The percutaneous absorption of the labeled compound was so scarce that no radioactivity was found in many blood samples taken intermittently after the application on rat skin. The clearance of the compound after oral administration was also very fast and the main excretion route was urine. The excretion to expiration, bile and feces was very little. More than 99% of the urinary radioactive substance was identified to be intact compound.

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As the pre-treatment for the determination of organic compounds in biological materials such as hair and hard tissues, freeze grinding method was found to be very useful and to have following advantages : (a) it can remarkably shorten the time necessary for grinding and extraction, (b) it is suitable for analysis of compounds unstable to heat, and (c) the size of powdered materials is small and relatively homogeneous. The particles are in the range of 5-60 μm. Methylmercury in the hair of monkeys given methylmercury chloride was determined by using this method. There were hardly any difference between the values measured just after grinding and those measured after soaking in 2N HCl solution for 24 hr. When the hair was cut with scissors, the values measured just after cutting were 1/2 to 1/4 of those measured after 24 hr soaking.

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Heavy metals in the bottom mud at the mouth of 91 rivers in Japan were analyzed and the data were published in a paper. In order to compare with these data, the distribution of heavy metals in the bottom mud up the river was studied in the present work. Samples of the mud were collected from the upper stream near the mountain, not polluted with waste water. It was concluded from these data that the concentration of the heavy metals among 193 mud samples up the river dispersed in a narrow limited range, compared with that of the river-mouth, i. e., the range of Ni and Cr concentration was 1-25ppm, that of Cu and Pb, 4-50ppm, Zn 10-200ppm, Cd 0.08-2.00 ppm, and Hg 0.0-1.0 ppm. The value of the mean of heavy metal concentration up the river decreased to 1/2 or 1/3 of those at the mouth of the river.

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