巻号一覧
44 巻, 11 号
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III.水解酵素含有複合体より尿素による抗菌因子の分離,ことに正常動物との比較金井 興美, 近藤 瑩子1969 年 44 巻 11 号 p. 351-355
発行日: 1969年
公開日: 2011/05/24
ジャーナル フリーThe lysosorne-rich fraction was separated from normal and BCG-revaccinated guinea pig lungs and was extracted with O.1 % Triton X-100. The extracts were fractionated by g elfiltration on Sepharose 2B column using 0.05M acetate buffer of pH 5.6 (Fig.1). The first protein peak associated with acid phosphatase and cathepsin activities from BCG-anim als (Fraction A) was found to be mycobactericidal, but the corresponding fraction f rom normal animals was not. These fractions were then treated with 2M urea and the extracts were fractionated by gel-filtration on Sephadex G-150 column (Fig.2). Fraction A from BCG anim als was divided into 2 protein peaks a and b by urea-treatment, the latter being separated from acid phosphatase activity. This small-molecular protein fraction was not obtained from no rmal animals.
Table 1 demonstrates the summarized data of the mycobactericidal effects of those subfractions on H87Rv tubercle bacilli suspended in 0.05M acetate buffer of pH 5.6. The bacilli of 0.1mg per ml were exposed to each sample of indicated protein concentrations. The urea-released Fraction b was found to be active in killing tubercle bacilli, but it required more amount of protein and longer exposure time to express its mycobactericidal activity than Fraction A and Fraction a. In the environment of pH 7.4, the activity of each fraction was reduced. Fraction b was, however, more active than Fraction a at this pH (Table 2). A curious observation was that, though Fraction A of normal animals was not mycobactericidal, its 90% acetonesoluble moiety was highly active in this respect just as the corresponding sample from BCGanimals of our previous report. It might be possible to suggest that the 90% acetone-soluble substance is a “created” mycobactericidal fraction.
When Fraction A of the normal animals was added to the corresponding fraction of the BCG-animals, the activity of the latter was reduced, which indicates a possible competition between the two fractions with respect to the contact with the bacilli. Fraction b appeared to be a small-molecular protein and it was heat-stable at 100°C and pH 5.6. However, lysozyme of the similar properties was not mycobactericidal in the same experimental conditions and in a much higher concentration (Table 4).抄録全体を表示PDF形式でダウンロード (832K) -
豊原 希一, 重松 昭世1969 年 44 巻 11 号 p. 357-360
発行日: 1969年
公開日: 2011/05/24
ジャーナル フリーChanges in the distribution of 14C-labelled INH in mouse body with the lapse of time was studied by combining the macroautoradiography (ARG) of whole body section of frozen mice and measuring radioactivity with the film scintillation counter. Mice were injected subcutaneously 20μCi of 14C-INH (carbonyl 14C, specific radioactivity 9.6mCi/mM), and at 1, 6 and 24 hours after injection each one mouse was killed and rapidly frozen in aceton-dry ice (-78°C). Then, the whole body of each mouse was sectioned in the thickness of about 20μm by Leitz 1300microtome in cryostat at the temperature of -20°C.
ARG of the whole body section was ca r ried out by the contact method using Sakura industrial X-ray film. After four weeks exposure X-ray film was developped and fixed. Distribution of 14C in the various organs was compared by the density of film.
As seen in photo.1, at one hour 14C-INH was distributed in the whole body including the subcutaneous tissue, muscles and markedly in the intestines, stomach, liver and brain.
After one hour 14C-INH was discharged so rapidly that at 6 hours 14C was ha r dly proved in the subcutaneous tissue and muscles but remained in the lung, liver, stomach, brain and kidney, and it was also transmitted to urine bladder. It is thought that 14C disappears from the circulating blood at six hours, as it cannot be found already in the heart.
At twenty-four hours significant photodensity was seen only in the liver, and its density was similar to that at six hours. The radioactive substance in the liver at 24 hours is not probably INH itself, but some labelled metabolites of INH retained in the liver.
Radioactivities of the whole body section and of each organ per 1 cm14 by film scintillation counter are shown in Fig.1, and the disintegrated rate calibrated from the count rate coincides well with that of ARG.
By combining the both methods, the more reliable informations on the distribution of labelled chemicals are obtained.抄録全体を表示PDF形式でダウンロード (1985K) -
1969 年 44 巻 11 号 p. 361-393
発行日: 1969年
公開日: 2011/05/24
ジャーナル フリーPDF形式でダウンロード (5990K)
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