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The susceptibility of Rous sarcoma virus (RSV) genomes integrated in mouse ascites sarcoma cells (SR-C3H/He cells) to DNase I and DNase II was investigated. Approximately half of the viral sequences were sensitive to DNase I and DNase II when 17% and 7.4% of the chromatin DNA was rendered acid soluble, respectively. The results suggest that newly acquired exogenous proviral sequences are integrated into both transcriptionally active and inactive regions of chromatin in cells lacking related endogenous viral sequences.

The localization of both the large T and small t tumor (T) antigens in cultured cells (Vn 12 cells) of hamster brain tumors induced with BK virus (BKV), a new human papovavirus, was studied by an enzyme labelled antibody method at both the light and electron microscopic levels. Under the light microscope, BKV T antigen was observed in the nucleus, except for the nucleoli, of cells in interphase, and under the electron microscope it was observed in the nucleus except for the nucleoli and nuclear membrane. BKV T antigen appears to be closely associated with nuclear chromatin as previously reported for simian virus 40 tumor antigen (SV40 T antigen). The intracellular localization of BKV T antigen was the same as that of SV40 T antigen. In metaphase, BKV T antigen seems to be distributed diffusely throughout the cytoplasm except for the chromosomes. In telophase, BKV T antigen transfers from the cytoplasm to the nucleus. The migration of BKV T antigen during the cell cycle is thought to be related to the function of T antigen.

Cancer cells obtained from human hepatocellular carcinoma nodules were subjected to primary culture, and a hepatoma cell line was established. The cell clumps obtained by needle puncture were plated directly in plastic tissue culture flasks without any cell dissociation procedures. Cell clusters became attached to flasks in 24 h with an efficiency of about 90%. No fibroblast outgrowth was observed. Primary cultured cells were composed of polygonally shaped epithelial cells with dense cytoplasm and one or more large nuclei. They excreted plasma protein biosynthetic markers of hepatocytes into the culture medium. Plasma protein synthesis of primary cultured hepatoma cells decreased as the age of the primary cultures increased. Cells seeded in September 1980 started to grow continuously after 5 months of cultivation. A new hepatocellular carcinoma cell line (designated as KG55T) was established from these growing cells. KG55T cells have been subcultured for more than 20 passages and form a monolayer of polygonal epithelial cells which pile up after they reach confluence. The cells had a doubling time of 50-60 h and a plating efficiency of 60-65%. Albumin, alpha 1-antitrypsin and alpha 2-macroglobulin syntheses and tyrosine aminotransferase activity were detected. At the 10th passage, KG55T cells were pseudotriploid (mode, 69), and 8q+ and 15q+ translocations were distinctive of this cell line. The morphological characteristics and the capacity for plasma protein synthesis of the primary cultured hepatoma cells and cells of the established hepatoma cell line were compared.

A selective and simultaneous staining method for nuclear apparatus and cytoplasmic membrane of some bacteria has been presented. Nuclear apparatus is stained with basic fuchsin after hydrolysis with I N HCl and cytoplasmic membrane is restained with Victoria blue 4R after treating with saturated mercuric chloride. By this method, the nuclear apparatuses of B. subtilis, Sal. typhi 57 and Staph. aureus were stained red, and the cytoplasmic membrane and septum bluish purple distinctly. Thus this staining method would be of a great advantage in displaying the cellular structures of the bacteria.

Die Verfasser erwahnen die Wichtigkeit der Myotonometrie und Myographie bei der Diagnostik und Auswahl der Therapie bei rheumatischen Erkrankungen. Auf Grund ihrer 15-jahrigen Erfahrung ist einer der richtigsten Wege der Rheuma-Prophylaxie die Behandlung durch die Verbesserung bzw. der besseren Oxygenversorgung der peripheren Zirkulation (Generelle Theorie der Oxygenversorgung der peripheren Zirkulation=G. T. O. P. C. ), auf, Grund der Oxygen-Insuffizienz-Theorie (O.I. T.) nach Szirmai. Nach diesem ist jeder Schmerz durch Hypoxaemie verursacht und das sogenannte "Rheuma" ist eine Erkrankung des ganzen Organismus und ist nur durch eine allgemeine Behandlung gtinstig beeinflussbar.

Protein synthesis of the liver in both normal and CCl4 intoxicatedguinea pigs has been examined in vitro by incubating liver slices with C14-glycine. It has been demonstrated that normal liver slices synthesize albumin in vitro, which in turn incorporates with Cl4-glycine and is finally liberated into the medium very rapidly. On the other hand, immunized lymph nodes, kidney, and spleen do not show any C14-glycine incorporation into albumin. The liver slices of CCl4. intoxicated animal revealed a marked decrease in C14-glycine incorporation into albumin. Observation on the subcellular fractions proved that the incorporation of C14-glycine into microsome fraction is severely arrested, and oxygen consumption of liver slices is only slightly reduced. With the observation on the liver slices incubated with DNP, the author attributes the effect of CCl4 on protein synthesis to the decreased ATP formation by the action of CCl4 as an uncoupler for oxidative phosphorylation.

It has been revealed that ribonuclease (RNase) can penetrate into living cells and inhibits amino acid incorporation into proteins resulting in the suppression of protein synthesis and growth of living cells. BHIDE, BRACHET¹, KAUFMAN and DAs have proven that the RNase penetrates into onion root-tip cells and induces a number of mitotic abnormalities. KIMOTO and others² also have revealed that RNase injection into mice results in the reduction of cytoplasmic basophilia with the morphologic change of endoplasmic reticulum and the disturbances in DNA synthesis as demonstrated histochemically on pancreatic exocrine cells and liver cells. But there is little information so far on the mechanisms of penetration of RNase into living cells. PILLERI³ and SCHUMAKER4 in Brachet's laboratory have demonstrated the uptake of RNase by pinocytosis in amoebae and cancer cells. This may suggest the penetration of RNase through the membrane of the endoplasmic reticulum in the cells whose RNase contents are low5, however it is reasonablly supposed that some phosphatase may be concerned with the permeability of RNase.

Activities of intracellular RNase of the liver cytoplasm, normal liver cells exposed to 3'-Me-DAB and heaptoma cells, have been studied in correlation with the contents of RNA and DNA and morphologic changes of the cells with or without treating RNase. The data showed that in hepatoma cells the intracellular acid RNase activity decreases with the decrease of RNA and unchanged DNA contents and alkaline RNase activity. Morphologic observation proved that hepatoma cells show a small low massed vesicular or vacuolar endoplasmic reticulum having ribosomes. For the exposure to RNase the hepatoma cells proved to be much less resistant comparing to normal liver cells. The former lost the granules and was destroyed in its endoplasmic reticulum, whereas the latter retained ribosomes and ER. From these experiments it has been speculated that acid RNase in the cell may be involved in RNA synthesis and alkaline RNase in RNA decompostition, though the effect of the difference in concentration in the case given RNase experimentally can not be neglected.

Der Autor berichtet uber verschiedene Beobachtungen, nach welche ein akutes Nierenversagen in der Aetiologie der aplastischen Anamie auch eine Rolle spielen kann.

The labeling of C14-glycine to pH-5 enzyme, PP32-ATP exchange by pH-5 enzyme, and the inhibitory action of p-chloromercuribenwate were examined. 1) The labeling of C14-glycine to pH-5 enzyme is inhibited by 10-4 mole of PCMB, and this inhibitory effect is reduced by addition of cysteine having 20 mole equivalent of PCMB. 2) The PP32-ATP exchange reaction is also reduced by addition of PCMB and inhibitory effect of PCMB is reduced by the addition of cysteine having 20 mole equivalent of PCMB. 3) These results show that the inhibition of the organic mercury compounds on the protein synthesis is responsible for attack the SH enzyme which catalyzes some amino-acid activation, and consequently influences activated amino-acid transfer reaction. 4) Paper electrophoretic pattern of pH-5 enzyme shows numerous peaks, each having the mobility between α2-globulin and γ-globulin.

Der Autor berichtet über seine Erfahrungen durch Beeinflussung der peripheren Zirkulation und NervenzusHinde mit Heacrin (HAH) bei der Behandlung der Facialis-Lähmungen verschiedener Aetiologie und über die diagnostische Möglichkeit, die mit dem von ihm entwickelten Myotonometer, Angio-Myograph, sowie mit Elektro-Hautthermometer. Mit seiner Oxygen-Insuffiziens-Theorie '(0. I. T.) und Behandlungsmethode der generellen Therapie durch Oxygenversorgung der peripheren Zirkulation (G.T.O.P.C.) hat der Autor die Erfolge erkHirt.

A case of postpartum necrosis of the anterior lobe of the pituitary, known as Sheehan's syndrome, is presented. The patient has done well on a combined replacement therapy with cortisone and thyroid. It is hoped that increased awareness will be given to this clinical entity.

The liver glucuronyl transferase (GT) activity and uridine diphosphate glucuronic acid (UDPGA) content in the patients with viral hepatitis were determined using 4-methyl umbelliferone (4-MU) as a glucuronide receptor. The results were as follows: 1. In acute viral hepatitis, the decrease in the GT activity was more remarkable in the later stage of the recovery. In chronic viral hepatitis, the GT activity was decreased in accordance with the increase in the degree of liver injury. Liver UDPGA content was significantly reduced only in postnecrotic cirrhosis. 2. The decrease or injury in the parenchymal liver cells caused a decrease in the liver GT activity. These quantitative reductions in the liver parenchyme were not the only factor for the alteration in the GT activity of the liver. The results of the present study suggested an involvement of a qualitative change in the liver GT activity in human liver injuries, especially in the early stage of acute viral hepatitis; namely, there might be even an activation of the liver GT other than the reduction resulting from the decrease in the liver parenchyme. 3. The decrease in the liver GT activity correlated significantly with the decrease in the salicylamide glucuronide formation in vivo, while the alteration in the liver UDPGA content failed to correlate with that in the glucuronide formation in vivo. It was suggested that the velocity of in vivo UDPGA production rather than the UDPGA content of the liver was as important a rate-limiting factor for the glucuronide formation in vivo as the liver GT activity.

1. An attempt has been made to find the diagnostic criteria for early gastric cancer. It is most important to detect the evidences or suspected features of the malignant growth in incipient stage in order to attain the radical cure by surgical operation. 2. Twelve patients with early gastric cancer (groups A and B) were selected out of 476 patients who had undergone gastrectomy during the past three years in the Okayama Saiseikai General Hospital. The other 6 patients in the "precancerous group" (group C) were also studied, who had abnormal epithelial proliferation in the resected stomach membrane during the same period. 3. The processes of discovery of early cancer have been described. Fairly precise diagnosis can be made in the mucosal carcinoma, but it is not in the ulcer-carcinoma. It was generally difficult to estimate the degree of the malignancy and the extension of the growth preoperatively. 4. The details of the diagnostic aids are as follows. i. Negative occult blood of stool does not always mean the definite diagnostic aid. ii. The malignant gastric change may occur even in non-anacidity. Further investigations should be followed up on gastric ulcer patients if malignant alteration is under the consideration. iii. Minor roentgenological findings, such as the absence or irregularity of mucosal folds, rigid and/or overlapped contour, localized absence or decrease of the peristaltic waves and absence or bow-shaped deformity of the angulus, are of important significance. Such changes should be minutely sought for by X-ray film examination. iv. On gastroscopy and gastrocamera photography, such changes as erosion or irregular granular thickening of the membrane with abnormal reddening and edematous appearance, irregularity of ulcer edge, uneven swelling on ulcer margin with reddening and unsharpness of the edge of adherent coat on ulcer floor, must be noted in the early gastric cancer. v. It is not safe to leave a patient having stomach ulceration under a mere conservative management because it is often quite difficult to dissolve the question of malignancy of the lesion with all sorts of examinations. vi. So far as clinical examinations have indicated malignancy, histological examination must be carried out immediately at the time of operation, even when malignant lesion is absent in inspection and palpation on the exposure of the stomach. vii. On the gross observation of the resected stomach, a particular attention must be paid to erosion, depression or atrophy, irregular granular thickening and abnormal reddening on the restricted areas of the mucosal surface.

An attempt was made to establish the method for the estimation of glucuronide formation in vivo using salicylamide and further to study the alteration in the glucuronide formation in liver disease. The results were as follows: 1. The method for the determination of free salicylamide separately from other conjugates of salicylamide in urine, without involving any hydrolysis of the other conjugates, was presented. When 1 g. of salicylamide was administered to the subjects with or without liver injuries, no free salicylamide was detected by the present method in the urine following the salicylamide administration. 2. The analytical method for the determination of salicylamide glucuronide was also devised by employing a hydrolysis with β-glucuronidase. The ratio of the salicylamide liberated by the enzymatic hydrolysis of the 10-hour urine following the administration of 1 g. of salicylamide to the total salicylamide excreted in the same urine was neither affected by the total recovery of the salicylamide nor by the urine volume. This ratio was thus used as a means of estimating the capacity of the glucuronide formation in vivo, although it was considered that the ratio might be affected to some extent by the competition between the glucuronide and other conjugate formations in vivo. 3. As a result of this salicylamide glucuronide excretion test, it was indicated that the in vivo formation of salicylamide glucuronide in the patients with postnecrotic cirrhosis was slightly decreased compared with that in normal controls.

A purified compound of the activator of glucuronide formation was isolated from a boiled extract of rat liver by charcoal adsorption, ethanol fractionation of barium salts, and finally paper and Dowex-l column chromatographies. The analytical data and the chemical properties of the compound sugggested that the endogenous activator of glucuronide formation in rat liver might be uridine diphosphate N-acetylglucosamine.

In order to see whether isovalthinuria can be induced in animals other than the cat, that was found to excrete isovalthine in normal urine as previously reported³, using rat, guinea pig, rabbit and dog as test animals, isovaleric acid was adminstered either orally or parenterally and their urine was analyzed for the presence of isovalthine. As the result it was found that the rat, guinea pig, rabbit and dog administered with isovaleric acid orally or parenterally all excreted isovalthine in their urine, which normally does not contain it.

Biological effect of the unsaturated fatty acid fraction from the X-ray irradiated rabbit liver (OX) on HeLa cells has been observed in vitro comparing with the effect displayed on the same strain cells by the unsaturated fatty acid fraction from the non-irradiated rabbits, which is extracted by the same method as in OX, The observations have proven that OX is a powerful cytotoxin in a concentration of 0.05-0.025 per cent and induces a severe cell degeneration and cell death, resulting in a marked arrest in the growth of the cells. The similar effect has been observed by unsaturated fatty acid fraction from the nonirradiated rabbits, but the effect was much less comparing to that of OX. Possible mechanism of the cell damage by OX has been discussed.

1. The objective of this investigation was to test the influence of mammary cancer tissue extract on the induction of various tumors by urethan. Three strains of female mice, Balb/c, Zb and Db, were used in this experiment. 2. It was found that urethan was a multipotential carcinogen in the induction of (a) lung tumor, ovarian hemorrhagic cyst, and hemorrhagic lesions in various tissue in Balb/c mice, (b) lung tumor, hepatoma, leukemia, mammary cancer, Harderian gland tumor and hemorrhagic lesions in various tissues such as spleen, mesenteric lymphnode, liver and ovary in Zb strain, and (c) lung tumor, mediastinal lymphoma, leukemia, hepatoma and hemorrhagic lesions in the liver and ovary in Db mice. 3. Mammary cancer tissue extract seemed to have a promotive effect on the generalization of the mediastinal lymphoma induced by urethan in Db female mice.

To verify whether the administered isovaleric acid does directly incorporate into urinary isovalthine or not, a young male dog was given labeled isovaleric acid-1-C14 (0.2 mc.) and the urinary isovalthine was analyzed by means of paper electrophoresis and column chromatography. Observations proved that isovalthine appeared in urine but showed no radioactivity while glutamic acid gave a strong activity. The possible metabolic pathway of isovaleric acid has been discussed.