While histochemical and morphological investigations of ceroid as well as biochemical analyses to its constituents have been published, there is as yet no report of histochemical study which aimed at analysis of the constituents of this acid-fast waxy pigment. The present study represents an attempt to determine the constituents of ceroid in the liver, uterus and aorta and of lipofuscin by means of the oxidation-Schiff techniques after pretreatment of tissue sections by oxidation with chloramine T and other, oxidizing reagents followed by treatment with methylation or 2,4-dinitrophenylhydrazine (DNPH) blocking. The findings obtained suggest that ceroid possesses a relatively well developed pigmental stroma rich in glycoprotein whose reactions are fairly exactly reflected as the reactions of ceroid, this property distinguishing ceroid from lipofuscin. Since the constituents of the stroma of ceroid are probably loosely bound to each other, such treatment as methylation preceded by oxidation of active groups of the pigmental constituent might give conversely rise to liberation of blocked active groups of other constituents of ceroid, revealing the result that the degree of the positive reactions of ceroid after such treatment was retained or at times even augmented. Ceroid, moreover, proved to give a more clearly positive reaction (owing partially to the pigmental stroma) than lipofuscin with the periodic acid-Schiff test after oxidation with chloramine T followed by methylation, whereas ceroid in the aorta (“hyaloceroid”) was devoid of such reactivity. It is noticeable that the positive reactions of hyaloceroid which a poorly developed stroma was markedly suppressed, insomuch as the histochemical invasion of the costituent probably brought about dissociation and even disintegration of the pigmental firmly-bound composition. In addition,“hyaloceroid”has not been deprived of its property of sudanophilia on a paraffin section by oxidation with acetic acid followed by treatment with methylation, thereby indicating that it differs not solely true ceroid (ceroid in the liver) but from the “pigment of vitamin E deficiency”and lipofuscin. This finding suggests that“hyaloceroid”belongs to“ceroid derived from fat”chiefly composed of polymerized unsaturated lipids. Aortic tissue specimens from rabbits with experimentally induced arteriosclerosis by prolonged feeding of a diet excessive in lanolin were examined as control materials containing no ceroid. On the other hand, the histochemical studies failed to demonstrate any difference in constituents between“pigment of vitamin E deficiency”and true ceroid.
Incidence of human breast cancer is high in developed countries and low in undeveloped countries with the notable exception of Japan. Among the factors responsible for the incidence of breast cancer, genetic, racial, hormonal and nutritional factors were thought to be important modifiers. Because of the paucity of information dealing with the relationship between mammary carcinogenesis and dietary protein, a further study of this problem seems justified. In this paper, to clarify the relationship between dietary protein and mammary carcinogenesis, the following experiments were undertaken. Part I of this study is concerned with the effect of arginine imbalanced diets and various proportions of casein diets on genesis and growth of mammary carcinoma in rats induced by 7,12-DMBA. The experimental animals were female Sprague-Dawley rats. The diets used in this experiment were as follows: Group I.50% casein diet Group II.20% casein diet Group III.8% casein diet Group IV.15% casein supplemented with 5% arginine Group V. CMF type of pellet (Oriental Yeast Co. ) These diets were given isocalorically every day. The rats were injected with 3 mg of DMBA at the age of 52 and 55 days. All animals were examined every day during the experimental period, and the onset and evolution of the tumors were recorded. The following results were obtained: 1. The normal body growth rates were not retarded on arginine imbalanced diets, and no ill effects of arginine were shown.
In the preceeding paper, it was shown that mammary carcinogenesis with 7,12-DMBA was significantly inhibited by arginine imbalanced diets given to the host, especially when 15% casein diet was supplemented with 5 % arginine. In this paper, to clarify the mechanism responsible for the inhibitory effect of excessive addition of dietary arginine on tumor growth, the influence of arginine treatment on nucleic acid synthesis in tumor tissue was analysed. Experimental animals employed in this series were female Sprague-Dawley rats with mammary carcinoma induced by 7,12-DMBA. Eight m moles/kg of arginine hydrochloride was injected intraperitoneally into rats bearing mammary carcinoma. One hour after the injection of arginine hydrochloride, experimental animals were received intraperitoneal injection of 3H-thymidine-6 or 14C-aspartic acid (U) respectively. Experimental animals were sacrificed either 60 minutes after the injection of 3H-thymidine-6 or 150 minutes after the injection of 14C-aspartic acid (U), and incorporation of radio activity into nucleic acid were analysed. The following results were obtained: 1. The incorporation of the 14C-aspartic acid (U) into the nucleic acid fraction of liver tissue showed no differences between the arginine treated and control groups, however, the incorporation of 14C-aspartic acid (U) into the nucleic acid fraction of tumor tissue were clearly inhibited in the arginine treated group. 2. The incorporation of 3H-thymidine-6 into the DNA fraction of liver tissue showed no differences between the arginine treated and control groups, whereas the incorpo- ration of 3H-thymidine-6 into the DNA fraction of tumor tissue caused marked inhibition in the arginine treated group. These results suggest that the influence of arginine treatment on nucleic acid syn- thesis in tumor tissue is clearly shown in the salvage pathway instead of the de novo pathway for pyrimidine biosynthesis. teriol.,107,589-591,1971.
Plasma ADH levels were studied during acute and chronic constriction of the inferior or superior caval vein in dogs. Mongrel dogs weighing 9.5 to 29 Kg were used. ADH was extracted from plasma according to Yoshida's method and bioassay was carried out using experimental diabetes insipidus rats. Plasma ADH level increased 2 to 5 folds 10 minutes after the caval vein obstruction with balloon inflation as the result of slight decrease in the venous return to the heart. In this case the left ventricular pressure did not change and the stretch receptor in the left atrium would be stimulated. Plasma ADH increased 8 to 30 folds when the venous return decreased markedly, the left ventricular pressure lowered significantly after balloon inflation and the baroreceptor in the carotid sinus or in the arterial system would be stimulated. There was no correlation between plasma ADH levels and peripheral venous pressure except in one case. In only 2 out of 11 dogs with chronic thoracic inferior caval vein constriction plasma ADH increased. In one case hyponatremia occurred one week after the constriction of caval vein. There was no correlation between plasma ADH levels and degree of ascites. ADH seems not to be a primary factor in occurrence of ascites or edema in these experimental models. But it will increase secondary to the change in distribution of the body fluid to keep the“effective circulating blood volume”.
The development of the trabecular meshwork and Schlemm's canal of one hundred and four albino rat eyes (from the 14th day of gestation to the 30th day after birth) were observed with electron microscope. The results obtained are as follows: 1) The appearance of the anterior chamber angle can be observed on the 17th day of embryonic life with light microscope, while the immature trabecular meshwork is seen on the 15th day of embryonic life with electron microscope. Following the development of mesenchymal cells which fill the future anterior chamber angle, immature trabecular meshwork is formed and each trabecular sheets become flat as they develop and remaining mesenchymal cells are arranged to form future cornea and iris. Thus the chamber angle is formed. Atrophic change of the mesoderm was not observed during the period of the anterior chamber formation. 2) During the period between the 15th day of gestation and the 1st day after birth the endothelial cells which covers the trabecular fibers shows no change in morphology of the cytoplasm. However, the core of the trabecular fibers gradually achieves their mature morphology. 3) In some places, the trabecular fibers are not completely covered by endothelial cells. 4) The endothelial cells of the inner wall of Schlemm's canal have no basal lamina. Giant vacuoles was observed in the cytoplasms. In some specimens giant vacuoles show their openings toward the basal side of the cells and pores were formed on the surface of the endothelial cells. As the endothelial cells of the inner wall of Schlemm's canal achieves their mature morphology the thickness of the endothelial cells decrease and pinocytotic vesicles which appeared on the surface of the cells increase in number.
To overcome the problem of non-specific staining presented by the ferric ferricyanide method for the protein-bound disulfide and sulfhydryl groups, an attempt to modify this method was made by means of the colloidal solution of ferric hydroxide used in MUller's method, instead of ferric sulphate or chloride. This modified method was then applied to the human hypothalamo-neurohypophyseal system to detect the disulfide groups in the neurosecretory substance. The materials used in this study were normal human brains and the brain from an autopsy case of Sheehan's postpartum necrosis. The staining methods used for comparison were the DDD method of Barrnett et al., Chveremont and Frederic's ferricyanide method, Gomori's neotetrazolium method, Bargmann's chrome-alum hematoxylin method, the performic acid-alcian blue staining, the performic acid-aldehyde fuchsin staining, and the periodic acid-Schiff reaction. By using the colloidal ferric hydroxide method, some distinct difference in the degree of disulfide-positive reaction found in the neurosecretory substance in the nucleus paraventricularis, nucleus supraopticus, infudibulum, and lobus posterior of the hypophysis could be observed between the brain of a case of Sheehan's postpartem necrosis and the normal human brains.
Ribosomal subunits were prepared from free and membrane-bound rat liver ribosomes. Molecular weights of the ribosomal proteins from the small and large subunit particles were determined from the relative mobilities in sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Analysis of the proteins of the large subunit showed that twenty-four bands can be resolved on 12.5 % SDS gel which correspond to molecular weight from 10,000 to 50,000 daltons. Analysis of the proteins of the subunit revealed fourteen bands in 12.5% SDS gel which correspond to molecular weight from 9,600 to 44,000 daltons. Densitometer tracing of the SDS acrylamide gels showed that there was no significant difference between free and membrane-bound ribosomal proteins from both the subunit particles.
A histochemical study was made of the small intestine with villous pseudomelanosis. Melanosis pigment was also examined on the purpose of comparing with villous pseudomelanosis pigment (Zottenpseudomelanosenpigment). The stroma of the latter means the pigment after sodium dithionite extraction of iron, while that of the former the one after bleaching with 3% hydrogen peroxide. Both of the two pigments may be considered to contain carboxylic groups of waterinsoluble protein, because they have their stroma stained positively by the nile blue sulphate (pH 2.7) staining. The aldehyde-fuchsin (AF)-positive stroma of villous pseudomelanosis pigment is apparently different from the AF-negative stroma of hemosiderin seen in hemorrhagic foci in that the former is considered rich in disulfide groups, because it's coloration with the peracetic acid-AF staining withstands blea ching with 6-hour methylation at 60°C, while melanosis pigment is regarded as containing sulfonic groups, because it's coloration with the AF staining withstands bleaching with 6-hour methylation at 60°C. From the histochemical point of view, therefore, villous pseudomelanosispigment may be said to be an unique pigment whose occurrence is thought to be in association with the iron absorbing and excretory function of the intestinal mucosa.