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See also AD470632.

Comparative study of cytochemical reactions in the blood cells, cornea and the neuroglia was made on 44 different vertebrates. Among the vertebrates examined, the peroxidase in the leukocytes was evenly distributed, while the phosphatase in the leukocytes was found sporadically. The strongest reaction of peroxidase in leukocytes was found in man, while that of alkaline phosphatase in leukocyte was in lower animals such as guinea pigs and salamanders. The alkaline phosphatase reaction occurred very intensely in the stratified epithelium of the cornea, while did not occur at all in the stroma which contains many lymph capillaries. The finding of this cytochemical reaction of the cornea was nearly the same in the animals examined. The cytochemical reactions in astrocytes, oligodendroglia and microglia in tissue cultures did not show any cytochemical specific natures respectively. The difference in the cytochemical reactions could not be found at all among animal species.

Attempt at identifying the cell types that make sources of the ERG components is continued. Of the three groups of responses recorded from single units in the frog's retina, only the Group-II responses were left suspected to be constituents of the ERG, but the present work excludes this possibility, since it is probable that the responses in question arise from the soma and/or dendrites of ganglion cells that contribute little to the ERG. The failure to re cord any constituents of the ERG in the frog led research in two directions. One is the effort to make intracellular recording possible from cell types other than those already studied. A technique of ''harpooning'' single cells with pipets at high momentary acceleration is being tested and will make a subject in the coming year. The other is to apply the conventional technique to materials other than the frog and see if the localization of ERG constituents is possible in these materials. Among several animals examined, the carp is subject to a more thorough test. Studied are properties of Group-III responses (S-potentials), and the spatial distribution of ERG components in the focal and non-focal regions of the retina.

The report consists of th following three parts. The first part is introductory, and treats of the classification of coastal types that are well developed in Japan, and tries to present the regionality of their distribution. The second part aims at elucidating the nature and processes of terrace surface formation, especially laying stress on the to the modes of crustal movements. Dislocation of earth's crust accompanying the earthquakes and their geomorphic manifestation on coastal districts are also inquired. In the last part, four areas that are studied by the present investigator and his collaborators are presented. They are (1) the Konsen uplands in eastern Hokkaido, (2) northern moiety of the Sanriku coast in northeastern Honshu, (3) dissected fans of western Tokai along the Pacific side of central Honshu, (4) the coalescing dissected fans of Toyama along the Japan Sea side of central Honshu. Emphasis is chiefly laid on the geomorphological features, and land use is treated subordinately in connection with them. (Author)

The ATPase activity was most prominent under the body surface or plasma membrane and in the posterior enlarged portion of toxonemes. It could be also recognized in mitochondoria, Golgi complex, cytoplasmic vacuoles and so-called dense bodies. The localization of H3-glucose after the sensitization of 30 minutes was distributed in the peripheral cytoplasm while it was transported to the more central portion especially on the surface of toxonemes after 60 minutes' sensitization. (Author)

The report is divided into seven sections. A brief exposition of the incidence matrix of the balanced incomplete block (BIB) design is given in section 1. The Hasse-Minkowski p-invariant of the rational congruence is discussed in section 2. In section 3, the necessary condition for existence of a symmetrical BIB design is given in terms of the Hasse-Minkowski p-invariant as the prototype of the non-existence proofs. In section 4, the properties of the association algebra of an association are presented with some examples of association schemes. Section 5 is devoted to the derivation of the necessary condition for existence of a regular and symmetrical partially balanced incomplete block (PBIB) design. The argument presented in section 5 is immediately carried over in section 6 to the derivation of the necessary condition for existence of a certain class of asymmetric PBIB designs. In section 7, the relationship algebra of a PBIB design is explained and some discussions of the non-existence proofs in connection with the relationship algebra are presented. (Author)

Intracellular recordings from single photoreceptors of Necturus maculosus and Gekko gekko reveal that the response to light is a hyperpolarization accompanied by a resistance increase. Passive hyperpolarization of the membrane by extrinsic current does not change the membrane resistance but makes the hyperpolarizing response to light larger. By applying a depolarizing current, the response is made smaller, or even inverted if the current is strong enough. It is suggested that the vertebrate photoreceptor is depolarized in darkness and repolarized in light. This new viewpoint accounts for the unusual polarity of the ERG component ascribed to the receptors. Some observations on single cells in the inner nuclear layer of the carp retina are also outlined in this report. (Author)

The effect of ascorbic acid on increasing tolerance to hypoxia has been proved. The noticeable findings have been found in our experiments; (1) ascorbic acid moves from organs to blood in hypoxia and (2) the proportion of oxydized ascorbic acid is much larger in the heart than any other organs in hypoxia. The explanation for these findings have not yet been given. However, these results suggest that there may be some important relationship between ascorbic acid and heart activity which was proved in our previous experiments to be the most significant factor limiting the altitude tolerance. (Author)

The responses used as indices of studying color reception are potential changes with latencies of several msec recorded from within single photoreceptors using micropipette electrodes having tip diameters less than 0.1 micron. Single carp cones are used for the study, but single octopus photoreceptors are also used to provide matrials for discussion of the significance of the receptor potential upon the flow of information within the receptor. The intracellular placement of the electrode is achieved by mounting the retina on a vibrating table which gives it a large acceleration against the slowly advancing electrode tip. The electrode penetration is signalled by the appearance of a resting potential of 30 - 40 mV. In single carp cones, responses to light are hyperpolarizing. The response spectra studied statistically are classified into three groups in terms of wavelengths of maximum sensitivity. These are in close agreement with those of the absorption spectra of single goldfish cones reported by Marks and MacNichol. The response of single octopus photoreceptors, studied as a control, is a graded sustained depolarization, as common in most invertebrate photoreceptors. The action spectrum calculated therefrom agrees well with the absorption spectrum of the receptor photopigment. Evidence is provided that each octopus photoreceptor is selectively sensitive to a certain plane of polarized light. (Author)

Nosema (=Encephalitozoon) cuniculi is found parasitic in variety of mammals and only a few cases of infection in man so far have been reported. As this parasite is an agent of encephalitis in animals, it is possible that it can be a cause of some of encephalitis-like disorders of unknown etiology of man. The main purpose of this work is to make clear how often man may be infected with this parasite. For the detection of the parasite, the spinal fluid of urine of suspected cases should be inoculated into animals or tissue culture. Most of laboratory animals, however, are very often infected with this parasite and what is the worse, their infections are mostly latent. Endoxan, an anti-tumor substance found to be effective in promoting the growth of N. cuniculi, was tested for the provocation of the latent infection. It was successful in some cases and not in others. Endoxan, therefore, cannot be used for the selection of uninfected animals. N. cuniculi grows in HeLa cell cultures. However, it was not possible to get a constant positive growth even though the parasites were inoculated into the culture. Much improvement must be accomplished before the tissue culture can be used for this purpose. Under these conditions, it was not possible to start the experiment to detect the parasite from human beings in this first year period. Morphological details of the parasite in successive developmental stages from the proliferative form to the mature spore were studied by electron microscope. (Author)

The objective of the research is to identify the Nosema parasite to be isolated from human beings. Work was directed toward the method of isolation and identification of the parasite by the establishment of a mouse colony free from Nosema infection and by tissue culture., See also AD-660 282.

Changes in stroke volume of the right and left heart were studied in relation to tolerance to altitude. It was concluded that the pulmonary circulation might be regarded as a limiting factor of importance for performance at altitude. Chronic experiments on rabbits with chronic implanted heart electrodes were performed with partial success. The animals can be kept alive about one month after the operation. The results of these chronic experiments showed that the contractile capacity of the right heart is strengthened, increasing the right heart stroke volume within one week period of training-exposure to hypoxic condition. This fact supports the view that the improvement of the right heart capacity might be involved in the establishment of acclimatization to altitude. Changes in ascorbic acid in different organs at altitude are being studied in order to elucidate the role of this substance in acclimatization or tolerance to altitude. The results obtained so far show that the well acclimatized or tolerant animals have a larger capacity for utilizing ascorbic acid at altitude than do unacclimatized or less tolerant animals. This suggests that the administration of large amount of ascorbic acid may improve the tolerance to hypoxia. (Author)

The metabolism of the brain is characterized by emphasis on carbohydrate as the energy source and the rate of the energy metabolism increases during the neural excitation. It is generally believed that the control steps in glycolytic pathway are those concerned with phosphorylation of glucose (hexokinase) and fructose-6-phosphate(phosphofructokinase). Although the data presented in this report are consistent with this consideration, they clearly indicate that, in addition to the above two, the step catalyzed by pyruvate kinase is rate-limiting and controlling for glycolysis in brain cortex slices. Pyruvate kinase in brain tissue seems to be strongly inhibited in some way which is related to the cellular structures and this inhibited activity is regulated by the ionic milieu. The changes in the intermediate levels of the glycolytic pathway in the brain cortex slices by the addition of high potassium in the incubation medium were characterized by decreases in glucose-6-phosphate and fructose-6-phosphate, and by increases of fructose diphosphate, triose phosphates and lactate. These changes were similar to those induced by ischemia in vivo brain. (Author)

Intracellular recording from single cones in the carp (Cyprinus carpio) is an attempt to clarify the mechanism of color information processing in vertebrate photoresceptors. As the only means to permit the intracellular recording, the retina detached from the pigment epithelium is jolted at a high acceleration against a vertically held, slowly advancing micropipette. The new model of jolting device is described. Whenever a potential identified as a single cone potential is found, the visible spectrum adjusted to equal quantum flux is scanned by means of a photostimulator especially built for this experiment and the single cone spectral response curve is obtained. Statistical analysis of 142 records selected out of hundreds, based solely on the greatest signal-to-noise ratio, reveals three groups of cones; the red cones (74%) with the average peaking wavelength at 611 millimicrons, green cones (10%) at 529 millimicrons, and blue cones (16%) at 462 millimicrons. These peaking wavelengths are in close accord with those of single cone absorption spectra measured by Marks and MacNichol in the goldfish by a microspectrophotometer. It is strongly suggested that Young's trichromatic theory applies at the photoreceptor level in the vertebrate. (Author)

Glutamic acid, the concentration of which in brain is higher than that of any other amino acid, increases the aerobic glucose metabolism of brain tissue in vitro. This effect was concluded to be due to the accumulated potassium ions in brain tissue, the accumulation of which was induced by glutamic acid. Ouabain inhibited the effect of glutamate on glycolysis as well as the accumulation of potassium ions. Aspartic acid inhibits the effect of glutamate on glycolysis but not the accumulation of potassium ions. Aspartic acid which was accumulated in brain tissue in vitro did not show any efflux almost at all, although glutamic acid showed significantly. Aspartic acid might be thought to be firmly attached to cellular components of brain tissue and thus to inhibit the uptake of exchangeable glutamate and potassium ions. Effect of high concentration of potassium ions on the aerobic glucose metabolism of brain tissue in vitro could be fully explained by its effect on pyruvate kinase activity in the glycolytic pathway. Pyruvate concentration after the incubation of brain slices in the addition of phosphoenolpyruvate as substrate was ten times higher in the presence of high concentration of potassium ions in the medium than in the absence. (Author)

The effect of intravenous administration of concentrated Na and K solutions, and cardiorespiratory stimulants on cerebral circulation, metabolism and electrical activity were investigated in cats. (1) Effect of concentrated cation solutions: (a) Increased Na(+) in the extracellular fluids of the brain induced the electrical and metabolic activation and increase in cerebral blood flow. Electroencephalogram showed the seizure or low voltage fast activity. Cerebral blood flow increased, brain PO2 and PCO2 increased, pH decreased, Na(+) usually increased and when seizure started Na(+) changed to decrease. (b) Intravenous administration of concentrated K solution resulted in the cardiac dysfunction and consequently showed a decrease in cerebral blood flow and metabolism, and electrical depression. After intracisternal administration of concentrated K solution, it appeared to show the metabolic and electrical activation. However, since there is a question about increase in concentration of K+ in the brain surface after intracisternal administration of K, further investigation with more appropriate method is necessary for obtaining the direct action of K+ on the brain. (2) Effect of intravenous administration of cardiorespiratory stimulants of aminophylline, dimorpholamine, nikethamide, vitacampher and lobeline were investigated. (Author), See also AD-438 200.

Mice which survived a prior infection with low virulent toxoplasmas (Beverley strain) tolerated a challenge infection with a highly virulent strain (RH strain). Of 35 mice which survived the challenge infection with RH, the brains of 22 of these mice were shown to contain living organisms. Vaccination with heat-killed Toxoplasma alone or in incomplete adjuvants seemed to have no effect in increasing the protection of the mice against a challenge RH infection. Vaccination with sonic disrupted Toxoplasma also failed to protect mice against a RH challenge. Passive immunization with anti-toxoplasma sera obtained from infected rabbits and pigs demonstrated very little effects in prolongation of the survival period of infected mice. Development of cellular immunity to some degree was indicated by the experiments employing implanted diffusion chambers in the peritoneal cavity of mice. The role of antibody in the formation of the cyst of Toxoplasma was investigated. The anti-metabolite 6-mercaptopurine was inoculated into infected mice in order to inhibit the normal immune response of the mice to the parasite. The results tend to indicate that antibody is not an essential prerequisite to cyst formation. (Author)

Experimental shigella infection in rabbit was carried out successfully by new technic and reproducible pathological change was observed. In this system, however no invasiveness of shigella into the tissue was recognized. This finding was supported by blocking method which was designed to avoid artificial transportation of shigella from the surface of the intestinal wall to the inside of the tissue. At the same time, it was proved by bacteriological technic that the multiplication of shigella in the intestines was produced on the surface of the intestinal mucous membrane rather than that in the rumen of the canal. Efficiency of the antagonism between resident and invader was influenced by the pre-culture condition of invader. That is, the bacteria in continuous flow culture was rejected easily compared with that of plate culture. Through biochemical analysis it was proved that bile acid was essential for inhibiting activity of bile to the establishment of pathological changes. However the existence of certain concentration of bile acid in the culture media used in continuous flow culture did not influence the antagonism between resident and invader. (Author)

A gigantic peroxidase granule was detected in the polymorphonuclear neutrophil leukocytes of the giant salamander, Megalobatrachus japonicus. The granule resembles the gigantism of the peroxidase granule that was recognized in human blood disease (Chediak and Higashi's disease). It is of interest that one of pathological characters of human leukocytes was found in an apparently healthy salamander. The pigmented leukocytes were found only in cold-blood animals such as reptiles and amphibia. These leukocytes could never be detected in man, mammals, birds and fishes. The pigmented leukocytes lack both peroxidase and PAS reacting substance suggesting that granules of the pigmented leukocytes are melanin injested in the blood forming organs. The peroxidase negative eosinophil leukocytes were detected in the liver tissue of Xenopus laevis Daudin. The lack of this enzyme in the eosinophil leukocyte suggests a profound philogenetical gap in comparative hematology. (Author)

This report has two sections: first, a detailed description concerning establishment of techniques for automatic recording of PO2, PCO2, pH, Na(+) and K(+), and second, of application of the techniques to studies of cerebral circulation and metabolism in vivo. PO2, PCO2, pH and Na(+) electrodes were constructed to our specifications. The PO2 electrode had a fine platinum cathode, surface of which covered with polyethylene membrane having a suitable thickness. The advantage was that the electrode could be calibrated easily and rapidly with gas at known oxygen tension, and was applicable for measurement of PO2 in absolute unit on the cerebral cortex. The pH electrode of McInnes type, the PCO2 electrode reported by Stow and the Na(+) electrode fabricated from Eisenmans NAS11-18 glass had been tested in vitro with satisfactory results. Application of these electrodes was made in the manner in which their sensitive surfaces were lightly placed on cerebral cortex of cats and their output currents were recorded together with systemic arterial blood proencephalogram and electrocardiogram on a graph. The validity of the technique can be confirmed by alternation of these parameters in an expected manner under various conditions of cats, such as high gaseous tension of oxygen and carbon dioxide in the blood due to inhalation of O2-CO2 gas mixture. (Author)

The phage mediated conversion (or lysogenic conversion) of salmonella 0(1) antigen by a sub 1 phage could be changed into the DNA mediated transformation when Sg cells are incubated with purified DNA extracted from a sub 1 phage and active particles of a sub 10s phage. As far as the resuspension of such mix infected Sg cell is directly examined before lysis for the agglutinability to the anti-0(1) factor serum, a small proportion of them is shown to be really converted into the one shared with 0(1) antigen. However the converted character is almost confined in the generation of such converted cells so that it is scarcely inherited into their progeny cells. On the other hand, such 0(1) antigen conversion could not be seen upon the mix infection with a sub 1 DNA together with a sub 2 phage particles which is incompatible with a sub 1 phage in the growth at the same host cells, or upon mix incubation with a sub 1 phage DNA alone. (Author)

Screening tests of various kinds of compounds were carried out with the purpose of obtaining new drugs for toxoplasmosis. Compounds tested were 66 pyrimidine derivatives, 14 guanidine derivatives, 11 triazine derivatives, 12 sulfone derivatives, 1 amidine derivatives and 4 sulfadrugs. Mice inoculated with 100,000 Toxoplasmas of RH strain were used for the tests. The compounds to be tested were given intraperitoneally for five days successively, starting one day before the inoculation. The dose was 1/15 amount of LD50 of each compound. The effect of the compound was evaluated by difference in survival days between the test and the control animals. None of the derivatives of pyrimidine, guanidine, triazine, amidine and sulfa-drugs so far tested was proved to be more effective than pyrimethamine. The average survival period of the animals treated with pyrimethamine was twice as long as that of the control groups. Among the sulfone derivatives, 4-Nitro-4'-formylamino-diphenylsulfone and 4-Nitro-4'-acetoamino-diphenylsulfone seemed to be a little more effective than pyrimethamine regarding prolongation of the survival period. 4-Nitro-4'-formylamino-diphenylsulfone was most effective, i.e., 2 out of 6 mice treated with oral administration of this derivative were considered to be cured. By these results it was expected that 4-Nitro-4'-formylamino-difenylsulfone might be effective in treatments of human toxoplasmosis. (Author)