Effects of NaNO2 and NaNO3 on the oxidative phosphorylation of mitochondria were studied, and following results were obtained. 1) The effect of NaNO2 on the oxidative phosphorylation is higher than NaNO3. 2) Relative activity of State 4 is activated by NaNO2 (1-10mM) added to reaction mixture. 3) The respiratory control index of mitochondria is decreased by NaNO2. 4) The effect of NO2 on oxidative phosphorylation in the borate buffer as reaction mixture is higher than that in the tris-HCL buffer. It is suggested that NO2 has a probability of the reaction on-NH2 site of mitochondria.
Thirty-six cases of human atrophic chronic gastritis were observed by transmission and scanning electron microscope. Generally, in the surface and neck mucous cells, vacuole formations and swelling of mitochondria were occasionally observed. Although, increase of young parietal cells were frequently noted, ultrastructural changes were hardly seen in the cytoplasm and the nucleus of both the parietal cells and the chief cells. Intestinal metaplastic mucosa was found in almost all cases of antral mucosa and in half cases of corporal mucosa of chronic gastritis. In the SEM observation, the gastric mucosa and the intestinal metaplastic mucosa were clearly bordered. In TEM observations, all types of normal small intestinal cells were found in the intestinal metaplastic mucosa, and there were no ultrastructural differences between normal intestinal cells and intestinal metaplastic mucosa cells. It was interesting that enterochromaffin cells, which were scarce in normal gastric mucosa and numerous in small intestine, were frequently observed in intestinal metaplastic mucosa. In the three cases of chronic gastritis, the peculiar type cells which had both endocrine cells granules and mucous granules were found and their origins were discussed.
Japanese Government has enforced statutory activities for the relief of the victims due to the adverse effects of legal vaccinations since October in 1970. The author collected and analyzed 985 cases applied for relief system run by the Government, and pointed out several problems arisen by the analysis. The results are as follows. 1) Concerning the distribution of the applicants classified by the time victims received vaccinations, the applicants decrease as the year of occurence of the victim goes back to the past. 2) As for kinds of vaccinations, more victims were observed among small pox, poliomyelitis and combined vaccines of diphtheria and pertussis (DP) than cholera and BCG vaccinations. 3) As for sex ratio, no significant difference was admitted as a whole, though a slight difference was observed among poliomyelitis and BCG. 4) About one third of the applicants died. The more they died, the shorten was the duaration from vaccination to the appearance of adverse effect. 5) There were differences of incidence among prefectures. According to the findings mentioned above, the author concluded that an effective surveillance system must be established for the complete relief activities.
A judgment committee was organized in Ministry of Health and Welfare when the relief system for the victim of adverse effects of vaccination was established. The committee has divided the applicants into three groups, that is, “the victim, ” “positive in denying the victim” and “questionable.” The author discussed the result of judgment by the committee and pointed out some questions of the system. The results of the analysis are as follows. 1) The committee identified only 5.8% of the applicants to the victim. 2) About two thirds of the applicants were decided as questionable and the others were screened to be positive in denying the victim. 3) As to termination of adverse effects, cases that recovered completely were apt to be decided as the victim. However, only a few cases of the deceased or the sufferers from late effect might be decided as the victim. 4) Medical certificates especially provided by medical doctors at public hospitals, were highly estimated by the committee. The author concluded that the committee should establish the clear concept of the adverse effect and the criteria for the judgment of their treatment based on social deliberation.
7, 12-Dimethylbenz(a)anthracene (DMBA) 20mg was administered to 55-days-old Sprague-Dawley rats with gastric intubation. Two or three rats were sacrificed daily during the first week following administration of DMBA, then, weekly up to the 20th week. Mammary glands of each different stage were examined by the whole mount method, light microscopy and electron microscopy to observe tumorigenesis of rats mammary tumors introduced by DMBA. 1. On the 4th day following administration of DMBA, mammary epithelial cells at the peripheral portion of the duct demonstrated dilatation of endoplasmic reticulum and Golgi apparatus. On the 2nd week, an evidence of hypersecretion was well established. 2. However, on the 4th week, mammary epithelial cells showed atrophy and their intracellular organelles were regressed. On the other hand, myoepithelial cells showed cytoplasmic extensions to surround mammary epithelial cells. Stroma around the ductules also showed marked proliferation. 3. On the 7th week, mammary glands, examined by the whole mount method, demonstrated hyperplasty to form small nodules, namely, hyperplastic alveolar nodules (HAN). Among some rats, stroma around the ductules kept growing to be fibroadenoma. 4. On the 9th week, a tumor became palpable, and its histology was tubular adenocarcinoma. 5. Electron microscopic findings of carcinoma were increase of nucleocytoplsmic ratio, irregularity of nucleus, hypertrophy of nuclear membrane and enlargement of intracellular space etc..
For the purpose to evalute the properties of rinderpest virus, scanning and transmission electron microscopic studies were conducted with MDCK cells inocu-lated with rinderpest virus by observing changes in the surface and intracellular structures in these cells. The results may be briefly presented as follows. (1) It is possible to distinguish readily multinucleated giant cells produced by rinderpest virus from normal cells because of their flat shape. Moreover, the microvilli on these multinucleated giant cell surface are markedly less than those on normal cells. Yet microvilli do not necessarily disappear completely. (2) Even when infected by the virus the cell that does not transform into a multinucleated giant cell does not show so much change exteriorly as compared with the normal cell except a little shrinkage. (3) As for observation on intracellular changes it was not possible to clarify any morphological correlation between intranuclear inclusion body and cytoplasmic inclusion body. (4) It was confirmed that virus particles are budding out from the infected cell surface as well as from microvilli surrounding the cell. (5) Particles of 150-340 nm in size arranged in a chain formation have been identified to be rinderpest virus by comparing the negatively stained picture of virus particles specimen isolated from the infected cells with purified virus particles negatively stained specimen in the same field by scanning microscopy. (6) Mature virus particles reveal the morphology that is irregular in shape and size by scanning electron microscopy, ultrathin sections, and negatively stained specimen, whose size ranged about 150-340 nm. In addition, the negatively stained picture shows the presence of spikes about 8 nm in size on the virus surface, and these can be observed nucleocapsids of about 17 nm in diameter with the lumen of 6 nm. And there can be seen also filamentous virus-like particles about 1 μm in length. However, by the scanning electron microscopy on the present stage it is impossible to detect such ultrastructures. (7) It has been demonstrated that the cell membrane where virus particles bud out tends to be electron dense membrane. (8) The halo observable around the inclusion body in hematoxylin-eosin stained specimen is an artifact.
A microcytotoxicity assay (MCA) and a 51Cr release cytotoxicity assay (CRA) for cell mediated immunity to the allogeneic tumor were done in parallel with same reagents used for both assays. Therefore, CRA was modified in which monolayer culture cells were used as target cells and MCA was devised in which the adherent surviving target cells after the interaction with effector cells were assessed by 51Cr uptake (so called 51Cr post-labeling assay... CPLA). Under standard conditions, CPLA detected a significant activity of weakly sensitized lymphocytes which was not detected by CRA. Reactivity of lymph node cells after the tumor transplantation was tested simultaneously in CRA, CPLA and the macrophage migration inhibition test (MIT). The data showed that CPLA detected immune reactivity for longer period than CRA, and as long as MIT. The lytic activity of lymph node cells 8 days after immunization was monitored by a long term CRA (40-hours' incubation). The cytotoxicity value for late 20 hr was remarkably higher than for early 20 hr. This in vitro activation of sensitized lymphocytes indicated a peak at 8 days and then declined, dissappeared at 14 days. When both assays were done under same condition, CPLA detected a moderate activity which was not detected by CRA. This result means that CPLA could detect immune responses which inhibit tumor cell growth without killing them. Both assays detected primarily a T lymphocyte-mediated activity. The CPLA also detected a weak non T-cell activity in anti-θ plus complement-treated alloimmune lymph node.
The effects of tumor-BCG mixture to the cellular cytotoxicity of the regional lymph node and the spleen were observed in BALB/c mice received allogeneic tumor cells. This tumor cells, when grafted subcutaneously, were normally enhanced in 40% of BALB/c mice and 100% of A/St mice, but failed to grow when injected as a mixture with tumor and BCG. By injection of tumor-BCG mixture, the development and decline of the cytotoxicity (51Cr release assay) to the tumor cells was accelerated, but the magnitude of the peak activity was diminished. Possible mechanisms by which the decreased activity induced of complete tumor suppression were examined. The result showed that immunized lymphocytes with non-immune macrophages exhibited a significatly increased cytotoxicity as compared with immunized lymphocytes in the abscence of macrophages.
The formaldehyde-induced fluorescence technique (Falck et al'62) allows for the microscopic visualization of intraneural monoamines. The recently introduced glyoxylic acid techique (Lindvall and Bjorklund'74) appears to offer a most advantageous approach to monoamine tract tracing. The technique allows the important finding that a marked degree of mixing of the various adrenergic components occurs in the central tract in the rat brain. However, the technique in its present form is difficult to apply to large mammals. Because, the catecholamine diffuse in the neural tissue during perfusion. In this report, the author shows that it is able to prevent this defect by adding chloralhydrate to the glyoxylic acid-formaldehyde solution. In this modified method, the fluorescence of catecholamine is enhanced and is easily classified in its color. For this reason, chloralhydrate-glyoxylic acid-formaldehyde method can apply to large mammals.
The distribution of catecholamine containing neurons and fibers was studied in the rabbit medulla-pontine-tegmentum by the chloralhydrate-glyoxylic acid-formaldehyde method. There are fundamental similarities between the rabbit and the other mammals on the catecholamine distribution. Most of the catecholamine fluorescent cell bodies in the rabbit are located within the nucleus locus coeruleus, subcoeruleus and substantia nigra in the pontine tegmentum. They are more extensive than those of the rat, monkey and more connective than those of the dog and cat. There are two main catecholamine fiber systems at the medulla level. One is ventrolateral catecholamine pathway originating from the area corresponding to A1 in the rat and the other is the dorsomedial catecholamine pathway originating from the area corresponding to A2 in the rat. These two pathways associate with the upper medulla oblongatae level. One catecholamine containing cell group extends toward dorsolateral in the nucleus reticularis lateralis of the rabbit, but it has not been found in the rat.
The hippocampal formation of the rat brain was studied by the Falck-Hillarp method and Karnovsky method after administration of dithizone, alloxan, and oxine. The purpose of this experiment was to clear the relation between cholinergic, adrenergic terminals and zinc in the mossy fiber synaps. The acetylcholinesterase and catecholamine reaction increased in oxine and dithizone administration. On the other hand, when dithizone and oxine were administrated, zinc decreased in the mossy fiber synaps. From this result, the relation between zinc in mossy fiber synaps and acetylcholinesterase, catecholamine in the hippocampus were discussed.
It is a very important problem in the field of public health among many Occidental countries to prevent against alcoholic addiction. Recently, even in Japan the consumption of alcoholic beverages has increased considerably and especially drinking habit among women and young people is becoming more popular than ever before. Such tendency has changed drinking trait of old traditional drinking pattern in this country which had been fixed for a long time. Also it suggests that will raise more important problems in the field of public health in Japan. There are several reports concerning with drinking trait in Japan but most of them were studied about special professional groups or residents of small area. Very few of population based studies have been reported especially in larger area on drinking trait. This study has been done from the point of view that a population based study in larger scale is needed because it is considered to be very important in the field of public health to clarify a relationship between drinking trait and alcoholic addiction in Japanese society at large. Method One thousand examinees among residents of Okayama prefecture were randomized by means of telephone books excluding those in urban area. A revised alcoholic addiction test (alcadd test) was mailed to these examinees. 519 answers gained were analysed. Results were as follows: (1) Rate of drinking alcoholic beverages are as high as 83.8%. Rate of habitual drinkers is 54.1% and 37.0% have the habit of drinking everyday. (2)There are differences of frequency of habitual drinkers among different age groups and also differences among occupational groups. (3) The higher score on the alcadd test he gets, the more frequently he drinks. (4) As for 32 of 38 items, there are demonstrated significant differences of the frequency of alcoholic response to the alcadd test among groups classified by frequency of drinking. (5) The persons of a group who drink everyday have high rate of response to regurality of and preference for drinking. On the other hand, those of a group who drink from four to six days per week have higher rate of response to problem drinking such as lack of controll and social manner and so on, rather than those who drink everyday. Conclusion (1) A revised alcadd test is usefull for an analysis of alcohol trait individually and as a group. (2) Alcohol trait has a close relationship with social and cultural background. The author, therefore, points out that the analysis of drinking trait should be necessary hereafter.
The first effort was made to document that elemental diet was satisfactory in maintaining normal growth in rats. A second effort was made to determine the maximum resectability of the intestine when rats were fed only by the elemental diet. These experiments were done in 80 ten-week-old female Wister rats. Results obtained are as follows (1) Eight rats fed only by the elemental diet through gastrostomy for 20 days gained more weight than pre-operative weight (P<0.02), and a positive nitrogen balance was obtained when rats were given with calories of 303 cal/kg/day and 8.25g/kg/day of amino acids. Also long term experiment of giving the elemental diet by mouth for 60 days resulted in increase of body weight (P<0.001), and there was no abnormality in hematological and pathological evaluations. (2) Body weight became heavier than that of before operation in the rats with 70% or 80% resection of the small intestine with P<0.001 and P<0.05, respectively. In the latter group, however, recovery from surgical assault were delayed, the pancreas and lung were often compromised and blood urea nitrogen as well as serum enzymes such as s-GOT and s-GPT were abnormally high. In 90% resection rats mortality was very high (85.7%). Therefore, it was concluded that the maximum resectality of the small intestine was 70% when rats were fed by elemental diet. (3)If the ileocecal valve was resected, pre-operative body weight was not resumed even in 70%-resection rats, indicating the importance of the valve. (4) When pyloroplasty was added in 70%-resection rats, gain in body weight was larger as compared with 70% resection group and non-resection group (P<0.005 and P<0.01, respectively). Abnormal findings in the liver, pancreas and lung were lesser and serum protein was elevated (P<0.001). These results show that elemental diet is effective tool of nutrition and exclusion of small intestine can be done safely as much as 70% However, improvement of administration and nutritional constitution may be able to endure much more extensive resection.
The assay of red cell superoxide dismutase (SOD) activity employing the method of nitroblue tetrazolium (NBT) reduction in aerobic xanthine oxidase system was studied. The results were as follows. 1, Determination of unit of SOD activity Assuming the range of NBT reduction is 100 percent, one unit is defined by the amount of SOD required to inhibit NBT reduction by 50 percent. However, the activity defined by this calculating method is lower than the real value. Because the maximal inhibition of NBT reduction in aerobic xanthine oxidase system by crude extract was in the range of 75 to 90 percent in this experiment, and differed from each sample. The activities calculated from the point of 50 percent inhibition and half maximal inhibition showed good correlation (correlation coefficient; 0.92). And to get 50 percent inhibition less samples are needed than to get the maximal inhibition. So, this calculating method could be used in the case of comparison of SOD activities. 2, Recovery of purified SOD 93 percent recovery of purified SOD added to the hemolysate before extraction was obtained. The activities were determined 5 times about the same sample to get the variation coefficient. The value was as low as 2 percent. 3, Interferrence by hemoglobin Methemoglobin formation from acatalasemic hemoglobin by SOD reaction was observed. And maximal inhibition of NBT reduction by hemolysate was low and varies widely. Using hemolysate for sample interferes the assay and the value of SOD activity is unreliable.
The levels of human erythrocyte superoxide dismutase activity were determined to establish a normal range classified by age. Blood samples were drawn from 11 little children aged 4 to 6, 47 young adults aged 20 to 35 who have no apparent diseases and 22 old people aged 70 to 100 living in a nursing home. The following results were obtained. 1, The distribution of SOD activity levels is 17.9±3.8 (m+SD) Units/mgHb in children, 14.8+3.3 in young adults and 11.4±3.0 in old people. 2, The data show SOD activity decreases with age. Statistically the mean value of each group is significantly differes from one another.
In vitro inhibition of cytoplasmic SOD by cyanide ions, nitriles, dimethyldithiocarbamate, paraquat and Gramoxon was determined. In this experiment NBT reduction in aerobic xanthine oxidase system was used. Inhibition by polluting agents against crude extract of SOD of human erythrocytes and purified SOD from bovine blood. The following results were obtained. 1, Potassium cyanide inhibits SOD reaction, giving 50 percent inhibition at 0.036mM in crude extract of human blood and 0.043mM in purified bovine blood SOD. 2, No inhibition was observed by nitriles up to 20mM both in crude extract and purified SOD. 3, Sodium thiocyanate gives no inhibition up to 40mM both in crude extract and purified SOD. 4, Dimethyldithiocarbamate inhibits SOD reaction giving 50 percent inhibition at 0.66mM in crude extract and 0.96mM in purified SOD. 5, Paraquat inhibits SOD reaction giving 50 percent inhibition at 0.06mM in crude extract and 1.09mM in purified SOD. And Gramoxon containing 24 percent paraquat and certain surfactants gives 50 percent inhibition as low as 0.0054mM in crude extract and 0.0084 in purified SOD.
The hippocamal formation of rat brain was studied by the silver sulfide method and was also observed by electron microscopy after the administration of dithizone, alloxan and oxine, respectively. In normal rat, the reaction products were found in the mossy fibers of granular cells of gyrus dentatus which ended to the dendrites of pyramydal cells present in h3, h4 and h5 area of the hippocampus. By electron microscopic observation, the mossy fiber endings of normal brain were filled with synaptic vesicles, and most of them ended to the dendritic spines of pyramidal cells. Zinc shown by silver sulfide method were not detected in the hippocampal formation from 1 to 3 hours after the administration of dithizone, and at the same time, the number of synaptic vesicles of the mossy fiber endings was decreased and the dendritic spines became to be smooth. Alloxan did not any effects of the amount of zinc and the number of synaptic vesicles in mossy fibers, while both of them decreased temporarily at 5 to 15 minutes after the administration of oxine. From these results, the correlation between zinc and synaptic vesicles in mossy fibers, and the mechanism of these drugs' action to zinc content and synaptic vesicles were discussed.
Serial oxygen uptake determination were done in 39 patients and 24 mongrel dogs during 20% hemodilutional cardiopulmonary bypass at normothermia. In patients, the time of the bypass ranged from 37 to 161 minutes. In dogs, the bypass time was standardized at 120 minutes. In patients, average of the mean perfusion index was 2.03 l/M2/min in the group I (BSA 1.2M2, n=13), 2.47 l/M2/min in the group II (0.8 BSA<1.2M2, n=6) and 2.72 l/M2/min in the group III (BSA<0.8M2, n=20). During total perfusion, the average of the mean arterial blood pressure was 75.7mmHg in group I, 53.6mmHg in group II, and 44.1mmHg in group III. The average of the mean oxygen saturation during perfusion were 60.5%, 64.2%, 56.7% and the average of the O2 consumption were 72.5ml/M2/min, 94.7ml/M2/min and 112.6ml/M2/min respectively. O2 consumption (during perfusion)/O2 consumption (predicted) ranged 0.22 to 0.96 in all patients and 0.52, 0.62, 0.63 respectively. The average of the O2 extraction rate during perfusion were 32.9%, 42.8% and 36.0% respectively. The average of hematocrit during perfusion was 24.4% in patients and 23.5% in dogs. In dogs, classified into 5 groups, group A perfused at rate 25ml/Kg/min had an average mean arterial blood pressure of 20.6mmHg, group B perfused at rate 50ml/Kg/min had 38.3mmHg, group C perfused at rate 75ml/Kg/min had 58.7mmHg, group D at rate 100ml/Kg/min had 67.7mmHg, group E at rate 125ml/Kg/min had 58.4mmHg. The average of the O2 consumption were 1.66ml/Kg/min (group A), 2.87 ml/Kg/min (group B), 4.10ml/Kg/min (group C), 3.62ml/Kg/min (group D), 5.91ml/Kg/min (group, E ). 1) Mean arterial blood pressure varied directly with flow rate in dogs, but not in patients. During perfusion mean arterial blood pressure was maintained at a lower level and mean central venous pressure at a higher level than at pre-perfusion value. 2) There is a linear relationship between flow rate and oxygen consumption both in patients and dogs. Oxygen consumption during perfusion were considerably lower than normal oxygen consumption. 3) Venous oxygen saturation during perfusion was lower than pre-perfusion value. There is a linear relationship between flow rate and venous oxygen saturation in dogs, but not in patients. 4) In patients, relationship between perfusion index and O2 consumption indicates that the perfusion index over than 2.7 l/M2/min will be desirable, and in dogs splanchnic pooling during perfusion suggested the flow rate 75-100ml/Kg/min will be desirable.
The author analysed the air pollution data of NO, NO2 and NOx (NO+NO2) measured by auto recorder in Mizushima. The results obtained were as follows. 1) NOx exhausted from factories in Mizushima Industrial District are carried by sea breeze (wind S-WSW, 1-6 m/sec.) and pollutes northern area of Mizushima as same as SOx. The sea breeze bings the NOx exhausted from factories to Matsue, closed to factories, Fukuda, about 3 km from factories, and also Kurashiki and Amagi, far from factories, as same as SOx. 2) NOx exhausted from cars is not negligible at four monitoring stations analysed, especially at Kurashiki.
The effects of n-paraffin on the oxidative phosphorylation of mitochondria and K+ compartmentation of mitochondria and erythrocyte membrane were studied and the following results were obtained. 1) The effects of n-paraffin on decoupling action of oxidative phosphorylation were observed in the order of C6, C7, C8>C9>C10>C11>C12>C13>C14>C15. 2) The effects of n-paraffin on release of K+ from the mitochondria were observed of C7>C8>C9>C10>C6>C11>C12>C13>C15. 3) The release of K+ by n-paraffin on the erythrocytes were also observed in the order of C6>C7>C8>C9>C10>C11>C12>C13>C14>C15. 4) The effects of n-paraffin, monocarboxylic acid and dicarboxylic acid of carbon number 6 and 8 on decoupling of oxidative phosphorylation of mitochondria were observed in the order of n-paraffin>monocarboxylic acid>dicarboxylic acid and those of carbon number 10 were observed in the order of monocarboxylic acid>dicarboxylic acid>n-paraffin.