The correlation among lymphoid infiltration (LI) in the primary tumor, cell-mediated immunity and the accumulative 5-year survival rate was studied in 515 patients with resected gastric cancer. As scales of cell-mediated immunity, the blastformation rate of peripheral lymphocytes against phytohemagglutinin (PHA) -M (Difco) and the PPD skin reaction were examined. The results were as follows: 1) LI of the primary tumor was found in 124 out of 515 patients (24.1%); patients with marked LI were 6.4%. 2) The blastformation rate in patients with positive LI was significantly higher than that in patients with negative LI (p<0.01). 3) The LI of the primary tumor tended to be found commonly in patients with gastric cancer in clinical stage I. 4) The blastformation rate decreased when the clinical stage of gastric cancer progressed, however, this trend was not seen in patients with positive LI regardless of the stage. 5) The accumulative 5-year survival rate in patients with positive LI was longer than that in the patients with LI negative (p<0.05). 6) There was no relationship among the degrees of LI in the primary tumor, the PPD skin reaction and lymphocyte count. In conclusion, LI of the primary tumor has a close relationship with the blastformation rate, and this phenomenon suggests that a local appearance of cell-mediated immunity in the cancer bearing body may occur.
Of patients with collagen disease, anti-lung tissue antibody (ALA) was extensively investigated in relation to the presence of pulmonary fibrosis. Relationship between ALA and other autoantibodies, e.q. rhuematoid factor (RF) and antinuclear factor (ANF) was also studied. From either dog or human lung tissue, crude extracts were obtained and used as antigen, after which ALA was assessed by tanned red cell hemagglutination. ALA was revealed in 16 cases (34.8%) of 46 cases of systemic lupus erythematosus (SLE), 9 cases (64.3%) of 14 cases of progressive systemic sclerosis (PSS), 9 cases (69.2%) of 13 cases of polymyositis plus dermatomyositis (PM-DM) and 10 cases (30.3%) of 33 cases of rheumatoid arthritis (RA). In the control group, only 2 cases (5.1%) of 39 cases ranging from 20 to 80 years old were positive. A significant correlation between the presence of pulmonary fibrosis and the level of ALA was demonstrated in patients with PSS and PM-DM, but not in cases with SLE and RA. ALA, however, did not correlate with the titer of RF and ANF. Antilung serum was prepared by injecting solubilized dog lung antigen into rabbits. The antibody absorbed with kidney antigen is bound to the lung, suggesting the existence of specific antigen in the lung. A linear deposition in the renal glomerulus of -globulin was seen in the lung and kidney of rabbits sensitized with lung antigen. These findings suggested the existence of an antigen common to lung and kidney.
Pulmonary lesions of systemic lupus erythematosus (SLE) were examined using chest X-ray film and studied clinically from various lines of investigation. 1) As intrathoracic lesions of SLE, there were 7 pleurisy cases (10%), 18 pulmonary fibrosis cases (24%), 6 pneumonia cases (8%), 2 pulmonary mycosis cases (3%) and 2 pulmonary tuberculosis cases (3%). 2) A streaky pattern of the chest X-ray film in pulmonary fibrosis of SLE was most common with nodular, reticular and cystic pattern occurring seldom. 3) The distribution of pulmonary fibrosis of SLE was predominant in the middle-lower lung field. 4) Pulmonary fibrosis of SLE did not correlate with antinuclear antibody, rheumatic facter, anti-DNA antibody, anti-lung antibody, serum complement titer and renal involvement. 5) The pulmonary lesions of SLE were compared with those of other collagen diseases (PSS, RA, PM-DM), Pleural lesions and pulmonary infections were observed in many cases while pulmonary fibrosis was seldom obseved and mild when obseved.
The uptake of metallic mercury by lactoperoxidase was measured. The results are described as follows: (1) The uptake of metallic mercury by lactoperoxidase in the presence of hydrogen peroxide was 10 times higher than that in the absence of hydrogen peroxide. (2) The uptake of metallic mercury by lactoperoxidase in the presence of L-dopa in the Lact. -H2O2 system decreased, and decreased further as the concentration of L-dopa increased. (3) The uptake of metallic mercury by lactoperoxidase in the Lact. -H2O2 system showed a tendency to increase, when the concentration of hydrogen peroxide increased. (4) The uptake of metallic mercury by lactoperoxidase increased with an increase in the lactoperoxidase concentration when the concentration of L-dopa and H2O2 in the Lact. -L-dopa-H2O2 system was constant. This uptake was similar in value to the increase changed by increase in the concentration of hydrogen peroxide.
Characteristcs of environmental health conditions in Manabe island were surveyed from the standpoint of public health. The results obtained were as follows: 1. Those employed in primary industries (fishing and farming) are greater in proportion in Manabe Island while those employed in secondary industries are higher in number in Kasaoka City excluding Manabe Island. 2. The population of aged people and the dependence population index in Manabe Island were higher than those in Kasaoka City, a trend thought to be characteristic of isolated islands in the Inland Sea. 3. The ratio of aged people and proportional mortality indicator (PMI) were higher in Manabe Island than that in Kasaoka City. 4. Heart disease, injury of extremities, liver disease, and parasytosis showed a higher incidence in people who belonged to national health insurance in Manabe Island than those of Kasaoka City excluding Manabe Island.
Questionaires on subjective symptoms of fatigue and Donaffio reaction tests were conducted with fishermen and farmers. The results were as follows: 1. Of symptoms described on questionaires, eyestrain, wish to lie-down, loss of memory, and stiff-shoulder were higher in frequency among male fishermen, and stiffshoulder was higher in frequency among female fishermen after work. Those having tired legs' wish to lie-down, and low back pain were of high frequency in both male and female farmers after work. Additionally in female farmers, stiff-shoulder was a frequent complaint. 2. Acute Donaggio values in urine showed a statistically significant increase in male fishermen. Actual Donaggio values and those values corrected for 1024 in urine showed statistically significant increases in female farmers.
Extracorporeal circulation (ECC) without donor blood priming in the pump oxygenator was applied in 23 cases, of which 16 cases needed no blood transfusion (Group Ia) and 7 cases required blood transfusion because of peri-and/or postoperative bleeding (Group Ib). In these two groups, hemodynamic study, blood gas study and chemical determinations were carried out during and after ECC and postoperatively for the initial three days, comparing the results with ten cases (Group II) of usual ECC with 20% hemodilution. The results were as follows: 1) The hematocrit value and hemoglobin level of Group Ia and Ib were significantly lower than those of Group II during ECC, but recovered to the same level as Group II four hours after ECC, and all blood in the pump oxygenator returned. 2) The parameters of hemodynamics, cardiac index (CI), stroke index (SI) and total peripheral vascular resistance (TPR), changed in a similar pattern post-operatively in Group Ia and II, whereas in Group Ib, CI and SI were markedly depressed during the first postoperative day but recovered by blood transfusion. 3) Oxygen transport, oxygen consumption and oxygen extraction ratio showed no significant differences among the three groups from four hours through three days postoperatively. 4) No remarkable differences of erythrocyte 2, 3 diphosphoglycerate (2, 3 DPG) and P50 in vivo were observed during ECC through three days postoperatively in all groups, proving that no hypoxemic state occurred during the period. 5) It was revealed that 2, 3 DPG had statistic linear correlation with P50 in vivo, oxygen transport, CI and venous oxygen content. These facts were thought to occur as a result of a physiological compensatory mechanism for increasing oxygen delivery. 6) Serum LDH increased markedly at the first postoperative day and remained at a high level even seven days after in all groups. In Group Ia and Group II, LDH isozyme 1+2/total increased from the second postoprative day, but LDH isozyme 5/total was slightly elevated as early as during ECC, though at the first postoperative day this fraction recovered to the preoperative level. In Group Ib, LDH isozyme 5/total was observed elevating markedly from four hours after ECC, while LDH isozyme 1+2/total decreased remarkably from the same time, but recovered to the normal range at the first postoperative day. It was thought that peripheral circulatory failure would persist and be improved by blood transfusion. It was concluded that ECC without donor blood prime was secure as the usual ECC with 20% hemodilution from the view points of hemodynamics, blood gas analysis, oxygen delivery and chemical determination. There were many advantages such as saving donor blood and relief of various complications of blood transfusion, when the hematocrit value must be kept above 20%.
The benzethonium chloride method of Iwata for the turbiditometric determination of minute amounts of protein reacted in alkali condition was improved for application to the measurement of urinary protein before and after physical loading. 1) In order to determine minute amounts of protein based on Iwata's method, an improved method was devised. A 0.2ml of urine was added to 3.9ml of a mixture of 33.85mol/l of EDTA 4Na and 0.5mol/l of NaOH. 2) The correlation coefficient of both modified Meulemans method and modified Iwata's one was R=0.96 and the regression equation was y=1.33x+0.17 where x was protein concentration obtained by modified Iwata's method using urine of students before Judo and basketball exercise. The correlation coefficient of both methods was R=0.98 and the regression equation was y=0.91x+0.27 using urine of students agter Judo and basketball exercise. The value obtained by both methods nearl coincided with each other. 3) Turbidity produced by modified Iwata's method was more stable after addition of reagents than that by modified Meulemans method. 4) A minute amount of urinary protein could be preserved and determined by modified Iwata's method up to 4days under freezing at -20°C.
An in vitro chemosensitivity test on permanent cell lines from adeno-, epidermoid and small cell carcinoma of the lung was performed using a double-layered soft agar cloning assay. Drugs tested in the present study were Adriamycin, 40497S, Mitomycin C, cis-Dichlorodiammineplatinum, Methotrexate and Vincristine. Adriamycin, 40497S, Mitomycin C and cis-Dichlorodiammineplatinum showed dose-dependent cytotoxity. Methotrexate and Vincristine did not show a significant cytotoxity when the cell lines were exposed for one hour. Differences in chemosensitivity to the six drugs examined among the three cell lines was relatively small e.g. the lethal dose 90% (LD90) of Adriamycin was 0.19 mcg/ml in adenocarcinoma, 0.46 mcg/ml in epidermoid carcinoma and 0.27 mcg/ml in small cell carcinoma. The lack of a difference in chemosensitivity in vitro might be attributed to the fact that the cell lines had been established from bronchogenic carcinomas which were refractory to intensive combination chemotherapy.
The extent of tumor dissemination at the time of diagnosis is an important predictor for survival of lung cancer patients. In vitro sensitivity to drugs is helpful for successful chemotherapy of advanced lung cancer patients. A major objective of the present study was to clarify the significance of direct cloning assay for the detection of metastatic sites as well as for the selection of sensitive drugs for individual patients. Tumor cell colony growth was evaluated in an enriched double-layered soft agar system. Colonies were successfully grown from 5 of 7 cytology positive effusions, and 4 of 5 histology/cytology positive bone marrow aspirates. Two of 17 histology/cytology negative bone marrow aspirates yielded colonies, whereas 5 cytology negative effusions yielded no colonies. Colony growth was observed in 3 of 5 tumor specimens obtained surgically. For in vitro chemosensitivity, tumor cells were exposed for 1 hour prior to plating. Although colonies were successfully grown from specimens of 5 patients, none of them were sensitive to the drugs tested. Three of the 5 patients were treated with intensive combination chemotherapy including therapy with drugs which had been defined to be resistant in the in vitro chemosensitivity test and no response was observed in these cases. In this sense, in vitro sensitivity correlated with in vivo sensitivity.
Granules of mast cells contain chemical mediators, histamine and SRS-A, and degranulation of the cell is induced by immunological reaction. To identify a relation between immunological reaction mediated by IgE or IgG and surface ultrastructures of mast cells, a probe was constructed for immunoglobulin receptors on the cell surface by covalently coupling antigen to latex particles (0.455±0.010 μm in diameter). Histamine release from mast cells and morphological aspects were examined using a technique for immune-scanning electron microscopy, when antigen was incubated with cells which were passively sensitized with antiserum. The following results were obtained: 1) IgE receptors were detected by the binding of numerous latex particles on the surface of the mast cell, in contrast with control experiments for passive sensitization with or without heat-inactivated anti-serum. Mast cells which showed ridge-and ruffledominant type or raspberry type, were ultrastructurally recognized in the immunological reaction, though control cells were of the microvilli-dominant type. 2) IgG-mediated reaction was recognized on the mast cell by marked release of histamine from cells and by a low degree of staining with toluidine blue. Surface ultrastructure of mast cells was also changed as those of the IgE-mediated reaction. Moreover, degranulation of mast cells was observed at the site of patch or cap formation of the receptor. In conclusion, these findings indicate that rat mast cells have both IgE and IgG receptors. Moreover, mast cells are activated and are induced to release these granules by the immunoglobulin-mediated reaction.
The eutaneous basophil hypersensitivity (CBH) reaction in guinea pig is similar but with some differences to delayed hypersensitivity. Distinctive features of the CBH reaction are that numerous basophils accumulate at the reaction site. Basophilic leucocytes, therefore, may play an important role as a cellular component in CBH. Morphological features of the CBH reaction over time were observed by light and electron microscopy. The following results were obtained: 1) The number of circulating basophilic leucocytes increased during the CBH skin reaction. 2) The CBH skin reaction turned red 12 hr after challenge of the antigen, and reached a maximum in size at 24 hr. Then it disappeared completely at 96 hr. At 20 min. after rechallenge of the antigen, a crimson color was displayed at the site of the CBH skin reaction. 3) Basophils, mononuclear cells and neutrophils were infiltrated into the site of the CBH reaction and these cells reached a maximum number 24 hr after challenge of the antigen. Mast cells and their granules recovered after 96 hr. 4) Degranulation of infiltrating basophils, induced especially with rechallenge of the antigen, was observed as halo-formation and extracellular release of granules by light and electron microscopy. In conclusion, these findings indicate that basophils in CBH reaction are sensitized with homocytotropic antibody, and that mast cells may have a factor for accumulation of basophilic leucocytes at the skin reaction site.
Changes in the structure and function of Ehrlich ascites tumor cell chromatin induced by extracting with 0.15 to 0.6 M NaCl were analyzed. H1 histone was selectively extracted from chromatin by treating with 0.6 M NaCl. The higher the salt concentration that was applied to the chromatin, the larger the amount of proteins that was extracted, and the higher the template activity and micrococcal nuclease sensitivity that were attained. The chromatin reconstituted from H1 histone and H1-depleted chromatin showed reduced template activity and reduced nuclease sensitivity. The size of nuclease-digested DNA fragments (nucleosome monomer) were identical regardless of the depletion of H1 histone and some non-histone proteins.
Simian virus 40 (SV40) chromatin and mature virion were extracted from formaldehyde-fixed SV40-infected CV-1 cells. Mature virion was eliminated from SV40 chromatin by sucrose density gradient centrifugation. Replicating and nonreplicating SV40 chromatin were separated on CsCl/ guanidine-HCl density gradients. Replicating SV40 chromatin was lower in density than the nonreplicating form. The analysis of chromatin proteins by polyacrylamide gel electrophoresis and silver staining revealed that replicating chromatin has less H1 histone than the nonreplicating form.
H1 histone was selectively extracted from calf thymus chromatin by treating with 0.6 M sodium chloride. The H1-depleted chromatin showed no activity for phosphorylation or dephosphorylation of H1 histone. Chromatin was reconstituted from the H1-depleted chromatin and H1 histone, which was extracted from calf thymus chromatin and purified after phosphorylation at serine 37 by cyclic AMP-dependent protein kinase. The rate of phosphorylation of H1 histone at serine 37 was estimated to be over 73%. The effects of phosphorylation of H1 at serine 37 in chromatin on structure and function of chromatin were studied using the reconstituted chromatin. When sodium chloride concentration in the reaction medium was increased from 0 to 80 m M, chromatin showed increased template activity and reduced nuclease sensitivity. The chromatin reconstituted from H1 and the H1-depleted chromatin showed reduced template activity and reduced nuclease sensitivity. The phosphorylation of H1 histone at serine 37 had no effect on template activity or sensitivity to micrococcal nuclease of the reconstituted chromatin.
The incidence of abnormal spermatozoa in normal and dichlorvos-treated Armigeres subalbatus was examined morphologically by electron microscopy. The mosquitos used in this study had been maintained in the laboratory for 160 generations after being collected in Kanagawa prefecture. For control mosquitos, ten seminal vesicles were removed from male adults 3 days after emergence, and sectioned at the middle. The numbers of all normal and abnormal spermatozoa in one side of the double-lumen of a cross-section were counted except for head nucleus parts, tail ends, and tangential sections which could not be identified as normal or abnormal. Normal spermatozoa had 1 flagellum and 2 mitochondrial cords, but abnormal ones had unusual numbers of them. Control mosquitos showed a 4, 37% incidence of abnormal spermatozoa as averaged from data on 10 seminal vesicles. For the insecticide treatment, living 4-th stage larvae which were exposed to 0.201 ppm of dichlorvos (DDVP) for 24 hours were put back into normal water without DDVP. The seminal vesicles from 10 male adults 3 days after emergence were examined likewise to control ones. The count was taken from both sides of the double-lumen of a cross-section. The average incidence of abnormal spermatozoa treated with DDVP was 19.1% which was 4.4 times higher than in control mosquitos. The total number of spermatozoa decreased in vesicles which were treated with DDVP. A large number of abnormal spermatozoa consisted of 1 flagellum and 1 or 3 mitochondria. The wall of seminal vesicles treated with DDVP became thicker than that of controls, and contained many vacuoles in the cytoplasm.