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Micromethods for estimation of bilirubin uridine diphosphate-glucuronyl and -xylosyl transferases in liver are described. With these methods, a liver specimen as small as 10 mg is sufficient for assay of either enzyme activity and 15 mg is sufficient for assay of both enzymes. Normal values for livers with minimal elevation of SGOT and minimal histological change were 0.260-0.400 U/g protein for glucuronyl transferase and 0.142-0.302 U/g protein for xylosyl transferase.

The case of a patient with repeated attacks of collapse induced by sublingual isosorbide dinitrate is reported. The patient was an 81 year-old female who was admitted to Yura Hospital because of attacks of precordial pain. Several minutes after the sublingual administration of isosorbide dinitrate (10 mg) for an anginal attack, she developed a sensation of general weakness, and thereafter because unconscious. Arterial blood pressure fell and became unmeasurable. Electrocardiograms recorded during the syncopal attack showed sinus tachycardia and significant elevation of ST-segment in right precordial leads. In response to a drip infusion of noradrenaline, arterial blood pressure returned to normal with recovery of consciousness. Two similar syncopal attacks induced by sublingual isosorbide dinitrate occurred in the next three days. These attacks were not due to augmentation of the vagal reflex. Decrease of venous return probably was the primary etiological factor.

Ninety-nine gastric cancer patients initially received levamisole at a daily dose of 150 mg for three consecutive days before operation. This therapy was repeated fortnightly (3-day administration followed by 11-day withdrawal period) for more than one month as long as possible and the survival rate up to 18 months was compared with thas of control patients. The 18 month survival rate of advanced Stage IV patients was significantly higher in patients receiving levamisole than that of control patients. The effects of levamisole in cases of advanced cancer have been discussed in relation to the literature available.

In view of comparatively-high incidence of papillary, well or moderately-well differentiated, transitional cell carcinomas in urology, and of frequent recurrence of carcinomas after appropriate procedures, we have undertaken the correlative study on urinary cytology with cystoscopy, pathology and the amount of fibrinogen and degradation products of fibrinogen and fibrin (FDP). Cytopathologically, 191 cases were confirmed histologically for cancers out of 442 cases studied cytologically; among these 442 classes IV and V were found in 136 cases, and 99 of 136 cases (72.8%) had cancers. Admittedly, we were unable to suspect tumors cytologically in 24.5% of the cases. As to histological gradings and cytology, 48.7% of classes I and II in well differentiated were found against 52.9% and 57.1% of classes IV and V in moderately-well and poorly differentiated carcinomas, respectively. Urinary FDP assay was quite promising; out of 19 cases with both positive cytology and FDP, 18 (94.7% had carcinomas histologically, whereas 12 of 56 cases (21.4%) with both negative cytology and FDP bore cancers. Subsequently, several proposals for improving the diagnostic accuracy were discussed. We concluded the importance of accumulating more cases by combining several diagnostic procedures, especially among well and moderately-well differentiated carcinomas.

Electron microscope observation of cultured human leukemic B cell, T cell and null cell lines and reverse transcriptase assay of the culture supernatants were all negative for the presence of C-type virus. Bat cell line, which propagates primate C-type viruses well, was cocultivated with the human leukemic cell lines, in the hope of amplification of virus if present. Three weeks after mixed culture, the culture supernatants were again examined for reverse transcriptase activity and the cells were tested for syncytia formation by cocultivation with rat XC, human KC and RSb cell lines. All these tests, except for the positive control using a simian sarcoma virus, were negative, suggesting that no C-type was produced from these human leukemic cell lines.

The effect of various hormones on cytological and biochemical properties of cultured hepatic cells were investigated in order to obtain long-term survival of the hepatocytes with adult liver functions in primary culture. Insulin supplementation of the culture medium enhanced the attachemnt efficiency of cells in primary culture without affecting either maintenance of morphological characters of epithelial cells or retention of liver-specific functions in cultured cells. A combination of dexamethasone and insulin was apparently effective in stimulating the formation of a monolayer of polygonal cells with granular cytoplasm and in maintenance of liver-specific functions for relatively longer periods. Supplementation with either dexamethasone or hydrocortisone alone enhanced tyrosine aminotransferase activities in cultured cells for at least 4 days postinoculation. These steroid hormones also allowed growth of small epithelial cells with clear cytoplasm and maintenance of increased albumin production for 8 days after inoculation. The roles of these hormones in the primary culture of isolated hepatic cells are discussed in the present paper.

The incidences, distribution and histopathological findings of N, N'-dimethylnitrosourea (DMNU)-induced brain microtumors in Sprague-Dawley rats were studied. Subcutaneous injections of DMNU in young adult rats one a week resulted in the induction of 122 gliomas in 38 animals with an incidence of 69% after a time lapse of between 157 and 246 days from the first injection. Of these tumors, 66 were classified as microtumors (diameter less than about 1 mm) by detailed light microscopy observation of serial sections. The microtumors were of 3 types: 55 oligodendrogliomas, 8 astrocytomas and 3 mixed gliomas. As the tumors became larger in size, anaplasia appeared, especially in the central part of the tumors. The microtumors developed randomly throughout the brain. It was concluded that, in adult rat brains, the target cells of DMNU were well differentiated glial cells which had already migrated from the matrix layer.

Normal development of the human stapes footplate was investigated in serial sections by light microscopy. Materials were obtained from 35 Japanese embryos from the 6th to 32nd week of embryonal age. Eighteen embryos up to 16 weeks of age (3.5mm to 105mm in crown-rump length) were examined, focusing particularly on the lamina stapedialis of the otic capsule. The present study showed that primordial formation of the lamina stapedialis appeared in 16mm embryo and that the lamina was completely formed and fused to the base of the annular stapes in a 35mm embryo. In a 50mm embryo, the adult form of stapes was found with a rim and annular ligament. The results, therefore, seemed to essentially agree with the theory of dual origin and development of the footplate proposed by Cauldwell and Anson, and teratogenic agents might affect any stage of the process producing anomalies,

The correlation between the systolic time interval and abnormal contraction in ischemic myocardium was studied in anesthetized open-chest dogs. A strain-gauge was sutured on the surface of the left ventricular wall perfused by the left anterior descending coronary artery (LAD) for measuring segment-length. The left ventricular stroke volume decreased progressively after occlusion of LAD. The left ventricular ejection time (LVET) was progressively shortened in close correlation with the elongation of segment-length at the onset of isometric relaxation in 20 seconds after LAD occlusion when early systolic myocardial contraction and isometric contraction time (ICT) were not affected. ICT was gradually prolonged and closely related with the lengthening of the early systolic segment-length, while LVET recovered toward the control level in spite of further decrease in stroke volume. A close relationship was observed between ICT/LVET and stroke volume (gamma = 0.76, P less than 0.01). The results suggested the possibility that LVET was normalized even when the left ventricular function was impaired, and ICT/LVET ratio was the most sensitive index of LV dysfunction.

Binding of bacterial endotoxin to platelets, erythrocytes, lymphocytes and granulocytes was examined by using diffusion dialysis. Platelets, erythrocytes, lymphocytes and granulocytes were fractionated from normal human blood and the binding of endotoxin (LPS: Lipopolysaccharide of E. coli) to each cell fraction was measured at 4 degrees C and the binding efficiency was expressed as a binding index (%d4degreesC +/- SD). The binding index for each cell fraction was as follows; 10.2 +/- 1.6 for platelets, 1.0 +/- 0.9 for erythrocytes, 4.3 +/- 1.6 for lymphocytes and 10.0 +/- 1.5 for granulocytes (n = 11) respectively. Since a platelet possesses a small cell surface area compared with other cells, it was clear that the endotoxin bound preferentially to platelets in vitro. The binding mechanism to the platelet cell surface was suggested to be direct binding of endotoxin to the receptor on platelet cell membrane rather than through an immunologically activated mechanism.

Endotoxin (lipopolysaccharide, LPS) and LPS antibody in the blood were studied in 61 cases of ulcerative colitis (U.C.) by radioimmunoassay. Lysozyme (LZM) concentration was also studied by the turbidimetric method. As a result, it was found that the blood LPS value as well as serum LZM concentration reflects the clinical observations. The case of endotoxemia in the active phase group showed a positive correlation between the LPS value and LZM concentration. LPS antibody which could not be detected in many cases of the active phase, had a high titer in cases of remission with a long history of the disease. These results would suggest that in U.C. with damaged intestinal mucosal barrier, LPS originating from intestinal flora enters into the blood and aggravates the disease and further that this invading LPS releases LZM into the blood. The same studies were performed on 7 cases of Crohn's disease and the same result was obtained.

Using in vivo and in vitro experimental models, the uptake and excretion of 67Ga-citrate in tumor cells and normal cells were studied. The time-lapse accumulation of 67Ga in the tumor of rats bearing Yoshida sarcoma reached its peak 24 h after the administration of 67Ga and gradually decreased thereafter. However, the excretion of 67Ga from the tumor was less than that from normal lung. For culture cells in vitro, the uptake of 67Ga increased with lapse of contact time between 67Ga and the cells, but there was no distinct difference between the results of tumor cells and normal skin fibroblasts. The excretion of 67Ga from the cells tended to decrease with prolongation of the contact time, the excretion from tumor cell being only about 10% after a contact time of 24 h. This indicated a significant delay in excretion in comparison with that of normal skin fibroblasts. This delay in the excretion of 67Ga may be an important factor in the tumor accumulation of 67Ga.

Lymphocytes were highly purified from synovial fluid and peripheral blood of 10 rheumatoid arthritis patients and assessed for responsiveness to PHA-P and Con-A. In all cases, both synovial and blood lymphocytes showed a marked reduction in response to these mitogens compared with normal blood lymphocytes. The factors responsible for this low T cell responsiveness are discussed.

The covalently closed form of circular duplex SV40 DNA was separated from the open and linear form of SV40 DNA by agarose gel electrophoresis in a large-scale gel system. The closed circular DNA was recovered from agarose gels by re-electrophoresing the gel slices. The recovery of DNA was about 70%. Electron microscopic analysis showed that the recovered DNA did not have doube- or single-stranded breaks. The recovered DNA can be used without further purification for electron microscopy, as a substrate for experiments using restriction endonuclease and as a template for in vitro RNA synthesis.

Euchromatin specimen prepared by the usual method formed large clumps and had various shapes under electron microscopy. A method of separation of the euchromatin specimen into chromatin fractions having relatively homogeneous form was examined and partial characterization of these fractions was carried out. The heavy euchromatin fraction was a large network of thin fibrils (about 100 A in diameter) and various thick fibers. The intermediate euchromatin fraction consisted of relatively homogeneous networks of thick knobby fibers (about 250 A in diameter). The light euchromatin fraction had metworks of thick fibers. These chromatin fractions were quantitatively prepared from sonicated nuclei of mouse ascites sarcoma cells. Twenty-one or twenty-two bands of non-histone proteins besides histones were detected in these chromatin fractions by SDS-polyacrylamide gel electrophoresis. There were significant differences in the electrophoretic patterns of non-histone proteins among these chromatin fractions.

Levels of erythrocyte superoxide dismutase (SOD) activitiy in a sample of Japanese people were determined. Blood samples were taken from new-born infants, preschool children, young and old people who had no apparent diseases and also from three anemic patients. Erythrocyte SOD activities in different age groups had a nearly normal distribution. Females had slightly lower activities than males, although the difference was statistically insignificant. The distributions of SOD activities were 12.6 +/- 2.7 (m +/- SD) unit/mg Hb in young people and 11.4 +/- 3.0 in old people, indicating that erythrocyte SOD activity falls with aging. Because of low concentration of hemoglobin, SOD activities of old people expressed as unit/ml blood were much lower than in young people. Three anemic patients had slightly lower SOD activity.

The effect of Cd2+ on the respiration of rat liver mitochondria was investigated. The uncoupling effect of Cd2+ was partially restored by the addition of Mg2+. The influence of Cd2+ on adenine nucleotide concentrations in the reaction mixture consisting of mitochondria and ATP was also studied using high performance liquid chromatography. In the presence of added Mg2+, a two-fold increase in AMP concentration was brought about by the addition of Cd2+. There was a concomitant decrease in ATP. In the prence of added ADP, an increase in AMP concentration was also brought about by addition of Cd2+. The results are discussed in relation to ATPase and adenylate kinase activity in mitochondria.

A mixture with essential and nonessential amino acids high in branched chain amino acids and low in aromatic amino acids (Fischer solution), and another synthetic mixture of branched chain amino acids containing 3 amino acids associated with the urea cycle (Hep-OU) were infused to control subjects and patients with severe hepatic disease. Alterations in serum aminograms, blood ammonia levels and electroencephalograms following the infusion were studied and compared with those obtained by a commercially available amino acid mixture. Short-term or continuous infusion of a commercially available amino acid solution to cirrhotic patients caused an increase in methionine, phenylalanine and tyrosine and a decrease in branched chain amino acids. These post-infusion results were similar to the patterns seen in hepatic encephalopathy. In cirrhotic patients, infusion of Fischer solution which contains small quantities of methionine and phenylalanine produced an increase in the concentrations of these 2 amino acids, probably because of impaired utilization by the injured liver. No marked alterations in serum aminograms, however, were observed in cirrhotic patients either immediately after, or 3 h after, the end of the Hep-OU infusion. Reduction of methionine, tyrosine and phenylalanine levels and elevation of the molar ratio of (valine+leucine+isoleucine) / (phenylalanine+tyrosine) were significant. The infusion of Hep-OU to patients with liver cirrhosis or subacute hepatitis resulted in clinical and neurological improvements and the restoration of the molar ratio of branched chain amino acids/aromatic amino acids.

The authors attempted to determine if the organic sulfur compounds usually contained in a crude oil could serve as a marker of oil pollution in fish. Eels (Anguilla Japonica Temminck et Schlegel) were maintained in a controlled laboratory environment of water with a suspension of crude oil. Gas chromatography-mass spectrometry and mass chromatography of eel flesh extract showed the presence of organic sulfur compounds of alkyl-(from mono- to pentametyl) benzothiophenes, dibenzothiophene and alkyl-(from mono- to trimethyl) dibenzothiophenes, and other organic sulfur compounds of alkyl-(from mono- to pentamethyl) naphthalenes.

The homovanillic acid (HVA) concentrations in the lumbar cerebrospinal fluid (CSF) were determined in 38 epileptic and 39 control patients. The mean concentration of HVA was 23.9 ng/ml +/- 2.8 SEM for the epileptic group and 30.2 ng/ml +/- 2.1 SEM for the control group, respectively. Thus, HVA was significandly reduced in the patients with epilepsy compared with the controls. The mean HVA in the female patients was higher than in the male patients in both groups but this failed to reach statistical significance. There was no apparent relationship between the degree of reduced HVA concentration and other clinical indexes of the epilepsy (age, type and frequency of seizures, and anticonvulsant medication). For the determination of concentration gradient of HVA three fractions of the spinal CSF were obtained from 11 patients. A pronounced gradient of HVA concentration was found with a ratio of 1 : 1.46 : 1.97 for the first, second and third fractions. This suggests that a standardized conditions for collecting CSF should be employed to study HVA levels in humans.