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The stability of rat (human) CRF in serum, urine and tissue incubation medium was examined using Sephadex gel filtration and CRF radioimmunoassay with anti-rat (human) CRF serum. Human serum after incubation with rat (human) CRF for 1 h at 37 degrees C showed two peaks of CRF immunoreactivity on a Sephadex G-50 fine column. Most of the immunoreactivity coeluted with the rat (human) CRF marker. When rat (human) CRF was incubated with rat liver, kidney or hypothalamus, only 3.1-14.9% of the CRF was recovered at the rat (human) CRF position on gel filtration, and two to four CRF-immunoreactive peaks appeared after the rat (human) CRF marker. When rat (human) CRF was incubated with human urine (pH 6.0) for 24 h at room temperature, one peak of CRF immunoreactivity coeluted with the rat (human) CRF marker on Sephadex gel filtration. The urine extracts of normal rats showed some small peaks of CRF-like immunoreactivity on the Sephadex column, with the main peak appearing after authentic CRF. These results suggest that rat (human) CRF is relatively stable in serum and urine, but is easily degraded by tissue enzymes, with the degraded CRF fragments being excreted in the urine.

A ten year follow-up study was made of 107 hips of 97 patients to find if derotation osteotomy affected the growth of the hip joint. The anteversion angles before and after treatment were measured in four groups classified according to treatment method: Pavlik harness, frog plaster, Colonna operation and derotation osteotomy. The difference in the angle was greatest in the derotation osteotomy group. However, no significant relation between the CE angles and the anteversion angles or the degree of derotation at the time of the follow-up study was found. The author concluded that the growth of the hip is not influenced by the extent of anteversion as much as was previously believed before.

Erythrocytes in human blood stored for 120 days were collected by centrifugation after dispersion in buffered physiological saline. The aged erythrocytes thus collected were incubated with inosine, adenine, glucose or other media, and their shapes and ATP levels were studied by scanning electron microscopy and a luciferine-luciferase method. The aged erythrocytes incubated in a mixture of adenine and inosine markedly regained their ATP levels, and also showed a marked transformation from spiked spherocytes to normal discocytes. Incubation with inosine alone restored ATP levels of the aged erythrocytes to some extent, but did not result in morphological rejuvenation. Incubation in a mixture of citrate and glucose caused morphological rejuvenation, though it restored ATP levels less effectively than incubation in inosine alone. Incubation with adenine alone neither restored ATP levels nor resulted in morphological rejuvenation of the stored erythrocytes.

A high performance liquid chromatographic (HPLC) method with both electrochemical detection (ECD) and ultraviolet spectrometric detection (UVD) was developed for the rapid and simultaneous measurement of estradiol (E2), estrone (E1), testosterone (T), 17 alpha-hydroxyprogesterone (17-OHP) and progesterone (P) in serum. These hormones were extracted with diethylether, and chromatographed on an octadecyl silane-silica (ODS) column with an eluent of a phosphate buffer solution - acetonitrile mixture (volume ratio 49:51). Estrogens were detected by ECD at +1.0 V vs. Ag/AgCl, and other hormones by UVD at 242 nm. With this method, the simultaneous determination of sex steroid hormones could be performed within approximately two hours with high precision. The hormones of 34 patients (39 menstrual cycles) undergoing human menopausal gonadotropin (HMG)-human chorionic gonadotropin (HCG) therapy were measured. It was concluded that the switch from HMG to HCG should be performed when the E2 level reaches 400 pg/ml for ovulation and 800 pg/ml for pregnancy. The occurrence of ovarian hyperstimulation syndrome (OHSS) can be predicted when the P level rises above 30 ng/ml on the 7th day after the switch. Moreover, conception may be indicated when the P level does not increase from the 7th to 14th day after the switch. In this way, this method proved to be useful for the monitoring of HMG-HCG therapy.

Candida albicans-induced histamine release from basophils was studied in 54 patients with bronchial asthma in comparison with the release caused by house dust and anti-IgE. The release of histamine induced by C. albicans and that induced by house dust were closely related to the serum levels of specific IgE antibodies as expressed by RAST scores. A correlation of C. albicans-induced histamine release with the release caused by anti-IgE was not generally observed. On the other hand, a close correlation was found between house dust- and anti-IgE-induced histamine release. It was suggested from these results that the differences between C. albicans- and house dust-induced histamine release might be due to the different antigenicity of the two allergens.

Problems with infusion therapy for correcting fluid and sodium imbalance in decompensated liver cirrhosis (DLC) were investigated by establishing the safety zone of Talbot et al. for parenteral fluid therapy in 4 DLC patients infused with over 900 ml of fluid each day for at least 9 days. The safety zone was different in each case. The safe infusion volume decreased and the safe electrolyte concentration shifted to a lower osmolality when there was ascites with renal failure than ascites without renal failure. Infusion therapy was performed without deterioration of the water and sodium balance in those patients whose infusion volume and fluid osmolality were in the safety zone. In contrast, ascites retention increased and peripheral edema appeared in patients whose infusion volume and osmolality were out of the safety zone. Therefore, the safety zone should be determined repeatedly during infusion therapy.

Under various conditions of culture and carcinogen treatment, the transformation of liver cells by 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) was studied. Primary liver cell (PLC) cultures from adult male rats and co-cultures with PLCs of ARL-D8 cells of a liver epithelial-like clear cell line from adult female rats were treated with 0.24 mM 3'-Me-DAB for 6 days. Four of 8 carcinogen-treated PLC cultures contained cells with marker chromosomes, and 3 of the 8 cultures contained gamma-glutamyltranspeptidase (GGT)-positive cells. Three of 5 carcinogen-treated co-cultures contained cells with marker chromosomes, and 2 of the 5 co-cultures contained GGT-positive cells. Pure cultures of ARL-D8 cells were treated for 6 or 12 days with 3'-Me-DAB (0.24 mM)-containing-medium perfused through the liver of adult male rats in situ. In the 6-day treatment, none of 5 carcinogen-treated cultures showed chromosomal abnormality or cytochemically exhibited GGT activity. However, in the 12-day treatment, 2 of the 5 carcinogen-treated cultures contained cells with marker chromosomes, and 2 of the 5 cultures contained GGT-positive cells. None of the control cultures exhibited chromosomal abnormality or GGT-positive cells. In summary, transformation markers increased in ARL-D8 cells when they were co-cultured with PLCs.

The onset of beta-methyl-digoxin action was investigated by the potentiation of the adenosine response in guinea pigs and rats, and compared with that of digoxin and dipyridamole. A number of i.v. infusions of adenosine were given to determine the mean control adenosine response and its 95% confidence limits. After oral administration of the drugs, successive infusions of adenosine were continued until a drug-induced potentiation of the adenosine response was observed. The time of appearance of the potentiated adenosine response was marked as the onset of action of the drugs. The onset of action in guinea pigs was 9 to 12 min for 0.2 to 0.4 mg/kg of beta-methyl-digoxin, 90 to 100 min for 0.2 mg/kg of digoxin and 25 min for 5 mg/kg of dipyridamole. The maximal potentiation was 48.8 to 53.8% at 18 to 21 min for beta-methyl-digoxin, 74.5% at 130 min for digoxin and 74.8% at 80 min for dipyridamole. Adenosine infused i.v. into rats produced heart block, as in guinea pigs. However, in rats, the adenosine response was not potentiated by beta-methyl-digoxin and digoxin. Dipyridamole at a dose as high as 200 mg/kg produced 25.8% potentiation at 36 min after oral administration to rats.

The changes in opioid receptors (Op-R) caused by repeated administration of pentazocine and the effect of concomitant clonidine were investigated. Binding of [3H] naloxone was markedly decreased in the absence of Na+, but was increased in the presence of Na+ in the diencephalon-mesencephalon of chronic pentazocine-treated rats. No significant changes were observed in the cerebral cortex of pentazocine-treated rats. The pentazocine-induced changes in Op-R were abolished by the concurrent use of clonidine, an alpha-adrenergic agonist, which has been shown to relieve the withdrawal symptoms of morphine. This result indicated that the behavioral action of clonidine can also be observed at the Op-R level.

The differences in physiological and safety conditions of one-man buses and two-man buses were examined from the view point of occupational fatigue. This survey consisted of a work load study which included a time study, study of subsidiary behavior, auditory task, memory test, Galvanic Skin Response (GSR) and physiological function tests and a self-administered questionnaire which involved items concerning safety and subjective fatigue complaints. The visual and postural restrictions in the one-man bus were greater than in the two-man bus. The mental capacity of the one-man bus drivers was found to be less. Greater mental fatigue and stress were observed in the one-man bus. More subjective fatigue complaints were observed in the one-man bus. More cases of near accidents were observed in the one-man bus. From these results it was concluded that the one-man bus caused bus drivers a greater mental and physical work load.

Levels of plasma cyclic AMP, serum immunoreactive insulin (IRI), serum c-peptide immunoreactivity (CPR) and blood sugar (BS) were determined 0, 15, 30, 45 and 60 min after a glucagon injection (0.01 mg per kg body weight) in normal controls, patients with acute hepatitis and liver cirrhosis. Plasma cyclic AMP responses to glucagon in liver disease patients varied widely in peak value, and only in patients with fulminant hepatitis and decompensated liver cirrhosis with poor prognosis was the response suppressed. The peak response of BS was found significantly later in liver cirrhosis patients than in normal controls. IRI and CPR responses to glucagon were lower in acute hepatitis patients than in normal controls and liver cirrhosis patients. IRI levels and their sum were also lower in acute hepatitis patients, although CPR levels were not significantly different. Thus, the ratio of the sum of CPR from 0 to 60 min to that of IRI was significantly higher in acute hepatitis, indicating impaired pancreatic secretion of insulin to glucagon stimulation as well as increased uptake of insulin by the liver in acute hepatitis.

In iron-induced epilepsy, partial section of the corpus callosum resulted in a decrease in rats showing dominant spike activity on the side of secondary focus and an increase in rats showing almost equal spike activity on the sides of primary and secondary focus in electrocorticograms (ECoGs). Changes in ECoG spike frequencies caused by sectioning the corpus callosum included an increase in unilateral spikes on the side of primary focus, a decrease in unilateral spikes on the side of secondary focus, and an increase in bilateral spikes appearing almost synchronously on both sides.

Polyamines have a close relationship with rapid cell proliferation. We measured polyamine levels in amniotic fluid, maternal plasma and urine during normal pregnancy. Plasma putrescine, spermidine and spermine gradually increased in the third trimester and reached the highest concentration at the end of pregnancy. There was a significant correlation between the level of these polyamines and the level of plasma estradiol and progesterone. In urine, putrescine and spermine increased with the progress of gestation and reached the highest level during the 8th to 10th months of gestation. In amniotic fluid, putrescine and spermidine concentrations were significantly high in the first trimester and decreased in the other trimesters, whereas spermine showed no significant change. Polyamine concentrations in maternal plasma and urine appear to reflect not only fetal metabolic changes but also the metabolic changes of the pregnant women, and to be influenced by several hormones which increase during pregnancy. Polyamines in amniotic fluid mainly reflect activated fetal metabolism and may be useful as biochemical indicators of fetal growth.

We examined the activity of peripheral blood monocytes in patients with autoimmune hemolytic anemia (AIHA) using an in vitro assay of monocyte-macrophage interaction with erythrocytes and an antibody-dependent cell-mediated cytotoxicity (ADCC) assay. The monocytes of AIHA patients in the hemolyzing period phagocytized autologous sensitized red cells and anti-D coated red cells more avidly than normal control monocytes. There was no significant relationship between phagocytic activity and ADCC activity. The activated monocytes phagocytized autologous sensitized red cells, but had no ADCC activity in a short time 51Cr release assay. Phagocytic activity of the patients' monocytes against autologous erythrocytes rapidly decreased after treatment with prednisolone even though the red cell sensitization with antibody remained almost the same as during the hemolyzing period. We postulated that the activation of monocytes in AIHA was due to the "arming" effect of anti-erythrocyte antibody, but we think that other mechanisms may also be involved in the activation of monocytes.

The effect of a lymphotoxin-like substance, OH-1, released by human acute lymphatic leukemia BALL-1 cells, on metastatic tumor proliferation was investigated in BDF1 mice with transplanted Lewis lung carcinoma cells. Mitomycin-C, cyclophosphamide and adriamycin were used as control agents. The effect of OH-1 on metastases, as determined by comparison of the numbers of pulmonary nodules and by 3H-thymidine labeling indices, was significant. Also, investigation of the effect of OH-1 on host immunity showed that, while the control preparations had considerable side effects, immunodepression and emaciation were not noted with OH-1. As to direct cytotoxicity, OH-1 is principally cytostatic in activity and effects cell progression delay in both the G1 and G2 phases.

The distribution of ferritin has been studied in many tissues, but has not yet been established on the cellular level. We investigated the cellular distribution of ferritin in the liver, spleen and bone marrow using the immunoperoxidase method, and compared it with that of hemosiderin. We also examined changes in the distribution of these proteins after phlebotomy and iron overload. In normal rats, ferritin was seen in centrilobular hepatocytes, Kupffer cells, macrophages in the red and white pulp of the spleen and central macrophages in bone marrow. Hemosiderin was observed almost exclusively in the red pulp and partly in tangible body macrophages of the white pulp. After phlebotomy, neither ferritin nor hemosiderin were detectable in these cells except for ferritin-positive cells in the white pulp, which showed little change after either phlebotomy or iron overload. In iron overloaded rats, both ferritin and hemosiderin increased in hepatocytes and reticulo-endothelial (RE) cells. Ferritin-positive cells in the liver were mainly located in the periportal area. These results indicated that hepatocytes and RE cells except for those in the white pulp may play an important role in iron storage, and that ferritin-positive cells in the white pulp may have a function other than iron reserve. They also suggested that the zonal distribution of ferritin-positive hepatocytes may be due to microcirculation in the hepatic lobules.

Twenty-five patients (30 hips) have had a total hip replacement using the cup supporter developed in our department. This report describes the follow-up findings on these patients. The mean period after hip replacement was 2 years and 8 months (range from 6 months to 6 years). The cup supporter was used in patients with rheumatoid arthritis with acetabular protrusions, central migration of the prosthesis after hemi-arthroplasty, revision operation for a defecting acetabular floor, primary acetabular protrusions (including osteoarthritis with acetabular protrusions) and traumatic fracture-dislocation of the hip. In five cases, autograft of bone was used in addition to the cup supporter for reinforcement of a thin acetabular floor. This combination appeared to provide good clinical results. The cup supporter was of value in revision operations due to loosening of the acetabular cup with severe acetabular protrusions.

Fulminant hepatic failure (FHF) was produced in rats with intraperitoneal injection of D-galactosamine. Control rats received only physiological saline. 15N-leucine (200 mg/kg of body weight) was injected into the rats via the tail vein. Arterial blood was drawn before and 5, 15, 30 and 60 min after the injection of 15N-leucine. 15N-amino acids were determined quantitatively by gas chromatography and mass spectrometry. The plasma 15N-leucine level decreased logarithmically in the same manner in both groups. This result suggests that leucine is mainly metabolized in extrahepatic tissues. The incorporation of 15N into plasma isoleucine and valine was not significantly different between the groups. Plasma alanine and glutamine concentrations increased in controls and decreased in FHF rates after the injection. The incorporation of 15N into plasma alanine in rats with FHF was significantly later than in controls. This result may suggest that undergoing hyperammonemia causes to form more glutamine from glutamate in extrahepatic sites as the same manner as for chronic hepatic failure. Additionally, insulin levels increased temporarily after the injection of leucine in both groups. This increase may play a role in the decrease in plasma isoleucine and valine concentrations after injection of leucine.

Three linear plots by which the liver's maximum removal rate (Rmax) of indocyanine green (ICG) and the Michaelis constant (Km) can be calculated were compared in a microcomputer simulation study. The widely-used Lineweaver-Burk plot (1/V vs. 1/S; V, ICG initial removal rate (mg/kg/min); S, ICG loading dose (mg/kg] presented the greatest bias and variance. There was no remarkable difference in bias between the S/V vs. S plot and the V vs. V/S plot, but the latter possessed a smaller variance. Therefore, the V vs. V/S plot was considered the best for estimating Rmax. The best combination of three ICG loading doses was 0.5, 2, and 5 mg/kg. This combination was selected by comparison of the Rmax estimated from three points with that estimated from six points (0.5, 1, 2, 3, 4 and 5 mg/kg).

A high performance liquid chromatographic (HPLC) method with electrochemical detection (ECD) was developed for the simultaneous measurement of estrone, estradiol, estriol and estetrol in serum. These hormones were extracted with diethylether, chromatographed on an silica-octadecyl silane (ODS) column with an eluent of phosphate buffer solution-acetonitrile-methanol (volume ratio 152:85:40), and detected by ECD at +1.0V vs. Ag/AgCl. In comparisons between the values measured by this method and radioimmunoassay, significant correlations were noted for estrone (r = 0.759, p less than 0.01), estradiol (r = 0.816, p less than 0.001) and estriol (r = 0.830, p less than 0.001). In clinical applications of this method, differences between cases of the normal and the anencephalic pregnancy in the thirty-eighth week of gestation were distinct not only in the individual estrogen, but also in the profile analysis of estrogens. With this method, all 4 serum estrogens above approximately 500 pg/ml could be measured within 2 h, and the method seemed to be clinically applicable.