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The in vivo effects of human placental extract (1-4 ml/kg) on hepatic lipid peroxidation, blood and liver glutathione (GSH) levels and several enzymes associated with the antioxidant defence mechanism; i.e., catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase, together with some blood biochemical responses were investigated in rats. At an optimal dose level (4 ml/kg), a single acute intraperitoneal administration of the extract caused a significant enhancement (49.9%; p < 0.001) of lipid peroxidation with a decline in GSH level both in blood (45.1%; p < 0.001) and liver (61.0%; p < 0.001) in comparison to control animals. Activities of catalase, glutathione peroxidase and glutathione reductase were inhibited in a dose-responsive way by the treatment with the extract which also increased the activity of glutathione S-transferase in a dose-dependent manner. The extract was found to be hepatotoxic in terms of elevation of serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, serum lactate dehydrogenase and blood methemoglobin concentration. Results of this study suggest the adverse consequences of the administration of the extract due to its substantial ability to alter normal cellular processes.

Antitumor activities of five platinum analogs, including cisplatin, carboplatin, 254-S, DWA2114R, and NK121, were compared using five human lung cancer cell lines and 19 tumor specimens obtained from lung cancer patients. The antitumor activity was evaluated by determining the ratio of the maximum tolerated dose of each drug to the 70% tumor growth inhibitory concentration in a colony assay. Cisplatin was the most potent agent, followed by 254-S and carboplatin. DWA2114R and NK121 were less potent than cisplatin and 254-S. Cross-resistance to adriamycin was also investigated using an adriamycin-resistant small cell lung cancer subline, SBC -3/ADM30. SBC-3/ADM30 was 1.7- to 4.0-fold more resistant to cisplatin, carboplatin, NK121, and DWA2114R, than was the parent line, SBC-3, and the subline was 2.0-fold more sensitive to 254-S. Using SBC-3, in vitro combination effects of etoposide and cisplatin, carboplatin, or 254-S were evaluated by the median-effect principle. Synergism was noted when cisplatin and etoposide were combined at a fixed molar ratio of 1:1. Combination of carboplatin and etoposide showed an additive effect. The combination of 254-S and etoposide was antagonistic at low concentrations, but was markedly synergistic at higher concentrations. These data suggested the efficacy of 254-S in the treatment of lung cancer.

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.

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).

The serum and urinary ferritin levels in 52 RA patients were measured by the 2-site immunoradiometric assay method. Serum ferritin levels in RA patients correlated with C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) but not with serum iron levels and hemoglobin concentrations, although they were within the normal range. High serum ferritin levels were associated with sera with hyper gamma-globulin and rheumatoid factors. In sequential studies, serum ferritin changed in parallel with ESR, CRP and disease activity in a majority of the patients. The urinary ferritin levels and u/s ratios in some RA patients were higher than control values. Higher values were found particularly in the group of patients under gold therapy but not in groups under other treatments.

Hepatitis B core antigen (HBc Ag) and hepatitis B surface antigen (HBs Ag) were detected in the liver tissue of a patient with chronic aggressive hepatitis by the immunofluorescent complement technique. The presence of anti-HBc was examined by the same method in 67 human sera previously tested for HBs Ag, anti-HBs and s-GPT levels. HBc Ag was localized mainly in the nucleus and sometimes in the cytoplasm of the hepatic cells. HBs Ag was found only in the cytoplasm. The focal area of HBc Ag positive hepatic cells seemed to correspond to the HBs Ag positive cells. Double staining demonstrated the simultaneous presence of HBs Ag and HBc Ag in individual cells. Anti-HBc positive serum was found in 46 (68.7%) cases. Forty-eight (71.6%) indicated a combination of HBs Ag and anti-HBc.

Phosphate-binding protein(s) was found in the inner mitochondrial membrane of calf heart by Sephadex G-200 and G-25 gel filtration. The binding activity was inhibited by N-ethylmaleimide and competed by a large amount of cold phosphate. The amount of phosphate bound to the fraction was 29 nmoles per mg of protein. Affinity chromatography with phosphate-bound Sepharose 4B confirmed the presence of phosphate-binding protein(s) in the active fraction of mitochondrial membrane fractionated by gel filtration.

Every cytologist in biology or medicine knows the so-called "Golgi-Complex", but no cytologist can state exactly the structure and the function of this complex. Nevertheless, in the last six years this" Golgi complex" in about 100 different cells has been seen in the electron microscope. That is the reason I have tried to make a comparative study of these fields. I would like to give a short review of this investigation here, having been kindly invited by Prof. S. SENO.

With the purpose to study sideroblasts as a means of diagnosing blood diseases and to pursue the metabolism of non-hemin iron in erythroblasts we investigated sideroblasts (erythroblasts containing iron granules stainable by Prussian blue) under various erythropoietic conditions in the human and rabbits, and obtained the following results: 1. In blood diseases the proportion of sideroblasts in the case of low erythropoietic condition is higher and in the case with accelerated erythropoietic condition and of iron deficiency it tends to be lower than that in normal persons. Further, obtaining sideroblastogram and sideroblast ratio (S. r.) from the classification of Types I, Ⅱ, and Ⅲ according to the iron granule content, it has been proven that abnormal conditions can be clearly distinguished from the normal, indicating that sideroblasts are closely associated with erythropoietic function. This is proven to be a far superior method for the diagnosis as well as for the prognosis of blood diseases than the determination of serum iron. 2. In experimental anemic rabbits the relationship of sideroblasts to the condition of erythropoietic function is still more clearly recognized, and it has been found that variations in the sideroblast count is dependent upon the condition of the equilibrium between the iron supply from serum iron and the iron utilization controlled by the erythropoietic function. 3. In addition, in the iron-treated rabbits under various erythropoietic conditions we have been able to confirm that there are a certain mechanism and a limitation to the iron intake by erythroblasts, and that erythroblasts take essentially three steps of metabolic processes, namely, intake, retention, and utilization of iron almost simultaneously, in the latter half of the maturation stage.

Affinity electrophoresis of human alpha-fetoprotein (AFP) in amniotic fluid from pregnant women between 6 to 42 weeks of gestation and in the serum of a yolk sac tumor was performed with concanavalin A (Con A), lentil lectin (LCA), erythroagglutinating phytohemagglutinin (E-PHA) and Allomyrina dichotoma lectin (allo A). Separated AFP bands were detected by sensitive antibody-affinity blotting. In the first trimester, amniotic fluid AFP showed elevated percentages of Con A-nonreacting AFP (AFP-C1) and LCA weakly-reacting AFP (AFP-L2) as previously reported. Additionally, high percentages of E-PHA strongly-reacting AFP (AFP-P5) and E-PHA-reacting AFP (AFP-P4) were observed. E-PHA-nonreacting AFP (AFP-P1), E-PHA weakly-reacting AFP (AFP-P3f), allo A-nonreacting AFP (AFP-A1) and asialo-AFP, AFP-A1 s, were present only in amniotic fluids from 6 to 17 weeks of gestation. With advancing gestation, percentages of AFP-C1, AFP-P4 and AFP-P5 decreased and AFP-L2, AFP-P3f, AFP-A1, and AFP-A1 s disappeared by the end of 18 weeks. The glycoforms of serum AFP of the yolk sac tumor resembled those of amniotic fluid AFP in the early gestational stages.

We studied the association of fatty liver with subcutaneous and visceral obesity in 46 male and 36 female patients with body mass index (BMI) over 22 kg/m2. The correlation coefficient between the ratio of the visceral adipose tissue to the subcutaneous adipose tissue (V/S) and the computed tomography (CT) number of the liver was -0.299 (P < 0.05) and that between the V/S ratio and the ratio of the CT number of the liver to that of the spleen (CT-L/CT-S) was -0.335 (P < 0.05) in the males. Partial correlation analysis after making correction for BMI showed an increased correlation coefficient of -0.485 (P < 0.05) between the V/S ratio and the CT-L/CT-S ratio in the males. The odds ratio in the males for CT-L/CT-S below 1.0 and V/S above 1.0 was 3.25 with a 95% confidence interval of 1.02 to 9.39. No such association between the V/S ratio and the CT-L/CT-S ratio was present in the female patients. Multiple regression analysis with serum level of alanine aminotransferase, a marker of fatty liver, as an independent variable revealed a partial regression coefficient of -17.7 for CT-L/CT-S (P < 0.05) in the males and -21.7 (P < 0.05) in the females, validating the CT-L/CT-S ratio as an index of fatty liver. The results indicate the association of fatty liver as determined by the CT-L/CT-S ratio with visceral obesity in males.

Marked IgE-mediated histamine release from rat mast cells sensitized in vitro with mouse antiserum occurs in the presence of added Ca++ and phosphatidylserine (PS), although a considerable degree of antigen-induced histamine release which may utilize intracellular or cell-bound calcium is also observed. The decay in the responsiveness to Ca++ of the sensitized cells stimulated by antigen in Ca++-free medium in the presence of PS is relatively slow, and maximum release is produced by Ca++ added 1 min after antigen. Histamine release also occurs when Ca++ is added after PS in the absence of antigen to the sensitized cells suspended in Ca++-free medium. Unlike the antigen-induced release, the intensity of this non-antigen-induced release varies depending on both mast-cell and antiserum pools. A heat-labile factor(s), which is different from antigen-specific IgE antibody and is also contained in normal mouse serum, is involved in this reaction. In the antigen-nondependent (PS + Ca++)-induced release, no decay in the responsiveness to Ca++ is observed after PS addition. Both the antigen-induced and non-antigen-induced release are completed fairly rapidly and are dependent of temperature, pH and energy.

For the purpose to reveal the role of R.E.S. for hemopoiesis and antibody formation, the R.E.S. of rabbits were severely blocked by the repeated intravenous injection of a vast amount of India ink, reaching 200 to 250 cc in total and the development of anemia and antibody formation by challenging egg albumin were observed while referring to the histologic changes in bone marrow, spleen and lymph nodes. The results were as follows: 1. The repeated intravenous injection of a vast amount of carbon particles induced a severe anemia. The anemia was always normo- or hyperchromic, showing not any disturbance in iron metabolism or hemoglobin formation. The data suggested that anemia is due to the arrest of reproduction of erythroblast or differentiation of the stem cells to erythroblasts, but not due to inhibition of the iron metabolism. 2. R.E.S. had no relation to the proliferation or the differentiation of granulocytes. 3. The functions of R.E.S. related to erythropoiesis and lymphopoiesis are affected by blocking independently of its phagocytic potency. In spite of a severe anemia, the phagocytic potency of R.E.S. could never be lowered and liver and spleen grew much larger in size and weight, showing that the phagocytic ability of R.E.S. is extremely resistant against such a blocking. 4. The serum antibody titer proved to be at the normal level, and no change of the antibody production in spite of heavy blocking of R.E.S. with India ink.

Almost all the umbilical lymphocytes showed more extensive blast cell formation than that of their mother's lymphocytes with PHA. Pathological conditions of mother in pregnancy and labor such as anemia, gestational toxicosis, difficult labor and asphyxia of babies, inhibited the normal response of both maternal and umbilical lymphocytes to PHA.

To investigate the role of vitronectin in the progression of diabetic nephropathy, plasma concentrations of vitronectin were measured by enzyme-linked immunosorbent assay in patients with diabetes mellitus and compared with normal control subjects. In diabetic patients with normoalbuminuria and microalbuminuria, plasma concentrations of vitronectin were significantly higher than those of control subjects. Plasma concentrations of vitronectin in diabetic patients with chronic renal failure were significantly lower than those with normal renal function. There was a significant positive correlation between plasma concentration of vitronectin and blood platelet counts. In the early stage of diabetic nephropathy, vitronectin may be increased caused by synthesis from activated platelets. With progression of diabetic nephropathy, plasma vitronectin may be decreased because of accumulation in sclerotic glomeruli and arteriosclerotic lesions. In conclusion, the plasma concentration of vitronectin appears to be an important marker for the progression of diabetic nephropathy.

To identify diffuse mucosal changes which may precede the development of colorectal cancer and a possible indicator for detecting high-risk populations, we immunohistochemically studied cell-cycle events in crypts of normal-appearing rectal mucosa of patients with colorectal adenoma and cancer using an in vitro labeling method with bromodeoxyuridine (BrdU). Biopsy specimens of endoscopically normal-appearing rectal mucosa were obtained during colonoscopy from 20 patients with colorectal adenocarcinoma, 20 with adenoma, and 15 without apparent colorectal diseases. The specimens were incubated with BrdU in vitro, and labeled S-phase cells were identified immunohistochemically using a monoclonal antibody to BrdU. Modification of the BrdU-labeling pattern in the normal appearing rectal mucosa, such as the presence of BrdU-labeled cells at the mucosal surface or in the upper one-fifth of the crypt column, was observed in 15 of the 20 patients with adenocarcinoma, 17 of the 20 patients with adenoma and 6 of the 15 controls. This upward shift in the frequency of proliferating cells in the crypt was significantly higher in the patients with colorectal adenoma and cancer than in the controls, and may be used to identify subjects at high risk for colorectal cancer.

The enzymatic conversion of azathioprine to 6-mercaptopurine was detected at pH 6.5 with rat liver supernatants, although the non-enzymatic reaction predominated at pH 7.0 and 7.5. Glutathione S-transferase may catalize this conversion. Activities of the enzyme in liver with both zathioprine and 1,2-dichloro-4-nitrobenzene as substrate decreased upon carbon tetrachloride-induced hepatic injury. These results may explain an ineffectiveness of azathioprine in patients with severe hepatic damage.

The cartilage-synovium junction of knees afflicted with rheumatoid arthritis was observed light microscopically using formalin-fixed, decalcified and immunohistochemically stained tissues. Decalcification had little or no influence on immunoreactivity for lysozyme and S-100 protein. All the specimens had pannus formation, which was classified into four types: A) cellular pannus with homogeneous cell pattern, B) cellular pannus of inflammatory cells, C) fibrous pannus with many fibrous bundles, D) fibrous pannus including round cells with scattered fibrous bundles. Type A pannus may be responsible for extensive cartilage degradation, and may occur at the first stage of pannus formation. Type B pannus may occur afterwards, and may be followed by type C pannus at a later stage. Type D pannus was found in two out of 19 specimens. Round cells in type D were positive for S-100 protein and lysozyme, and were probably chondrocytes. The findings indicated that chondrocytes were responsible for cartilage degradation and pannus formation.