Evidence for a common binding site for omeprazole and N-ethylmaleimide in subunit A of chromaffin granule vacuolar-type H(+)-ATPase.

Vacuolar-type H(+)-ATPase from adrenal chromaffin granules was found to be sensitive to omeprazole, a known gastric H+/K(+)-ATPase inhibitor, the concentration required for 50% inhibition being 80 microM freshly-prepared and 12 microM acid-treated reagent. ATP and ADP protected the enzyme from inhibition by omeprazole. The activity of the inhibited enzyme was restored by the addition of reduced glutathione.

Inhibition of intracellular transport of newly synthesized prolactin by bafilomycin A1 in a pituitary tumor cell line, GH3 cells.

We have investigated the effects of bafilomycin A1, a specific inhibitor of vacuolar type H(+)-ATPase, on the secretion and intracellular distribution of prolactin in cultured rat pituitary tumor cells (GH3 cells). Pulse-labeling the cells with L-[35S]methionine for 5 min and subsequently incubating in chase medium containing 1 microM bafilomycin A1 showed inhibition of basal secretion of labeled prolactin. The inhibition of the secretion by the drug was clearly observed when it was added within 7.

Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) after chemotherapy in patients with non-Hodgkin's lymphoma; a placebo-controlled double blind phase III trial.

To assess the clinical and hematopoietic effects of rhGM-CSF, a placebo-controlled double blind multicenter phase III study was undertaken in patients with non-Hodgkin's lymphoma receiving cytotoxic chemotherapy. Sixty-two patients who had granulocytopenia (< 1 x 10(3)/microliters) after the first cycle of chemotherapy with cyclophosphamide, adriamycin, vincristine, and prednisolone were enrolled. After the second cycle of chemotherapy with the same regimen, patients randomly received either rhGM-CSF (125 micrograms/m2/day) or placebo for 14 days (rhGM-CSF; 31 patients and placebo; 31 patients).

Immunological detection of F1-ATPase gamma-subunit in rat liver peroxisomes.

The gamma-subunit of F1-ATPase was detected in highly purified rat liver peroxisomes by immunoblot analysis using an antiserum against purified mitochondrial F1-ATPase. Other subunits of F1-ATPase such as alpha, beta and delta were not detected in the peroxisomes. The polypeptide corresponding to the gamma-subunit in peroxisomes was also recognized by affinity purified antibody against F1-ATPase gamma-subunit.

Energy-dependent accumulation of neuron blockers causes selective inhibition of neurotransmitter uptake by brain synaptic vesicles.

The effects of neuron blockers on neurotransmitter accumulation in synaptic vesicles were investigated. Upon addition of ATP, brain synaptic vesicles accumulated chlorpromazine, haloperidol, and propranolol against concentration gradients of more than 100-fold. Bioenergetic analysis indicated that the transmembrane pH gradient (delta pH) established by the vacuolar-type H(+)-ATPase is a direct driving force for these uptakes.

Uptake of the neurotoxin, 4-methylphenylpyridinium, into chromaffin granules and synaptic vesicles: a proton gradient drives its uptake through monoamine transporter.

Energy dependence for uptake of 4-methyphenylpyridinium (MPP+), a neurotoxin causing Parkinsonism-like symptoms, by adrenal chromaffin granule membrane vesicles and brain synaptic vesicles was studied. The compound was actively taken up by the chromaffin vesicles dependent on hydrolysis of ATP with a Km value of 22 microM and maximum velocity of 2.9 nmol/min/mg protein.

[The relationships of sex hormone binding globulin (SHBG) with high density lipoprotein cholesterol and fasting insulin in healthy men].

The relationships of sex hormone binding globulin (SHBG) with high density lipoprotein cholesterol (HDL-C) and fasting insulin (IRI) were investigated in healthy employed men (n = 235). In simple correlation analysis, the SHBG was positively related to total cholesterol and HDL-C, and negatively related to IRI. There were no relationships among SHBG, triglyceride and fasting blood glucose.

[Two cases of intrahepatic biloma during hepatic arterial infusion chemotherapy proved by CT-Scan].

Hepatic arterial infusion chemotherapy using 5-FU is effective for colorectal cancer, but there are increasing reports of complications, for example, liver abscess, bile duct necrosis and sclerosing cholangitis. We encountered two cases of liver cell necrosis around the Glisson's sheath which were discovered by CT-scan. At the first operation, liver metastasis was present and a resection of the main tumor and metastatic lesion was performed.

[Two cases of improved quality of life with intra-arterial infusion of CDDP or unresectable gallbladder and pancreatic cancer].

The prognosis of patients with advanced gallbladder and pancreatic cancers is poor. Intra-arterial infusion chemotherapy with CDDP was imported in patients with unresectable gallbladder cancer (First case) and pancreatic cancer (Second case). The values of CEA and CA 19-9 were reduced after chemotherapy in the first case, but the CA 19-9 level was not reduced in the second case.

Prediction of the ability to purge clonogenic B cell lymphoma from normal BM in vitro by heat: their survival curves correspond to a curve reflecting mortality in humans.

To develop new purging regimens for ABMT the ability to predict potential for purging of tumor cells from BM is important. Since the sensitivity of human B cell lymphoma to hyperthermia is not known, we examined its effect on the growth of B cell lymphoma cell lines (Raji and Daudi) in vitro to evaluate potential for purging clonogenic tumor cells from normal marrow by heat, using a limiting dilution assay to measure log depletion of tumor cells in a 20-fold excess of normal BM. When exposed to heat (42-43 degrees C) for 120 min, both clonogenic Raji and Daudi cells were dramatically reduced (a 4-to-6 log reduction) with time, whereas at 42 degrees C over half and at 43 degrees C 10% of normal granulocyte-macrophage progenitor cells survived for the same time period.

[Surgical treatment of primary cardiac tumors].

Thirty-four cases of primary cardiac tumors, consisted of 11 men and 23 women, aged between 26 and 80 years have been operated on in our institution. A total of 30 myxomas and four malignant tumors were resected. The clinical presentations included congestive heart failure in 25 patients, tachyarrhythmia in 6, chest pain in 4, and embolism in 10.

Characterization of human autoantibodies reactive to gastric parietal cells.

Human sera from autoimmune gastritis patients containing autoantibodies to gastric parietal cells were analyzed by immunological methods. Enzyme linked immuno-sorbent assay demonstrated that all nine sera reacted with pig gastric vesicles enriched in H+/K(+)-ATPase (gastric proton pump). Immunoblotting experiments indicated that the alpha subunit of the H+/K(+)-ATPase was the major antigen in the vesicles with two of the sera reacting strongly.

Effect of granulocyte-macrophage colony-stimulating factor on chemotherapy-induced granulocytopenia in patients with malignancies.

To investigate the effect of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) on cytotoxic chemotherapy-induced granulocytopenia, we performed an open nonrandomized clinical trial in 46 patients with malignancies receiving cytotoxic chemotherapy regimen. Twenty-six patients who received two cycles of the identical chemotherapy regimen and had granulocytopenia less than 1 x 10(9) cells/l after the first cycle of chemotherapy were eligible for this study. They received 60, 125 or 250 micrograms/m2/day of rhGM-CSF randomly.

Gliosarcoma of the posterior cranial fossa: MRI findings.

We report the MR findings of a biopsy-proven gliosarcoma of the posterior cranial fossa. Multiple homogeneously enhancing lesions had shaggy margins and broad-based dural attachments, which may reflect the gliomatous and sarcomatous element of this tumour.

Combination therapy with rhGM-CSF and rhEpo for two patients with refractory anemia and aplastic anemia.

Because GM-CSF possesses burst-promoting activity (BPA) and megakaryocyte colony-stimulating activity (Meg-CSF) as well as stimulating activity on granulocyte-macrophage progenitors, and erythropoietin (Epo) has thrombopoietin-like activity, the combination therapy of GM-CSF and Epo seems to be more effective for stimulating erythropoiesis and thrombocytopoiesis in patients with pancytopenia. For this reason, the combination therapy of recombinant human GM-CSF (rhGM-CSF) and rhEpo was performed in two patients with refractory anemia (RA) and aplastic anemia (AA). Epo-unresponsive anemia was remarkably improved by adding rhGM-CSF to Epo and the effect lasted for 1 1/2 months in a patient with RA, but severe anemia occurred again immediately after the discontinuation of Epo.

Phase II study of recombinant human granulocyte-macrophage colony-stimulating factor in myelodysplastic syndrome and aplastic anemia.

As phase II study, we treated 18 patients with myelodysplastic syndrome (MDS) and 37 patients with aplastic anemia (AA) with recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) for 14-28 days. Administration of rhGM-CSF resulted in a dose-dependent increase in circulating granulocyte counts, which was statistically significant in patients with AA. There were no consistent changes in monocyte and lymphocyte counts.

Secondary structural changes of metmyoglobin and apomyoglobin in anionic and cationic surfactant solutions: effect of the hydrophobic chain length of the surfactants on the structural changes.

Secondary structural changes of metmyoglobin and apomyoglobin were examined in solutions of sodium alkylsulfates with hydrocarbon numbers of 8 and 12, and alkyltrimethylammonium bromides with hydrocarbon numbers of 10, 12, 14, and 16. The relative proportion of alpha-helical structure was estimated by the curve-fitting method of circular dichroic spectrum. The helical proportions of metmyoglobin and apomyoglobin were 82 and 63%, respectively.

Effect of casein, casein phosphopeptides and calcium intake on ileal 45Ca disappearance and temporal systolic blood pressure in spontaneously hypertensive rats.

Paracellular 45Ca absorption and temporal systolic blood pressure (SBP) measurements were recorded in spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats fed on casein (C) and soya-bean-protein isolate (S) diets, containing 20 (H), 5 (H) and 0.5 (L) g Ca/kg. Similar measurements were also taken in SHR rats only fed on C-M and S-M diets supplemented with 30 g caseinophosphopeptides (CPP)/kg.

The role of V-ATPase in neuronal and endocrine systems.

Synaptic vesicles have important roles in the neural transmission at nerve terminals: the storage and the controlled exocytosis of neurotransmitters. At least two different factors are responsible for the concentration process: the vacuolar-type H(+)-ATPase (V-ATPase), establishing an electrochemical gradient of protons, and specific transport systems for transmitters. We will discuss our recent progress on the energy-transducing systems in synaptic vesicles: (1) structural aspects of V-ATPase; (2) energy coupling of transport of transmitters; (3) reconstitution of transporters; (4) effects of neurotoxins and neuron blocking agents; (5) function of synaptic-vesicle-like microvesicles from endocrine tissues.

In vitro purging of clonogenic leukemic cells from human bone marrow by heat: simulation experiments for autologous bone marrow transplantation.

In order to apply a simple purging method by heat to autologous bone marrow transplantation (ABMT), we have revaluated the ability to purge clonogenic leukemic cells from the simulated marrow mixture of normal marrow cells and leukemic cell lines (HL-60, Molt-3 and HEL) in vitro by heat, using two different clonogenic assays for normal granulocyte-macrophage progenitors (CFU-GM) and leukemic cell lines. It appeared that in vitro hyperthermia (42 degrees C for 120 min) is able to selectively remove clonogenic leukemic cells from simulated tumor cell-normal marrow mixtures even when leukemic cell concentrations are increased up to 3 x 10(6) cells/ml in vitro, and results in a 4-6 log destruction of clonogenic leukemic cells/ml according to a limiting dilution assay, while leaving half of normal CFU-GM surviving. The hyperthermic purging of clonogenic leukemic cells was not affected in the presence of normal marrow cells in vitro.