Retinoid status and RARs expression.
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Animals↗
Biomedical subjects
H Mano
Publications and source records attributed to H Mano.
Fetal human immunodeficiency virus type 1 infection of different organs in the second trimester.
Human immunodeficiency virus type 1 (HIV-1) infection in utero was examined by isolating the virus and detecting the HIV-1 DNA sequence from different fetal tissues. The brain, thymus, lung, liver, spleen, and placenta tissues from fetuses (10-23 weeks of gestation) born to HIV-1-infected asymptomatic mothers were examined. HIV-1 was isolated from 2 of 7, 1 of 7, and 1 of 7 cocultures of splenic, thymic, and trypsin-resistant cells from the liver and placenta, respectively, with peripheral blood mononuclear cells; 20-30% and 40-60% of splenic and of thymic cells were CD4+ lymphoid cells and 40-80% of trypsin-resistant cells were mononuclear phagocytes. The HIV-1 DNA sequence was detected in 4 of 7, 3 of 7, 1 of 7, 1 of 7, 2 of 7, and 2 of 6 samples from the spleen, thymus, brain, lung, liver, and placenta, respectively, using the polymerase chain reaction. In one case, the intensity of the HIV-1 DNA sequence appeared to be correlated with the success of viral isolation. We indicate that fetal HIV-1 infection may frequently occur in the second trimester and the cells responsible for the infection may be CD4+ lymphoid cells and mononuclear phagocytes.
Adult↗
Cytogenetic analysis of human bile for mutagenicity and co-mutagenicity.
Cytogenetic analysis was used to test whether or not human bile induced chromosome abnormalities in lymphocytes grown in culture. Bile was obtained from gallbladders resected for various reasons such as cholecystitis, cholelithiasis, polypus and cancers of the biliary tract, stomach and pancreas. After adding human bile to a final concentration of 25 microliters/ml or 12.5 microliters/ml, the culture medium was incubated at 37 degrees C for 72 hr. Air-dried slides were stained with conventional Giemsa and the numerical and structural chromosome abnormalities were scored. Positive and negative controls in terms of chromosome abnormalities were established by using 0.03 micrograms/ml mitomycin C (MMC) and 0.9% normal saline, respectively. Cytogenetic analysis was successfully performed in 6 out of 10 bile samples (60.0%). Bile alone did not induce numerical or structural chromosome abnormalities. Structural abnormalities increased significantly in the 25 microliters/ml bile + 0.03 micrograms/ml MMC group, compared with the 0.03 micrograms/ml MMC group: 36.0% vs. 20.7% in the chromatid-type gaps and breaks, 27.8% vs. 22.7% in the chromosome-type gaps and breaks, and 8.3% vs. 3.2% in the exchange-type abnormalities. It is likely that the interaction between bile and MMC is synergistic rather than additive.
Adult↗
Selenium contents in human gallbladder bile.
The selenium contents in human gallbladder bile were analyzed. Thirty-seven subjects were studied; 22 patients with cholelithiasis in Niigata Prefecture and 15 patients (13 with cholelithiasis and 2 with gallbladder polypus) in Kochi Prefecture. Five ml of bile was withdrawn with a syringe from the gallbladder during the operation and stored at -20 degrees C until analysis. For the analysis by gas chromatograph with an electron-capture detector, 0.2 ml of sample was used. The mean selenium contents in bile were 269 +/- 39.0 (mean +/- S.D.) ng/ml for the subjects in Niigata and 285 +/- 84.4 ng/ml in Kochi; without significant difference. Of 37 samples analyzed, the mean content was 276 +/- 61.0 ng/ml.
Adult↗
Establishment of hybridomas secreting monoclonal antibodies to placental alkaline phosphatase and development of an enzyme immunoassay for its determination.
We established seven hybridomas secreting murine IgG monoclonal antibodies (MoAbs) to placental alkaline phosphatase (PLAP). The seven hybridomas were designated (1) 7C6, (2) 6G10, (3) 5B9, (4) 6D5, (5) 6B5, (6) 11G6 and (7) 3E10, respectively. The characteristics of these hybridomas were evaluated by radioimmunoassay (RIA) with 125I-PLAP. Their reactivity with the intestinal alkaline phosphatase, one of the alkaline phosphatase isozymes, was (1) 0.04, (2) 0.2, (3) 1.4, (4) 1.8, (5) 0, (6) 4.0 and (7) 6.2(%), respectively. None of them showed signs of cross-reactivity with the liver-type alkaline phosphatase, also one of the alkaline phosphatase isozymes, within a PLAP concentration of 2,000 IU/l. The subtype of 5B9 was IgG1, and that of the others was IgG2a. We then used 7C6, to develop a sensitive, specific and convenient enzyme immunoassay (EIA) for the determination of PLAP, and assayed sera from patients with various gynecologic diseases. The incidence of increased PLAP was 6.4% in patients with benign diseases, 21.5% in cervical cancer, 36.4% in endometrial carcinoma, and 39.5% in malignant ovarian tumors. The specificity for malignant diseases seemed to be higher than that of CA125. Among endometrial carcinomas, well-differentiated adenocarcinoma had the highest incidence of an increased concentration. Among malignant ovarian tumors, serous cystadenocarcinoma, endometrioid carcinoma, dysgerminoma and Krukenberg's tumor showed a higher incidence than the other types.
Alkaline Phosphatase↗
A novel protein-tyrosine kinase, tec, is preferentially expressed in liver.
To identify protein-tyrosine kinases which play an important role in the process of hepatocarcinogenesis, we have screened a murine liver cDNA library with v-fps kinase domain as a probe. Using low stringency screening, we could isolate cDNAs of a putative protein-tyrosine kinase, tec (tyrosine kinase expressed in hepatocellular carcinoma). Nucleotide sequences of the cDNAs show that the C-terminal domain of its predicted protein has significant homology with that of the members of the src family. The tec gene is expressed mainly in liver and faintly in heart, kidney and ovary. Northern analysis further shows that in 2 out of 4 cell lines of human hepatocellular carcinoma (HCC) the tec gene is highly expressed compared to normal human liver. This is the first report showing a protein-tyrosine kinase which may be specifically involved in the cell growth of hepatocytes or in the step of hepatocarcinogenesis.
Amino Acid Sequence↗
Invasive character of malignant endothelial cells in vinyl-chloride-induced liver angiosarcoma.
Two cases of vinyl-chloride-induced angiosarcoma of the liver were investigated with special reference to the invasive character of malignant endothelial cells. Histological findings of the liver were similar in both cases. The liver angiosarcoma was multicentric and consisted of three portions: a peripheral parenchymal area, an intermediate mottled area, and a central cavernous area. Microscopically the intermediate area showed a mixture of fibrosis and cell infiltration of various patterns. At the periphery, yellowish brown parenchymal tissues remained and were always associated with sinusoidal changes, from non-cellular dilatation to trabecular growth of the small round cells. Under electron microscope, the small round cells showed direct contact with red blood cells and hepatocytes. Thus, the sinusoidal small cell infiltration is thought to be an initial lesion of angiosarcomatous transformation. Because none of the liver is free from these sinusoidal changes, partial hepatectomy would not be indicated even in a patient with few cavernous lesions.
Endothelium↗
Paraneoplastic syndrome of hypercalcemia and leukocytosis caused by squamous carcinoma cells (T3M-1) producing parathyroid hormone-related protein, interleukin 1 alpha, and granulocyte colony-stimulating factor.
Previously we reported that a clonal squamous cell carcinoma cell line (T3M-1) derived from a lower jaw cancer of a patient with marked leukocytosis and hypercalcemia produced factors containing a potent bone-resorbing activity (BRA) (Mr 15,000-20,000) and a colony-stimulating activity. To elucidate the pathogenesis of this humoral hypercalcemia, BRA and colony-stimulating activity in both the conditioned medium and cells were characterized. The conditioned medium, when eluted at neutral pH, contained colony-stimulating activity and thymocyte proliferation-stimulating activity, the latter of which comigrated with BRA. Upon elution with acetic acid (pH 2.0), the conditioned medium contained no interleukin 1-like activity but potent parathyroid hormone-like activity, which comigrated with BRA. Northern blot hydridization analysis revealed that T3M-1 cells produced constitutively mRNA for parathyroid hormone-related protein and granulocyte colony-stimulating factor. Furthermore, primer extension analysis revealed that the cells also produced mRNA for interleukin 1 alpha (IL-1 alpha). Since parathyroid hormone-related protein and IL-1 alpha (osteoclast-activating factor) synergistically increase the concentration of serum calcium, and since IL-1 alpha (hemopoietin 1) potentiates granulocyte colony-stimulating factor-induced granulocytopoiesis, we speculate that parathyroid hormone-related protein, granulocyte colony-stimulating factor, and IL-1 alpha are synergistically involved in a paraneoplastic syndrome of hypercalcemia and leukocytosis, at least in some patients with solid tumors.
Animals↗
Mutations of N-ras oncogene in myelodysplastic syndromes and leukemias detected by polymerase chain reaction.
By using polymerase chain reaction and differential hybridization, point mutations of N-ras oncogene were investigated among 43 patients with hematopoietic malignancies. Six cases were revealed to carry mutational N-ras oncogenes. One case with acute myelocytic leukemia was found to carry a mutation at codon 13, although this case was in complete remission.
Adolescent↗
Analysis of the mechanism of influenza B virus inactivation by guinea pig serum.
Normal guinea pig serum lacking detectable antiviral antibody inactivated influenza B virus via the classical complement pathway. This virus inactivation appeared to result from the steric hindrance of HA activity by the association with the virus of serum proteins presumed to be complement components. Trypsin digestion of the associated proteins fully restored the HA activity but not infectivity. It was found that the virus underwent minor disruption of the envelope and degradation of M1 protein and genomic RNA.
Animals↗
Relationship between an activated N-ras oncogene and chromosomal abnormality during leukemic progression from myelodysplastic syndrome.
The relationship between chromosomal abnormality and oncogene activation was investigated during leukemic progression in two patients with myelodysplastic syndrome (MDS). Both patients had partial or complete deletion of chromosome 5 in metaphase cells obtained throughout the progression to leukemia. Analysis with specific oligonucleotide probes revealed that bone marrow cells containing an activated N-ras oncogene proliferated in a dominant manner during the process of leukemic conversion in both patients. These observations suggest that the chromosomal abnormality may precede activation of the N-ras gene in these patients, and that both the chromosomal abnormality and the activated N-ras oncogene contribute to the development of leukemia.
Acute Disease↗
Constitutive expression of the granulocyte-macrophage colony-stimulating factor gene in human solid tumors.
We detected constitutive expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene in 3 human solid tumors by Northern blot analysis. Two of them were also found to secrete the GM-CSF protein by colony forming unit-culture assay. Southern blot analysis of each tumor DNA showed no gross rearrangement of the GM-CSF gene. This is the first report that demonstrates expression of the GM-CSF gene in solid tumors at the mRNA level.
Colony-Forming Units Assay↗
Visualization of moving fluid: quantitative analysis of blood flow velocity using MR imaging.
A new method for the measurement of blood flow using magnetic resonance imaging has been developed. The flow velocities are calculated from the distances that the fluid has moved. The distances are directly visualized by a new pulse sequence. In a phantom study, the measured flow rates showed very good correlation with actual flow rates of up to 20 l/min (3 m/sec). In a volunteer study, pulsatile flow velocities of a large artery were measured with electrocardiographic gating. The flow pattern of a cardiac cycle at the abdominal aorta is similar to that revealed by other methods of measurement, such as Doppler ultrasound. This method allows reasonably accurate quantitative analysis of blood flow in the large arteries.
Blood Flow Velocity↗
A new radioimmunoassay for human chorionic gonadotropin using monoclonal antibody.
A new radioimmunoassay (RIA) for human Chorionic Gonadotropin (hCG) was developed using murine monoclonal antibody to the beta-subunit of hCG (beta-hCG). The IgG fraction of the monoclonal antibody which did not react with 125I-beta-hCG was purified from hybridoma ascites, and covalently coupled to Sepharose 4B. This solid-phase antibody was incubated with standard hCG or serum sampled for 48 hours. The reaction medium was then removed by centrifugation and 125I-beta-hCG and anti-beta-hCG rabbit polyclonal antibody were added to the precipitate. The alcohol precipitation method was used for separating "bound" and "free" forms in the second reaction. The sensitivity for hCG in this assay system was 0.5 mIU/ml serum and the cross-reactivity with human Luteinizing Hormone (hLH) was 0.4%. This assay system was shown to be clinically applicable. Serial serum samples from two patients with trophoblastic disease were assayed and minute amounts of hCG, which could not be determined by conventional assay methods, could be assayed by this new RIA.
Adult↗
Radioimmunoassay of placental alkaline phosphatase in ovarian cancer sera and tissues.
A specific radioimmunoassay for human placental alkaline phosphatase has been developed using the 125I-labeled enzyme, highly purified with a fast protein liquid chromatography system and an absorbed rabbit antiserum. The sensitivity of this assay was 0.2 U/L. Serum levels of over 0.2 U/L were found in 27% of ovarian cancer patients, and most of these elevated enzyme levels occurred with more advanced stages of the disease. On the other hand, almost all ovarian cancer tissue contained detectable levels of the enzyme. Serous adenocarcinoma, endometrioid adenocarcinoma, and dysgerminoma had particularly large amounts. Placental alkaline phosphatase was more frequently detected in tissue than in the serum of ovarian cancer, and therefore may be a useful target in immunodetection and immunotherapy and in studying the histopathology of ovarian cancer.
Adenocarcinoma↗
Radioimmunodetection of human choriocarcinoma xenografts by monoclonal antibody to placental alkaline phosphatase.
Placental alkaline phosphatase (PLAP)-specific monoclonal antibody (MAb) 11-D-10, which did not react with other isoenzymes of alkaline phosphatase (AP), was raised by a hybridoma technique. MAb 11-D-10 was radiolabeled and administered to athymic mice bearing human choriocarcinoma containing PLAP. This antibody was found to be more specifically localized in tumor tissue as compared to normal tissues. The tissue-to-blood ratio (T/B ratio) of MAb 11-D-10 in tumor tissue increased from 1.38 at 2 days to 2.51 at 5 days after administration. On the other hand, the T/B ratios of isotype control non-immunized IgM in tumor tissue were 0.72 and 0.87 at 2 days and 5 days after administration, respectively. 131I-labeled MAb 11-D-10 was administered to athymic mice bearing choriocarcinomas of various sizes and various PLAP contents to examine the effect on the radioimage of the differences in tumor size and PLAP content. Tumors less than 0.3 cm in diameter could be imaged clearly by gamma-scintigraphy without blood pool image subtraction. The strength of the radioimage correlated fairly well with PLAP content.
Alkaline Phosphatase↗
Purification of placental alkaline phosphatase and its monoclonal antibody.
Monoclonal antibodies (MAbs) to placental alkaline phosphatase (PAlP) were raised by using the hybridoma technique. Spleen cells from immunized mice were fused with the mouse myeloma line NS-1 using 50% polyethylene glycol and cultured in a selection medium. Antibodies were screened by the enzyme immunoassay using immobilized solid-phase antigen and anti-mouse immunogloblin Fab' (rabbit)-beta-D-galactosidase complex. Four double-cloned hybridomas were obtained. The cross-reactivity with different kinds of alkaline phosphatases of the raised MAbs was examined. At first, MAb 11-D-10, with which placenta was stained but liver and small intestine were not stained in indirect immunofluorescence, was selected. Then, the cross-reactivity of MAb 11-D-10 was further investigated by immunoblotting (Western blotting). MAb 11-D-10 reacted with PAlP but did not react with hepatic and intestinal alkaline phosphatase at all. The binding activity of 125I-labeled MAb 11-D-10 with different choriocarcinoma cell lines (SCH, BeWo, NaUCC-1, NaUCC-2, and NaUCC-3) was highest in SCH, then in the order of BeWo, NaUCC-3, NaUCC-1 and NaUCC-2, and correlated to the PAlP content of the cells (SCH greater than BeWo greater than NaUCC-3 greater than NaUCC-1 greater than NaUCC-2). Also the intensity of indirect immunofluorescence of the above cell lines with MAb 11-D-10 correlated with the PAlP content of each cell.
Alkaline Phosphatase↗
Radioimmunolocalization of monoclonal antibodies to human chorionic gonadotropin in choriocarcinoma transplanted into athymic mice.
Radioimmunodetection of monoclonal antibodies to human Chorionic Gonadotropin (hCG) or its beta-subunit in choriocarcinoma tissue was investigated in athymic mouse model system. Antibody to hCG exhibited a higher localization index (L.I.) in tumor tissue than those in other organs, but the tissue/blood ratio of this antibody in tumor tissue was not so high as expected (0.38 and 0.5 on Day 2 and 5 respectively). On the other hand, antibody to beta-subunit showed the same L.I. in tumor tissue as those in other organs. Gel filtration of serum of mouse given an antibody to hCG showed that the antibody was forming immune complexes in the blood stream. Autoradiography showed many grains in the capillary vessels and intercellular spaces in the tumor tissue. We concluded that a monoclonal antibody to hCG localized in choriocarcinoma tissue via a mechanism of retention of immune complexes in the tissue, and that there may be a limitation to the clinical application of localization of antibodies to secretory products such as hCG of tumor cells from their low specific localization.
Animals↗