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Biomedical subjects

A Fuse

Publications and source records attributed to A Fuse.

At least 55 records · Page 3Linked to original sources
PubMed1991

[The hemodynamics during hemodilution and its influence on the liver functions after hepatectomy for hepatocellular carcinoma with liver cirrhosis].

The hemodynamics during hemodilution occurred after hepatectomy for hepatocellular carcinoma with liver cirrhosis and its influences on the liver functions were studied. The hematocrit value gradually decreased about 10% until the 4th postoperative day owing to hemodilution after hepatectomy. While anemia progressed, cardiac index inversely increased. Under such a condition, oxygen consumption was maintained so that acidosis did not develop. Arterial blood ketone body ratio was also kept within a normal range except for a case whose hematocrit value decreased to 17.1%. Although the escaped hepatic enzymes such as GOT and GPT increased in the serum after hepatectomy, hemodilution was not responsible for their increase. While total bilirubin increased in the severe hemodiluted group, the increase was not due to hemodilution but caused by blood transfusion. The protein synthesis of the liver measured by rapid turnover protein levels in plasma was depressed after surgery, and this depression prolonged to the 14th postoperative day in the group whose hematocrit value decreased below 20%. These results suggest that it is better to keep hemodynamics without blood transfusion unless the hematocrit value decrease below 20%, and also better to maintain the hematocrit above 20% for liver regeneration after hepatectomy.

Adult↗
PubMed1989

Detection of the human papillomavirus 6b E2 gene product in genital condyloma and laryngeal papilloma tissues.

We have previously reported that the E2 open reading frame of human papillomavirus type 6b (HPV6b) was expressed in Bombyx mori cell line Bm-N as a fusion protein with baculovirus polyhedrin. Here we show that antibody against this E2-polyhedrin fusion protein recognized a putative gene product as a 49-kDa protein in some genital condyloma and laryngeal papilloma tissues, which were associated with HPV6 and HPV11, by immunoblotting analysis. The antibody reacted also with antigens in the nucleus of cells in the middle and upper layers of these papilloma tissues.

Animals↗
PubMed1989

Characterization of bovine papillomavirus type 1-transformed clones which show distinct transformed phenotypes.

Seventeen independent cell clones were isolated from C127 cells transformed by bovine papillomavirus type 1 (BPV-1). Transformants showed differing degrees of expression of the transformed phenotype as monitored by saturation density, doubling time, growth in medium with a low serum concentration and colony-forming efficiency in soft agar. The degree of expression of the transformed phenotype did not correlate with either the BPV-1 copy number or levels of BPV-1-specific RNA in the transformed cell clones. A characteristic transformed cell clone, T1c, showed the lowest degree of expression of the transformed phenotype but contained the highest copy number of BPV-1 DNA and the highest level of BPV-1-specific mRNA. When we analysed different transformants by two-dimensional gel electrophoresis, we found that a set of six proteins changed quantitatively. Changes in the expression of these proteins were most consistent in clones expressing the greatest number of parameters of transformation, e.g. clone T4a. These data indicate that changes in the expression of cellular genes may correlate with the degree of expression of the transformed phenotype.

Animals↗
PubMed1989

Establishment of a human leukaemic cell line (CMK) with megakaryocytic characteristics from a Down's syndrome patient with acute megakaryoblastic leukaemia.

A new megakaryoblastic cell line (CMK), which also exhibits erythroid and myeloid markers, was established from a Down's syndrome patient suffering from acute megakaryoblastic leukaemia. The CMK cells were found to be positive in reactions with anti-platelet antibodies (anti-glycoproteins IIb/IIIa and Ib, and Plt-1). Platelet peroxidase (PPO) reactivity was found to be associated with the nuclear envelope and the endoplasmic reticulum but not with the Golgi apparatus. Some cells possessed cytoplasmic granules with the characteristics of alpha-granules and demarcation membranes. Karyotyping revealed near-tetraploidy (modal chromosome number of 95; ranging 87-98) and a translocation der(17)t(11;17), also found in the original leukaemic cells, confirming that the cells were derived from the patient's malignant blasts. The CMK cells were also found to be positive in reaction with anti-glycophorin A antibody, as well as with anti-myeloid antibodies (MY4, MY7 and MY9). Treatment of CMK cells with phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) greatly enhanced the reactivity with anti-platelet antibodies, increased the number of cells in which cytoplasm was dissociated into numerous segments and suppressed the reactivity with anti-glycophorin A. The proliferation of CMK cells was stimulated by interleukin-3 (IL-3) and granulocyte-macrophage colony stimulation factor (GM-CSF). This cell line should be a useful tool for analysing the basis of the afferent association between megakaryoblastic leukaemia and Down's syndrome, as well as for further study of megakaryocytic differentiation.

Bone Marrow↗
PubMed1989

Growth and differentiation of a human megakaryoblastic cell line, CMK.

Recently, a human megakaryoblastic cell line, CMK, was established from the peripheral blood of a megakaryoblastic leukemia patient with Down syndrome. Using this cell line, we studied the proliferation and differentiation of megakaryocytic cells in the presence of highly purified human hematopoietic factors and phorbol 12-myristate-13-acetate (PMA). In a methylcellulose culture system, interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitated colony formation by CMK cells in a dose-dependent manner. The maximum stimulating doses of these factors were 10 and 200 U/mL, respectively. These concentrations were comparable to those that stimulate activity in normal hematopoietic cells. In contrast, granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF), and erythropoietin (EPO) had no effects on the colony formation of CMK cells. In a liquid culture system, 20% of the CMK cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hematopoietic factors, whereas 40% of the cells expressed GPIIb/IIIa with the addition of IL-3 and GM-CSF. EPO also slightly enhanced expression of GPIIb/IIIa. On the other hand, PMA inhibited growth of CMK cells and induced most of them to express the GPIIb/IIIa antigen. Furthermore, PMA induced CMK cells to produce growth activity toward new inocula of CMK cells. This growth factor (GF) contained colony-stimulating activity (CSA) in normal bone marrow (BM) cells. The activity was believed to be attributable mainly to GM-CSF, since 64% of this activity was neutralized by anti-GM-CSF antibodies and a transcript of GM-CSF was detected in mRNA from PMA-treated CMK cells by Northern blot analysis. These observations suggest that GM-CSF, as well as IL-3, should play an important role in megakaryocytopoiesis.

Blotting, Northern↗
PubMed1989

[Experimental studies on influences of intermittent hemihepatic vascular occlusion].

The influences of intermittent hemihepatic vascular occlusion in rats were investigated. The restoration of tissue energy level during 10 min of reperfusion after more than 40 min of occlusion was significantly lower than after occlusion within 30 min. over 40 min of occlusion which resulted in a significant increase of tissue free fatty acids during occlusion showed marked decrease of total adenine nucleotides (TAN) in the liver and marked increase of serum TAN in the hepatic vein after reperfusion. It was presumed that these findings were caused by wash-out of adenine-nucleotides from liver cell to the hepatic vein after reperfusion correlated with degradation of cell membrane. Thus, less than 30 min of occlusion was safe in hemihepatic vascular occlusion. Three times intermittent occlusion--repeats of 30 min of occlusion and 10 min of reperfusion--resulted in a same tissue energy level as once 40 min of occlusion. Furthermore, there was no difference in serum ornithine carbamoyltransferase, tissue oxygen consumption and ICG test during 48 hrs after reperfusion between three times intermittent occlusion and once 40 min of occlusion. These findings indicated that repetition of short time occlusion was useful in order to safely prolong total ischemic time in hemihepatic vascular occlusion.

Adenine Nucleotides↗
PubMed1988

Expression of human papillomavirus type 6b E2 gene product with DNA-binding activity in insect (Bombyx mori) cells using a baculovirus expression vector.

Human papillomavirus type 6b (HPV6b) has been shown to be a major etiologic agent of genital condylomas. The E2 gene, one of the early genes, has been shown to activate the enhancer element in trans in cells transformed with bovine papillomavirus type 1a (BPV1a) but the E2 gene product of any HPV has not been identified. The E2 gene of HPV6b was inserted into the polyhedrin gene of a Baculovirus, Bombyx mori nuclear polyhedrosis virus (BmNPV), 156 bp downstream from the translational start codon, and transferred into BmNPV by allelic replacement in a cotransfected Bombyx mori cell line, Bm-N. The predicted 46-kDa protein was produced by a recombinant virus in the infected Bm-N cells at a high level under the control of the polyhedrin promoter. We obtained the antibody against the putative E2-polyhedrin fusion protein by immunizing a rabbit with this protein. This protein reacted with the antibodies against polyhedrin and the fusion protein. This protein did specifically bind to the upstream regulatory region of the HPV6b and BPV1a genomes. This DNA binding activity was blocked by the antibody against this protein.

Animals↗
PubMed1988

Expression of the E2 open reading frame of papillomaviruses BPV1 and HPV6b in silkworm by a baculovirus vector.

To produce and characterize the nature of the E2 protein of papillomaviruses, we have developed a system to express the DNA sequence containing an open reading frame (ORF) as a fusion protein in cultured insect cells and silkworms. The DNA fragments of the E2 ORF predicted from the DNA sequence of bovine papillomavirus type 1 and human papillomavirus type 6b were linked to the N-terminal part of polyhedrin gene of the Baculovirus Bombyx mori nuclear polyhedrosis virus (BmNPV) vector. Hybrid proteins composed of polyhedrin protein (52 amino acids) and the E2 proteins of BPV1 (410 amino acids) or HPV6b (346 amino acids) were efficiently produced in B. mori cells and silkworm larvae infected with recombinant viruses. The amount of E2 fusion proteins produced by recombinant viruses was comparable to that of polyhedrin produced by wild type BmNPV. The hybrid proteins were immunologically reactive to antiserum against polyhedrin.

Animals↗
PubMed1988

Purification and characterization of human renal dehydropeptidase I.

Dehydropeptidase I from human kidney was purified over 100-fold. The purified enzyme had an isoelectric point of 4.75, apparent molecular weights of 135,000 by gel filtration and of 66,500 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and an optimal pH of 7.4. Human renal dehydropeptidase I hydrolyzed imipenem, carpetimycins A and B, and Sch 29,482.

Anti-Bacterial Agents↗
PubMed1987

Retrovirus produced by a lymphoid cell line from an infant with acute lymphoblastic leukemia.

A lymphoid cell line CK-a was established from peripheral blood of an infant with acute lymphoblastic leukemia of non-T, non-B cell type with mediastinal tumor. The CK-a cells were positive for surface immunoglobulins, Epstein-Barr virus-specific nuclear antigen, HLA-DR and Leu 12 antigens, and negative for sheep erythrocyte-rosette-receptor, and Leu 1, 2, 3 and 4 antigens. Budding particles were detected in electron micrographs of ultrathin sections of the CK-a cells. In the culture media of CK-a cells, particles with a buoyant density of 1.16 g/ml and labeled with [3H]uridine and [35S]methionine but not with [3H]thymidine were found to carry reverse transcriptase activity which preferred Mg2+ to Mn2+. Enveloped particles of 80 to 120 nm in diameter were detected in the fractions at 1.16 g/ml by electron microscopy. Thus, the particles had properties compatible with a definition of Retroviridae, and were tentatively named CK virus (CKV). The genome size of CKV RNA determined by agarose-acrylamide composite gel electrophoresis was 6.1 +/- 0.2 kb. Immune electroblotting assay detected antibody reactive with a CKV protein with a molecular weight of 67,000 in the serum of the patient, but not in sera of an adult T cell leukemia patient and healthy controls. No syncytia were formed by mixed cultures of CK-a and XC cells.

Antibodies, Viral↗
PubMed1987

The c-sis gene expression in cells from a patient with acute megakaryoblastic leukemia and Down's syndrome.

The c-sis gene expression in leukemia cells from a patient with acute megakaryoblastic leukemia and Down's syndrome was studied. The leukemia blasts were identified as megakaryoblasts by the platelet peroxidase reaction and the reactivity against antiplatelet monoclonal antibodies. Leukemia cells obtained from peripheral blood or bone marrow specimens before and after initiation of chemotherapy were analyzed for c-sis gene expression by the RNA-DNA dot blot hybridization. Although the level varied, the mRNA of the c-sis gene was detected in all megakaryoblastic leukemia cells obtained at different clinical stages. The c-sis mRNA level in cells obtained after relapse was higher than that obtained before the initiation of therapy. The 25S c-sis mRNA was detected in a megakaryoblastic leukemia cell line established from this patient. The role of the expression of the c-sis gene in acute megakaryoblastic leukemia cells is discussed.

Acute Disease↗
PubMed1986

Increased resistance to the anticellular effect of interferon in an ultraviolet light-resistant human cell line, UVr-1.

Interferon (alpha, beta and gamma) susceptibility was tested in a human cell line, UVr-1, a u.v. light-resistant variant of RSa cells; the latter have high sensitivity to both u.v. lethality and the cell proliferation inhibition (anticellular) effect of human interferon (HuIFN) preparations. UVr-1 cells were less sensitive than the parental RSa cells to the inhibitory effects of HuIFN preparations, as measured by cell proliferation and the incorporation of [3H]deoxythymidine and [3H]deoxyadenosine into acid-insoluble cellular material. Nevertheless, UVr-1 cells exposed to HuIFN showed almost the same enhanced levels of antiviral activity and pppA(2'p5'A)n synthetase activity as similarly treated RSa cells. Further, UVr-1 cells had much the same binding capacity for 125I-labelled HuIFN-alpha A. Thus, it seems likely that the variant has an increased resistance to the anticellular effect but not to the antiviral effect of HuIFN preparations. UVr-1 cells showed no significant difference from RSa cells in u.v.-induced DNA repair synthesis. However, when a comparison was made between the susceptibility of normal fibroblasts and fibroblasts from patients with Cockayne's syndrome, characterized by an altered u.v. sensitivity but no alteration of DNA repair replication synthesis, the Cockayne's syndrome fibroblasts, CCK-3 and CCK-4, were more susceptible to HuIFN-beta as judged by cell proliferation and deoxythymidine incorporation tests.

2',5'-Oligoadenylate Synthetase↗
PubMed1986

The effects of glycolipids and carbohydrates on bilirubin cytotoxicity in vitro.

We have studied the inhibitory effects of various glycolipids including gangliosides and carbohydrates on bilirubin cytotoxicity towards human glia cells using a 51Cr release assay. All cytotoxic activity was abolished when 100 microM bilirubin was incubated with 40-80 microM gangliosides (GM1, galactosyl-N-acetylgalactosaminyl-(N-acetylneuraminyl) galactosylglucosylceramide; GM2, N-acetylgalactosaminyl-(N-acetylneuraminyl)galactosylglucosylceramide; GM3, N-acetylneuraminylgalactosylglucosylceramide, and GD1a, N-acetylneuraminylgalactosyl-N-acetylgalactosaminyl-(N-acetylneura minyl) galactosylglucosylceramide). On the other hand, inhibition by asialoglycolipids was less effective than that by gangliosides, but became more effective with increasing numbers of sugars per molecule, the efficacy of inhibition being as follows: globoside greater than trihexosyl ceramide greater than lactosyl ceramide greater than glucosyl ceramide. Glucosyl ceramide and the carbohydrates of neuramine lactose or N-acetylneuraminic acid (NANA) did not have inhibitory effects. These results suggest that the terminal acidic group of NANA and the numbers of sugar chains in glycolipids play important roles in the inhibition of bilirubin cytotoxicity.

Bilirubin↗