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Biomedical subjects

Makoto Takeuchi

Publications and source records attributed to Makoto Takeuchi.

64 records · Page 4Linked to original sources

Establishment of the T-cell large granular lymphocyte leukemia cell line MOTN-1 carrying natural killer-cell antigens.

A novel interleukin-2 (IL-2) dependent leukemia cell line MOTN-1 was established from the peripheral blood of a 63-year-old woman with T-cell large granular lymphocyte (LGL) leukemia in chronic phase. Primary peripheral blood leukemia cells were CD3+, CD5+, CD7+, CD56+, CD94+, CD161+, TcRalphabeta+, and HLA-DR+. The immunoprofile of the established cell line MOTN-1, however, showed CD3-, CD5-, CD7+, CD56+, CD94+, CD159+, CD161+, TcRalphabeta- and HLA-DR+; the MOTN-1 cells were cytoplasmatically positive for CD3varepsilon and the products of the T-cell receptor (TcR) genes beta and gamma. While the TcRbeta and TcRgamma genes were rearranged, the TcRdelta gene was found to be deleted. DNA fingerprinting and chromosome analysis identifying the t(2;6)(q?23;q?21) and t(12;18)(q13;q?22) alterations demonstrated the authenticity and the malignant nature of the cell line. The scientific significance of MOTN-1 lies in (1) the rarity of this type of leukemia cell lines, (2) the co-expression of various T- and natural killer (NK)-cell-associated markers, and (3) its unique chromosomal aberrations.

Antigens, CD↗

Can t(8;21) oligoblastic leukemia be called a myelodysplastic syndrome?

The new World Health Organization (WHO) classification of hematologic malignancies has incorporated t(8;21) myelodysplastic syndromes (MDS) according to the French-American-British classification into the category of acute myeloid leukemia (AML) with t(8;21)(q22;q22), while our knowledge about clinicopathological features of t(8;21) oligoblastic leukemia is still limited. We present our experience with 12 patients meeting the FAB diagnostic criteria of MDS and having t(8;21), who were compared to 43 t(8;21) AML patients. The MDS and AML patients shared most hematomorphologic, immunophenotypic, and clinical features, whereas the differences lay along myeloid maturation. The MDS patients had higher percentages of circulating neutrophils and marrow myeloid cells beyond promyelocytes than the AML patients. The incidence of Auer rods in mature neutrophils in MDS was significantly higher than that in AML, and furthermore, the neutrophils in MDS more commonly contain t(8;21) than in AML. Our findings support the rationale for the WHO classification, and future studies on large patient populations should help clarify whether the spontaneous differentiation potential could be actively associated with a hematological manifestation of t(8;21) leukemias.

Adult↗

A novel L3-acute lymphoblastic leukemia cell line (BALM-27) carrying cytoplasmic immunoglobulin 8 chain but lacking expression of cell surface immunoglobulin chains.

A human acute lymphoblastic leukemia (ALL) cell line, BALM-27, was established from the peripheral blood specimen of a patient with B-cell ALL L3 type (ALL-L3) at diagnosis. As with the original leukemia cells, the established line was negative for all cell surface immunoglobulin (Ig) chains, but carrying only cytoplasmic Ig delta heavy chain. Southern blot analysis of the various Ig chain genes demonstrated homozygous deletion of the Jkappa gene, germ line configuration of the Jlambda and rearrangement of IgJH genes. Cytogenetic analysis of both primary leukemic bone marrow and BALM-27 cells showed the der(8;15)(q10;q10) chromosomal alteration, in addition to the t(8;22)(q24;q11) abnormality which is highly associated with ALL-L3 and Burkitt's lymphoma. The established cell line BALM-27 represents a rich resource of abundant, accessible, and manipulable cell material for analyzing the unique expression of Ig chain and for investigating the pathogenesis of B-cell malignancy. The scientific significance of BALM-27 lies in (1) the rarity of this type of leukemia cell lines, and (2) its unique chromosomal aberrations.

Aged↗

Identification of an endo-beta-N-acetylglucosaminidase gene in Caenorhabditis elegans and its expression in Escherichia coli.

We report the identification, molecular cloning, and characterization of an endo-beta-N-acetylglucosaminidase from the nematode Caenorhabditis elegans. A search of the C. elegans genome database revealed the existence of a gene exhibiting 34% identity to Mucor hiemalis (a fungus) endo-beta-N-acetylglucosaminidase (Endo-M). Actually, the C. elegans extract contained endo-beta-N-acetylglucosaminidase activity. The putative cDNA for the C. elegans endo-beta-N-acetylglucosaminidase (Endo-CE) was amplified by polymerase chain reaction from the Uni-ZAP XR library, cloned, and sequenced. The recombinant Endo-CE expressed in Escherichia coli exhibited substrate specificity mainly for high-mannose type oligosaccharides. Man(8)GlcNAc(2) was the best substrate for Endo-CE, and Man(3)GlcNAc(2) was also hydrolyzed. Biantennary complex type oligosaccharides were poor substrates, and triantennary complex substrates were not hydrolyzed. Its substrate specificity was similar to those of Endo-M and endo-beta-N-acetylglucosaminidase from hen oviduct. Endo-CE was confirmed to exhibit transglycosylation activity, as seen for some microbial endo-beta-N-acetylglucosaminidases. This is the first report of the molecular cloning of an endo-beta-N-acetylglucosaminidase gene from a multicellular organism, which shows the possibility of using this well-characterized nematode as a model system for elucidating the role of this enzyme.

Acetylglucosaminidase↗

Production in yeast of alpha-galactosidase A, a lysosomal enzyme applicable to enzyme replacement therapy for Fabry disease.

A mammalian-like sugar moiety was created in glycoprotein by Saccharomyces cerevisiae in combination with bacterial alpha-mannosidase to produce a more economic enzyme replacement therapy for patients with Fabry disease. We introduced the human alpha-galactosidase A (alpha-GalA) gene into an S. cerevisiae mutant that was deficient in the outer chains of N-linked mannan. The recombinant alpha-GalA contained both neutral (Man(8)GlcNAc(2)) and acidic ([Man-P](1-2)Man(8)GlcNAc(2)) sugar chains. Because an efficient incorporation of alpha-GalA into lysosomes of human cells requires mannose-6-phosphate (Man-6-P) residues that should be recognized by the specific receptor, we trimmed down the sugar chains of the alpha-GalA by a newly isolated bacterial alpha-mannosidase. Treatment of the alpha-GalA with the alpha-mannosidase resulted in the exposure of a Man-6-P residue on a nonreduced end of oligosaccharide chains after the removal of phosphodiester-linked nonreduced-end mannose. The treated alpha-GalA was efficiently incorporated into fibroblasts derived from patients with Fabry disease. The uptake was three to four times higher than that of the nontreated alpha-GalA and was inhibited by the addition of 5 mM Man-6-P. Incorporated alpha-GalA was targeted to the lysosome, and hydrolyzed ceramide trihexoside accumulated in the Fabry fibroblasts after 5 days. This method provides an effective and economic therapy for many lysosomal disorders, including Fabry disease.

Carbohydrate Sequence↗

Pretreatment of leishmania homologue of receptors for activated C kinase (LACK) promotes disease progression caused by Leishmania amazonensis.

A cDNA coding Leishmania homologue of receptors for activated C kinase (LACK), which was known to play an important role in the early phase of Leishmania infection, was molecularly cloned from Leishmania amazonensis promastigote by using reverse transcription and nested polymerase chain reaction, and was sequenced. The L. amazonenis LACK cDNA showed 97.3 to 99.3% homology and its deduced amino acid sequence showed 98.7 to 99.7% identity in comparison with LACK sequences from five other species. The amino acid sequences in the immunodominant peptide region were completely conserved among Leishmania spp. tested. Intravenous pretreatment of the recombinant L. amazonensis LACK into BALB/c mice showed progressive lesion development compared to PBS (-) injected control mice, suggesting the important role of LACK in the early phase of L. amazonensis infection.

Amino Acid Sequence↗

Focal tuberculous lymphadenitis in an HIV-1 infected patient.

A 41-year-old man was admitted to the hospital because of focal swelling of the left supraclavicular lymph nodes. Eighteen months prior to admission, he had been diagnosed with human immunodeficiency virus type 1 (HIV-1) infection and was started on highly active antiretroviral therapy (HAART). He responded well to HAART with an increase in CD4+ cell count and improvement in symptoms. However, one year after the initiation of HAART, he developed progressive enlargement of left supraclavicular lymph nodes. An excisional lymph node biopsy was performed for diagnosis, which revealed tuberculous lymphadenitis. Rifabutin, isoniazid, and ethambutol were initiated for treatment.

AIDS-Related Opportunistic Infections↗

P wave signal averaged electrocardiography in patients undergoing the Fontan operation.

OBJECTIVES: Supraventricular arrhythmias are one of the most common and fatal sequelae of the Fontan operation. P wave triggered signal averaged electrocardiography was performed in patients undergoing the Fontan operation to evaluate the presence of atrial degeneration, and to clarify which factors affected the development of atrial arrhythmias. METHODS: P wave triggered signal averaged electrocardiography was recorded in 14 patients after the Fontan-type operation (conventional atriopulmonary connection in 5 and total cavopulmonary connection in 9) and 15 healthy controls. The duration and area of the filtered P wave, and the signal magnitudes (M20, M30) at 20 Hz and 30 Hz obtained from the frequency domain analysis of the P wave (M20, M30) were evaluated and compared with the hemodynamic data. RESULTS: The duration and area of the filtered P wave, M20 and M30 in patients after atriopulmonary connection were significantly greater than in those after total cavopulmonary connection and the control subjects (p < 0.05). M20 was significantly greater in patients after total cavopulmonary connection than in the control subjects. Right atrial volume in patients after atriopulmonary connection was significantly (p < 0.001) larger than in patients after total cavopulmonary connection (p < 0.05). There were no significant differences in other indices including atrial pressure between the two groups. CONCLUSIONS: Our results suggest that the substrate for atrial arrhythmias such as atrial myocardial degeneration and fibrosis is frequently present in patients after the Fontan operation, especially after atriopulmonary connection. Thus, the enlarged right atrium may be involved in the presence of a substrate for atrial arrhythmias. The developmental risk for late atrial arrhythmias seems to be present even in patients after total cavopulmonary connection.

Adolescent↗

Congenital cervical rhabdomyosarcoma arising in one fetus of a twin pregnancy.

OBJECTIVE: To describe a huge congenital cervical rhabdomyosarcoma. METHODS: We were recently confronted with a case of fetal solid neck mass arising in one fetus of a twin pregnancy. Prenatally, the cervical tumor was consistent with teratoma, but it was diagnosed histologically as a rhabdomyosarcoma. Genetic amniocentesis showed a mosaic pattern consisting of 46,XY/46,XY,t(2;8)(q35;q21.2). RESULTS: EXIT procedure was proposed to the parents but declined. The twin with huge cervical tumor died in utero at 35 weeks' gestation due to hydrops fetalis. CONCLUSION: Fetal cervical rhabdomyosarcoma is an extremely rare condition that has not been previously reported, but should be considered in the presentation of fetal solid neck mass.

Adult↗

Successful treatment of a patient with synchronous advanced non-small cell lung cancer and acute myeloid leukemia by a combination of gefitinib, low-dose cytarabine and aclarubicin.

There are few reports describing simultaneous occurrence of acute leukemia and lung cancer. We describe here an 83-year-old woman who simultaneously developed advanced adenocarcinoma of the lung and acute myeloid leukemia. She could not receive intensive chemotherapy due to poor performance status. This patient was treated with a combination of gefitinib, low-dose cytarabine and aclarubicin. This combination could be safely administered in the elderly patient with poor performance status and was effective for both lung cancer and acute myeloid leukemia.

Aclarubicin↗