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Biomedical subjects

K Kita

Publications and source records attributed to K Kita.

At least 127 records · Page 7Linked to original sources

Granulocyte colony-stimulating factor and its receptor in acute promyelocytic leukemia.

Expression of granulocyte colony-stimulating factor (G-CSF) receptor (G-CSFR) and in vitro proliferative response to G-CSF were investigated by quantitative immunofluorescence and [3H] thymidine uptake, respectively, in a series of acute myeloid leukemias (AML). The results indicated that G-CSFR was detected at high levels in acute promyelocytic leukemia (APL) cells, in comparison with other types of AML. Moreover, APL cells were also seen to predominantly proliferate in response to G-CSF. Based on these observations, we administered recombinant human G-CSF to a patient with APL in the third relapse that was resistant to both cytotoxic agents and all trans retinoic acid, in an attempt to sensitize the leukemic cells to cell-cycle-dependent agents. Complete remission was achieved. The finding that APL cells are exquisitely responsive to G-CSF supports the view that G-CSF is useful for augmentation of their vulnerability to cell-cycle specific agents.

Adult↗

Enhancement of XPG mRNA expression by human interferon-beta in Cockayne syndrome cells.

Using PCR-differential display, we have searched for genes expressed specially in human interferon (HuIFn)-beta-treated Cockayne syndrome (CS) fibroblast cells. Eighteen expressed genes induced by HuIFN-beta were identified, the sequences of seven of which were highly homologous to previously cloned sequences. The cDNAs of six of these seven clones were similar to expression tagged sequences from unknown genes in databases and the remaining one was identical to the cDNA of the xeroderma pigmentosum XPG gene. These results, together with our previous finding of increased resistance to ultraviolet (UV) cell-killing of CS cells pretreated with HuIFN-beta prior to UV irradiation suggest that XPG might be one of the genes possibly involved in the HuIFN-beta-induced UV-resistance.

Cells, Cultured↗

Frequent expression of human Thy-1 antigen on pre-B cell acute lymphoblastic leukemia with t(9;22).

Thy-1 (CDw90) is a phosphatidylinositol-anchored protein, and is expressed on human pluripotential hematopoietic stem cells. The expression pattern of this antigen on leukemia cells is still controversial. In this study, 72 adult patients with pre-B cell acute lymphoblastic leukemia (pre-B ALL) were examined for the expression pattern of Thy-1 by using indirect immunofluorescence and reversed transcription polymerase chain reaction (RT-PCR) methods. Twelve cases were judged positive on the basis of conventional immunophenotype criteria. Thirteen cases showed a weak clonal shift on the fluorogram, even though their positive percentages were from 6.7% to 14.9%. Thy-1 gene transcripts were detected in all of the 13 cases showing a weak clonal shift. The study of antibody binding capacity, which was calculated by the mean fluorescence intensity of the test sample on the basis of a calibration curve using standard beads, showed that cases with more than 150 sites/cell could be positive. Thy-1 positivity in pre-B ALL was not associated with the expression of B-cell differentiation antigens. Thy-1 expression was significantly higher in pre-B ALL cases with karyotypic abnormalities than in those with normal karyotype (p = 0.0071). The t(9;22) abnormality was found in 18 of the 25 Thy-1+ cases. Simultaneous expression of transcriptional factors, GATA-2 and SCL, was frequently detected in the Thy-1+ cases. bcr-abl and GATA-2 are thought to play important roles in the proliferation of immature hematopoietic cells. Indeed, cell-cycle analysis showed that the cell population in the S/G2/M phase of the present Thy-1+ cases was less than that in the Thy-1- cases (p = 0.001770). Our data suggest that Thy-1 expression indicates the proliferative status of the leukemia cells, not their phenotypic immaturity.

Adolescent↗

Biphasic expression of CD4 in acute myelocytic leukemia (AML) cells: AML of monocyte origin and hematopoietic precursor cell origin.

In 227 of 495 (45.9%) Japanese adult patients with acute myelocytic leukemia (AML), leukemic cells expressed CD4. Incidence of CD4 expression in each FAB subtype was as follows: M1 37.4%, M2 33.7%, M3 35.4%, M4 65.0%, and M5 78.3%. The typical expression pattern of myelomonocytic differentiation antigens and cytokine receptors in CD4+ AML was CD34lowCD33high CD11bhighGM-CSFRhigh. AML cases with 11q23 abnormalities and with inv(16) were frequently CD4-positive. These data collectively indicate that CD4 expression in AML cells is associated with monocytic characteristics. However, CD4+CD34high AML cases appear to have unique immature characteristics including low expression of myelomonocytic differentiation antigens (ie CD33 and CD11b), and accumulation of chromosome abnormalities (ie t(8;21) in CD4lowCD34high AML and chromosome 7 abnormalities in CD4highCD34high AML). We speculate that these leukemia subsets originate from CD4+ hematopoietic precursor cells, therefore then should be considered separately from most of the CD4+ AML as represented by CD34lowCD33high CD11bhighGM-CSFRhigh. Overall survival of patients with CD4+ AML in our series was worse than that of those with CD4 AML (P = 0.0202).

Adult↗

The expression of co-stimulatory molecules and their relationship to the prognosis of human acute myeloid leukaemia: poor prognosis of B7-2-positive leukaemia.

We examined the expression of co-stimulatory molecules on leukaemic cells of 52 adult patients with acute myeloid leukaemia (AML) (34 men and 18 women) and analysed the relationship between these expressions and the patient's prognosis. B7-1 was not expressed in any of the 23 patients investigated, whereas B7-2 was expressed in 26/52 patients (50.0%). B7-2 was expressed in all AML patients with monocytic morphology (M4 or M5) and in 16/42 cases without monocytic morphology. CD54 was expressed in 28/ 37 patients examined (75.7%), and CD58 was expressed in all of the AML patients except one (M 7). The overall survival of the 26 B7-2-positive leukaemia patients (1-24 months, median survival 11.5 months) was significantly shorter than that of the 26 B7-2-negative leukaemia patients (1-71+ months, median 35.1 months) (P=0.0080). In addition, the B7-2-positive patients exhibited significantly shorter disease-free survival periods compared to the B7-2-negative patients (P=0.021). There was no significant difference in age, sex, haematological data and complete remission rate between the B7-2-positive and B7-2-negative patients. Our results indicated that B7-2 is one of the most crucial factors in the prognosis of adult acute leukaemia and can be expected to have an important role in tumour immunity.

Acute Disease↗

Refeeding increases hepatic insulin-like growth factor-I (IGF-I) gene expression and plasma IGF-I concentration in fasted chicks.

1. We examined the influence of refeeding after 2 d of fasting on plasma insulin-like growth factor-I (IGF-I) concentration and hepatic IGF-I gene expression in chickens at 6 weeks of age. 2. Hepatic IGF-I mRNA was measured by ribonuclease protection assay and plasma IGF-I concentration was determined by radioimmunoassay. 3. Plasma IGF-I concentration decreased following fasting, increased to the level of fed controls after 2 h of refeeding but then fell back to the level of fasted chickens after 6 h of refeeding. 4. Fasting reduced hepatic IGF-I mRNA concentrations to less than half of those in the fed controls. Refeeding increased IGF-I mRNA sharply at 2 h after refeeding, but by 6 h after refeeding they had taller back again to levels significantly lower than at 2 h. 5. A significant correlation between plasma IGF-I concentration and hepatic IGF-I gene expression was found, suggesting that when chicks are refed after 2 d of fasting, the short-term increase in plasma IGF-I concentration may be partly regulated by the alteration in hepatic IGF-I mRNA.

Animals↗

Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower.

This paper describes the synthesis of 14C-labeled glycosphingolipids using the reverse hydrolysis reaction (condensation) of sphingolipid ceramide N-deacylase. It was found that 50-70% of 14C-fatty acids were incorporated into various lyso-glycosphingolipids when a mixture of lyso-glycosphingolipids and fatty acids was incubated at 37 degrees C with 1 mU of the enzyme for 20 h in 1 ml of 25 mM phosphate buffer, pH 6.0-7.0, containing 0-0.1% Triton X-100. The optimum concentration of lyso-glycosphingolipids was 100-400 microM depending on the species of lyso-form when [14C]stearic acid was used at the concentration of 100 microM. Free 14C-fatty acids and lyso-glycosphingolipids were separated from the synthesized 14C-glycosphingolipids by using a Sep-Pak Plus Silica and a Sep-Pak CM or a QMA cartridge, respectively. After treatment of 14C-glycosphingolipids with endoglycoceramidase or sphingolipid ceramide N-deacylase, digestion products were clearly separated from the parent glycosphingolipids on TLC and determined using an image analyzer with a sensitivity 100 times higher than that using non-radiolabeled substrates. Using this method, we found endoglycoceramidase activity in a seaflower, Condylactis sp., for the first time.

Amidohydrolases↗

Stress induced urinary incontinence in patients with spinocerebellar degeneration.

OBJECTIVES: To examine the pathophysiology of "stress induced urinary incontinence" (urinary incontinence evoked by abdominal straining) in patients with spinocerebellar degeneration. METHODS: Micturitional symptoms of 184 patients with spinocerebellar degeneration who were admitted to hospital were studied repeatedly. Urodynamic studies were made in symptomatic patients, and consisted of uroflowmetry, measurement of residual urine, urethral pressure profilometry, medium fill water cystometry, and external sphincter EMG. RESULTS: Twenty nine (15.8%) patients with spinocerebellar degeneration showed stress induced urinary incontinence. Twenty of the 29 patients had detrusor overactivity, low compliance detrusor, or residual urine, resembling urgency and overflow types of incontinence (complicated form). The other nine had none of these findings (pure form), but showed decreased maximum urethral closure pressure in four, absence of bulbocavernosus reflex in two, absence of voluntary sphincter contraction in one, incompetent urinary storage even at the first sensation in two, and high amplitude and polyphasic neurogenic changes in three of five patients studied, indicative of neurogenic sphincter dysfunction. CONCLUSIONS: Stress induced urinary incontinence in spinocerebellar degeneration had various underlying mechanisms. Some of the patients only showed evidence of pudendal denervation, which can cause external sphincter weakness and may reflect lesions of the sacral Onuf's nucleus and the pudendal nerve. Urodynamic studies are necessary to evaluate stress induced urinary incontinence in patients with spinocerebellar degeneration, to prescribe appropriate therapies.

Adolescent↗

Escherichia coli transformant expressing the glucose dehydrogenase gene from Bacillus megaterium as a cofactor regenerator in a chiral alcohol production system.

Escherichia coli JM109 (pGDA2) overexpressing the glucose dehydrogenase (GDH) gene from Bacillus megaterium IWG3 was examined for use as a cofactor regenerator. In the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate by E. coli JM109 (pKAR) which is an aldehyde reductase-overproducing transformant, E. coli JM109 (pGDA2) can act as an NADPH regenerator with NADP+ and glucose, similarly to commercially available GDH.

Alcohols↗

Expression of the extracellular levansucrase and invertase genes from Zymomonas mobilis in Escherichia coli cells.

We investigated the expression and localization in Escherichia coli of sucZE2 and sucZE3, encoding Zymomonas mobilis extracellular levansucrase and invertase, respectively, and lacking a typical N-terminal secretion signal. Levansucrase and invertase were expressed efficiently under the lac and tac promoters in E. coli cells, and some of the levansucrase produced was localized in the periplasmic space. The sucZE2 expression was not lethal to E. coli in the presence of 5% sucrose, and led to the accumulation of levan from sucrose.

Amino Acid Sequence↗

Successful treatment of aplastic anemia with G-CSF and high dose erythropoietin.

We report the successful treatment of pancytopenia with G-CSF and high dose erythropoietin (Epo) in an elderly patient diagnosed with aplastic anemia (AA). Furthermore this effect is dose dependent for Epo in vivo. Detection of apoptosis by gel electrophoresis shows that high dose Epo protects bone marrow mononuclear cells from spontaneous apoptosis in vitro. These findings may explain some of the mechanisms of aplastic anemia.

Aged↗

Mutagenicity of microcystin-LR in human RSa cells.

Microcystin-LR (MCLR) is a potent cyclic heptapeptidic hepatotoxin produced by bloom-forming cyanobacteria. The mutagenicity induced by this compound in cultured human RSa cells was found by determination of ouabain-resistant (OuaR) mutation, with the highest frequency at the concentration of 15 microgram/ml. Moreover, base substitution mutations at K-ras codon 12 in genomic DNA, assessed by polymerase chain reaction (PCR) and differential dot-blot hybridization using digoxigenin-labeled probes, were detected in RSa cells 6 days after exposure to MCLR (7.5-15 microgram/ml).

Cells, Cultured↗

Activation of MAP kinases by 5-fluorouracil in a 5-fluorouracil-resistant variant human cell line derived from a KT breast cancer cell line.

To investigate the mechanism of the acquired resistance of human cells to an anticancer drug, 5-fluorouracil (5-FU), a drug-resistant clone, KTFU-4, was isolated from a human KT breast carcinoma cell line, treated with ethylmethanesulfonate and then with 5-FU. The viability of the KT cells, analyzed using an MTT assay, was suppressed by 5-FU in a dose-dependent manner, while that of the KTFU-4 cells was enhanced by it at concentrations between 0.1 and 1.0 microgram/ml. Treatment of KTFU-4 cells with 5-FU resulted in increased amounts of activated phosphorylated ERK1/2 and p38 MAP kinases, but not in the parent KT cells. It is thus possible that 5-FU stimulated the proliferation of KTFU-4 cells by activating a signal transduction pathway leading to cell growth.

Antimetabolites, Antineoplastic↗

[TAL1 gene analysis in T-cell malignancies].

Site-specific recombination of the TAL1 gene was analyzed by Southern blotting and polymerase chain reaction (PCR) in 44 cases of childhood T-cell acute lymphoblastic leukemia (T-ALL), 20 cases of childhood T-cell non-Hodgkin's lymphoma (T-NHL) and 35 cases of adult T-cell malignancies. This recombination was found in 10 (22.7%) of 44 childhood T-ALL patients, but in none of the T-NHL or adult T-cell malignancies. Recombination of the TAL1 gene was therefore suggested to be specific for childhood T-ALL. The immunophenotypic features of the 10 T-ALL patients with this recombination were CD1-, CD2+, CD4-, CD7+, CD10-, and they had a significantly better outcome than other T-ALL cases without the recombination. The PCR technique revealed minimal residual disease (MRD) in 2 patients. One showed persistent MRD, while in the other MRD was recognized only at initial diagnosis. Further investigation is needed whether T-ALL with this recombination constitutes a distinct clinical subgroup among childhood T-ALL patients.

Adolescent↗

Abnormal DNA synthesis activity induced by X-rays in nevoid basal cell carcinoma syndrome cells.

DNA synthesis activity was examined in fibroblasts and isolated nuclei derived from patients with nevoid basal cell carcinoma syndrome (NBCCS) upon exposure to X-ray and ultraviolet (UV). The DNA synthesis activity in NBCCS fibroblasts increased after X-ray irradiation, i.e., to twice that on mock-irradiation, while it decreased in healthy donor-derived fibroblasts. The DNA synthesis activity in isolated nuclei of X-ray irradiated NBCCS fibroblasts also increased, i.e., more than twice that on mock-irradiated. In the experiments using synchronized cells, DNA synthesis activity showed the most marked increase when the fibroblasts at S phase were irradiated with X-rays. In contrast, UV-irradiated NBCCS fibroblasts showed no such increase in DNA synthesis. These results revealed that DNA synthesis is abnormally induced in X-ray irradiated NBCCS cells and that this abnormality might be related with the tendency of tumorigenesis in NBCCS patients after exposure to X-ray.

Ataxia Telangiectasia↗

Partial reconstitution of mammalian phosphoribosylpyrophosphate synthetase in Escherichia coli cells. Coexpression of catalytic subunits with the 39-kDa associated protein leads to formation of soluble multimeric complexes of various compositions.

Rat liver phosphoribosylpyrophosphate (PRPP) synthetase exists as complex aggregates composed of 34-kDa catalytic subunits (PRS I and PRS II) and homologous 39- and 41-kDa proteins termed PRPP synthetase-associated proteins (PAPs). While a negative regulatory role was indicated for PAPs, the physiological function of PAPs is less well understood. We attempted to prepare recombinant 39-kDa PAP (PAP39) and to reconstitute the enzyme complex. Free PAP39 was poorly expressed in Escherichia coli, while expression of protein fused with glutathione S-transferase was successful. The purified fusion protein had no PRPP synthetase activity, and bound to dissociated PRS I and PRS II, with a similar affinity. A free form of PAP39 prepared from the fusion protein formed insoluble aggregates. The enzyme complex was then partially reconstituted in situ by coexpression of PAP39 with PRS I or PRS II in E. coli cells. This coexpression led to formation of soluble complexes of various compositions, depending on the conditions. When the relative amount of PAP39 was higher, specific catalytic activities, in terms of the amount of the catalytic subunit, were lowered. PAP39 complexed with PRS I was more readily degraded by proteolysis than seen with PRS II, in vivo and in vitro. These results provide additional, strong evidence for that PAP39 has no catalytic activity in the enzyme complex, but does exert inhibitory effects in an amount-dependent manner, and that composition of the enzyme complex varies, depending on the relative abundance of components present at the site of aggregate formation.

Adenosine Diphosphate↗

cDNA sequence of a translational elongation factor Ts homologue from Caenorhabditis elegans: mitochondrial factor-specific features found in the nematode homologue peptide.

The cDNA for a homologue of elongation factor Ts which probably functions in mitochondria has been sequenced from a nematode Caenorhabditis elegans. The deduced amino acid sequence (316 amino acids long) has a possible transit peptide sequence at the amino terminus and several common specific features for mammalian mitochondrial EF-Ts. The amino acid identities in the protein from C. elegans compared with those of bovine mitochondria and Escherichia coli are 29.5% and 24.0%, respectively. The C. elegans sequence was classified as a long EF-Ts (ca. 280 amino acids long) similar to peptides from mammalian mitochondria and eubacteria other than Thermus and cyanobacteria (except Spirulina platensis), rather than short EF-Ts (ca. 200 amino acids long) as those of Thermus, cyanobacteria and plastids.

Amino Acid Sequence↗