Biomedical subjects

Y Maeda

Publications and source records attributed to Y Maeda.

At least 199 records · Page 11Linked to original sources

PubMed2000

Altered T cell development in human thymoma is related to impairment of MHC class II transactivator expression induced by interferon-gamma (IFN-gamma).

Thymoma is known to contain CD4+CD8+ T cells, indicating that neoplastic epithelial cells of thymoma have a function as thymic cortical epithelium. However, it has been shown that there is an impairment of CD4+ T cell development in thymoma and that IFN-gamma-induced HLA-DR expression on cultured thymic epithelial cells (TEC) derived from thymoma is decreased when compared with the normal thymus. MHC class II transactivator (CIITA) is known to play a critical role in IFN-gamma-induced MHC II expression. In this study, we attempted to elucidate whether CIITA is responsible for the impaired up-regulation of MHC II molecules in response to IFN-gamma in thymoma TEC. A quantitative reverse transriptase-polymerase chain reaction examination revealed that the induced level of CIITA was significantly lower in thymoma TEC than in normal TEC. The induced levels of invariant chain (Ii) and HLA-DR in thymoma TEC were correlated with CIITA expression. The proportion of CD3+ cells in the CD4+CD8- subset in thymoma was also correlated with CIITA expression. A gel mobility shift assay however, revealed translocation of STAT1 to the nucleus in thymoma as well as normal TEC. Intercellular adhesion molecule-1 was up-regulated in the thymoma TEC to a level similar to normal TEC in response to IFN-gamma. These results indicate that impaired up-regulation of HLA-DR in response to IFN-gamma results from insufficient induction of CIITA, but not from the signal from IFN-gamma receptor to the nucleus. The abnormal regulation of HLA-DR expression caused by impaired induction of CIITA may affect CD4+ T cell development in thymoma.

Adolescent↗

PubMed2000

Rapid patterning in 2-D cultures of Dictyostelium cells and its relationship to zonal differentiation.

Rapid patterning has been observed in confined 2-D cultures of Dictyostelium discoideum Ax-2 cells as an outer dark zone and a inner light zone. The width of outer zone was usually approximately100 microm, irrespective of the size of cell masses under atmospheric conditions. The width of the outer zone, however, changed depending on external O2 concentrations and reached up to 250 microm at 100% O2. A clear regional difference in tetramethyl rhodamine methyl ester (TMRM) staining was noticed between the outer zone and the inner zone: the inner zone was more strongly stained with TMRM than the outer zone, which faced the air. Using inhibitors of oxidative phosphorylation (dinitrophenol (DNP) or NaN3) and a specific inhibitor of CN-resistant respiration (benzohydroxamic acid (BHAM)), it has been demonstrated that the outer zone is basically formed by the O2 threshold for oxidative phosphorylation, while the inner cells mainly perform cyanide-resistant respiration. When cells around the early mound stage (just before prestalk and prespore differentiation) were cultured as 2-D cell masses, ecmA-expressing cells (pstA cells), ecmB-expressing cells (pstB cells) and D19-expressing cells (prespore; psp cells), arose in a position-dependent manner in the outer zone. In the inner zone, cell motility seemed to be markedly impaired and neither prestalk nor prespore differentiation occurred. In addition, once-differentiated prespore cells were found to dedifferentiate rapidly in the inner zone. The reason for dedifferentiation as well as for failure of cells to differentiate in the inner zone is discussed with reference to O2 radicals.

Animals↗

PubMed2000

The survey of sleeping time of junior high school students: a study on the sleep questionnaire.

We investigated the questionnaire study on sleep of junior high school students to estimate the effect of excessive study hours on their sleep time in 1998. A total of 501 students answered the questionnaire. It was noted that most of the students sleep for 6-8 h on weekdays. The majority of the students of the junior high school felt that their sleep is insufficient on weekdays.

Adolescent↗

PubMed2000

A new periodogram using one-way analysis of variance for circadian rhythms.

A new periodogram was proposed using one-way analysis of variance (ANOVA), termed ANOVA periodogram, in order to reveal a precise significant periodicity. Thirty 3-day complex computer-simulated time series with known periodicity (24 h) and three 2-h data-missing occurring periodically (23 h, 20 min) were used to compare the ANOVA periodogram with Enright's one. In results, the ANOVA periodogram was superior to Enright's periodogram in the accuracy of assessing the major periodicity.

Algorithms↗

PubMed2000

Japanese couples' childbirth experiences in Michigan: implications for care.

BACKGROUND: Subtle cultural differences in the childbirth experience for expatriate Japanese couples living in southeast Michigan can sometimes cause conflicts between them and American health caregivers. The purpose of this study was to examine Japanese couples' perceptions and experiences of prenatal care and childbirth in a region of the United States, and to explore the implications for providing culturally competent care. METHODS: In this qualitative study, in-depth interviews of 11 Japanese couples (n = 22) were conducted at an outpatient primary care clinic in southeast Michigan by a team of researchers fluent in the Japanese language and knowledgeable about the culture. Participants also completed a short questionnaire. RESULTS: The major themes that emerged comprised perceptions related to a language barrier, ultrasonography, prenatal vitamin supplementation, episiotomy, epidural anesthesia, and practitioner-patient relationship. Throughout these six themes it was evident that Japanese health care professionals had difficulty accepting health care that was different from the care provided in their home country. CONCLUSION: The most striking finding of this study was the difficulty for health caregivers to provide culturally competent care. Although the program has strongly invested in health professionals and support staff who were fluent in Japanese and knowledgeable about Japanese culture, conflicts still occurred.

Adult↗

PubMed2000

Potassium-adsorption filter for RBC transfusion: a phase III clinical trial.

BACKGROUND: Within the past 2 years, three cases of cardiac arrest just after rapid transfusion of RBCs preserved for over 7 days after 15-Gy irradiation were found. This severe complication caused transient hyperkalemia. To prevent potassium (K(+)) overload by RBC transfusion at the bedside, a K(+)-adsorption filter made of sodium polystyrene sulfonate was developed. STUDY DESIGN AND METHODS: After in vitro and animal safety and efficacy tests, a Phase III clinical trial was conducted with 65 patients given transfusions via the newly developed filter (filter group) and 37 patients in whom the filter was not used (control group) and transfusions were given at twice the usual flow rate (20 mL/min). RESULTS: More than 85-percent (94.4+/-3.8%) removal of K(+) in RBCs in mannitol-adenine-phosphate (MAP) that had been preserved for more than 14 days or that were used 3 days after 15-Gy irradiation (calculated K(+): 3.8+/-1.3 mEq/bag) was achieved in 82 of 83 bags of MAP RBCs in the filter group, with 79.6 percent removed in the other, even in rapid transfusions. RBC recovery 1 day after transfusion, determined by increments in RBCs, Hb, and Hct, were 24 and 0.4 x 10(4) per microL, 0.7 and 0.3 g per dL, and 1.6 and 0 percent, respectively, in the filter and control groups. No adverse transfusion reactions, such as hypotension, anaphylactoid reactions, or asthma-like attacks, were observed, except for one case of urticaria in the filter group. Mild fever (within 1 degrees C) after transfusion was observed in both groups. Serologic markers of hemolysis rose slightly in both groups, but there was no significant difference between the two groups. CONCLUSION: The newly developed K(+)-adsorption filter is useful, especially in a rapid transfusion setting.

Adenine↗

PubMed2000

Carcase characteristics of a heavy Japanese quail line under introgression with the roux gene.

1. The roux plumage sex-linked recessive gene may be used for early sexing of Japanese quail in crossbreeding production systems with wild-type and roux lines. However, associated effects of the gene on carcase and meat composition need to be assessed. 2. Quail carcases from pure Line K males and females (100% K), a heavy meat line which was used as the recipient line for the introgression of the roux gene, and from same-age roux or wild-type females from the second generation of introgression (75% K) were dissected. The effects of sex, line and plumage colour on carcase components and on protein and lipid contents of various tissues were estimated. 3. Expected sex differences in carcase weights were obtained, with marginally higher values for females. However, weights of parts and carcase yield (ratio of empty carcase weight without head, neck and feet over live body weight) were similar for both sexes in pure Line K which had a 68% carcase yield. Lipid contents in leg meat were higher in males (3.1%) than in females (2.7%). 4. The roux gene did not seem to have any major impact on carcase parts or composition. However, in roux birds, leg skin was marginally higher in lipids and pectoralis major lower in proteins than in wild-type ones.

Animals↗

PubMed2000

Long-term egg production and heterosis in quail lines after within-line or reciprocal recurrent selection for high early egg production.

1. Four Japanese quail lines were developed using 13 generations of reciprocal recurrent (lines AA and BB) or within-line selection (lines DD and EE) for high egg number until 98 d of age. In these lines and their reciprocal crosses, egg production, egg weight and mortality were monitored for up to 21 months to evaluate the impact of selection method and line origin on long-term production and heterosis. 2. Both hen-housed total egg number (EN) and hen-day egg laying rate (ELR) were consistently lower in recurrent lines than in lines under within-line selection: after 13 months of test, the difference was -47.1 eggs for EN and -20% for ELR%, whereas mortality was similar at around 20%. 3. Line EE was the best pure line for early egg production (55.2 eggs at 98 d of age) and also gave eggs which were consistently about 2 g larger than those from line DD but it had the highest overall mortality (78.5%) of all lines. On average, line DD laid 399 eggs (EN) over the whole experiment, 84 more than Line EE. 4. Crossbreds from the 2 types of pure lines and line DD had similar patterns and rates of egg production during the whole experiment, as indicated by the similarity of the curves adjusted for those lines by using the monomolecular model for EN and the modified compartmental model for ELR. 5. However, heterosis was higher from recurrent selection lines and it increased more with time than did heterosis from within-line selection lines.

Animals↗

PubMed2000

Gaa1p and gpi8p are components of a glycosylphosphatidylinositol (GPI) transamidase that mediates attachment of GPI to proteins.

Many eukaryotic cell surface proteins are anchored to the membrane via glycosylphosphatidylinositol (GPI). The GPI is attached to proteins that have a GPI attachment signal peptide at the carboxyl terminus. The GPI attachment signal peptide is replaced by a preassembled GPI in the endoplasmic reticulum by a transamidation reaction through the formation of a carbonyl intermediate. GPI transamidase is a key enzyme of this posttranslational modification. Here we report that Gaa1p and Gpi8p are components of a GPI transamidase. To determine a role of Gaa1p we disrupted a GAA1/GPAA1 gene in mouse F9 cells by homologous recombination. GAA1 knockout cells were defective in the formation of carbonyl intermediates between precursor proteins and transamidase as determined by an in vitro GPI-anchoring assay. We also show that cysteine and histidine residues of Gpi8p, which are conserved in members of a cysteine protease family, are essential for generation of a carbonyl intermediate. This result suggests that Gpi8p is a catalytic component that cleaves the GPI attachment signal peptide. Moreover, Gaa1p and Gpi8p are associated with each other. Therefore, Gaa1p and Gpi8p constitute a GPI transamidase and cooperate in generating a carbonyl intermediate, a prerequisite for GPI attachment.

Acyltransferases↗

PubMed2000

Genome structure of Ri plasmid (3). Sequencing analysis of the vir region of pRi1724 in Japanese Agrobacterium rhizogenes.

The entire genome of the pRi1724 (217.6-kb) in the mikimopine type Agrobacterium rhizogenes strain MAFF03-01724 has been completely sequenced. The vir region covering 30.2-kb has found to be composed of 21 genes resembling virH1, virA, virB1-11, virG, virC1-2, and virD1-5. The structural organization of the pRi1724 vir operons in this study is exactly the same as that of the previously reported vir operons of other Ri or Ti plasmids, although the size of some ORFs showed little variations among the plasmids. We also found virE3 gene in the pRi1724 (1), but different from Ti plasmids, virE1 and virE2 that are also important for the virulence do not exist in the vir region of pRi1724.

Bacterial Proteins↗

PubMed2000

Engineering subtilisin E for enhanced stability and activity in polar organic solvents.

We examined the effect of a novel disulfide bond engineered in subtilisin E from Bacillus subtilis based on the structure of a thermophilic subtilisin-type serine protease aqualysin I. Four sites (Ser163/Ser194, Lys170/Ser194, Lys170/Glu195, and Pro172/Glu195) in subtilisin E were chosen as candidates for Cys substitutions by site-directed mutagenesis. The Cys170/Cys195 mutant subtilisin formed a disulfide bond in B. subtilis, and showed a 5-10-fold increase in specific activity for an authentic peptide substrate for subtilisin, N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide, compared with the single-Cys mutants. However, the disulfide mutant had a 50% decrease in catalytic efficiency due to a smaller k(cat) and was thermolabile relative to the wild-type enzyme, whereas it was greatly stabilized relative to its reduced form. These results suggest that an electrostatic interaction between Lys170 and Glu195 is important for catalysis and stability in subtilisin E. Interestingly, the disulfide mutant was found to be more stable in polar organic solvents, such as dimethylformamide and ethanol, than the wild-type enzyme, even under reducing conditions; this is probably due to the substitution of uncharged Cys by charged surface residues (Lys170 and Glu195). Further, the amino-terminal engineered disulfide bond (Gly61Cys/Ser98Cys) and the mutation Ile31Leu were introduced to enhance the stability and catalytic activity. A prominent 3-4-fold increase in the catalytic efficiency occurred in the quintet mutant enzyme over the range of dimethylformamide concentration (up to 40%).

Bacillus subtilis↗

PubMed2000

Adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes.

Vascular endothelial growth factor (VEGF) is an angiogenic growth factor that stimulates endothelial cell proliferation, increases endothelial permeability, and promotes collateral vessel formation. We transferred human VEGF gene into rat cardiac myocytes using adeno-associated virus (AAV) vectors and investigated whether VEGF secreted from the transduced cardiac myocytes promoted proliferation of endothelial cells. We produced VEGF-expressing AAV vectors (AAV-VEGF) by the adenovirus-free method. Immunoblotting revealed VEGF protein expression in AAV-VEGF-transduced rat cardiac myocytes. More than 60% of cardiac myocytes were stained positively on immunohistochemical staining using anti-VEGF antibody. Concentration of VEGF in the culture medium of AAV-VEGF-transduced myocytes was increased in a vector dose-dependent manner, and VEGF secretion from the transduced myocytes persisted for > or = 14 days. Thymidine incorporation into human vascular endothelial cells was significantly increased by incubation with the conditioned medium from AAV-VEGF-transduced myocytes. This increased thymidine uptake was significantly inhibited by anti-VEGF antibody. We demonstrated here that AAV-mediated VEGF gene transfer into cardiac myocytes induces the secretion of functional VEGF.

Animals↗

PubMed2000

Inhibition of vascular endothelial growth factor activity by transfection with the soluble FLT-1 gene.

Vascular endothelial growth factor (VEGF)-induced angiogenesis is involved in the etiology of some cardiovascular diseases. The soluble form of VEGF receptor, FLT-1 (sFLT-1), is a potent antagonist of VEGF. Therefore, we investigated whether transfection with the sFLT-1 gene could inhibit VEGF-induced angiogenesis. Human embryonic kidney (HEK)-293 cells were transfected with plasmids containing VEGF and sFLT-1 (pCMV-VEGF and pCMV-sFLT-1) by the calcium-phosphate co-precipitation method. VEGF- and/or sFLT-1-transfected HEK-293 cells were incubated for 24 h, and then conditioned medium was collected. The effects of conditioned medium on angiogenesis were tested by incorporation of [3H]thymidine into human umbilical vein endothelial cells (HUVECs). Expression of VEGF protein was determined by Western blotting. The conditioned medium from sFLT-1 gene-transfected HEK-293 cells significantly inhibited recombinant VEGF-induced increase in [3H]thymidine incorporation by HUVECs. VEGF gene-transfected HEK-293 cells secreted VEGF protein into conditioned medium. This conditioned medium increased [3H]thymidine incorporation by HUVECs, which was significantly inhibited by co-transfection of sFLT-1 gene with VEGF gene. These observations suggested that sFLT-1 gene transfer could inhibit VEGF-induced DNA synthesis of vascular endothelial cells.

Blotting, Western↗

PubMed2000

Identification of critical amino acid residues in human immunodeficiency virus type 1 IN required for efficient proviral DNA formation at steps prior to integration in dividing and nondividing cells.

Human immunodeficiency virus type 1 integrase (HIV-1 IN) is thought to have several putative roles at steps prior to integration, such as reverse transcription and nuclear transport of the preintegration complex (PIC). Here, we investigated new functional aspects of HIV-1 IN in the context of the viral replication cycle through point mutagenesis of Ser, Thr, Tyr, Lys, and Arg residues conserved in IN, some of which are located at possible phosphorylation sites. Our results showed that mutations of these Ser or Thr residues had no effect on reverse transcription and nuclear transport of PIC but had a slight effect on integration. Of note, mutations in the conserved KRK motif (amino acids 186 to 189), proposed previously as a putative nuclear localization signal (NLS) of HIV-1 IN, did not affect the karyophilic property of HIV-1 IN as shown by using a green fluorescent protein fusion protein expression system. Instead, these KRK mutations resulted in an almost complete lack of viral gene expression due to the failure to complete reverse transcription. This defect was complemented by supplying wild-type IN in trans, suggesting a trans-acting function of the KRK motif of IN in reverse transcription. Mutation at the conserved Tyr 143 (Y143G) resulted in partial impairment of completion of reverse transcription in monocyte-derived macrophages (MDM) but not in rhabdomyosarcoma cells. Similar effects were obtained by introducing a stop codon in the vpr gene (DeltaVpr), and additive effects of both mutations (Y143G plus DeltaVpr) were observed. In addition, these mutants did not produce two-long terminal repeat DNA, a surrogate marker for nuclear entry, in MDM. Thus, the possible impairment of Y143G might occur during the nuclear transport of the PIC. Taken together, our results identified new functional aspects of the conserved residues in HIV-1 IN: i) the KRK motif might have a role in efficient reverse transcription in both dividing and nondividing cells but not in the NLS function; ii) Y143 might be an important residue for maintaining efficient proviral DNA formation in nondividing cells.

Amino Acid Sequence↗

PubMed2000

Involvement of both the V2 and V3 regions of the CCR5-tropic human immunodeficiency virus type 1 envelope in reduced sensitivity to macrophage inflammatory protein 1alpha.

To determine whether C-C chemokines play an important role in the phenotype switch of human immunodeficiency virus (HIV) from CCR5 to CXCR4 usage during the course of an infection in vivo, macrophage inflammatory protein (MIP)-1alpha-resistant variants were isolated from CCR5-tropic (R5) HIV-1 in vitro. The selected variants displayed reduced sensitivities to MIP-1alpha (fourfold) through CCR5-expressing CD4-HeLa/long terminal repeat-beta-galactosidase (MAGI/CCR5) cells. The variants were also resistant to other natural ligands for CCR5, namely, MIP-1beta (>4-fold) and RANTES (regulated upon activation, normal T-cell expressed and secreted) (6-fold). The env sequence analyses revealed that the variants had amino acid substitutions in V2 (valine 166 to methionine) and V3 (serine 303 to glycine), although the same V3 substitution appeared in virus passaged without MIP-1alpha. A single-round replication assay using a luciferase reporter HIV-1 strain pseudotyped with mutant envelopes confirmed that mutations in both V2 and V3 were necessary to confer the reduced sensitivity to MIP-1alpha, MIP-1beta, and RANTES. However, the double mutant did not switch its chemokine receptor usage from CCR5 to CXCR4, indicating the altered recognition of CCR5 by this mutant. These results indicated that V2 combined with the V3 region of the CCR5-tropic HIV-1 envelope modulates the sensitivity of HIV-1 to C-C chemokines without altering the ability to use chemokine receptors.

Amino Acid Sequence↗

PubMed2000

Effect of a novel anti-allergic agent, HSR-609, on antigen-induced airway hyperresponsiveness in mice.

The effects of a newly synthesized anti-allergic agent, HSR-609, on allergic airway hyperresponsiveness and airway inflammation have been studied in sensitized mice. The effects were compared with those of two histamine H(1) receptor antagonists, cetirizine and terfenadine, and prednisolone. Three inhalations of antigen caused an increase in leukocytes (including eosinophils) with increases in IL-5 in BALF and airway hyperresponsiveness to acetylcholine in BALB/c mice. All drugs were orally administered once a day for 10 days from the day before the first inhalation of antigen. HSR-609 (10 mg/kg) and prednisolone (5 mg/kg) significantly inhibited the antigen-induced airway hyperresponsiveness, whereas cetirizine (10 mg/kg) or terfenadine (100 mg/kg) did not affect this airway response. At the same time HSR-609 inhibited the antigen-induced eosinophilia and IL-5 production in BALF. Prednisolone also showed an inhibitory effect on the airway eosinophilia and IL-5 production but not cetirizine and terfenadine in the same experiments. In addition, HSR-609 (p.o.) dose-dependently suppressed the accumulation of eosinophils elicited by antigen-stimulated D10G4.1 cells, a murine Th2 clone, in peritoneal cavity lavage fluid in AKR/J mice. These results suggest that HSR-609 inhibits allergic airway hyperresponsiveness to acetylcholine probably because of the inhibition of Th2-dependent eosinophilia caused by IL-5. In addition, effects of HSR-609 were different from those of cetirizine and terfenadine concerning the inhibition of antigen-induced airway hyperresponsiveness in mice.

Acetylcholine↗

PubMed2000

Effects of aspirin-like drugs on nitric oxide synthesis in rat vascular smooth muscle cells.

The purpose of this study was to investigate the effects of aspirin-like drugs on nitric oxide (NO) synthesis in rat vascular smooth muscle cells (VSMCs). We measured the accumulation of nitrite, a stable oxidation product of NO, and the expression of inducible NO synthase (iNOS) mRNA and protein in rat cultured VSMCs. Sodium salicylate, aspirin, and indomethacin dose-dependently enhanced nitrite production by interleukin (IL)-1beta-stimulated VSMCs at therapeutic plasma concentration ranges. Increased nitrite production by aspirin-like drugs was accompanied by increased iNOS mRNA and protein accumulation in VSMCs. Addition of IL-1beta activated nuclear factor kappaB (NF-kappaB) in VSMCs, but sodium salicylate did not affect IL-1beta-induced NF-kappaB activation. The nonselective lipoxygenase (LO) inhibitor nordihydroguaiaretic acid inhibited sodium salicylate-induced nitrite production, whereas the selective 5-LO inhibitor caffeic acid did not influence production of nitrite. The 12-LO product 12-HETE dose-dependently enhanced nitrite production by IL-1beta-stimulated VSMCs, whereas the 15-LO product 15-HETE did not. Our study demonstrates that aspirin and the aspirin-like drugs, sodium salicylate and indomethacin, increase NO synthesis in IL-1beta-stimulated VSMCs by upregulation of iNOS transcription via a 12-LO pathway. These effects were independent of NF-kappaB activation. In addition to the direct inhibition of platelet function, aspirin-like drugs may contribute to the reduction of atherothrombotic risk in myocardial ischemia via enhancing NO production by VSMCs.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

PubMed2000

A case of adult-onset Still's disease complicated by non-Hodgkin's lymphoma.

Left cervical lymphadenopathy developed in a 50-year-old male who had a history of adult-onset Still's disease for the preceding 18 months. Still's disease is characterized by rash, fever, and leukocytosis. Lymphadenopathy has been reported in about 60% of the patients, and most histopathologic studies have shown non-specific reactive hyperplasia. However, in this case, an open biopsy of the cervical node revealed a histology of diffuse large B-cell lymphoma. The B-cell malignant lymphoma that developed may have resulted from a sequential progression of a previous stage of benign lymphoproliferative lesion. Our case suggests that the pathophysiology of adult-onset Still's disease involves the stimulation of lymphoid systems to the point of progression towards lymphoma. Malignant lymphoma should be added to the list of life-threatening complications which, although rare, are associated with this disease.

Humans↗