Biomedical subjects

M Wada

Publications and source records attributed to M Wada.

At least 163 records · Page 9Linked to original sources

PubMed2000

[T1-weighted vs. short-TE-long-TR images: usefulness for knee MR examinations of ligament and meniscal lesions].

The purpose of this study was to compare short-TE-long-TR images with T1-weighted images in knee MR examinations. Sagittal MR images of the knee were obtained in 31 patients with knee pain. T1-weighted images were obtained by the spin-echo technique (TR/TE = 350/15), and short-TE-long-TR images by fast spin-echo (TR/TE = 1300/15) with an echo-train length of 5. Contrast-to-noise-ratios (CNRs) of the anterior cruciate ligament and synovial space, meniscus and articular cartilage, and meniscal lesion and normal meniscus were compared between short-TE-long-TR images and T1-weighted images. On each of the three examinations, short-TE-long-TR images provided significantly higher CNRs than T1-weighted images. It was concluded that short-TE-long-TR images can be a useful alternative to T1-weighted images in evaluating the anterior cruciate ligament and meniscal lesions.

Adolescent↗

PubMed2000

MDR1 gene overexpression and altered degree of methylation at the promoter region in bladder cancer during chemotherapeutic treatment.

Overexpression of the multidrug resistance 1 (MDR1) gene is closely associated with the clinical outcome of hematopoietic malignancies, but the alteration of its expression during chemotherapeutic treatment and the precise mechanism underlying MDR1 gene overexpression in solid tumors remains unclear. We determined the expression and degree of methylation at the promoter of the MDR1 gene in bladder cancer. The mRNA levels of the MDR1 gene were found to be markedly enhanced, 3.5- to 5.7-fold higher in bladder cancers after chemotherapeutic treatment than those in untreated primary tumors. The MDR1 gene was overexpressed in recurrent tumors in 89% of patients who showed rerecurrence, whereas overexpression was observed in 25% of the patients without re-recurrence. A statistically significant inverse correlation existed between MDR1 expression and the methylation of 5'CpG sites at the promoter in patients with bladder cancer after chemotherapeutic treatment, with the degree of methylation at several CpG sites, rather than other specific sites, involved in this regulation. Consistent with the increase in MDR1 expression, the frequency of patients with a hypermethylated promoter decreased to 50 and 17% after intravesical and systemic chemotherapy, respectively. Thus, overexpression of the MDR1 gene might be a prognostic marker for intravesical recurrence, whereas methylation of the promoter region negatively regulates MDR1 expression and the appearance of multidrug resistance mediated by P-glycoprotein in bladder cancers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2000

[Limits of nucleic acid amplification tests for diagnosis of smear negative pulmonary tuberculosis].

STUDY OBJECTIVES: To assess the usefulness of commercial kits of nucleic acid amplification test (NAAT) for diagnosis of smear negative (SN) pulmonary tuberculosis. DESIGN AND PATIENTS: Retrospective study of patients who were diagnosed as, or suspected of pulmonary tuberculosis during 3 years from January 1996 to December 1998 in Fukujuji Hospital which has 100 beds for tuberculosis patients. MEASUREMENTS AND RESULTS: 145 smear-negative culture-positive pulmonary tuberculosis patients are entered to our analysis. The DNA-based amplification test kit (Amplicor Mycobacterium Tuberculosis Test (AMPL), Roche Diagnostic Systems, Basel, Switzerland) detected 39.2% (20/51, 95% confidence interval (CI): 25.8-52.6%) of smear-negative culture-positive (SNCP) pulmonary tuberculosis cases. The RNA-based amplification test kit (Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (AMTDT), Gen-Probe Inc., San Diego, Calif., USA) detected 40.5% (15/37, 95% CI: 24.7-56.3%) of SNCP pulmonary tuberculosis cases. For both NAATs (AMPL and AMTDT), between two groups with and without the NAAT at diagnosis of SNCP pulmonary tuberculosis, there was statistical difference in culture-positive rate (proportion of positivity in sputum culture tests at diagnosis), but no statistical difference in maximum number of colony of Mycobacterium tuberculosis (MTB). When stratified for the culture-positive rate, adjusted sensitivity for SNCP patients was 44.2% (AMPL) and 40.4% (AMTDT) respectively. On the other hand, among 245 patients with sputum AMPL positive results during the 3 years, 8 were smear-negative culture-negative (SNCN), only one out of these 8 cases was judged as true active tuberculosis without treatment. Among 89 patients with sputum AMTDT positive results, 7 were SNCN, and 3 out of them were judged as true active tuberculosis without treatment. CONCLUSION: Usefulness of commercial NAAT kits (AMPL and AMTDT) to diagnosis SN pulmonary tuberculosis is limited in the point of sensitivity.

DNA, Bacterial↗

PubMed2000

Olfactory evoked responses in Alzheimer's disease.

In Alzheimer's disease, a rapid adaptation was recognized by suprathreshold odorant stimulation. This adaptation in Alzheimer's disease may be caused by dysfunction of the mechanism in adaptation of a quickly recovering central process.

Alzheimer Disease↗

PubMed1999

Structural characterization of the human interleukin-13 receptor alpha1 gene promoter.

Human cancer cells have been found to express a large number of IL-13 receptors. We have previously shown that mRNA encoding one of these receptors, IL-13Ralpha1, is increased in cisplatin-resistant cells and is upregulated in tumor cells cultured with cisplatin. To understand the molecular mechanism of IL-13Ralpha1 gene expression, we cloned approximately 52 kbp of the IL-13Ralpha1 gene and sequenced the first exon and about 1 kbp of the upstream DNA. The first exon is 211 bp and contains 88 bp of coding sequence, while the first intron is about 13 kbp in length. The promoter region, which is GC rich, was found to lack both TATA and CCAAT boxes. Transient expression assays revealed that transcription of the IL-13Ralpha1 gene was significantly higher in cisplatin-resistant cells than in parental, cisplatin-sensitive cells. Deletion analysis of the IL-13Ralpha1 promoter identified a 70-bp core promoter region upstream of the transcription initiation site. Electrophoretic gel mobility shift assays showed that a synthetic IL-13Ralpha1 oligonucleotide (nt -40 to nt -15) bound a nuclear factor from cisplatin-resistant cells to a significantly greater degree than the equivalent factor from parental cells. This oligonucleotide was found to contain a palindromic sequence with a BstEII recognition site at its center. This palindromic sequence functions to mediate upregulation of IL-13Ralpha1 promoter in cisplatin-resistant cells and deletion or disruption of this sequence also resulted in severe reduction of the promoter activity. These findings suggest that IL-13Ralpha1 expression is upregulated at the transcriptional level in cisplatin-resistant cells. The characterization of both the IL-13Ralpha1 promoter and the transcription factors binding to it may contribute to our understanding of IL-13Ralpha1 regulation in cancer cells.

Antineoplastic Agents↗

PubMed1999

Facilitation of the cellular uptake of a triplex-forming oligonucleotide by novel polyamine analogues: structure-activity relationships.

The inefficient uptake of oligodeoxynucleotides, including that of TFO, through the cell membrane is a limiting factor in developing gene therapy approaches for cancer and other diseases. To develop a new strategy for oligonucleotide delivery into the nucleus, we synthesized a series of novel polyamine analogues and examined their effects on the uptake of a 37-mer [32P]-labeled TFO, targeted to the promoter region of c-myc oncogene. We used MCF-7 breast cancer cells to investigate the efficacy of polyamines on the internalization of the TFO. The uptake of TFO was enhanced by complexing it with several unsubstituted polyamine analogues at 0. 1-5 microM concentrations, with up to 6-fold increase in TFO uptake in the presence of a hexamine, 1,21-diamino-4,9,13, 18-tetraazahenicosane (H2N(CH2)(3)NH(CH2)(4)NH(CH2)(3)NH(CH2)(4)NH(CH2)(3)NH(2) or 3-4-3-4-3). TFO uptake increased with the cationicity of the polyamines; however, bis(ethyl) substitution and structural features of the methylene bridging region had significant effects on TFO uptake. The majority of labeled TFO was recovered from the nuclear fraction containing genomic DNA. Electrophoretic mobility shift assay revealed enhanced binding of TFO to a target duplex containing promoter region sequence of c-myc oncogene. Treatment of MCF-7 cells with the TFO complexed with 0.5 microM 3-4-3-4-3 suppressed c-myc mRNA level by 65%, as determined by Northern blot analysis. These data indicate a novel approach to deliver oligodeoxynucleotides to the cell nucleus, and suppress the expression of target genes, and provide new insights into the mechanism of oligonucleotide transport in living cells.

Breast Neoplasms↗

PubMed1999

Purification and characterization of a novel enzyme, L-threo-3-hydroxyaspartate dehydratase, from Pseudomonas sp. T62.

L-threo-3-Hydroxyaspartate dehydratase (L-threo-3-hydroxyaspartate hydro-lyase), which exhibited specificity for L-threo-3-hydroxyaspartate (K(m)=0.74 mM, V(max)=37.5 micromol min(-1) (mg protein)(-1)) but not for D-threo or D, L-erythro-3-hydroxyaspartate, was purified from a cell-free extract of Pseudomonas sp. T62. The activity of the enzyme was inhibited by hydroxylamine and EDTA, which suggests that pyridoxal 5'-phosphate and divalent cations participate in the enzyme reaction. The NH(2)-terminal amino acid sequence showed significant similarity to the Saccharomyces cerevisiae YKL218c gene product, a hypothetical threonine dehydratase. However, the purified enzyme showed no threonine dehydratase activity.

Amino Acid Sequence↗

PubMed1999

Type-II major-surface-glycoprotein family of Pneumocystis carinii under the control of novel expression elements.

Pneumocystis carinii are coated by abundant surface proteins, named MSGs for major surface glycoproteins, that undergo antigenic variation. There are many MSG genes on the chromosomes but the MSG expression site is restricted to a unique site called type-I UCS, and silent MSG genes can be translocated to the expression site by DNA recombination. We have reported previously the existence of alternative UCS elements named type-II UCS by MSG transcript analyses (J. Infect. Dis., 171, 1563-1568 [1995]). In this study we characterized the features of type-II UCS and MSG genes that are under the control of type-II UCS (hence designated the type-II MSG family) by cDNA and genomic analyses. Contrary to type-I UCS, type-II UCS elements are located at most of the P. carinii chromosomes and are directly attached to type-II MSG genes. Type-II MSG genes consist of two subtypes in coding sizes of 2.3 kb and 3.3 kb. Antibody raised against the synthetic type-II UCS polypeptide detected products in molecular masses ranging from 90 to 100 kDa. Although the cloned type-II MSG sequences are significantly diverse, cysteine residues that are rich and highly conservative in type-I MSGs are also strictly conserved in type-II MSG proteins, showing the importance of multiple cysteines in the structural integrity of MSGs in the cell surface of P. carinii.

Amino Acid Sequence↗

PubMed1999

Enhanced transport of anticancer agents and leukotriene C4 by the human canalicular multispecific organic anion transporter (cMOAT/MRP2).

We established stable human canalicular multispecific organic anion transporter (cMOAT/MRP2) cDNA transfectants, CHO/cMOAT from non-polarized Chinese hamster ovary (CHO)-K1 and LLC/cMOAT from polarized pig kidney epithelial LLC-PK1. Human cMOAT was mainly localized in the plasma membrane of CHO/cMOAT and in the apical membrane of LLC/cMOAT. The ATP-dependent uptake of leukotriene C4 (LTC4) into CHO/cMOAT membrane vesicles was enhanced compared with empty vector transfectants. Km values in CHO/cMOAT membrane vesicles were 0.24 microM for LTC4 and 175 microM for ATP. Drug sensitivity to vincristine and cisplatin in human cMOAT cDNA transfectants decreased, but not to etoposide. Cellular accumulation of vincristine and cisplatin in human cMOAT cDNA transfectants decreased, but not of etoposide. The uptake of LTC4 into CHO/cMOAT membrane vesicles was inhibited by exogenous administration of vincristine or cisplatin, but not that of etoposide. Moreover, this inhibition was more enhanced in the presence of glutathione. These consequences indicate that drug resistance to vincristine or cisplatin appears to be modulated by human cMOAT through transport of the agents, possibly in direct or indirect association with glutathione.

Adenosine Triphosphate↗

PubMed1999

LOV (light, oxygen, or voltage) domains of the blue-light photoreceptor phototropin (nph1): binding sites for the chromophore flavin mononucleotide.

Phototropism, the bending response of plant organs to or away from a directional light source, is one of the best studied blue light responses in plants. Although phototropism has been studied for more than a century, recent advances have improved our understanding of the underlying signaling mechanisms involved. The NPH1 gene of Arabidopsis thaliana encodes a blue light-dependent autophosphorylating protein kinase with the properties of a photoreceptor for phototropism. NPH1 apoprotein noncovalently binds FMN to form the holoprotein nph1. The N-terminal region of the protein contains two LOV (light, oxygen, or voltage) domains that share homology with sensor proteins from a diverse group of organisms. These include the bacterial proteins NIFL and AER, both of which bind FAD, and the phy3 photoreceptor from Adiantium capillus-veneris. The LOV domain has therefore been proposed to reflect a flavin-binding site, regulating nph1 kinase activity in response to blue light-induced redox changes. Herein we demonstrate that the LOV domains of two nph1 proteins and phy3 bind stoichiometric amounts of FMN when expressed in Escherichia coli. The spectral properties of the chromopeptides are similar to the action spectrum for phototropism, implying that the LOV domain binds FMN to function as a light sensor. Thus, our findings support the earlier model that nph1 is a dual-chromophoric flavoprotein photoreceptor regulating phototropic responses in higher plants. We therefore propose the name phototropin to designate the nph1 holoprotein.

Arabidopsis↗

PubMed1999

Induction of cell-cycle arrest and apoptosis by a novel retinobenzoic-acid derivative, TAC-101, in human pancreatic-cancer cells.

In this study, we investigated the effect of a novel retinobenzoic acid, 4-[3,5-bis (trimethylsilyl) benzamido] benzoic acid (TAC-101), on the growth of 4 human pancreatic-cancer cell lines; BxPC-3, MIAPaCa-2, CFPAC-1 and AsPC-1. TAC-101 significantly inhibited the proliferation of BxPC-3 and MIAPaCa-2 cells in a time- and concentration-dependent manner, but not the proliferation of AsPC-1 cells. Furthermore, the anti-proliferative effects of TAC-101 on BxPC-3 and MIAPaCa-2 cells were stronger than those of all-trans retinoic acid. Flow-cytometric analyses indicated that treatment of BxPC-3 with TAC-101 strongly induces cell-cycle arrest at the G1 phase. The cell-cycle arrest induced by TAC-101 was accompanied by reduction of retinoblastoma-gene product (RB) phosphorylation and an increase of 2 cyclin-dependent kinase (CDK) inhibitors, p21(WAF1/Cip1) (p21) and p27Kip1 (p27). TAC-101 also caused a decrease in cyclin A and thymidylate synthase, which are E2F-regulated gene products. No changes were observed in the expression of cyclin D1, cyclin E on CDK2. In addition, Hoechst staining, gel electrophoresis and flow-cytometric analysis indicated that a marked reduction in the number of BxPC-3 cells with TAC-101 was related to the induction of apoptosis. Our results suggest that TAC-101 inhibits the growth of certain pancreatic-cancer cells by means of G1-phase cell-cycle arrest resulting from the reduction of RB phosphorylation and the up-regulation of p21 and p27 as well as the induction of apoptosis. TAC-101 may therefore be a useful agent for new therapeutic strategies focusing on inhibition of pancreatic-cancer-cell proliferation.

Antineoplastic Agents↗

PubMed1999

Sensitive high-performance liquid chromatographic determination with fluorescence detection of phenol and chlorophenols with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride as a labeling reagent.

A sensitive HPLC method for the determination of phenol and chlorophenols was developed. The fluorescence labeling reaction of phenols with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) was completed in 30 min at 60 degrees C. The separation of DIB-derivatives of five representative phenols, i.e., phenol, o-, p-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, was achieved within 35 min with an ODS column using isocratic elution. The detection limits of these DIB derivatives at a signal-to-noise ratio (SIN) of 3 were in the range of 0.024 to 0.08 microM (0.12-0.45 pmol/20 microl injection). Twelve kinds of DIB derivatives with phenols containing mono-, di-, tri-, tetra- and penta-chlorophenol were also well separated within 208 min by changing the elution conditions. The derivatives were stable for at least for 24 h when they were placed at room temperature in the dark. The proposed method was applied to the assay of human urine samples and free and total phenol were determined. The relative standard deviations (RSDs) of the proposed method for within and between-day assay were <7.0% and <14.2%, respectively. The average concentrations of free and total phenol found in urine (n=6) were 4.3+/-2.5 and 29.5+/-14.0 microM, respectively.

Benzoates↗

PubMed1999

Chondroid chordoma: fine-needle aspiration cytology with histopathological, immunohistochemical, and ultrastructural study of two cases.

Chondroid chordoma is a controversial and confusing entity that was originally described by Heffelfinger et al. (Cancer 1973; 32:410-420) as a biphasic malignant neoplasm possessing elements of both chordoma and cartilaginous tissue. Fine-needle aspiration (FNA) cytology of chondroid chordoma has not been described. The aim of our investigation was to characterize the chondroid area of chondroid chordoma and to compare the FNA features with those of well-differentiated chondrosarcoma. Clival and cervical spine chondroid chordomas were studied with light microscopy, immunohistochemistry, and electron microscopy. Chondroid chordomas demonstrated an epithelial nature by immunohistochemistry and ultrastructural studies. The FNA smears showed low cellularity, with loosely arranged or dispersed round cells in a myxoid background. Although the smears were similar to those of well-differentiated chondrosarcomas, they showed a positive reaction for epithelial markers. These findings reveal that chondroid chordoma is a variant of chordoma which possesses a hyaline matrix. Immunohistochemical demonstration of epithelial markers is useful to distinguish it from chondrosarcoma. Diagn. Cytopathol. 1999; 21:335-339.

Biomarkers, Tumor↗

PubMed1999

Simple and rapid high-performance liquid chromatography analysis of propentofylline and its main metabolites in serum using a direct injection technique.

A rapid, simple and reliable high-performance liquid chromatography (HPLC) column-switching method with UV detection (270 nm) for the simultaneous determination of propentofylline and its metabolites in human and rat sera was developed. The method involves direct injection of serum onto an HPLC column, which contains a shielded hydrophobic stationary phase for the separation of analytes from proteins in serum, and then loading the analytes onto a short octadecylsilylated silica gel (ODS) column using a switching valve. Propentofylline and its three metabolites in serum were separated from the serum components within 30 min after the injection. The detection limits (S/N = 3) of analytes spiked in human and rat sera ranged from 0.08 to 0.57 nmol/mL, and the net volume of serum used was 20 microL. The relative standard deviations for within- and between-day variations using rat serum were less than 4.3 and 5.6%, respectively. The method was used to determine propentofylline and its main metabolites in rat serum after a single intravenous dose of propentofylline (5 mg/kg).

Animals↗

PubMed1999

Enantiomer-specific high-performance liquid chromatography with fluorescence detection of methamphetamines in abusers' hair and urine.

Enantiomer-specific high-performance liquid chromatography with fluorescence detection using 4-(4,5-diphenyl-1H-imidazol-2-yl)-benzoyl chloride as a fluorescence labeling reagent was applied to determine methamphetamine and its metabolites in abusers' hair and urine. Hair samples were segmentally analyzed based on 1 cm long segments. In four hair samples, only the S(+)-enantiomers of methamphetamine and its N-demethylated metabolite, S(+)-amphetamine were detected. Satisfactory correlation (r = 0.901) between the results of high-performance liquid chromatography-fluorescence and those of gas chromatography-nitrogen phosphorous detection was obtained (n = 19). In an abuser's urine sample, the S(+)- and R(-)-enantiomers of methamphetamine, amphetamine and para-hydroxymethamphetamine were detected. The degree of N-demethylation of S(+)-methamphetamine into the corresponding metabolite of amphetamine was significantly higher than that of the R(-)-enantiomer.

Chromatography, Gas↗

PubMed1999

The human multidrug resistance protein 2 gene: functional characterization of the 5'-flanking region and expression in hepatic cells.

The human multidrug resistance protein 2 (MRP2), also termed as the canalicular multispecific organic anion transporter (cMOAT), is a member of the adenosine triphosphate-binding cassette transporter superfamily. In the liver, MRP2 mediates the multispecific efflux of various types of organic anions, including glucuronate, sulfate, and glutathione conjugates, across the canalicular hepatocyte membrane to the bile. To investigate how the MRP2 gene is expressed in liver cells, the 5'-flanking region of the human MRP2 gene was isolated from a human placental genomic library. Sequence analysis of the MRP2 promoter showed a number of consensus binding sites for both ubiquitous and liver-enriched transcription factors. Transfection of human hepatic HepG2 cells with a series of 5'-deleted promoter luciferase constructs identified a putative silencer element localized in the -1,659/-491 region and a liver-specific positive regulatory element localized in the -491/-258 region. This latter region contained the liver-abundant transcription factor CCAAT-enhancer binding protein beta (C/EBPbeta). The transcriptional activity of the promoter construct containing a mutation in the C/EBPbeta binding site was significantly decreased in HepG2 cells. This study suggests that C/EBPbeta (-356 to -343) may regulate the liver expression of the MRP2 gene.

ATP Binding Cassette Transporter, Subfamily B↗

PubMed1999

Behavioral and hormonal basis of polygynous breeding in male bush warblers (Cettia diphone).

Plasma levels of testosterone and corticosterone were measured in free-living male bush warblers captured on their breeding ground at different times of the breeding season. Their territoriality was also estimated from their singing response to song playbacks. The pattern of change detected in the levels of plasma testosterone was different from that of "typical" monogamous species but similar to that of polygynous species. In "typical" monogamous species, plasma testosterone levels elevated during territory settlement and courtship behavior and then declined to low, stable levels during incubation. In bush warblers, plasma levels of testosterone were already high (1-2 ng/ml) upon arrival in late March and peaked (2. 5-4 ng/ml) in early June. They then decreased but relatively high levels were maintained until early August. In late August the testosterone concentration was 0.03 ng/ml or less. Plasma levels of corticosterone also showed a seasonal change, being highest in May to July and declining in late August. Territoriality showed clear seasonality, reflecting the levels of circulating testosterone. Upon arrival, latency periods for responses to song playback were long and singing activity was rather low but this behavior was soon stabilized and a high degree of territoriality was maintained to late August. These results suggest that high levels of circulating testosterone and corticosterone allow males to pursue a polygynous breeding strategy, to hold a territory, and to maintain breeding activity for a prolonged period, characteristics which are likely to be adaptations to dense bushes with high rates of predation and brood parasitism of this species.

Animals↗

PubMed1999

Treatment for empyema with bronchopleural fistulas using endobronchial occlusion coils: report of a case.

We report herein the case of a woman with bronchopleural fistulas treated with the endobronchial placement of vascular embolization coils. She was referred to our hospital to undergo lavage of a postoperative empyema. She had undergone an air plombage operation for pulmonary tuberculosis 9 years previously. However, bronchopleural fistulas occurred postoperatively and she had to continue the use of a chest drainage tube since then. Lavage of her empyema space with 5kE of OK-432 (Picibanil: Chugai) plus 100 mg minocycline was performed once every 2 weeks for 3 months, and the purulent discharge from the empyema remarkably decreased. Thereafter, the bronchopleural fistulas were occluded endobronchially by the placement of vascular embolization coils. Soon after the procedure, air leakage from the fistulas was stopped and the drainage tube was removed 2 days later. The patient remains well without any additional treatment at 20 months after this treatment. As treatment for empyema with bronchopleural fistulas, it would be worth trying to lavage the empyema space with OK-432 until it is cleaned out and to plug the fistulas by the endobronchial placement of embolization coils, before such radical operations as thoracoplasty and space-filling of the empyema are considered.

Bronchial Fistula↗