Biomedical subjects

J Kawai

Publications and source records attributed to J Kawai.

At least 37 records · Page 2Linked to original sources

PubMed1999

Cloning of a novel gene specifically expressed in clonal mouse chondroprogenitor-like EC cells, ATDC5.

We cloned a full-length cDNA encoding a novel mouse protein, A-C2, by differential display method using mouse embryonic fibroblast C3H10T1/2 cells and mouse chondroprogenitor-like EC cells, ATDC5. The deduced amino acid sequence of A-C2 consisted of 106 amino acids with no significant homology to the sequences previously reported. Northern blot analysis showed two major bands of 2.1 and 1.8 kb sizes. Expression of A-C2 mRNA was exclusive to ATDC5 cells at their undifferentiated stage. None of ATDC5 cells at their differentiated stage and adult mice tissues examined expressed A-C2 gene.

Amino Acid Sequence↗

PubMed1999

Effects of transforming growth factor-beta signaling on chondrogenesis in mouse chondrogenic EC cells, ATDC5.

Cellular condensation of chondroprogenitors is a distinct cellular event in chondrogenesis. During this process, N-cadherin mediates cell-cell interactions responsible for the initial stage of cellular condensation and subsequently fibronectin contributes to cell-matrix interactions mediating a progression of chondrogenesis. We previously showed that chondrogenesis in mouse chondrogenic EC cells, ATDC5, was induced, at a high incidence in the presence of insulin, through formation of cellular condensation. In this study, we took advantage of the sequential progression of chondrogenesis in ATDC5 cells and evaluated, in vitro in these cells, the role of endogenous transforming growth factor (TGF)-beta in chondrogenesis. ATDC5 cells expressed TGF-beta2 mRNA at a cellular condensation stage. The treatment of undifferentiated ATDC5 cells with anti-TGF-beta32 neutralizing antibody inhibited the accumulation of Alcian blue stainable proteoglycan in a dose-dependent manner. Transfection of a dominant-negative mutant of mouse TGF-beta type II receptor to undifferentiated ATDC5 cells completely inhibited cellular condensation. Moreover, exogenously administered TGF-beta2 upregulated the expression of fibronectin and type II collagen (a phenotypic marker gene of chondrogenesis) mRNAs and downregulated that of N-cadherin mRNA in time- and dose-dependent manners. These results indicate that TGF-beta stimulates chondrogenesis via initiation of cellular condensation by transition from an initial N-cadherin-contributing stage to a fibronectin-contributing stage during processes of chondrogenesis in ATDC5 cells.

Animals↗

PubMed1999

Comparison of a 1.0 molar and a 0.5 molar formulation of gadobutrol in dynamic MR imaging of the liver in rats with hepatocellular carcinoma.

The efficacy of a 1.0 molar formulation of gadobutrol was compared with that of a 0.5 molar formulation in dynamic magnetic resonance (MR) imaging of the liver in rats with hepatocellular carcinoma (HCC). Seven nodules histologically identified as moderately differentiated HCC underwent dynamic study. In the normal parenchyma surrounding these tumors, the 0.5 molar and 1.0 molar formulations yielded similar enhancement patterns, with maximum enhancement value of approximately 35% at 20 to 40 s after injection. In the tumors, the 0.5 molar formulation induced approximately 65% enhancement, whereas the 1.0 molar formulation, used at the same dose as the 0.5 molar formulation, induced significantly higher enhancement until 10 min after injection with maximum enhancement of approximately 90%. These findings indicate that the 1.0 molar formulation could be more efficacious than the 0.5 molar formulation in dynamic MR studies of moderately differentiated HCC even when injected at the same dose.

Animals↗

PubMed1999

Phase separation with multiple length scales in polymer mixtures induced by autocatalytic reactions.

A ternary polymer blend with two components photo-cross-linked independently in its miscible region undergoes phase separation, exhibiting morphology with multiple length scales. Contrary to the case of thermally induced phase separation, the morphology exhibits a unimodal-->multimodal transition. It is shown that these multiple length scales are caused by the inhomogeneous freezing kinetics of the cross-linking process. This inhomogeneity arises from the autocatalytic feedback driven by the couplings between concentration fluctuations and the photo-cross-linking reactions.

Journal Article↗

PubMed1999

[Evaluation of mitral valve lesions in patients with infective endocarditis by three-dimensional echocardiography].

Recognition of the involved lesions is extremely important in mitral valve repair for infective endocarditis. Transesophageal echocardiography (TEE) is more sensitive for the detection of lesions than transthoracic echocardiography, but localization of the lesions is sometimes difficult by TEE. Three-dimensional (3D) TEE provides images of the mitral valve similar to the view from the left atrium. This study evaluated the value of 3D echocardiography for the diagnosis of involved lesions in 12 patients who underwent surgery for mitral regurgitation due to infective endocarditis. The location of the lesion in the mitral valve was classified as the medial, central and lateral portions of the anterior leaflet, and the medial, middle and lateral scallops of the posterior leaflet, respectively. In all patients, the involved sites were confirmed at operation. The sensitivities of 3D TEE for detecting the lesions at the medial, central and lateral portions of the anterior leaflet, and the medial, middle and lateral scallops of the posterior leaflet were 100%, 78% and 67%, and 100%, 100% and 100%, respectively. The specificities were 90%, 100% and 78%, and 100%, 100% and 100%, respectively. The lesions diagnosed by 3D TEE coincided with lesions confirmed at operation in 23 (92%) of 25 lesions. 3D TEE is useful for the assessment of the involved lesion of the mitral valve in patients with infective endocarditis.

Echocardiography, Three-Dimensional↗

PubMed1999

[Usefulness of tissue harmonic imaging for the detection of left ventricular endocardial border].

Assessment of segmental wall motion is one of the most challenging tasks in echocardiography. One major limitation is impairment of echocardiographic regional wall motion by incomplete endocardial definition. The newly developed tissue harmonic imaging method may improve the detection of left ventricular endocardial border. This study examined the impact of native tissue harmonic imaging on endocardial border definition. Fundamental and harmonic imaging were compared for detecting the endocardial border in 96 segments of 16 patients (age 54 +/- 8 years). Visualization of endocardial border was better with harmonic imaging than with fundamental mode in 49% (47 of 96 segments). Thus, tissue harmonic imaging has a significant impact on endocardial border definition.

Cardiomyopathies↗

PubMed1999

[Effects of imaging parameters on automated cardiac flow measurement using color Doppler echocardiography].

The color Doppler echocardiographic technique has been developed for automated cardiac flow measurement (ACM). This study evaluated the effect of imaging parameters on stroke volume measurement. Cardiac output derived from the ACM method was compared with that obtained from pulsed wave Doppler in 36 patients (26 men and 10 women, mean age 54 +/- 8 years) in whom clear two-dimensional and color Doppler images of the left ventricular outflow tract were obtained. The effects of frame rate, color gain and moving target indicator (MTI) filter on cardiac output were evaluated in 13 patients (8 men and 5 women, mean age 49 +/- 6 years). Using ACM at a frame rate of 30 Hz, optimal color gain setting and high-frequency MTI filter (cutoff frequency: 915 Hz), there was an excellent correlation in cardiac output between the ACM and pulsed wave Doppler methods (stroke volume: r = 0.91, SEE = 0.32 l/min). Using ACM at a frame rate of 30, 22 and 15 Hz, the differences in stroke volume were 4.4%, 5.2% and 8.6%, respectively. When color gain was reduced, left ventricular stroke volume reduction was 12.1% (-2 dB), 18.9% (-4 dB). In contrast, there was no significant change in stroke volume measurement when color gain was increased. There was a significant decrease in stroke volume using the low-frequency MTI filter [cutoff frequency: 467 Hz (-35.6%)] and medium-frequency MTI filter [cutoff frequency: 703 Hz (-13.4%)]. Color Doppler imaging parameters are extremely important for automated assessment of cardiac output.

Automation↗

PubMed1999

Automated filtration-based high-throughput plasmid preparation system.

Current methods of plasmid preparation do not allow for large capacity automated processing. We have developed an automated high-throughput system that prepares plasmid DNA for large-scale sequencing. This system is based on our previously reported filtration method. In this method, cell harvesting, alkaline lysis, and plasmid purification occur in a single 96-well microtiter plate from which sequence-ready DNA samples are collected. The plates are designed to allow all reagents to be injected from above the wells and the spent reagents to be aspirated from below. This design has enabled us to build a linear process plasmid preparation system consisting of an automated filter plate stacker and a 21-stage automated plasmid preparator. The 96-well plates used are outfitted with glass-filters that trap Escherichia coli before the plates are stacked in the automated stacker. The plates move from the stacker to each of the 21 stages of the preparator. At specific stages, various reagents or chemicals are injected into the wells from above. Finally, the plates are collected in the second stacker. The optimal throughput of the preparator is 40,000 samples in 17.5 hr. Here, we describe a pilot experiment preparing 15,360 templates in 160 specially designed 96-well glass-filter plates. The prepared plasmids were subjected to restriction digestion, DNA sequencing, and transcriptional sequencing.

Base Sequence↗

PubMed1998

Improved restriction landmark cDNA scanning and its application to global analysis of genes regulated by nerve growth factor in PC12 cells.

Restriction landmark cDNA scanning (RLCS) is a novel method by which more than 1000 genes can be simultaneously and quantitatively displayed as two-dimensional gel spots. Here we present an adaptation that allows an individual spot to correspond to a unique gene species without redundancy in more than two gel patterns. Using this improved RLCS, we examined global changes on the gene expression of PC12 cells before and after treatment with nerve growth factor. Among a total of 3000 spots, 21 (0.70%) and 91 (3.03%) spots newly appeared and became more intense with treatment. On the other hand, 15 (0.50%) and 44 (1.47%) spots disappeared, becoming less intense with treatment. These observations suggest that approx. 6% of the detected PC12 genes are up-(3.73%) or down-(1.97%) regulated when the cells differentiate to neuronal cells. In comparison with the results obtained using the expressed-sequence-tag approach, previously reported by Lee et al. (Proc. Natl. Acad. Sci. USA 92 (1995) 8303-8307), RLCS should be useful for quantitatively examining the global change of differentially expressed genes of various expression levels.

Animals↗

PubMed1998

Mn Lalpha,beta and F Kalpha resonant X-ray fluorescence spectroscopy of MnF(2).

Mn Lalpha,beta and F Kalpha(1,2) (K-L(3,2)) X-ray fluorescence spectra of MnF(2) were measured when the excitation X-ray energy was higher and lower than the threshold energies. Monochromated synchrotron radiation was used for primary excitation. The resonance fluorescence of Mn Lbeta and the multiply ionized F Kalpha(3,4) satellites (KL-L(2)) were observed.

Journal Article↗

PubMed1998

Material analysis end-station of the Hyogo-ken beamline at SPring-8.

Plans to construct surface-analysis equipment which will be placed on beamline BL24XU of SPring-8 are presented. There are three experimental hutches in BL24XU, which are available simultaneously by using diamond monochromators as beam splitters. The purpose of the surface-analysis equipment is the simultaneous measurement of fluorescent and diffracted X-rays in grazing-incidence geometry. The instrument is equipped with a solid-state detector (SSD) and a flat position-sensitive proportional counter (PSPC) combined with analysing crystals for X-ray fluorescence (XRF) analysis. A curved PSPC and the goniometer that mounts the SSD used for XRF are also installed for X-ray diffraction. X-ray fluorescence holography and polarized X-ray emission spectroscopy modes are available, so three-dimensional images of atomic configurations and also the anisotropic structure of materials will be studied.

Journal Article↗

PubMed1998

Human and mouse chromosomal mapping of Stac, a neuron-specific protein with an SH3 domain.

Stac is a novel neuron-specific protein consisting of a cysteine-rich domain and an SH3 domain. We determined the chromosomal location of both the mouse and the human Stac genes (Stac and STAC). By linkage analysis using a mouse recombinant inbred (RI) strain panel (BXD), it was determined that Stac is located between D9mit15 and D9mit20 at the distal region of chromosome 9, around which two neurological disease genes, du and tip, have been found. This result was also supported by analysis using an interspecific backcross panel BSS. The mouse chromosomal location around Stac was syntenic with the human chromosome around 3p21-p23, where the gene for cerebellar ataxia (ADCA type II) was recently mapped. By radiation hybrid mapping, STAC was assigned to human chromosome 3p22-p24, and it was found to be distinct from the ADCA type II locus.

Animals↗

PubMed1998

Properties, sequence, and synthesis in Escherichia coli of 1-aminocyclopropane-1-carboxylate deaminase from Hansenula saturnus.

The plant hormone ethylene is generated from a unique precursor, 1-aminocyclopropane-1-carboxylate (ACC). In previous studies, ACC deaminase, which degrades ACC to alpha-ketobutyrate and ammonia, was found in four strains of Pseudomonas, characterized, and sequenced. To verify the wider distribution of ACC deaminase in microorganisms, we purified and sequenced ACC deaminase from the yeast Hansenula saturnus. The purified enzyme was active toward ACC, D-serine and dl-coronamic acid, indicating the same stereospecificity as the Pseudomonas enzyme, but unlike the bacterial enzyme it was not active toward beta-chloro-D-alanine and O-acetyl-D-serine. Analyses of peptides from proteolytic digests of the purified and modified ACC deaminase covered more than 90% of its amino acid sequence and showed a blocked N-terminal residue as N-acetylserine. A cDNA encoding the ACC deaminase was isolated from H. saturnus cells incubated in alpha-aminoisobutyrate medium, and sequenced. The yeast enzyme has 441 amino acid residues, of which 60 to 63% are identical to those of reported Pseudomonas enzymes. The open reading frame encoding ACC deaminase was subcloned into pET-11d and expressed in Escherichia coli BL21 (DE3) as an active enzyme.

Amino Acid Sequence↗

PubMed1998

Ovarian mucinous cystadenocarcinoma with malignant mural nodules.

A case of ovarian mucinous cystadenocarcinoma with malignant mural nodules is reported. The patient was a 28-year-old Japanese female (gravida 0, para 0) with a 2 year history of increasing abdominal fullness and edema of the lower extremity. Physical examination showed a large mass in the abdomen. An abdominal ultrasound and computed tomography demonstrated a multilocular cyst with a solid component. Lymph node and distant metastases were not found. These tests were followed by surgery. The resected right ovarian tumor measured 25 x 22 x 18 cm. On cut sectioning, it was multilocular with multiple mural nodules. Microscopically, the cyst wall was lined with papillary infoldings of atypical mucinous epithelium (intestinal type). Nuclear stratification, cribriform and back-to-back glandular patterns and stromal invasion were observed. In addition, the solid area of the mural nodules showed spindle or polygonal cells with increased mitotic activity including atypical mitoses. Nuclear pleomorphism was marked. Necrosis and hemorrhage were also present. Reticulin stain showed these pleomorphic cell clusters circumscribed by reticulin fibers and these cells were immunoreactive for vimentin and p53.

Adult↗

PubMed1998

[Automated cardiac output measurement by color Doppler echocardiography].

Recently, automated cardiac flow measurement (ACM) has been developed for measuring the volume flow rate of blood flow through the left ventricular outflow tract. Measurements of left ventricular cardiac output by the ACM method were compared with those by the thermal dilution method with a Swan-Ganz catheter in 27 patients (16 men and 11 women; aged 44 +/- 3 years) in whom clear two-dimensional and color Doppler images of the left ventricular outflow tract were obtained. The total time required for left ventricular stroke volume calculation by both ACM and pulsed Doppler methods was measured in 10 patients (six men and four women; aged 41 +/- 2 years). There was an excellent correlation in the measurements of cardiac output between the ACM and thermal dilution methods (y = 0.77x + 0.77, r = 0.84, SEE = 0.4 l/min). The total time required for left ventricular stroke volume calculation by the ACM method was significantly shorter than that by the pulsed Doppler method (92 +/- 10 vs 177 +/- 30 sec, p < 0.01). The ACM method is simple, quick, and accurate for the automated assessment of cardiac output.

Adult↗

PubMed1997

A chronological evaluation of experimental brain infarct by diffusion-mapping and magnetization transfer contrast imaging.

BACKGROUND AND PURPOSE: There is a complex system of evolving physiochemical processes in the ischemic brain. The evaluation of this chain of processes is a major challenge of recent stroke studies. Two magnetic resonance imaging techniques: diffusion-weighted (DW) and magnetization transfer contrast (MTC) imaging were introduced in experimental studies and were shown to have sensitivity for different stages of brain infarct. MATERIALS AND METHODS: We used a reproducible middle cerebral artery (MCA) occlusion model in rat to examine infarcts of different time courses. Magnetic resonance T2-weighted (T2). DW, and MTC imaging were performed 3 h, 1 d, 3 d, 5 d, 2 w, 3 w, and 4 w after MCA occlusion. Haematoxylin/eosin (HE) stained sections, which revealed sub regions within infarct lesions, were compared to T2, DW, diffusion-mapping and MTC-mapping images. RESULTS: On DW images 3 hours post occlusion lesions were detected as an area with high signal intensity, while T2 imaging does not raise significant contrast of the lesion at this early stage. Three sub regions of the lesion having different ADCs, are discernible on diffusion-mapping images from 1 day to 7 days. The decrease of MTC effect was measured within the infarct lesion from core to marginal zone, from 7 days to 4 weeks. CONCLUSIONS: Diffusion-mapping imaging may help to examine brain infarction from 3 hours to 5 days. MTC imaging recognizes infarcts 7 days after the onset. Following this stage, MTC-mapping images may provide a quantitative method to assess infarct size.

Animals↗

PubMed1997

Apparent diffusion coefficient (ADC) and magnetization transfer contrast (MTC) mapping of experimental brain tumor.

Brain tumor tissue contains different pathological areas, such as tumor cell rich parts, necrotic tissues, and cyst. Furthermore, both neovascularization and edema formation progress along with the tumor progression. In this study we employed diffusion weighted (DW) and magnetization transfer contrast (MTC) imaging to chronologically investigate the biological characteristics of a rat glioma. RG-2 glioma cells were implanted stereotactically into the right hemisphere of male Wistar rats. MR images were taken 1, 2 and 3 weeks after inoculation. Apparent diffusion coefficient (ADC) and MTC values were calculated as follows; ADC = -ln (SI-DW/SI-T2)/1096, MTC = 1-SI-MTon/SI-MToff. Each mapping image was made based on the calculated average values of four pixels. The spatial signal changes and the real values were compared to the histological findings. The apparent increase of ADC was noted in the parenchyma adjacent to tumor suggesting the progression of edema. The tumor itself had similar or slightly increased ADC. Cystic and necrotic components appeared 2 weeks after implantation and they showed significantly higher ADC than those calculated in the contralateral putamen. On the other hand, MTC was slightly decreased in the parenchyma adjacent to the tumor, markedly within the tumor, and maximally in the cystic and necrotic area suggesting accumulation of macromolecules such as growth factors, cytokines, and serum albumin.

Animals↗

PubMed1997

[Evaluation of left ventricular ejection fraction using an automated contour tracking method: comparison with equilibrium radionuclide angiography].

A recently developed contour tracking method provides automated edge detection of left ventricular endocardium without a region of interest. The accuracy of this automated contour tracking (ACT) method in the evaluation of left ventricular ejection fraction was assessed in 20 patients (16 males and 4 females, mean age 58 years) using both the ACT method and equilibrium radionuclide angiography. Ejection fractions measured by the ACT method correlated well with those measured by equilibrium radionuclide angiography (y = 0.8x + 11.9, r = 0.91, SEE = 4.0%, p < 0.01). The ACT method provides an accurate estimate of left ventricular ejection fraction.

Echocardiography↗