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Biomedical subjects

N Fujita

Publications and source records attributed to N Fujita.

At least 19 recordsLinked to original sources

Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 min region.

A contiguous 111,402-nucleotide sequence corresponding to the 0 to 2.4 min region of the E. coli chromosome was determined as a first step to complete structural analysis of the genome. The resulting sequence was used to predict open reading frames and to search for sequence similarity against the PIR protein database. A number of novel genes were found whose predicted protein sequences showed significant homology with known proteins from various organisms, including several clusters of genes similar to those involved in fatty acid metabolism in bacteria (e.g., betT, baiF) and higher organisms, iron transport (sfuA, B, C) in Serratia marcescens, and symbiotic nitrogen fixation or electron transport (fixA, B, C, X) in Azorhizobium caulinodans. In addition, several genes and IS elements that had been mapped but not sequenced (e.g., leuA, B, C, D) were identified. We estimate that about 90 genes are represented in this region of the chromosome with little spacer.

Bacterial Proteins

Measurement of right ventricular mass in normal and dilated cardiomyopathic ventricles using cine magnetic resonance imaging.

The accurate quantification of right ventricular (RV) mass has eluded conventional imaging modalities. Accordingly, cine magnetic resonance imaging was used for quantification of RV as well as left ventricular (LV) mass in 10 normal subjects and in 10 patients with dilated cardiomyopathy with an LV ejection fraction less than 0.40. Hearts were imaged with 10 mm thick short-axis slices from apex to base with a short echo delay time of 5 ms. Each slice was partitioned into 3 sections: RV free wall, ventricular septum and LV free wall, for calculation of end-diastolic and end-systolic mass and LV:RV free wall ratio. RV end-diastolic mass in normal subjects was 45 +/- 8 g, which was similar to the values determined in previously published postmortem studies, mean 46 g (range 23 to 68). The value determined in patients with dilated cardiomyopathy was higher (50 +/- 11 g), but this difference was not significant. LV:RV free wall ratio in cardiomyopathy (3.6 +/- 1.0) was greater than in normal subjects (2.4 +/- 0.3), because of the greater LV free wall mass in dilated cardiomyopathy, where LV free wall end-diastolic mass was 173 +/- 40 g vs 107.1 +/- 19.9 g in normal subjects (p less than 0.05). RV mass measurements had 6.4 +/- 3.6% interobserver and 7.3 +/- 6.1% intraobserver variability. There were no significant differences between end-diastolic and end-systolic mass measurements. Thus, cine magnetic resonance imaging can reproducibly calculate RV mass. The values in normal subjects correspond to previously reported postmortem values for a population without heart disease.

Adult

Identification of Escherichia coli proteins cross-reacting with antibodies against region 2.2 peptide of RNA polymerase sigma subunit.

Antisera against a synthetic tetradecameric peptide with the sequence DLIQEGNIGLMKAV, which is present in region 2.2 of both sigma 70 and sigma 32 subunits of Escherichia coli RNA polymerase, cross-reacted with more than 10 E. coli proteins including these two sigma subunits. Four major species of these cross-reacting proteins (SCRPs) were purified. N-Terminal amino acid sequence analysis revealed that one of them (SCRP-27A) was an as yet unidentified protein while the other three (SCRP-34, SCRP-27B and SCRP-23) were thioredoxin reductase, ribosomal protein S2, and alkyl hydroperoxide reductase, respectively. Immunological competition experiments with various fragments of this sigma region 2.2 peptide indicated that the anti-sigma peptide serum contained at least three different species of antibodies. All the four SCRPs analyzed here reacted with an antibody against a C-terminus-proximal epitope.

Amino Acid Sequence

[MR imaging of hilar cholangiocarcinoma--comparative study with CT].

Magnetic resonance (MR) images of 18 cases of hilar cholangiocarcinoma were evaluated to compare the effectiveness of Gd-DTPA with that of high dose contrast enhancement computed tomography (HCE-CT) in detecting the primary tumor. The primary tumor was demonstrated as having slightly low intensity compared with liver parenchyma and high intensity compared with the dilated bile duct on T1 weighted images. In contrast, MRI using Gd-DTPA, which was carried out in five cases, revealed intense enhancement of the tumor. As the differentiation between cholangiocarcinoma and dilated bile duct was difficult, it was concluded that the use of Gd-DTPA improves the efficacy of MRI in diagnosing cholangiocarcinoma. Gd-DTPA was also effective in differentiating the growth pattern of the tumor: the infiltrating type was demonstrated as thickening of the wall of the bile duct, the polypoid type as a soft tissue mass in the bile duct. Contrast MRI study is effective for the detection of cholangiocarcinoma. It is also expected to be effective in the staging diagnosis of cholangiocarcinoma.

Adenoma, Bile Duct

Promoter selectivity of the stationary-phase forms of Escherichia coli RNA polymerase and conversion in vitro of the S1 form enzyme into a log-phase enzyme-like form.

Upon growth transition of Escherichia coli cells from exponential to stationary phase, RNA polymerase is converted into at least three different forms (S1, S2 and S3), which could be separately isolated by phosphocellulose column chromatography (Ozaki et al., 1991 (2)). Here, the promoter selectivity of these three stationary-phase enzymes was examined using an in vitro mixed transcription system and an E. coli promoter collection. These altered forms of RNA polymerase showed different recognition properties of promoters from that by the log-phase holoenzyme (L1). One of the stationary-phase RNA polymerases, S1, was found to be converted in vitro into an enzyme like the log-phase form following incubation with nucleotides or pyrophosphate. The conversion was indicated by not only the shift of elution position from a phosphocellulose column but also the change in the promoter selectivity. These results may suggest that RNA polymerase is interconvertible between different forms with different promoter selectivity by interaction with a phosphorylated compound(s).

Adenosine Triphosphate

Immunophenotype and DNA content of myeloma cells in primary plasma cell leukemia.

To clarify the biological characteristics of the myeloma cells in primary plasma cell leukemia (PCL), we studied the immunological phenotype and DNA content of cells from five patients with primary PCL as compared to those from the patients with typical multiple myeloma (MM). In two of five patients with PCL myeloma cells had B-cell-associated antigens (B1, J5) and surface immunoglobulin in addition to plasma cell associated antigens, suggesting that these cells are immunologically immature as compared to mature plasma cells. Concerning the DNA content of myeloma cells, two of four patients had hypodiploid while two had diploid myeloma cells. In contrast, 24 of the 37 (65%) patients with typical MM had hyperdiploid and 1 had hypodiploid myeloma cells. These observations suggest that the myeloma cells in PCL are immunologically heterogeneous. The increased incidence of hypodiploidy in PCL may explain its relatively poor prognosis as previously shown in those with typical MM and hypodiploid DNA content.

Adult

Separation of diffusion and slow flow effects by use of flow rephasing and dephasing.

Spin-echo signal intensity alterations due to diffusion and slow flow are investigated in connection with modified Stejskal-Tanner pulse sequences for 2D Fourier MR imaging, one for flow rephasing and the other for flow dephasing gradient waveforms. The theoretical considerations and experimental results concerning the diffusion coefficient measurements of slowly flowing material by these sequences are summarized as follows: (a) By using the flow dephasing sequence with different diffusion and flow-sensitive gradients, slow flow effects can be distinguished from diffusion effects based on the quantitative difference between the diffusion- and flow-sensitive gradient amplitude dependences of these processes. (b) By using the combination of the flow rephasing and dephasing sequences with the same diffusion sensitivity, slow flow effects can be distinguished from diffusion effects directly based on their qualitative difference. By use of the latter approach, several in vivo images are also presented, which mainly represent macroscopic motions of spins, including slow flow such as tissue perfusion.

Acetone

Follow-up MRI in dural arteriovenous malformations involving the cavernous sinus: emphasis on detection of venous thrombosis.

Six patients with a dural arteriovenous malformation (dural AVM) involving the cavernous sinus were followed up with magnetic resonance imaging in order to assess change in the lesions. Spin-echo (SE) imaging of three patients in whom the AVM appeared to have closed at least 1 month earlier (two of them spontaneously, and one after external carotid artery embolization) showed neither apparent flow void in the involved cavernous sinus nor evidence of venous thrombosis. SE images of the other three patients who had not been cured by external carotid artery embolization (two of whom were examined within a week of treatment), detected persisting arteriovenous shunts, including high-flow cortical venous drainage, seen as flow void. Two-dimensional time-of-flight MR angiography (2D TOF MRA) was performed simultaneously in three patients. Whereas shunting blood and the normal cavernous sinus were of high intensity, presumed thrombosed cavernous sinuses were isointense with stationary brain tissue. SE imaging can confirm the resolution of arteriovenous shunts, but poorly delineates very acute and chronic thrombosis of the draining veins. In contrast, 2D TOF MRA directly demonstrates flowing blood, permitting the diagnosis of venous thrombosis; it should be included in follow-up of a dural AVM involving the cavernous sinus when venous thrombosis is suspected.

Adult

Effects of bilateral jugular vein ligation on intracranial pressure and cerebrospinal fluid outflow resistance in cats.

In order to study the effect of jugular venous outflow obstruction on intracranial pressure and cerebrospinal fluid (CSF) reabsorption capability, changes in epidural pressure (EDP) and CSF outflow resistance (Ro) were examined following bilateral jugular vein ligation in cats. EDP increased significantly (P less than 0.01) immediately after ligation from the control value of 4.9 +/- 0.5 mmHg (mean +/- SEM) to 15.9 +/- 0.9 mmHg. Thereafter, EDP gradually decreased back toward the control value. The pressure level had decreased to 6.7 +/- 0.5 mmHg by 20 minutes after ligation. The mean Ro was significantly (P less than 0.01) higher in the ligation group (200.4 +/- 9.7 mmHg/ml/min) that in the non-ligation group (120.0 +/- 9.9 mmHg/ml/min). These results suggest that bilateral jugular vein ligation impairs CSF reabsorption.

Absorption

Alveolar hydatid disease of the liver: computed tomography and transabdominal ultrasound with histopathological correlation.

The appearances of alveolar hydatid disease of the liver (AHDL) on computed tomography (CT) and ultrasound (US) were retrospectively compared with histopathological appearances in 67 patients with 100 separate lesions. The radiological features were correlated directly with the pathological specimens obtained from each patient. We conclude that the CT appearances are more specific, but that US has a role to play in mass screening in endemic areas, and intraoperatively.

Adolescent

Visual space perception and visually directed action.

The results of two types of experiments are reported. In 1 type, Ss matched depth intervals on the ground plane that appeared equal to frontal intervals at the same distance. The depth intervals had to be made considerably larger than the frontal intervals to appear equal in length, with this physical inequality of equal-appearing intervals increasing with egocentric distance of the intervals (4 m-12 m). In the other type of experiment, Ss viewed targets lying on the ground plane and then, with eyes closed, attempted either to walk directly to their locations or to point continuously toward them while walking along paths that passed off to the side. Performance was quite accurate in both motoric tasks, indicating that the distortion in the mapping from physical to visual space evident in the visual matching task does not manifest itself in the visually open-loop motoric tasks.

Adult

CD7 expression in malignant pleural mesothelioma.

CD7 antigen was found to be expressed on malignant mesothelioma arising from the right pleura in a 15-year-old girl not only by immunostaining using monoclonal antibodies, but also by Northern blot analysis. The level of expression in this tumor was comparable to those in T-cell lines, Jurkat and CCRF-CEM. Cytogenetic analysis of the tumor showed hypodiploidy (n = 43). CD7 has been regarded as one of the hematopoietic cell markers selectively expressed on the majority of T cells and multipotential stem cells. To our knowledge, this is the first report of a non-hematopoietic tumor expressing CD7.

Adolescent

Mapping the cAMP receptor protein contact site on the alpha subunit of Escherichia coli RNA polymerase.

The C-terminal region (amino acid residues 236-329) of the Escherichia coli RNA polymerase alpha subunit carries the contact site I for positive transcription factors. For detailed mapping of the contact site for the cAMP receptor protein (CRP), we made a library of mutant rpoA by polymerase chain reaction (PCR) mutagenesis, such that each should carry a single mutation on average and exclusively in the C-terminal half of the rpoA gene, and then screened this library for mutants with decreased expression of the lacZ gene. Reconstituted holoenzyme containing the mutant alpha subunits transcribed galP1 but not lacP1 in vitro in the presence of cAMP-CRP. DNA sequence determination of several 'Lac-' mutant rpoA genes revealed that all had mutations clustered within a short segment near the C-terminus of alpha, between amino acid residues 265 and 270. A cluster of contact sites appear to exist within the contact site I region, each comprising of about five amino acids and responding in molecular communication with a different transcription factor(s).

Bacterial Proteins

Production of basic fetoprotein in human peripheral lymphocytes during blastic transformation.

To clarify the significance of basic fetoprotein (BFP) in lymphocytes, we investigated whether BFP is produced in lymphocytes during blastic transformation. Peripheral blood lymphocytes obtained from 14 adults were cultured under the stimulation of lectins. The concentration of BFP in the culture medium (extracellular BFP) was estimated serially. The incorporation of [6-3H]-thymidine was assayed simultaneously. The intracellular BFP was measured by dual flow cytometry for DNA and BFP. A lymph node was studied immunohistochemically. Serum BFP was measured in four cases of lymphocytic leukaemia. In two cases, dual staining was performed. The intracellular BFP of the mitogen-stimulated lymphocytes was increased within 24 h. The extracellular BFP was increased exponentially from 72 h. The extracellular BFP at 96 h did not correlate with the [3H]-thymidine incorporation. The intracellular BFP increase began in G1 phase. Immunostaining showed that the B cells also produced BFP. The serum BFP level in leukaemia was high in 1 of 4 cases and the leukaemic cells in two cases showed high intracellular BFP content. These observations indicate that BFP is produced in activated human lymphocytes and in lymphocytic leukaemic cells. The production of BFP during blastic transformation will be a useful new in vitro model for studying the biological role of BFP, and BFP labelling may offer some new possibilities for the study of lymphocytes.

Adult

Novel type of clonally involved cytoplasmic immunoglobulin-negative cells in multiple myeloma: flow cytometric study.

In order to identify myeloma cells and their precursor cells, we studied bone marrow samples from 39 patients with multiple myeloma, including 2 with plasma cell leukemia, using dual parameter cytometry of both DNA and cytoplasmic immunoglobulin (Cyt-Ig). Myeloma cells from 25 patients (64.1%) were aneuploid and those from the remaining 14 patients were diploid. In 25 patients with aneuploid myeloma, we found a small component of cells which had aneuploid DNA content without Cyt-Ig in addition to myeloma cells showing aneuploidy with Cyt-Ig. These aneuploid Cyt-Ig-negative cells were considered to be clonally involved because they were aneuploid, and detected predominantly in patients with stage III myeloma. This suggests that these cells may contribute to disease progression. The characteristics of these cells are described and the relationship between these cells and myeloma precursor cells is discussed.

Aneuploidy