Biomedical subjects

T Watabe

Publications and source records attributed to T Watabe.

At least 19 recordsLinked to original sources

PubMed1995

Disruption of BMP signals in embryonic Xenopus ectoderm leads to direct neural induction.

Bone morphogenetic proteins (BMPs), which have been implicated in the patterning of mesoderm, are members of the transforming growth factor-beta (TGF-beta) superfamily. We have investigated the roles of Xenopus BMP-7 (XBMP-7) and BMP-4 (XBMP-4), and activin (another TGF-beta-related molecule) in early development by generating dominant-negative versions of these growth factors. Mutations were generated by altering the cleavage sites that are required for maturation of the active dimeric forms of XBMP-7, XBMP-4, and activin. These mutant constructs, designated Cm-XBMP-7, Cm-XBMP-4, and Cm-activin, result in polypeptides that allow for dimerization of the subunits, but are incapable of maturation. Expression of Cm-XBMP-7 and Cm-XBMP-4, but not Cm-activin, in the ventral marginal zone of the Xenopus embryo results in the development of a secondary axis, similar to that seen by ectopic expression of the truncated BMP receptor. These results suggest that the cleavage mutants interfere with BMP signaling during mesodermal patterning. We also found that expression of Cm-XBMP-7 or Cm-XBMP-4 in animal cap ectoderm directly induces neuroectoderm. The neural induction was specific for Cm-XBMP-7 and Cm-XBMP-4 because ectopic expression of Cm-activin or Vg-1 did not mimic the same phenotype. Molecular study of neural patterning by Cm-XBMP-7 and Cm-XBMP-4 revealed that only anterior neuroectodermal markers are expressed in response to these Cm-XBMPs. These results suggest that the BMPs are involved in the specification of ectoderm in Xenopus development, and that neural induction requires the removal of BMP signals in the ectoderm. We propose that neural induction occurs by a default mechanism, whereby the inhibition of BMP signaling is required for the conversion of ectoderm to neuroectoderm in the developing Xenopus embryo.

Activins↗

PubMed1995

A subfamily 2 homo-dimeric glutathione S-transferase mYrs-mYrs of class theta in mouse liver cytosol.

A homo-dimeric subfamily 2 glutathione (GSH) S-transferase (GST) mYrs-mYrs of the class theta was isolated from mouse liver cytosol and purified to homogeneity. The first 28 N-terminal amino acid sequence of the GST was completely identical to that of rat subfamily 2 GST Yrs-Yrs of the class theta. GST mYrs-mYrs cross-reacted with anti-rat GST Yrs-anti-sera but not with anti-sera raised against rat GSTs Ya-Ya (alpha), Yb1-Yb1 (mu), and Yp-Yp (pi) and represented more than 95% of the mouse liver cytosolic GST activity to scavenge the reactive sulfate ester 5-sulfoxymethylchrysene of the potent carcinogen 5-hydroxymethylchrysene. The mouse class theta GST had little activity toward 1-chloro-2,4-dinitrobenzene and was unretainable on GSH and an S-hexyl-GSH affinity columns. GST mYrs-mYrs had a much higher GSH peroxidase activity toward fatty acid hydroperoxides than did the other classes of mouse GSTs.

Amino Acid Sequence↗

PubMed1995

Functional conservation of the Wnt signaling pathway revealed by ectopic expression of Drosophila dishevelled in Xenopus.

Wnt genes encode secreted growth factors that exhibit potent effects on both embryonic and postembryonic development in vertebrates and invertebrates. Recently, the dishevelled (dsh), shaggy/zeste-white 3, and armadillo genes have been shown to participate in Wnt (wingless; wg) signaling in Drosophila. Vertebrate genes that have sequence similarities to all of these Drosophila genes have been identified. To determine whether these structurally conserved components of insect wg signaling represent a functionally conserved Wnt signaling pathway in vertebrates, we investigated the role of Drosophila dsh in Xenopus Wnt signaling. Xenopus embryos ectopically injected with Drosophila dsh mRNA developed duplicated axes similar to those seen in embryos injected with Wnt mRNAs. The involvement of dsh function in the Wnt signaling pathway in Xenopus was demonstrated using two assays which are specifically sensitive to Wnt signaling: synergistic induction of dorsal mesoderm with bFGF and the specific induction of a Wnt-responsive reporter gene. These findings support the notion that the intracellular response to the Wnt signal has been conserved during evolution to such an extent that its components may be interchanged between distantly related species.

Animals↗

PubMed1995

Comparative response of the immature and mature ovine fetus to corticotrophin-releasing hormone (CRH).

The aim of this study was to address the possibility that the low concentrations of adrenocorticotrophin (ACTH) seen in the ovine fetus between 90 and 120 days of gestation could be attributed to an alteration in the sensitivity or responsiveness of the fetal pituitary to corticotrophin-releasing hormone (CRH), a key regulator of ACTH secretion. Chronically cannulated ovine fetuses at Days 104-108 (n = 11, representing fetuses from this 90-120-day period) and Days 138-142 (n = 6) of pregnancy received graded doses of ovine CRH (0.8, 1.6, 3.8 and 7.6 micrograms h-1 for 60 min each, given consecutively and in ascending order) or isotonic saline (n = 4 at both age groups studied). Arterial blood samples were taken concurrently for analysis of plasma immunoreactive CRH, ACTH and cortisol throughout the infusion to assess the pituitary-adrenal response. Regression lines describing the relationship between log.PCRH and log.PACTH were calculated for both age group studied. A significant (P < 0.001) rightward shift in the log.PCRH/log.PACTH regression line for the Day 104-108 group was found, suggesting that the ovine fetus at this age is less sensitive or responsive to exogenous oCRH than the mature Day 138-142 fetus. This decreased responsiveness could explain the low concentrations of endogenous ACTH seen during the 90-120-day period.

Adrenocorticotropic Hormone↗

PubMed1995

[Contrast-enhanced 3D MR angiography of the chest and abdomen with breath-holding using phase reordering].

This report presents the feasibility of phase-recordered contrast-enhanced three-dimensional MR angiography in 32 consecutive patients with vascular abnormalities in the chest and abdomen. To suppress motion artifacts due to respiratory corruption, a phase-reordering technique was introduced so that the low frequency components of the phase data were obtained first during the imaging period. Image quality and degree of motion suppression were assessed by four radiologists independently without information on breath-holding time. Abnormalities were detected in 30 cases (93.8%), and their extent was correctly assessed in 28 cases (87.5%). More confident assessment was possible in abnormalities of the pulmonary vessels and thoracic aorta than in those of the abdominal aorta and portal venous system. With phase reordering, more than 20 seconds of breath-holding ensured image quality sufficient to correctly assess the vascular abnormalities. While this technique is easy and requires only single breath-holding, it can provide excellent MRA without slice-to-slice spatial misregistration.

Aortic Aneurysm, Abdominal↗

PubMed1995

[MR imaging of temporomandibular joint (TMJ): mandibular fracture and traumatic disk injury].

Using an 1.5 Tesla superconducting MR unit and surface coil, 12 mandibular fracture patients were evaluated for TMJ disk change. Sagittal and coronal images of the TMJ with a slice thickness of 2mm were obtained with FISP 2D or 3D. TR = 30 msec, TE = 12 msec, and flip angle = 40 degrees were applied. Among 12 patients, FISP 3D revealed increased signal intensity of the disk in 70% of cases. Traumatic fluid collection at or adjacent to the TMJ showed high intensity. The disk was displaced anteromedially regardless of the site of fracture.

Adolescent↗

PubMed1995

[Breath-holding three-dimensional MR imaging: evaluation of phase reordering for motion artifact suppression].

To suppress image degradation due to respiratory corruption during breath holding, a method of reordering of phase encoding was introduced in three-dimensional MR imaging. Without trade-offs in SNR, CNR, and examination time, the method provided a significant reduction of respiratory motion-induced artifacts both in phantom and in vivo experiments. More than twenty seconds of breath holding ensured acceptable image quality regardless of respiratory motion thereafter. Signal intensity change during data acquisition altered the obtained image contrast compared with that of standard sequential linear phase encoding. This technique can be readily implemented on standard two-and three-dimensional pulse sequences and will improve image quality when a patient cannot hold his/her breath during the whole imaging period.

Artifacts↗

PubMed1994

[Three-dimensional MR angiography of aorta with Gd-DTPA infusion and breath-holding].

Using the rapid three-dimensional gradient echo technique (FLASH 18 degrees 10/4) and Gd-DTPA administration, we performed MR angiography (MRA) of the thoracic and abdominal aorta during single or double breath-holding periods. Basic in vitro and in vivo dynamic studies showed that infusion of a relatively low concentration (74 mg Gd/ml) Gd-DTPA solution provided a strong and persistent flow signal for a considerable length of time. Eleven of 12 cases clearly demonstrated MRAs of the aorta and its main branches without respiration-induced motion artifacts, although visualization of secondary and more peripheral branches depended on their size, direction and injection-imaging timing. Although limited spatial resolution and pulsatility-induced ghost artifacts remained as problems to be solved, the ease of this method, short examination time and excellent arteriograms with no need for venous or fat suppression make it clinically acceptable and very promising.

Adult↗

PubMed1994

Isolation and characterization of the gene encoding rat class theta glutathione S-transferase subunit yrs.

The rat gene encoding the class theta glutathione S-transferase (GST) subunit Yrs has been isolated from a rat lambda EMBL4 DNA library and completely sequenced. The gene spans approximately 4 kb comprising 5 exons separated by 4 introns, the smallest number of the gene components among known rat genes encoding GST subunits. Sequence analysis of the 5' flanking region of the GST Yrs gene indicated the absence of both TATA and CAAT boxes. However, potential binding sites for the transcription factors SP1, PU1, PEA3, and AP-2 were present in this region. Primer extension and 5'-rapid amplification of cDNA ends studies demonstrated the existence of multiple transcription initiation sites distributed over 58 bp in the upstream sequence from the translation initiation codon. The present study provides the first evidence for the primary structure of the gene encoding a class theta GST subunit.

Animals↗

PubMed1993

Transport pathways for macromolecules in the aortic endothelium: I. Transendothelial channels revealed by three-dimensional reconstruction using serial sections.

Three-dimensional organization of structures labeled by horseradish peroxidase as a tracer molecule in rat aortic endothelium was examined to elucidate the transport pathways for macromolecules. Aortic endothelium was divisible morphologically into four distinct parts, i.e., the parajunctional region (JR), peripheral region (PR), organelle region (OR), and nuclear region (NR). Almost all vesicles, intercellular clefts, and phagosomes were labeled by horseradish peroxidase. Peroxidase-positive vesicles tended to gather in the PR, occasional JR, and the upper part of the NR. Ultrathin serial micrographs revealed the transcellular channels composed of vesicles in the PR and JR, and the paracellular channels composed of the intercellular cleft without constrictions of tight and gap junctions. Transcellular channels were subdivided into three morphologically different types, where not only vesicles but invagination of abluminal membrane and intercellular cleft occasionally participated in their formation. Three-dimensional reconstructions from three series of consecutive electron micrographs revealed that almost all peroxidase-positive vesicles were connected directly or indirectly with the cell surface. These results indicate that the transcellular and paracellular channels are the transport pathways for macromolecules in the aortic endothelium and may suggest that the "shuttle" hypothesis is unsuitable to explain transport system for macromolecules because it postulates the existence of many free vesicles in the cytoplasm.

Animals↗

PubMed1992

[2D MR angiography of the aortic aneurysm--effectiveness of oblique saturation pulses for selective angiogram].

2D time-of-flight MR angiography was performed in 6 cases of thoracic aortic aneurysm. Oblique saturation pulses were used to suppress the signals of the pulmonary artery and SVC, providing excellent selective MR aortograms. 3 dimensional extension of the aneurysm and its relation with cervical branches were easily assessed. It could be possible to replace invasive aortography by this technique.

Aorta, Thoracic↗

PubMed1992

Vitreous, retinal and subretinal hemorrhages associated with von Willebrand's syndrome.

Two patients with von Willebrand's syndrome had intraocular hemorrhages without trauma or neovascularization. A 13-year-old Japanese girl complained of sudden spotted vision in her left eye. Vitreous, peripapillary retinal, and subretinal hemorrhages were seen by ophthalmoscopy in the affected eye. In addition, a 19-year-old Japanese man had repeated vitreous hemorrhage and epistaxis. Also noted was gyrate atrophy of the choroid and retina. Analysis of the coagulation-fibrinolysis system revealed decreased activities of Factor VIII and von Willebrand's factor in both patients. The amount of von Willebrand's factor antigen also was decreased. These results suggested that the patients had von Willebrand's syndrome. The bleeding tendency in the disorder appeared to be responsible for the pathogenesis of the intraocular hemorrhages seen in these patients.

Adolescent↗

PubMed1992

Giant air cell of the petrous apex: a possible cause of facial hypalgesia.

A giant air cell of the left petrous apex was found in a 23-year-old man with ipsilateral facial hypalgesia. The size of the giant air cell depicted on computed tomography was 1.5 x 2.0 x 2.0 cm. A coronal T1-weighted magnetic resonance image showed that the trigeminal nerve was compressed superomedially by a large signal void area that was probably a result of excessive pneumatization of the petrous apex. It is suggested that the facial hypalgesia was caused by the compression by the giant air cell of the petrous apex on the trigeminal nerve.

Adult↗

PubMed1991

Molecular cloning and amino acid sequencing of rat liver class theta glutathione S-transferase Yrs-Yrs inactivating reactive sulfate esters of carcinogenic arylmethanols.

A cDNA containing the entire coding sequence for the subunit protein of rat liver class theta glutathione S-transferase (GST) Yrs-Yrs was isolated from a rat liver lambda gt11 cDNA library. The cDNA, designated GST theta-1, consisted of 1,258 bp which had an open reading frame of 732 bp encoding a polypeptide of 244 amino acid (AA) residues, including the leading AA Met to be removed on expression. The authenticity of the cDNA structure was supported by matching its deduced AA sequence with N-termini of Yrs and peptides obtained thereof by tryptic digestion as well as by CNBr cleavage. The deduced AA sequence of the subunit Yrs (M.W. 27,311) had only a weak homology (19-23%) with those of rat liver classes alpha, mu, and pi GST isozymes. Thus, the first evidence for the molecular cloning of the class theta GST was provided.

Amino Acid Sequence↗

PubMed1991

Occurrence of cholesterol 7 alpha- and 7 beta-hydroperoxides in rat skin as aging markers.

Evidence for presence of cholesterol 7 alpha- and 7 beta-hydroperoxides in rat skin was presented for the first time. The 7-hydroperoxides in rat skin were reduced with sodium borohydride and trimethylsilylated for identification with the authentic compounds by gas chromatography/mass spectrometry. A content of cholesterol 7-hydroperoxides in rat skin, determined by high performance liquid chromatography with a chemiluminescence detector, highly correlated with the age of rats (r = 0.874; between 1 and 45 weeks old), indicating that cholesterol 7 alpha- and 7 beta-hydroperoxides were good markers for aging.

Aging↗

PubMed1991

Reductive metabolism of nitro-p-phenylenediamine by rat liver.

Reductive metabolism of the hair dye constituent, nitro-p-phenylenediamine (2-nitro-1,4-diaminobenzene, NPDA), and its acetylated metabolite, NPDA N4-acetate, was investigated with rat liver subcellular fractions, microsomes and cytosol. Under anaerobic conditions, these compounds were reduced to their corresponding amines by these fractions. The microsomal nitro-reducing activity was retarded completely by air and strongly by carbon monoxide. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) functioned more effectively than reduced nicotinamide adenine dinucleotide (NADH) as an electron donor in the microsomal reduction of the nitro compounds, and flavin mononucleotide (FMN) gave rise to a marked enhancement in the microsomal activity, especially when added to an anaerobic incubation mixture containing both NADH and NADPH. The cytosolic nitro-reducing activity was attributed to xanthine oxidase, aldehyde oxidase and other unknown enzyme(s), based on the results of cofactor requirements and inhibition experiments.

Animals↗

PubMed1991

Ontogeny of corticotrophin-releasing factor (CRF) in the ovine fetal hypothalamus: use of multiple CRF antibodies.

In previous studies, using one particular antibody, immunohistochemical localization of corticotrophin-releasing factor (CRF) in ovine fetal brain was not possible before 90 days of gestation (term is approximately 150 days), although radioimmunoassay of hypothalamic extracts, using a different antibody, had shown CRF to be present from 63 days. The purpose of this study was to use a variety of CRF antibodies in both immunohistochemistry and radioimmunoassay to determine the presence and concentration of the CRF peptide as early in gestation as possible, and to determine whether more than one molecular size of CRF is detectable at any time in gestation. Seven different antibodies were used on hypothalamic tissue or extracts from seven adult sheep and 37 fetuses from 48 to 140 days of gestation. With one ovine CRF antibody (provided by Dr W. Vale, Salk Institute) immunohistochemical detection of CRF-labelled neurones and nerve fibres of the paraventricular nucleus and median eminence was possible from 49 days. The antibody with the greatest sensitivity in radioimmunoassay was one raised against human CRF, Ab-code R1 (provided by Dr E. Hillhouse, University of Newcastle upon Tyne). The hypothalamic contents of CRF (ng/whole hypothalamus) were 0.28 +/- 0.06 (mean +/- S.E.M.) (n = 4), 9.0 +/- 0.6 (n = 5), 14.3 +/- 0.6 (n = 5), 30.0 +/- 3.4 (n = 4) in fetuses at 48-50, 100-109 and 139-140 days of gestation and in adult sheep respectively. At all ages only one peak of CRF-like activity, which co-eluted with synthetic ovine CRF, was observed after chromatography of hypothalamic extracts, and assays performed with three different antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

DNA polymerase action blocked by adenine adducts induced by 5-hydroxymethylchrysene sulfate.

Modification of M13mp10 single-stranded DNA with 5-hydroxymethylchrysene (5HCR) sulfate, the ultimate carcinogenic metabolite of 5-methylchrysene, resulted in formation of N6[(chrysen-5-yl)methyl]adenine and N2[(chrysen-5-yl)methyl]-guanine at the ratio of 2.7:1. Measurement of DNA synthesis using this modified template and E.coli DNA polymerase I (Klenow fragment) demonstrated that increasing levels of adducts caused a progressive decline in replication. Analysis of reaction products on DNA-sequence gels revealed DNA elongation to be arrested exclusively at adenine adducts in -AAAGGA- and -AACA- sequences.

Adenine↗