Search PubMed⌕ Search

Biomedical subjects

T Kawakami

Publications and source records attributed to T Kawakami.

At least 37 records · Page 2Linked to original sources
PubMed2002

Localization of matrix proteins of hard tissue in osteochondromas.

We examined 10 cases of osteochondroma by means of histopathological, histochemical and immunohistochemical techniques. The surface of the masses was covered with a cartilage tissue showing positive immunohistochemical reaction for collagen type 2, and the deep region was composed of spongy bone, showing positive immunohistochemical reaction for collagen type 1 and osteocalcin. Between the cartilage and spongy bone, which is a metaphysis-like region, a chondroidal pattern appeared in the matrix of hypertrophy cartilage. In these regions, both type 1 (and osteocalcin) and 2 collagens were immunohistochemically detected. Although there is still no direct evidence, we believe that the cells involved in so-called "chondroid bone" temporally express cartilage phenotypes and then transform directly into bone-forming cells that survive in the chondroid bone until the tissue is resorbed and remodeled. Our examination results suggest that bone formation in osteochondromas, at least in some regions, occurs through transchondroid bone formation.

Adolescent↗
PubMed2002

Development of growth-associated protein-43 and neurotrophin receptor immunoreactivities in the rat urinary bladder.

Little is known about how the prenatal innervation of the urinary bladder develops. The objective of this study was to define the timing and pattern of innervation of the developing rat bladder using growth-associated protein-43 (GAP-43) immunohistochemistry. In addition, we examined the development of two different classes of transmembrane receptor proteins, the Trk family of tyrosine kinases and neurotrophin receptor p75 immunoreactivities. GAP-43-positive nerves were first detected in the bladder at embryonic day (E) 16. They were growing rapidly to the bladder dome and covered the whole bladder by E18. The density of GAP-43-containing nerves increased in the muscle layer by postnatal day (P) 0. Both Trk receptor-positive and p75 neurotrophin receptor-positive fibers were also first seen at E16. The number of Trk immunoreactive nerves reached a peak at E18, and then decreased over the following days. In contrast, p75-labeled fibers were abundant at E18-P14. There were few GAP-43 or neurotrophin receptor-positive fibers in the adult. GAP-43 immunohistochemistry provided us with a picture of innervation in the developing rat bladder. Furthermore, the demonstration of neurotrophin receptors positive fibers in the prenatal and early postnatal bladders suggests that neurotrophins may contribute to the development of the peripheral nervous system in the urinary bladder.

Animals↗
PubMed2002

Pressure-induced transition from a charge-disproportionated antiferromagnetic state to a charge-uniform ferromagnetic state in Sr(2/3)La(1/3)FeO3.

The perovskite oxide Sr(2/3)La(1/3)FeO3 has been found to switch its electronic ground state drastically at 23 GPa through measurements of 57Fe Mössbauer spectroscopy and powder x-ray diffraction up to 56 GPa. In the low-pressure region a first-order transition from a charge-uniform paramagnetic metallic phase to a charge-disproportionated (3Fe(11/3+)-->2Fe(3+)+Fe5+) antiferromagnetic insulating phase occurs at 207 K at 0.1 MPa and 165 K at 21 GPa, typically. Above 25 GPa, however, a charge-uniform ferromagnetic (and most probably metallic) phase persists below 300 K. This switching occurs at a lattice volume of V(23 GPa)/V(0)(0.1 MPa) = 0.89.

Journal Article↗
PubMed2002

Cell death induced by a caspase-cleaved transmembrane fragment of the Alzheimer amyloid precursor protein.

The Alzheimer amyloid precursor protein (APP) is a transmembrane protein whose abnormal processing is associated with the pathogenesis of Alzheimer's disease. Activated caspases cleave APP and generate its carboxyl-terminally truncated fragment (APPdeltaC31). We have previously reported that overexpression of wild-type APP induces caspase-3 activation and apoptosis in postmitotic neurons. We now report that APPdeltaC31 potentially plays pathophysiological roles in neuronal death. Adenovirus-mediated overexpression of wild-type APP695 induced activation of caspase-3 and accumulation of APPdeltaC31 in postmitotic neurons derived from human NT2 embryonal carcinoma cells, whereas an APP mutant lacking the Abeta(1-20) region induced neither caspase-3 activation nor APPdeltaC31 generation. Inhibition of caspase-3 suppressed the generation of APPdeltaC31 in APP-overexpressing neurons. Forced expression of APPdeltaC31 induced apoptotic changes of neurons and non-neuronal cells, but failed to activate caspase-3. The cytotoxicity of APPdeltaC31 was also dependent on the Abeta(1-20) region. These results suggest that accumulation of wild-type APP activates neuronal caspase-3 to generate APPdeltaC31 that mediates caspase-3-independent cell death.

Adenoviridae↗
PubMed2002

Caffeine test in predicting flutamide-induced hepatic injury in patients with prostate cancer.

The caffeine test measures the activity of cytochrome p450 (CYP1A2) which is a major enzyme involved in the activation of flutamide. The usefulness of this test in predicting flutamide-induced hepatic injury in patients with prostate cancer was examined. The subjects were: (1). five patients whose aspartate aminotransferase (AST) or alanine aminotransferase (ALT) level rose to 100 IU/l or higher following the start of flutamide (moderately injured group); (2). four patients whose AST and ALT levels were higher than normal but less than 100 IU/l (mildly injured group); and (3). two patients whose hepatic function remained normal (normal group). The subjects were each given canned coffee to drink. Urinary caffeine (137X), paraxanthine (17X) and 1, 7-dimethyluric acid (17U) levels were measured 4-5 h later. The metabolite ratio, (17U+17X)/137X, was calculated to serve as an indicator of CYP1A2 activity. The metabolite ratio for the moderately injured group (3.98+/-1.56) and the mildly injured group (5.55+/-1.42) were lower than that for the normal group (9.56). The results suggest that a decrease in CYP1A2 activity is involved in the onset of flutamide-induced hepatic injury, and that the caffeine test seems to provide a useful means of its prediction.

Aged↗
PubMed2002

Importance of preanalytical handling of samples for measurement of cardiac troponin T in coronary effluent from isolated rat hearts.

The isolated, buffer-perfused heart is probably the most widely used model in experimental heart research, and the coronary effluent is often analysed for markers of myocardial injury. Adsorption to surrounding materials may be a serious problem of protein measurements in solutions with low protein concentrations. The aims of the present study were to investigate the importance of the preanalytical phase when measuring cardiac troponin T (cTnT) in a buffer perfusate and to investigate whether addition of albumin to the effluent might increase recovery of cTnT and improve the assay. Coronary effluent was collected in tubes of different materials and in tubes with 40 g/L bovine albumin, and then frozen. cTnT was analysed at different time points after withdrawal from the freezer. cTnT was 2.3-119 times higher in effluent with albumin. In effluent without albumin, cTnT concentration declined to 2% of the initial concentration after two episodes of freezing and thawing. The cTnT loss could not be prevented by using polystyrene or siliconized glass, but was partially inhibited in effluent with albumin. Furthermore, creatine kinase and lactate dehydrogenase levels were higher in effluent with albumin. The within-series coefficient of variation for cTnT was markedly improved when using effluent with albumin.

Adsorption↗
PubMed2002

Changes in the peptidergic innervation in the carotid body of rats chronically exposed to hypercapnic hypoxia: an effect of arterial CO2 tension.

The abundance of neuropeptide Y (NPY)-, vasoactive intestinal polypeptide (VIP)-, substance P (SP)-, and calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibers in the carotid body was examined in chronically hypercapnic hypoxic rats (10% O2 and 6-7% CO2 for 3 months), and the distribution and abundance of these four peptidergic fibers were compared with those of previously reported hypocapnic- and isocapnic hypoxic carotid bodies to evaluate the effect of arterial CO2 tension. The vasculature in the carotid body of chronically hypercapnic hypoxic rats was found to be enlarged in comparison with that of normoxic control rats, but the rate of vascular enlargement was smaller than that in the previously reported hypocapnic- and isocapnic hypoxic carotid bodies. In the chronically hypercapnic hypoxic carotid body, the density per unit area of parenchymal NPY fibers was significantly increased, and that of VIP fibers was unchanged, although the density of NPY and VIP fibers in the previously reportetd chronically hypocapnic and isocapnic hypoxic carotid bodies was opposite to that in hypercapnic hypoxia as observed in this study. The density of SP and CGRP fibers was decreased. These results along with previous reports suggest that different levels of arterial CO2 tension change the peptidergic innervation in the carotid body during chronically hypoxic exposure, and altered peptidergic innervation of the chronically hypercapnic hypoxic carotid body is one feature of hypoxic adaptation.

Animals↗
PubMed2001

Nitrate leakage from deciduous forest soils into streams on Kureha Hill, Japan.

Nitrate leakage from deciduous forest soils into streams was investigated for two adjacent hills. Many of the streams on Kureha Hill, located in Toyama City, Japan, have extremely high nitrate concentrations. The nitrate concentration of Hyakumakidani, one of the streams on Kureha Hill, averaged 158 microeq l(-1) and reached 470 microeq l(-1) during an episodic event. In contrast, the streams on Imizu Hill, adjacent to Kureha Hill, had low concentrations, below 15 microeq l(-1). Even during an episode, the nitrate concentrations increased to no more than 75 microeq l(-1). Both areas have similar blown forest soils, C/N ratios in O horizons, and vegetation consisting primarily of deciduous trees. However, soil incubation experiments, which lasted for 4 weeks, revealed that the nitrification rates in the surface soils of Kureha Hill were much higher than in the soils of Imizu Hill.

Agriculture↗
PubMed2001

Helical structure of phospholamban in membrane bilayers.

The regulation of calcium levels across the membrane of the sarcoplasmic reticulum involves the complex interplay of several membrane proteins. Phospholamban is a 52 residue integral membrane protein that is involved in reversibly inhibiting the Ca(2+) pump and regulating the flow of Ca ions across the sarcoplasmic reticulum membrane during muscle contraction and relaxation. The structure of phospholamban is central to its regulatory role. Using homonuclear rotational resonance NMR methods, we show that the internuclear distances between [1-(13)C]Leu7 and [3-(13)C]Ala11 in the cytoplasmic region, between [1-(13)C]Pro21 and [3-(13)C]Ala24 in the juxtamembrane region and between [1-(13)C]Leu42 and [3-(13)C]Cys46 in the transmembrane domain of phospholamban are consistent with alpha-helical secondary structure. Additional heteronuclear rotational-echo double-resonance NMR measurements confirm that the secondary structure is helical in the region of Pro21 and that there are no large conformational changes upon phosphorylation. These results support the model of the phospholamban pentamer as a bundle of five long alpha-helices. The long extended helices provide a mechanism by which the cytoplasmic region of phospholamban interacts with residues in the cytoplasmic domain of the Ca(2+) pump.

Amino Acid Sequence↗
PubMed2001

PKCbeta modulates antigen receptor signaling via regulation of Btk membrane localization.

Mutations in Bruton's tyrosine kinase (Btk) result in X-linked agammaglobulinemia (XLA) in humans and X-linked immunodeficiency (xid) in mice. While targeted disruption of the protein kinase C-beta (PKCbeta) gene in mice results in an immunodeficiency similar to xid, the overall tyrosine phosphorylation of Btk is significantly enhanced in PKCbeta-deficient B cells. We provide direct evidence that PKCbeta acts as a feedback loop inhibitor of Btk activation. Inhibition of PKCbeta results in a dramatic increase in B-cell receptor (BCR)-mediated Ca2+ signaling. We identified a highly conserved PKCbeta serine phosphorylation site in a short linker within the Tec homology domain of Btk. Mutation of this phosphorylation site led to enhanced tyrosine phosphorylation and membrane association of Btk, and augmented BCR and FcepsilonRI-mediated signaling in B and mast cells, respectively. These findings provide a novel mechanism whereby reversible translocation of Btk/Tec kinases regulates the threshold for immunoreceptor signaling and thereby modulates lymphocyte activation.

3T3 Cells↗
PubMed2001

NEDD8 recruits E2-ubiquitin to SCF E3 ligase.

NEDD8/Rub1 is a ubiquitin (Ub)-like post-translational modifier that is covalently linked to cullin (Cul)-family proteins in a manner analogous to ubiquitylation. NEDD8 is known to enhance the ubiquitylating activity of the SCF complex (composed of Skp1, Cul-1, ROC1 and F-box protein), but the mechanistic role is largely unknown. Using an in vitro reconstituted system, we report here that NEDD8 modification of Cul-1 enhances recruitment of Ub-conjugating enzyme Ubc4 (E2) to the SCF complex (E3). This recruitment requires thioester linkage of Ub to Ubc4. Our findings indicate that the NEDD8-modifying system accelerates the formation of the E2-E3 complex, which stimulates protein polyubiquitylation.

Anaphase-Promoting Complex-Cyclosome↗
PubMed2001

Peptide bond formation mediated by 4,5-dimethoxy-2-mercaptobenzylamine after periodate oxidation of the N-terminal serine residue.

[reaction in text] A thiol linker-attached peptide was prepared from a nonprotected peptide via an N(alpha)()-alpha-oxoacyl peptide. Selective oxidation of the N-terminal serine with sodium periodate gave the N(alpha)-glyoxyloyl peptide, reductive amination of which with 4,5-dimethoxy-2-(triphenylmethylthio)benzylamine gave an N(alpha)-4,5-dimethoxy-2-mercaptobenzyl glycyl peptide after removal of the trityl group. The N(alpha)-4,5-dimethoxy-2-mercaptobenzyl peptide can be condensed with a peptide thioester, and the linker is removable. This strategy provides a useful method for the synthesis of peptides using recombinant proteins.

Amino Acid Sequence↗
PubMed2001

On-demand droplet spotter for preparing pico- to femtoliter droplets on surfaces.

A droplet spotter for on-demand generation of pico- to femtoliter droplets was developed to meet the requirement for high-density spotting of chemicals on a surface. Our droplet spotter involves applying a approximately 1000-V and approximately 10-ms pulse voltage to the tip of a capillary tube (o.d. approximately 18 microns; i.d. approximately 11 microns) supplied with water or a dye solution. The capability of the spotter was demonstrated by preparing a microarray of dye molecules. The microarray was prepared by spotting approximately 30-fL droplets of a dye solution on a surface at the density of one spot per 20 x 20 micron 2.

Journal Article↗
PubMed2001

Application of chemical selective cleavage methods to analyze post-translational modification in proteins.

Three chemical specific cleavage reactions, one for the carboxyl side of aspartyl peptide bonds, one for the carboxyl side of asparaginyl peptide bonds and another for the amino side of seryl/threonyl peptide bonds have been recently established. Additionally, these reactions simultaneously react on several post-translationally modified groups in peptides or proteins. The modified groups cover the external modifications N-formyl, N-acetyl, N-pyroglutamyi residues and C-terminal-alpha amide, as well as the internal modifications such as O-acetyl serine, phosphorylated serine/tyrosine, sulfonylated tyrosine, glycosylated serine/threonine and glycosylated asparagine. These three cleavage reactions relate to key amino acids for modifications, deamidation for asparagine, phosphorylation and acetylation for serine, and glycosylation for asparagine, serine and threonine. The chemical reactions on these modifications change the peptide mapping pattern, and information from these reactions may contribute characterization and location of post-translational modified groups in the protein.

Animals↗
PubMed2001

Polypeptide synthesis using an expressed peptide as a building block for condensation with a peptide thioester: application to the synthesis of phosphorylated p21Max protein(1-101).

An expressed peptide proved to be useful as a building block for the synthesis of a polypeptide via the thioester method. A partially protected peptide segment, for use as a C-terminal building block, could be prepared from a recombinant protein; its N-terminal amino acid residue was transaminated to an alpha-oxoacyl group, the side-chain amino groups were then protected with t-butoxycarbonyl (Boc) groups, and. finally, the alpha-oxoacyl group was removed. On the other hand, an O-phosphoserine-containing peptide thioester was synthesized via a solid-phase method using Boc chemistry. These building blocks were then condensed in the presence of silver ions and an active ester component. During the condensation, epimerization at the condensation site could be suppressed by the use of N,N-dimthylformamide (DMF) as a solvent. Using this strategy, a phosphorylated partial peptide of the p21Max protein, [Ser(PO3H2)2.11]-p21Max(1-101), was successfully synthesized.

Basic-Leucine Zipper Transcription Factors↗
PubMed2001

The molt-inhibiting hormone in the American crayfish Procambarus clarkii: its chemical synthesis and biological activity.

The molt-inhibiting hormone of the American crayfish Procambarus clarkii (Prc-MIH), a 75-residue polypeptide containing three disulfide bridges, was synthesized by chemical ligation of two peptides, i.e., synthetic Prc-MIH(1-39) and Prc-MIH(40-75)-NH(2), and by subsequent folding to form the native disulfide-containing peptide molecule. The synthetic peptide was comparable to the natural Prc-MIH in inhibiting ecdysteroid secretion by in vitro bioassay and shared features with the natural Prc-MIH in some biochemical analyses. These results indicate that the chemical ligation method can be used for the synthesis of Prc-MIH. Furthermore, it was demonstrated that synthetic Prc-MIH has hyperglycemic activity, although the activity was weaker than that of the authentic crustacean hyperglycemic hormone in the American crayfish. To examine the structural requirement of the Prc-MIH for eliciting biological activity, an antibody raised against the C-terminal region (residues 55-75) and two synthetic peptides, i.e., a core region (residues 1-54) containing three disulfide bridges and the C-terminal region, were utilized. It is suggested that Prc-MIH exerts its activities through coordination between the core region and the C-terminal region.

Animals↗
PubMed2001

Evaluation of an internal thoracic artery as a coronary artery bypass graft by intercostal duplex scanning ultrasonography.

OBJECTIVES: Although angiography is often used to determine whether the internal thoracic artery is appropriate as a coronary bypass graft, but use of duplex scanning ultrasonography for this purpose is not yet widespread. METHODS: The internal diameter and flow of the internal thoracic artery were measured using intercostal duplex scanning in 100 patients during April 1995. The ultrasonographic device (sonos 2000, Hewlett Packard) used had a linear probe delivering a frequency of 7.5 MHz. Bilateral internal thoracic arteries and their blood flow were imaged clearly in all subjects. Diameter was compared by angiography and duplex scanning ultrasonography in 20 patients. RESULTS: The average internal diameter of internal thoracic artery was 2.19 +/- 0.46 mm (right) or 2.13 +/- 0.32 mm (left) in men and 2.05 +/- 0.44 mm (right) or 2.09 +/- 0.42 mm (left) in women. The gender difference was statistically significant (p = 0.05). The maximum systolic blood flow velocity through the internal thoracic artery was 0.85 +/- 0.34 m/s (right) or 0.84 +/- 0.36 m/s (left) in men and 0.87 +/- 0.28 m/s (right) or 0.82 +/- 0.28 m/s (left) in women. The average internal thoracic arterial blood flow (F) was 54.6 +/- 29.0 ml/min (right) or 50.9 +/- 28.8 ml/min (left) in men and 56.8 +/- 38.2 ml/min (right) or 58.2 +/- 33.4 ml/min (left) in women. Duplex scanning ultrasonography using an intercostal approach enables easy imaging of bilateral internal thoracic arteries and visualizes entire internal thoracic artery structure by simply changing the probe position. CONCLUSION: Intercostal duplex scanning ultrasonography is thus recommended for reliable evaluation of the internal diameter and blood flow of the internal thoracic artery.

Aged↗