Biomedical subjects

T Kondo

Publications and source records attributed to T Kondo.

At least 19 recordsLinked to original sources

PubMed1992

Domains for G-protein coupling in angiotensin II receptor type I: studies by site-directed mutagenesis.

To delineate domains essential for G-protein coupling in angiotensin II type 1 receptor (AT1), we mutated the receptor cDNA in the putative cytosolic regions and determined consequent changes in the effect of GTP analogs on angiotensin II (Ang II) binding and in inositol trisphosphate production in response to Ang II. Polar residues in targeted areas were replaced by small neutral residues. Mutations in the second cytosolic loop, carboxy terminal region of the third cytosolic loop or deletional mutation in the carboxyl terminal tail simultaneously abolished both the GTP-induced shift to the low affinity form and Ang II-induced stimulation of inositol trisphosphate production. These results suggest that polar residues in the second cytosolic loop, the carboxy terminal region of the third cytosolic loop, and the carboxy terminal cytosolic tail are important for G-protein coupling of AT1 receptor.

Angiotensin II↗

PubMed1992

Phospholipase D mimics platelet-derived growth factor as a competence factor in vascular smooth muscle cells.

Recent studies have suggested the importance of phosphatidylcholine (PC) metabolism in growth factor-stimulated cells. In these cells, PC is hydrolyzed not only by PC-specific phospholipase C but also by phospholipase D (PLD). In the present investigation, we show that the simple addition of PC-hydrolyzing PLD from Streptomyces chromofuscus to the culture medium of vascular smooth muscle cells elicits choline release into the medium accompanied by the formation of phosphatidic acid. In the presence of ethanol, this treatment elicits a formation of phosphatidylethanol (PEt) at the expense of phosphatidic acid. Furthermore, we show here that exogenous addition of S. chromofuscus PLD induces a marked DNA synthesis in quiescent vascular smooth muscle cells. This DNA synthesis induced by S. chromofuscus PLD is, like platelet-derived growth factor (PDGF)-elicited DNA synthesis, largely dependent on the presence of insulin. In addition, S. chromofuscus PLD-induced PEt formation and DNA synthesis were not affected by protein kinase C down-regulation, whereas PDGF-induced PEt formation and DNA synthesis were significantly inhibited. These observations strongly suggest that protein kinase-dependent activation of PLD is involved in mitogenic signal in PDGF-stimulated cells and that exogenously added PLD acts as a competence factor in the same way as PDGF.

Animals↗

PubMed1992

The regulation of the murine Hox-2.5 gene expression during cell differentiation.

The mouse Hox-2.5 gene containing a Drosophila Antennapedia-type homeobox sequence is expressed in a spatially and temporally restricted manner during embryogenesis. We found that the mouse embryonal carcinoma cell line P19 expresses Hox-2.5 during differentiation by the treatment with retinoic acid (RA). Expression of the Hox-2.5 gene was not detected in undifferentiated P19 cells, but detected 72 hours after treatment with RA. In order to analyze this inductive response, we first identified the Hox-2.5 transcription initiation site and a possible promoter region. Subsequently, we prepared constructs containing various Hox-2.5 DNA fragments fused to a firefly luciferase reporter gene and transfected these into undifferentiated or differentiating P19 cells. These studies have demonstrated that a region -279 to +15 with respect to the transcription initiation site has a differentiation-responsive promoter activity. Deletion analysis suggests that the sequences responsible for this induction are located in several distinct domains within the 294 bp promoter region. Two of the possible differentiation-responsive elements were identified by analysis of DNA-protein interactions, and in vivo competition assays lend support to the notion that these regions are involved in the differential expression of Hox-2.5 promoter activity.

Animals↗

PubMed1992

Platelet-derived growth factor AA homodimer stimulates protein synthesis rather than DNA synthesis in vascular smooth muscle cells from spontaneously hypertensive rats but not from normotensive rats.

Platelet-derived growth factor (PDGF) AB and BB isoforms were potent mitogens for cultured vascular smooth muscle cells from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). PDGF-AA promotes protein synthesis in a dose-dependent manner in SHR cells, whereas DNA synthesis was stimulated only slightly. However, this isoform did not activate either DNA or protein synthesis in WKY cells. PDGF-AA stimulated tyrosine phosphorylation of its receptor protein and phospholipase C-gamma 1 in SHR cell but not in WKY cells. These results indicate that vascular smooth muscle cell of SHR is uniquely responsive to PDGF-AA, presumably due to abnormality in receptor expression, in its hypertrophic response.

Animals↗

PubMed1992

Diacylglycerol formation from phosphatidylcholine in angiotensin II-stimulated vascular smooth muscle cells.

In cultured vascular smooth muscle cells (VSMC), angiotensin II (Ang II) induces a biphasic diacylglycerol (DAG) formation peaking at 15 sec and 5 min. Although it has been well established that the first peak is produced by the hydrolysis of inositol 4,5-bisphosphate (PIP2), the origin of the second DAG peak has never been examined in detail. In the present paper, we provide evidence that the second peak of DAG formation in Ang II-stimulated VSMC originates mainly from PC.

Angiotensin II↗

PubMed1992

Gly145 to Arg substitution in HBs antigen of immune escape mutant of hepatitis B virus.

A Japanese child born to an HBeAg-positive carrier mother received anti-HBs immunoglobulins and a plasma-derived HBs vaccine with a poor anti-HBs-antibody response. The child, who is now 3 years old, is presently suffering from chronic hepatitis with unusual serological findings that are positive for HBsAg, anti-HBs and HBeAg, since being infected with a measles virus at 12 months of age. The nucleotide sequences of the S region of HBV DNA obtained from the patient, the mother and an HBeAg-positive brother were completely identical except for one nucleotide at position 587 (mother and brother: guanosine, patient: adenosine), giving an amino acid change: Gly - greater than Arg at position 145 of the major HBs protein.

Amino Acid Sequence↗

PubMed1992

Follistatin is a developmentally regulated cytokine in neural differentiation.

Activin acts mitogenically on P19 cells as well as being inhibitory of the differentiation of retinoic acid-treated P19 cells and some neuroblastoma cell lines. Here, we show some lines of evidence that follistatin, an activin-binding protein, is also involved in neural differentiation. Counteracting the activity of activin, addition of follistatin suppresses the anchorage-independent growth of P19 cells in soft agar and stimulates neurite outgrowth of a neuroblastoma cell line, IMR-32 cells. While activin does not seem to be expressed significantly, follistatin is demonstrated in the conditioned medium of these cells. Furthermore, the expression of follistatin in P19 cells is subject to dynamic fluctuations in response to retinoic acid treatment. These neural cells may produce follistatin in a cell stage-specific manner in order to interact with exogenously derived activin.

Activin Receptors↗

PubMed1992

Functional regulation of osteoblastic cells by the interaction of activin-A with follistatin.

A high number of 125I-activin-A binding sites (an apparent Kd of 260 pM and 5,600 sites/cell) were observed on MC3T3-E1 cells, a well characterized osteoblastic cell line. Activin-A has a mitogenic effect on these cells, with the greatest influence being observed on cells in an undifferentiated state, as well as a suppressive effect on the alkaline phosphatase activity. Northern and ligand blotting analyses revealed that these osteoblastic cells produce follistatin, which was down-regulated by retinoic acid treatment. Because follistatin is an activin-A-binding protein, we suggest that activin-A modulates the function of osteoblastic cells by being regulated by follistatin during differentiation.

Activins↗

PubMed1992

Hydrolysis of phospholipid monolayers by phospholipase D at the oil/water interface under the control of the potential drop across the monolayer.

A new method has been proposed for measuring the enzymatic hydrolysis of phosphatidylcholine (PC) monolayers formed at the polarized nitrobenzene(NB)/water(W) interface under the precise control of the potential drop across the interface. As a probe for the hydrolysis, the method utilized the capacitance (Cd1) of the monolayer. Phospholipase D (EC. 3.1.44., PLD) converted L-alpha-dipalmitoylphosphatidylcholine (DPPC) in the monolayer to L-alpha-dipalmitoylphosphatidic acid (DPPA), leading to a drastic decrease in Cdl. This change in Cdl was sensitive enough to monitor the course of enzymatic hydrolysis of the PC monolayer by PLD. The rate of the hydrolysis was markedly dependent on the potential drop across the interface. When the potential of the aqueous phase with respect to that of the NB phase (delta W0 phi) was -140 mV, no hydrolysis was observed, whereas at delta W0 phi = 60 mV the hydrolysis proceeded promptly.

Electrochemistry↗

PubMed1992

Mouse rRNA gene transcription factor mUBF requires both HMG-box1 and an acidic tail for nucleolar accumulation: molecular analysis of the nucleolar targeting mechanism.

RNA polymerase I requires at least two nucleolar transcription factors, UBF and SL-1, for ribosomal RNA gene (rDNA) transcription. UBF requires SL-1 for the formation of a stable initiation complex on the rDNA promoter region. We have determined the region of mouse UBF (mUBF) required for nucleolar targeting. Although mUBF has a nuclear localization sequence, this sequence alone is not sufficient for mUBF to accumulate in the nucleolus. Deletion analyses show that mUBF requires a wide region except for the N-terminal 101 amino acids for nucleolar targeting. Deletion of either the HMG-box1, a region crucial for rDNA binding, or the acidic tail, a region that may interact with SL-1, results in the loss of nucleolar targeting. We show by DNA affinity analysis that the HMG-box1 is absolutely necessary for mUBF to bind to the upstream control element of the rDNA. We also show that mUBFs with various internal deletions retain both nucleolar targeting and DNA binding ability. A clear correlation was demonstrated between the DNA binding and nucleolar targeting ability. These results suggest that UBF is transferred to the nucleus by its NLS and is sequestered in the nucleolus by its specific and stable binding to the rDNA promoter via HMG-boxes and the acidic tail.

Animals↗

PubMed1992

Enhanced rectal absorption and reduced local irritation of the anti-inflammatory drug ethyl 4-biphenylylacetate in rats by complexation with water-soluble beta-cyclodextrin derivatives and formulation as oleaginous suppository.

To improve the rectal delivery of ethyl 4-biphenylylacetate (EBA), a prodrug of the anti-inflammatory drug 4-biphenylylacetic acid (BPAA), the use of highly water-soluble 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) and heptakis(2,6-di-O-methyl)-beta-cyclodextrin (DM-beta-CyD) was investigated and compared with the use of the parent beta-cyclodextrin (beta-CyD). Among the three beta-CyDs, HP-beta-CyD was best at improving the rectal bioavailability of EBA in rats after single and multiple administrations of oleaginous suppositories (Witepsol H-5) containing the complexes. To gain insight into the enhancing effect of beta-CyDs, the absorption behaviors of EBA (observed by monitoring BPAA as an active metabolite of EBA) and beta-CyDs themselves were examined in vitro, in situ, and in vivo. The in situ recirculation study revealed that the complexed form of EBA was less absorbable from the rectal lumen in the solution state, but this disadvantageous effect of beta-CyDs was compensated in part by the inhibition of the bioconversion of EBA to BPAA. When beta-CyDs were coadministered with EBA in vivo, however, rather high amounts of HP-beta-CyD (approximately 26% of dose) and DM-beta-CyD (approximately 21% of dose), compared with beta-CyD (approximately 5% of dose), were absorbed from the rat rectum. Thus, the enhancement of rectal absorption of EBA in vivo can be explained by the facts that the hydrophilic beta-CyDs increased the release rate of EBA from the vehicle and stabilized EBA in the rectal lumen and that the drug was partly absorbed in the form of the complex.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Application of superfine fiberscope for endovasculoscopy, ventriculoscopy, and myeloscopy.

In the past three years, we have used a superfine fiberscope for endovasculoscopy, ventriculoscopy, and myeloscopy. Flexible superfine fiberscope, 0.75 mm in outer diameter, could visualize various intravascular findings. In in-vivo canine experiment, sequential changes of thrombus produced by endothelial abrasion by needle or balloon in the canine carotid artery can be clearly seen. And thrombolysis by focal arterial injection of tissue plasminogen activator was sequentially observed. In the clinical study, we could evaluate the stenotic lesions of the subclavian and vertebral arteries before and after balloon angioplasty. During ventriculoscopy, a 2 mm of mini-caliber fiberscope was introduced under ultrasound monitoring, and provided clear visualization of intraventricular tumours. In myeloscopy, draining veins of arteriovenous malformations and nerve roots of the cauda equina could be clearly seen by flexible fiberscope. From these results, it can be said that the superfine fiberscope provides clear and useful visualizations of the interior of vessels, ventricles, and the intrathecal area of the spinal canal. The new applications of this superfine fiberscope for minimally invasive neurosurgery may bring about a marked improvement of therapeutic results.

Animals↗

PubMed1992

A possible association between basic amino acids of position 13 of DRB1 chains and autoimmune hepatitis.

Fifty-one patients with autoimmune hepatitis have been studied for HLA association by conventional serology and also by modified polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotyping. HLA-DR4 was significantly associated with autoimmune hepatitis (46 of 51 patients, 90.2%). DNA typing of the DRB1 gene for 43 DR4-positive patients by using the PCR-RFLP technique revealed that of 43 patients, 33 had DRB1*0405 (Dw15), five had DRB1*0406 (DwKT2), four had DRB1*0403 (Dw13a), two had DRB1*0401 (Dw4), two of 43 had DRB1*0407 (Dw13b) and one had DRB1*0408 (Dw14b). Thus, there was no significant difference in Dw frequencies between DR4-positive patients and DR4-positive healthy subjects. These findings suggest that the DR4-specific sequence (Val 11 and His 13 at amino acid positions 11 and 13, respectively), but not particular Dw-associated DR4 sequence, in the first domain of the DRB1 chain contributes to susceptibility to autoimmune hepatitis among Japanese. Interestingly, all five of the DR4-negative patients had the DR2 specificity (DRB1*1502 or 1601). Taken together, these results imply that the basic amino acids at position 13, which is present only on the DR2 and DR4 B1 molecules (Arg on DR2 and His on DR4), are most important for determining the predisposition to autoimmune hepatitis.

Amino Acid Sequence↗

PubMed1992

Mitochondrial complex I and II activities of lymphocytes and platelets in Parkinson's disease.

Mitochondrial Complex I deficiencies have been described not only in the brain but also in the skeletal muscle and platelets in Parkinson's disease (PD). We report activities of Complex I, II, III, and IV in lymphocytes and platelets in 20 patients with PD and age-matched controls. A small but a significant decrease in the platelet Complex I activity was found in PD (9.14 +/- 1.86 units/mg protein) compared with that in the control (12.37 +/- 2.66 units/mg protein) (P = 0.0002). The lymphocyte Complex I activity was about the same between PD and the control. The activity of Complex II was slightly diminished in both platelets and lymphocytes in PD. Rather small decrease in the platelet Complex I activity in PD may be clinically non-significant. But it may indicate the presence of a qualitatively similar abnormality in platelets as in the nigro-striatal neurons. The cause for decrease in the Complex II activity is unknown at this moment. Further studies seem necessary.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed1992

Peritoneal absorption of pancreatic enzymes in dogs.

To elucidate peritoneal absorption of pancreatic enzymes, plasma levels of amylase, lipase, and trypsinogen were measured after the intraperitoneal injection of 10 ml human pancreatic juice in dogs. Plasma pancreatic amylase, lipase, and trypsinogen were determined using the immunoassay specific to the corresponding human pancreatic enzyme to exclude cross-reaction with the endogenous enzyme activities of the canine plasma. Plasma immunoreactivity of human amylase persistently rose during 24 h after the injection, whereas elevation of plasma amylase enzyme activity become significant only at 24 h. Increase of plasma lipase was not remarkable. Significant increase of the immunoreactivity was observed only at 24 h but there was no significant increase of the enzyme activity during this period. Plasma trypsinogen immunoreactivity peaked at 2 h remained significantly elevated during 24 h. The transperitoneal absorption of pancreatic enzymes in dogs was confirmed using intraperitoneal injection of human pancreatic juice combined with immunoassay specific to human pancreatic enzymes.

Amylases↗

PubMed1992

Molecular form of pancreatic elastase 1 in human plasma.

In order to study the molecular forms of immunoreactive pancreatic elastase 1 (IRE) in human plasma, we investigated the characteristics of proelastase 1, elastase 1, and their alpha 1-protease inhibitor (alpha 1-PI) complexes on anion-exchange (Mono Q) chromatography, gel (Superose 12) chromatography, and enzyme immunoassay systems with monoclonal antielastase 1 antibodies of different affinities for alpha 1-PI-elastase 1 and alpha 1-PI-proelastase 1 complexes. The rate of complex formation between proelastase 1 and alpha 1-PI was dependent on temperature of incubation and concentration of alpha 1-PI. At 37 degrees C, 90% of proelastase 1 was bound to alpha 1-PI during 2 h of incubation of purified human proelastase 1 with human alpha 1-PI. Similar results were also obtained by 2 h of incubation of proelastase 1 with human plasma at 37 degrees C. Therefore, under physiological conditions, neither free proelastase 1 nor elastase 1 can be detected in the human blood stream, and most IRE exists as the complex of proelastase 1 with alpha 1-PI in human plasma.

Chromatography, Ion Exchange↗

PubMed1992

Differences in the transport systems between cementocytes and osteocytes in rats using microperoxidase as a tracer.

Microperoxidase (MP) tracer was injected intravenously into rats to investigate any differences in transport pathways of tissue fluids in the lacunae and canaliculi of cementum and bone. Light microscopically, in deep cementum lacunae, pericellular spaces contained a large amount of MP, while close to the cementum surface, the spaces contained scarcely any. In bone, MP was detected throughout all pericellular spaces. MP was detected intracellularly as granular reaction products in most cementocytes and osteocytes. Electron microscopically, MP was found in the pericellular spaces of cementum and bone lacunae, particularly on collagen fibrils and amorphous material. MP deposits were also intense along the plasma membrane of cementocytes in the deep cementum and along the innermost edge of the deep cementum matrix and bone matrix. In uptake of MP by cementocytes, although extracellular tracer was deposited extensively along the plasma membrane of the deeply positioned cementocytes, uptake by these deep cementocytes was less than that of those close to the surface. However, in bone, most osteocytes showed uniform uptake. These results suggest that the transport pathways for tissue fluids in cementum are in the pericellular spaces, but that cementum has an uneven circulation of tissue fluid. In cementum, although there seems to be a well-developed canalicular system to transport tissue fluid into the deep regions, the deep cementocytes had less endocytotic ability than those close to the surface.

Animals↗

PubMed1992

Immunohistochemical study of nerve fibres with substance P- or calcitonin gene-related peptide-like immunoreactivity in the junctional epithelium of developing rats.

The beginning of innervation in the junctional epithelium of maxillary first molars was examined in gingival tissues from 19 to 32-day-old rats. Substance P- or calcitonin gene-related peptide (CGRP)-like immunoreactivity was demonstrated by the avidin-biotin peroxidase complex method. In 19-day-old rats, nerve fibres with substance P- or CGRP-like immunoreactivity were seen in the connective tissue and oral epithelium, but not in the reduced enamel epithelium, which would be transformed into the junctional epithelium. In 21-day-old rats, the fibres with substance P- or CGRP-like immunoreactivity formed a plexus in the oral sulcular epithelium and thin varicose fibres were seen for the first time entering the adjacent reduced enamel epithelium. These fibres also penetrated the middle portion of the reduced enamel epithelium, but did not reach the cuboidal reduced ameloblasts. More nerve fibres had CGRP-like immunoreactivity than substance P-like immunoreactivity. In 23-day-old rats, many fibres with both immunoreactivities were seen in the basal layers of the junctional epithelium, but only a few were seen in its superficial layers. In 28-32-day-old rats, numerous fibres with both immunoreactivities were distributed in the whole junctional epithelium and showed a similar pattern of innervation. For all immunoreactive fibres, the density in the middle portion in the junctional epithelium was the highest. The nerve plexus was formed in the basal layers and some fibres with a varicose appearance were found in the superficial layers.

Animals↗