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Biomedical subjects

R Kuroda

Publications and source records attributed to R Kuroda.

At least 73 records · Page 4Linked to original sources
PubMed1999

[Primary racemose hemangioma of bronchial artery].

A 38-year-old woman was admitted to the hospital because of massive hemoptysis. A chest X-ray film disclosed an infiltrative shadow. A bronchoscopic examination revealed a small, pulsatile, papillary protrusion in the orifice of the middle lobe bronchus, and massive bleeding from the protrusion was observed during the examination. Bronchial arteriography showed convolution and mild hypervascularization of the right bronchial artery. A middle lobectomy was performed, and the protrusion was histologically shown to be a primary racemose hemangioma of the bronchial artery.

Adult↗
PubMed1999

Characterization and application of thin film pressure sensors.

Data regarding intra-joint loads during range of motion is essential to understanding normal joint mechanics and pathology. The investigators configured and characterized the response of a 440 N range, 0.076 mm thick commercial thin film sensor to monitor joint loads through a range of motion. Following preconditioning, static and dynamic tests were performed to evaluate the sensor response under varied environmental conditions. Both tests utilized a fixture to align the sensor and applied load. Under conditions that included dry, wet, folded and kinked configurations, a static load of 100 N was applied and the sensor output monitored up to thirty minutes from the time of loading. A load of 400 N at 50 N/s was applied to the sensor to determine dynamic characteristics and calibration curves under the conditions described for static tests in addition to curved, hard and soft contact surfaces and sensor overload. Static kinked data was significantly different from the dry, wet and folded conditions. Dynamic data showed that inter-package variability was not significant but that differences between sensor packages and sensor configuration conditions were significant. To investigate applicability of these sensors to the field of orthopaedics, a cadaveric knee was instrumented with sensors to examine the role of the meniscus in load transmission and distribution across the knee. The sensors were placed bilaterally below each meniscus on the anterior, posterior and center of the tibial plateau. Sensor data were obtained at these locations during manually flexed range of motion for the intact, re-attached and lateral menisectomized conditions of the knee specimen.

Analysis of Variance↗
PubMed1998

Charged amino acids at the carboxyl-terminal portions determine the intracellular locations of two isoforms of cytochrome b5.

Outer mitochondrial membrane cytochrome b5 (OMb), which is an isoform of cytochrome b5 (cyt b5) in the endoplasmic reticulum, is a typical tail-anchored protein of the outer mitochondrial membrane. We cloned cDNA containing the complete amino acid sequence of OMb and found that the protein has no typical structural feature common to the mitochondrial targeting signal at the amino terminus. To identify the region responsible for the mitochondrial targeting of OMb, various mutated proteins were expressed in cultured mammalian cells, and the subcellular localization of the expressed proteins was analyzed. The deletion of more than 11 amino acid residues from the carboxyl-terminal end of OMb abolished the targeting of the protein to the mitochondria. When the carboxyl-terminal 10 amino acids of OMb were fused to the cyt b5 that was previously deleted in the corresponding 10 residues, the fused protein localized in the mitochondria, thereby indicating that the carboxyl-terminal 10 amino acid residues of OMb have sufficient information to transport OMb to the mitochondria. The replacement of either of the two positively charged residues within the carboxyl-terminal 10 amino acids by alanine resulted in the transport of the mutant proteins to the endoplasmic reticulum. The mutant cyt b5, in which the acidic amino acid in its carboxyl-terminal end was replaced by basic amino acid, could be transported to the mitochondria. It would thus seem that charged amino acids in the carboxyl-terminal portion of these proteins determine their locations in the cell.

Amino Acid Sequence↗
PubMed1998

Penetration of cisplatin into mouse brain by lipopolysaccharide.

We investigated the penetration of cisplatin into the mouse cerebral cortex-rich region (CCR) induced by lipopolysaccharide (LPS). With the injection of cisplatin into mice 3 h after the LPS treatment, platinum was detected in the CCR during the 7 days after the injection, while platinum was not detected in the CCR of cisplatin-injected mice without LPS pretreatment and of mice simultaneous treated with cisplatin and LPS. The N(G)-nitro-L-arginine methyl ester dose-dependently lowered the platinum level. A dose of 5 mg/kg of aminoguanidine reduced the increase in the platinum level of the LPS-treated mouse, and platinum was no longer detected at doses of 20 mg/kg in the aminoguanidine-injected group. At doses of 500 mg/kg aminoguanidine, however, no effect was seen on the platinum level of the CCR induced by LPS. Regarding indomethacin, the injection of 5 mg/kg resulted in a decrease in the platinum content of the CCR, but not undetectable level. These results suggest that LPS increases the penetration of cisplatin into the mouse brain, and platinum may be accumulated in the CCR. Nitric oxide and prostaglandins contribute to the penetration of platinum into the cerebral cortex.

Animals↗
PubMed1998

Cross tolerance to environmental stress and endotoxin.

Mild DIC-like phenomena develop in rodents exposed to environmental stress, induced by prolonged fluctuation in ambient temperature, known as SART (specific alternation of rhythm in temperature) stress, the symptoms being essentially similar to those produced by endotoxin. The aim of the present study was to investigate whether common mechanisms are involved in the stress-induced DIC-like phenomena and endotoxaemia. The mortality in mice after intraperitoneal injection with endotoxin at 8.3, 16.7 and 33.3 mg/kg increased in a dose-dependent manner. In the mice exposed to stress for 7 days, the death rate following endotoxin at 8.3 mg/kg was markedly lower than that in unstressed mice, although no difference was detected between the two groups following endotoxin at 16.7 and 33.3 mg/kg. Conversely, repeated administration of endotoxin at a low dose, 0.5 mg/kg, during exposure to stress for 7 days inhibited the stress-induced thrombocytopenia and hypofibrinogenemia. These findings indicate the development of cross tolerance to prolonged environmental stress and endotoxin, implying involvement of a common mechanism.

Animals↗
PubMed1998

Increased vascular permeability by a specific agonist of protease-activated receptor-2 in rat hindpaw.

The present study examined the effect of intraplantar (i.pl.) administration of a selective agonist of protease-activated receptor (PAR)-2, SLIGRL-NH2(PP6-NH2), on vascular permeability in rat hindpaw. PP6-NH2, administered i.pl. at 10-100 nmol per paw, enhanced vascular permeability and caused oedema formation in rat hindpaw. SLIGRL (PP6-OH) and trypsin, by i.pl. administration, also elicited an increase in vascular permeability, although i.pl. administration of the mixture of constituent amino acids of PP6-OH at an equivalent dose did not. The PP6-NH2-induced increase in vascular permeability was abolished by repeated pretreatment with compound 48/80 to deplete bioactive amines in mast cells. These findings suggest that the activation of PAR-2 induces acute inflammation, at least partially, via mast cell degranulation in rat hindpaw.

Administration, Topical↗
PubMed1998

[Pulmonary disease after massive inhalation of Aspergillus niger].

A 60-year-old man was admitted to the hospital because of fever, coughing, and dyspnea that developed after he entered a silo that had been filled with chips of wood in the preceeding 3 months. A chest X-ray film revealed bilateral ground-glass shadows. Histologic study of the lung showed a multifocal acute process; the alveoli and interstitial areas contained many fungal hyphae and spores. Cultures from both bronchoalveolar-lavage-fluid and the chips in the silo revealed Aspergillus niger. Serologic reactions were negative to 10 antigens known to induce hypersensitivity pneumonitis. Furthermore, the patient's serologic reaction to the extracts of fungi obtained from the bronchoalveolar-lavage-fluid was negative. The patients recovered quickly without steroid therapy. We believe that this patient's diseases was "organic dust toxic syndrome".

Aspergillosis↗
PubMed1997

Studies on the mechanism for Cai-transients in sea urchin zygotes caused by refertilization and external application of sperm extract.

Sea urchin zygotes can be refertilized when they are deprived of the fertilization membrane and the hyaline layer. We have earlier reported that a transient increase of the intracellular Ca2+ concentration (Cai-transient) is induced in zygotes refertilized by sperm or treated with a sperm extract (spex) (M. Osawa et al., 1994, Dev. Biol. 166, 268-276). We investigated quantitative characteristics of the Cai-transient induced by sperm and spex, using a Ca2+ indicator, Indo-1. When sperm or spex was applied to zygotes, the peak value of the Cai-transient was 1.16 or 0.69 microM, respectively. Although these values were lower than the peak value of 1.95 microM measured during normal fertilization, the entire time courses of the three types of Cai-transients were similar. The Cai-transients during fertilization is known to be caused both by the IP3-induced Ca2+ release (IICR) and by a mechanism independent of IICR. The Cai-transients during refertilization and fertilization were not inhibited by an IP3 receptor inhibitor, heparin or by a G-protein inhibitor, GDPbetaS. However, heparin delayed the time courses of both Cai-transients. These results suggest that there may be two signal transduction pathways operating during refertilization, one dependent and the other independent of IICR. By contrast, both heparin and GDPbetaS inhibited the spex-induced Cai-transient. The IP3 content in spex-treated zygotes increased, and the spex-induced Cai-transient occurred even in the absence of external Ca2+. Cai-transient was not observed when spex was injected into zygotes. These data suggest that spex induces IICR in zygotes by activating certain cell surface receptors coupled to G-proteins.

Animals↗
PubMed1997

New renin inhibitors containing novel analogues of statine.

Solid-phase methodology has been used to synthesize a series of peptides based on the N-terminal sequence of human angiotensinogen in which statine (Sta) or the novel analogues (3S,4S)-3,4-diamino- or (3R,4S)-3,4-diamino-6-methylheptanoic acid (Ads or R-Ads) and (3S,4S)-4-amino-3-aminomethyl- or (3R,4S)-4-amino-3-aminomethyl-6-methylheptanoic acid (Amd or R-Amd) replace either residue 10 or both residues 10-11 at the P1-P1' cleavage site. The synthesis of these novel analogues of statine together with biological results on the inhibition of human and rat renin by peptides derived from them is reported. The absolute stereochemistry of the (3S,4S) Ads was determined by an X-ray crystallographic analysis of its N gamma-Boc, B beta-Z, R(+)-1-methyl benzamide derivative. Peptide Boc-His-Pro-Phe-His-Sta-Val-Ile-His-NH2 (VI) is the best inhibitor of human renin containing Sta at position 10. However, peptides containing Ads and Amd gave better rat renin inhibitors than the corresponding Sta-containing peptides. Peptides Boc-His-Pro-Phe-His-Ads-Val-Ile-His-NH2 (VII) having Ads at position 10 had an IC50 of 12 nM against rat renin. Although Sta has come to be accepted as an isosteric replacement for a dipeptide unit rather than for a single amino acid residue, in our series of inhibitors Sta is more effective when replacing only the amino acid at position 10 in the natural angiotensinogen sequence. None of the peptides gave any effect in vivo in a hypertensive rat model.

Amino Acid Sequence↗
PubMed1996

Visual evoked potentials to a faint light: signal propagation analyzed with peak latency and topographic mapping.

Accurate localization for the process of recognition of a light stimulus is yet to be determined. We studied 19-channel VEP from nine healthy volunteers, evoked by physiologically faint light less than 200 Cd/m2, using a light emitting diode (5 mm phi, 0.3 degrees, energized for 5 msec). VEP was bandpass filtered from 0.16 to 120 Hz, and analyzed from 20 msec pre-stimulus to 184.2 msec post-stimulus. The grand average VEP suggested an initial positive peak at 115.8 msec at the frontal poles and at 136 msec over the occipital areas. An initial negative peak was noted at 156 msec at the frontal poles and at 179.2 msec over the occipital areas. This might suggest that the potentials evoked had propagated from the frontal poles to the occipital areas, taking about 20 msec. However, the topographic maps contained little evidence for such a propagation, but rather indicated the waxing and waning of positive or negative extremes.

Adult↗
PubMed1996

In situ topology of cytochrome b5 in the endoplasmic reticulum membrane.

Cytochrome b5 is tail-anchored in the ER membrane and is composed of three functionally different portions; amino-terminal heme-containing catalytic, central hydrophobic membrane-anchoring, and carboxy-terminal ER-targeting portions [Mitoma, J. and Ito, A. (1992) EMBO J. 11, 4197-4203]. In situ topology of cytochrome b5 in the ER-membrane was studied using immunofluorescence microscopy. Antibodies were raised against the hydrophilic portion (anti-b5) and the carboxy-terminal seven amino acid residues (anti-peptide) of cytochrome b5 and used for detection of the cytochrome in COS cells which expressed the rat cytochrome. Anti-b5 antibody detected the cytochrome in a reticular staining pattern characteristic of the ER, even when the cell plasma membrane was permeabilized with Streptolysin O. The anti-peptide displayed a fluorescence signal only with Triton-permeabilized cells in which the antibody was able to penetrate into the ER lumen. In a double immuno-staining of the cell using the antipeptide antibody and the antibody against protein disulfide isomerase, both antibodies showed the same staining pattern in the presence of either Triton X-100 or Streptolysin O. The results indicate that the carboxy-terminal hydrophilic stretch is exposed to the luminal side. Cytochrome b5 was tagged with c-myc peptide at the carboxy-terminal end and the topology of the c-myc peptide was analyzed by the same method. Anti c-myc monoclonal IgG detected the tagged cytochrome b5 only after Triton treatment of the fixed cells, suggesting that the addition of c-myc peptide to the carboxy-terminal end does not affect insertion or orientation of the cytochrome in the ER membrane.

Animals↗
PubMed1995

Myristoylation of endothelial cell nitric oxide synthase is important for extracellular release of nitric oxide.

Endothelial cell nitric oxide synthase (NOS) is known to have a N-myristoylation consensus sequence. Such a consensus sequence is not evident in the macrophage, smooth muscle and neuronal NOS. A functional role for this N-terminal myristoylation is not clear yet. In the present study, we examined the effect of N-terminal myristoylation on the NOS activity determined by the conversion of L-[3H]arginine to L-[3H]citrulline and extracellular NO release determined by nitrite production in the conditioned medium from the COS-7 cells transfected with wild-type bovine aortic endothelial cell (BAEC) NOS cDNA or non-myristoylated BAEC-NOS mutant cDNA. NOS activity of wild type BAEC-NOS in COS-7 cells was localized in the particulate fraction and that of mutant NOS was in the cytosolic fraction. In contrast, nitrite production from COS-7 cells transfected with wild type BAEC-NOS cDNA was greater than that of mutant cDNA in a time dependent and a concentration dependent manner. These results suggest that membrane localization of NOS with myristoylation facilitates extracellular transport of NO and leads to enhanced NO signaling on the vascular smooth muscle cells and the intravascular blood cells including neutrophils, macrophages and platelets.

Animals↗
PubMed1995

Novel DNA photocleaving agents with high DNA sequence specificity related to the antibiotic bleomycin A2.

We have designed and synthesized a series of novel DNA photocleaving agents which break DNA with high sequence specificity. These compounds contain the non-diffusible photoactive p-nitrobenzoyl group covalently linked via a dimethylene (or tetramethylene) spacer to thiazole analogues of the DNA binding portion of the antibiotic bleomycin A2. By using a variety of 5' or 3' 32P-end labeled restriction fragments from plasmid pBR322 as substrate, we have shown that photoactive bithiazole compounds bind DNA at the consensus sequence 5'-AAAT-3' and induce DNA cleavage 3' of the site. Analysis of cleavage sites on the complementary DNA strand and inhibition of DNA breakage by distamycin A indicates these bithiazole derivatives bind and attack the minor groove of DNA. A photoactive unithiazole compound was less specific inducing DNA breakage at the degenerate site 5'-(A/T)(AA/TT)TPu(A/T)-3'. DNA sequence recognition of these derivatives appears to be determined by the thiazole moiety rather than the p-nitrobenzoyl group: use of a tetramethylene group in place of a dimethylene spacer shifted the position of DNA breakage by one base pair. Moreover, much less specific DNA photocleavage was observed for a compound in which p-nitrobenzoyl was linked to the intercalator acridine via a sequence-neutral hexamethylene spacer. The 5'-AAAT-3' specificity of photoactive bithiazole derivatives contrasts with that of bleomycin A2 which cleaves DNA most frequently at 5'-GPy-3' sequences. These results suggest that the cleavage specificity exhibited by bleomycin is not simply determined by its bithiazole/sulphonium terminus, and the contributions from other features, e.g. its metal-chelating domain, must be considered. The novel thiazole-based DNA cleavage agents described here should prove useful as reagents for probing DNA structure and for elucidating the molecular basis of DNA recognition by bleomycin and other ligands.

Base Sequence↗
PubMed1995

Frontal cingulotomy reconsidered from a WGA-HRP and c-Fos study in cat.

A recent positron emission tomography (PET) study demonstrated that the anterior cingulate cortex (area 24), in addition to SI and SII cortices, was activated by painful stimuli. In order to elucidate the participation of relay nuclei in the ascending pain pathway to area 24, we performed a regrograde labelling study with WGA-HRP injection into area 24 in cats. Area 24 was found to receive pain-related thalamic inputs from the intralaminar nuclei including the central medial nucleus, midline nuclei, modiodorsal nucleus and possibly the submedial nucleus. We then examined the expression of Fos protein in CNS induced by formalin injection into the face in cats. Fos positive neurons were demonstrated in areas 23 and 24, the anterior limbic area, insular cortex, midline and paraventricular nuclei in the thalamus, paraventricular nucleus and other areas in the hypothalamus, and in many nuclei in the brainstem in both the formalin-injected group and the control group (anesthesia only). Labelled regions appeared to correspond to stress-related sites. The sole difference from the control group was the expression of Fos in the coronal gyrus and in the trigeminal caudalis nucleus in the experimental group. Although more Fos positive cells were observed in area 24 in experimental than in control cats, the difference was not significant. Our findings suggest that the demonstrated response of area 24 on PET scan represents stress- and emotion-related events rather than pain. Surgical intervention into the anterior cingulate cortex including cingulotomy thus appears to relieve stress and emotion associated with chronic pain, but not pain itself.

Animals↗
PubMed1995

c-fos expression after formalin injection into the face in the cat.

Expression of the c-fos gene in CNS induced by formalin injection into the face as pain stimulus was examined in cats. Fos-positive neurons were demonstrated in the anterior cingulate, anterior insula and other areas of the cerebral cortex bilaterally, midline thalamic nuclei, hypothalamus, and brainstem in both the formalin-injected and control group (anesthesia only). Most of these labeled regions appeared to correspond to stress- and anesthesia-related sites. The difference from the control was the finding of Fos-positive cells in the SI, possible SII and trigeminal subnucleus caudalis in the experimental group. Almost significant increase in Fos-positive cells was also observed in areas 24, 23 and the anterior agranular insular cortex in experimental cats. Our findings appear to be compatible with recently reported PET findings in man, except laterality. For full appreciation of the complex pain experience, these diverse areas of the brain appear to be activated.

Animals↗
PubMed1995

Inhibition of endothelial nitric oxide synthase activity by protein kinase C.

Nitric oxide (NO) is an important molecular messenger accounting for endothelium-derived relaxing factor. Recently, NO synthase (NOS) from cultured endothelial cells has been purified and molecularly cloned. To evaluate the effect of phosphorylation by protein kinase C (PKC) and cyclic AMP-dependent protein kinase (PKA) on endothelial constitutive NOS catalytic activity, we incubated purified endothelial NOS with PKC or PKA. Endothelial NOS was stoichiometrically phosphorylated by PKC and PKA. In intact bovine aortic endothelial cells (BAECs), NOS was phosphorylated by stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA). NOS activity measured by the conversion of [3H]arginine to [3H]citrulline in homogenates of BAECs treated with TPA or phorbol 12,13-dibutyrate was reduced by 30%, whereas dibutylyl cyclic AMP did not affect NOS activity. Moreover, we measured NO release from cultured BAECs by a chemiluminescence method to examine the effect of PKC and PKA on endothelial NOS activity. In cultured BAECs, ATP gamma S and A23187 induced NO release in time- and dose-dependent manners. Phorbol esters such as TPA and phorbol 12,13-dibutyrate dose dependently inhibited NO release stimulated by A23187 as well as ATP gamma S. Reduction of NO release by TPA was almost completely prevented by pretreatment with staurosporine, an inhibitor of PKC. NO release by A23187 was increased in PKC-downregulated BAECs. In contrast, dibutylyl cyclic AMP or 8-bromo cyclic GMP had no effect on NO release from BAECs induced by A23187 or ATP gamma S. These results indicate that phosphorylation of NOS by PKC is associated with a reduction of its catalytic activity in vascular endothelial cells.

Amino Acid Oxidoreductases↗