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Biomedical subjects

S Kawabata

Publications and source records attributed to S Kawabata.

At least 181 records · Page 10Linked to original sources

[A case of multiple broncholithiasis caused by mucus retention].

The patient was a 57-year-old male with long-standing bronchiectasis who developed severe respiratory failure and died in 1991. Autopsy revealed multiple broncholithiasis in both lungs, but no calcified lymph nodes in the hilar region. Since histological examination of the broncholiths showed only stratified structures but no tissue structure, most likely cause was considered to be calcification of mucus in the bronchi. Analysis of the stone components revealed 78% calcium and 22% protein. This patient represents a case of multiple broncholithiasis caused by mucus retention, which is thought to be very rare.

Adult↗

Molecular characterization and expression of the cell-associated glucosyltransferase gene from Streptococcus mutans.

A gene encoding cell-associated glucosyltransferase (CA-GTase) was cloned from Streptococcus mutans MT8148 into Escherichia coli DH5 alpha by using a low-copy-number plasmid, pMW119. After screening of a gene library with the oligonucleotide probe designed on the basis of a partial amino acid sequence of CA-GTase, a recombinant plasmid, pSK6, that had a 5.6 kb insert carrying the CA-GTase gene was selected. The gene product (recombinant CA-GTase) of pSK6 was expressed by using a lac promoter in pMW119. Western blotting revealed that rCA-GTase reacted with antibody to CA-GTase. rCA-GTase was found to synthesize water-insoluble glucans. Southern blotting indicated that the MT8148 chromosome contained another gene which was homologous to pSK6. A plasmid harboring this gene (pSK16) was also isolated from the gene library, the gene product of pSK16 exhibited GTase activity but ten times lower than that of pSK6.

Base Sequence↗

A microsomal endopeptidase from liver with substrate specificity for processing proproteins such as the vitamin K-dependent proteins of plasma.

The microsomal fraction of rabbit liver contains an endopeptidase that cleaves synthetic peptides that mimic the amino acid sequences of the processing sites of many proproteins, including the vitamin K-dependent proteins. The endopeptidase (M(r) 69,000) was extracted from liver microsomes with 1% Lubrol and purified about 2,700-fold. The substrate employed for isolation and characterization of the enzyme was the decapeptide acetyl-Ala-Arg-Val-Arg-Arg-Ala-Asn-Ser-Phe-Leu (prothrombin peptide), in which hydrolysis occurred on the carboxyl side of the paired Arg-Arg residues. The purified enzyme, whose activity was enhanced 1.8-fold by 0.1 mM CoCl2, has a Km = 80 microM and Vmax = 21,000 nmol.min-1.mg-1 and a pH optimum of 8.7. Proteolytic cleavage of decapeptide substrates was dependent on an arginine residue at positions P1 and P4. The enzyme was completely inhibited by EDTA and 1,10-phenanthroline as well as by p-chloromercuriphenylsulfonic acid and Hg2+. Inhibitors of serine proteases and cysteine proteases had no effect. Based on the substrate preference, the endopeptidase appears to be a good candidate for the enzyme responsible for the precursor processing of the vitamin K-dependent proteins and a number of other proproteins that are synthesized via the secretory pathway in liver and other tissues.

4-Chloromercuribenzenesulfonate↗

Telescience testbed examination aboard Japanese Experiment Module (JEM): life and material science experiments.

A telescience ground testbed experiment was conducted by the National Space Development Agency of Japan (NASDA) at the Tsukuba Space Center in March 1991. The objectives of the ground testbed experiment were to extract scientists' requirements for a communication method, to evaluate the influence of transmission delay and capacity on experiment operations, and to evaluate performance and functions of the system for the testbed experiment. The microscopic operations experiment, the image furnace experiment and the onboard training experiment were selected as typical ground testbed experiments. In these experiments, motion video transmission at 320 kbps was acceptable for observing the experiments and communicating between the principal investigator and the payload specialist. In the microscopic operations experiment, motion video transmission at 1.5 Mbps or more was required for detailed observation. A 4-second transmission delay (roundtrip) was allowable for mutual communication.

Aerospace Medicine↗

[Prediction formulas for pulmonary function tests expressed in linear and exponential form for healthy Japanese adults].

We studied 21 parameters of pulmonary function in 172 healthy Japanese adult smokers and non-smokers ranging from 18 to 83 years of age. Prediction formulas for each parameter were calculated in both linear and exponential form using multiple regression analysis with regard to sex, age, height and weight. The exponential form was superior to the linear form for V25, V50, PEF, FVC, VC, FEV1.0, FEV1.0%, TLC, DLCo and DL/VA, namely, the parameters whose predictive value decreases with aging. In particular for V25, only the exponential form of the predictive formula could be applied for subjects with advanced age. For PMI, LCI, IDI, pulmonary N2 clearance delay, and single breath delta N2, indices for unevenness of intrapulmonary gas distribution, there has been no previous report about their predictive formulas and ours is the first. These predictive formulas all showed increasing values with aging, and showed higher predictive values in females than in males, except for delta N2. These parameters require further study with respect to their normal value ranges and differences between males and females.

Adolescent↗

Amyloid plaques, neurofibrillary tangles and neuronal loss in brains of transgenic mice overexpressing a C-terminal fragment of human amyloid precursor protein.

Alzheimer's disease (AD) affects more than 30% of people over 80 years of age. The aetiology and pathogenesis of this progressive dementia is poorly understood, but symptomatic disease is associated histopathologically with amyloid plaques, neurofibrillary tangles and neuronal loss primarily in the temporal lobe and neocortex of the brain. The core of the extracellular plaque is a derivative of the amyloid precursor protein (APP), referred to as beta/A4, and contains the amino-acid residues 29-42 that are normally embedded in the membrane-spanning region of the precursor. The cellular source of APP and the relationship of its deposition to the neuropathology of AD is unknown. To investigate the relationship between APP overexpression and amyloidogenesis, we have developed a vector to drive expression specifically in neurons of a C-terminal fragment of APP that contains the beta/A4 region, and have used a transgenic mouse system to insert and express this construct. We report here that overexpression of this APP transgene in neurons is sufficient to produce extracellular dense-core amyloid plaques, neurofibrillary tangles and neuronal degeneration similar to that in the AD brain.

Alzheimer Disease↗

Effect of cholecystokinin receptor antagonists, MK-329 and L-365,260, on cholecystokinin-induced acid secretion and histidine decarboxylase activity in the rat.

To elucidate the regulatory mechanism of acid secretion by cholecystokinin (CCK) in vivo, we compared the effects of CCK and gastrin on acid secretion and histidine decarboxylase (HDC) activity. We also examined the effects of MK-329, a specific antagonist for pancreatic-type CCK receptor, and L-365,260, a specific antagonist for gastrin-type CCK receptor, on the action of CCK. Graded doses of CCK or gastrin were intravenously infused into conscious rats with gastric fistula. Gastrin-17 I infusion up to 10 nmol/kg/h resulted in dose-related increases in acid secretion. CCK-8 infusion also caused an increase in acid secretion. However, it reached a peak with 0.3 nmol/kg/h CCK-8 and attenuated with higher concentrations of CCK-8. This attenuating effect of a higher dose of CCK was reversed by MK-329, but not by L-365,260. Both CCK and gastrin were potent in increasing fundic HDC activity, and the effect of CCK on HDC activity was significantly inhibited by L-365,260, but not by MK-329. Taken together, the present study suggests that CCK and gastrin stimulate histamine formation via a gastrin-type CCK receptor, and the attenuating action of CCK with higher concentrations on acid secretion in vivo is mediated by a pancreatic-type CCK receptor.

Animals↗

Involvement of substance P in hyperalgesia induced by intrathecal galanin.

Previously we have demonstrated that an intrathecal injection of galanin (GAL) decreases the nociceptive threshold for mechanical stimulation without effect on thermal nociceptive responses. The present experiments were conducted to determine whether substance P (SP) would be involved in such a decrease in the nociceptive threshold produced by GAL. An intrathecal injection of anti-SP monoclonal antibody inhibited the nociceptive threshold-decreasing effect of intrathecal GAL (0.1 nmol/rat). This antibody significantly suppressed the contractile action of SP (3 nM) on the longitudinal muscle and that of neurokinin A (3 nM) to a lesser degree. Binding of [125I]Tyr8-SP to this antibody was inhibited by SP in a concentration-dependent manner in the range 0.1-33 nM without suppression by GAL at a concentration of 3300 nM. In addition, an intrathecal injection of the anti-SP monoclonal antibody increased the nociceptive threshold for mechanical stimulation in carrageenin-inflamed rats without effect on thermal nociceptive behaviors. The capsaicin (0.5 microM)-evoked release of immunoreactive SP from dorsal-half slices of the spinal cord was increased by galanin (1 microM, but not 0.1 microM) without effects on basal release. An intrathecal injection of GAL did not produce aversive responses (biting, licking and scratching) at doses of 0.1 and 1 nmol/rat. GAL (0.1 nmol/rat) did not affect biting/licking behaviors evoked by SP (1 nmol/rat), but inhibited SP-evoked scratching behavior. These results suggest that the nociceptive threshold-decreasing action of intrathecal GAL is at least in part mediated by SP, and that GAL may act on primary afferent terminals to increase the release of SP evoked by stimulation.

Animals↗

Clinical and bacteriologic evaluation of OPC-7251 in patients with acne: a double-blind group comparison study versus cream base.

Twenty-eight patients with acne were assigned to 4 weeks of treatment with OPC-7251 (a new fluoroquinolone derivative) 1% cream or the cream base in a double-blind manner to evaluate the antibacterial effect of the drug on resident bacteria in the hair follicles and to evaluate clinical response. Propionibacterium acnes was isolated from 21 of the 28 acne patients. When the number of P. acnes was compared before and after treatment, the posttreatment P. acnes count in the OPC-7251 1% cream group was significantly (p = 0.000) reduced compared with that in the cream base group. OPC-7251 1% cream was also significantly (p = 0.019) superior to the cream base in terms of clinical response. P. acnes and Staphylococcus epidermidis isolated from the acne lesions were selected for their susceptibility to various antibacterial agents. The minimal inhibitory concentration of OPC-7251 against P. acnes and S. epidermidis was 0.10 to 0.20 and 0.024 to 0.10 micrograms/ml, respectively, which indicates that the drug has a potent antibacterial effect.

Acne Vulgaris↗

Intrathecal injections of galanin and its antiserum affect nociceptive response of rat to mechanical, but not thermal, stimuli.

To ascertain the involvement of galanin in nociceptive transmission in the spinal dorsal horn, we examined the effects of intrathecal injections of porcine galanin and its antiserum on behavioral nociceptive responses of the rat, using the paw-pressure and paw-radiant heat tests. An intrathecal injection of antiserum against porcine galanin reversed the decreased nociceptive threshold for mechanical, but not thermal, stimulation of the carrageenin-treated hind paw of rat, with no effect on the nociceptive threshold of the non-inflamed hind paw. Rat galanin as well as porcine galanin suppressed the binding of [125I]porcine galanin to the antiserum, a finding suggesting that the antiserum can bind rat galanin. An intrathecal injection of porcine galanin (0.1 and 1 nmol, but not 0.01 nmol) decreased the mechanical nociceptive threshold of the rat hind paw with no effect on thermal nociception. These results suggest that galanin present in the dorsal horn is involved in the facilitation of mechanical, but not thermal, nociceptive transmission.

Animals↗

Oral passive immunization against dental caries in rats by use of hen egg yolk antibodies specific for cell-associated glucosyltransferase of Streptococcus mutans.

The effect of polyclonal egg yolk immunoglobulin G antibodies (yIgG) raised against whole cells, cell-free (CF) glucosyltransferase (GTase), or cell-associated (CA) GTase of serotype c Streptococcus mutans was examined in terms of in vitro inhibition of virulence-related factors of S. mutans and protection of S. mutans-infected rats against the development of dental caries. Hens (18 weeks old) were immunized with formalin-treated whole cells, purified CF-GTase, or CA-GTase together with Freund's complete adjuvant. In addition, yIgG to surface protein antigen was used in some in vitro experiments for comparison with other antibodies. yIgG was purified by ammonium sulfate precipitation followed by DEAE-Sephacel column chromatography or fractional precipitation with ethanol. Purified yIgG was found to be a 220-kDa protein, which was dissociated into heavy and light chains upon addition of 2-mercaptoethanol. yIgG to whole cells and surface protein antigen gave a heavy aggregation of S. mutans organisms, while yIgG to CF- and CA-GTase specifically inhibited the enzymatic activity of the respective GTase. yIgG to CA-GTase and whole cells was found to clearly suppress the adherence of S. mutans cells to a glass surface. Specific-pathogen-free Sprague-Dawley rats that had been infected heavily and repeatedly with S. mutans and fed diet no. 2000 developed severe dental caries, while rats fed diet 2000 containing greater than or equal to 0.1% yIgG to CA-GTase showed a statistically significant reduction in dental plaque accumulation and caries development. Administration of yIgG to CF-GTase and whole cells failed to protect against caries. These results clearly suggest that yIgG to S. mutants CA-GTase specifically inhibited a virulence factor of this organism, i.e., insoluble glucan-synthesizing CA-GTase, resulting in a significant reduction in the development of dental caries.

Administration, Oral↗

The structure of (xylose)2glucose-O-serine 53 found in the first epidermal growth factor-like domain of bovine blood clotting factor IX.

We reported the presence of a new trisaccharide composed of two xylose and reducing terminal glucose residues linked to serine residues of bovine blood clotting factors VII and IX (Hase, S., Kawabata, S., Nishimura, H., Takeya, H., Sueyoshi, T., Miyata, T., Iwanaga, S., Takao, T., Shimonishi, Y., and Ikenaka, T. (1988) J. Biochem. (Tokyo) 104, 867-868). The present paper describes the detailed structural analysis of the trisaccharide. Glycopeptides were prepared from bovine factor IX by digestion with Pronase followed by purification by column chromatography. The trisaccharide was released from the protein by the beta-elimination reaction with hydrazine, and the reducing end of the sugar chain was tagged with 2-aminopyridine. The fluorescent pyridylamino derivative of the trisaccharide was purified by gel filtration and reversed-phase high performance liquid chromatography. The glycopeptides and pyridylamino-trisaccharide thus obtained were subjected to methylation study, 500-MHz 1H nuclear magnetic resonance spectroscopy, and periodate oxidation. Glucose and xylose belong to the D series by high performance liquid chromatography on a chiral column. From the results, the structure of the trisaccharide is proposed as: D-Xyl p alpha 1-3-D-Xyl p alpha 1-3-D-Glcp beta 1-O-Ser-53.

Animals↗

A new trisaccharide sugar chain linked to a serine residue in the first EGF-like domain of clotting factors VII and IX and protein Z.

Recently, we determined the complete amino acid sequence of bovine factor VII (Takeya, H. et al. (1988) J. Biol. Chem. 263, 14868-14877). In the course of the studies, we found an unknown serine derivative at position 52 in the first epidermal growth factor-like domain of factor VII. A pentapeptide isolated from the S-aminoethylated factor VII contained Ser-52, which could not be identified with a gas-phase sequencer. The same results were also obtained for a pentapeptide containing Ser-53 of factor IX and protein Z. Component sugar analysis revealed that the peptide contained 1 mol of glucose and 2 mol of xylose. This sugar component was also confirmed by high-resolution fast atom bombardment mass spectrometric analysis of the pentapeptide. The trisaccharide was released from the peptides by means of beta-elimination reaction and its reducing end was identified as pyridylamino-glucose. These results indicate the existence of a (Xyl2)Glc-Ser structure in factors VII, IX and protein Z. Similar results were obtained for human factors VII, IX and protein Z. This is the first report of a (Xyl2)-Glc-Ser structure in glycoproteins to our knowledge. The presence of the unique trisaccharide structure in factors VII, IX and protein Z leads us to anticipate its biological role in the tissue factor pathway.

Amino Acid Sequence↗

Monoclonal antibody (VII-M31) to bovine factor VII: a specific epitope in the gamma-carboxyglutamic acid domain.

A murine monoclonal antibody (designated VII-M31) directed against bovine factor VII was prepared and characterized. Antibody VII-M31 inhibited the activations of both factors IX and X catalyzed by factor VIIa in the presence of tissue factor, phospholipids, and Ca2+. It possessed a strong affinity for factor VII in the presence of 5 mM Ca2+ (Kd = 1.12 x 10(-10)M). The immunoblotting test of other bovine proteins with the antibody, such as prothrombin, factor X, factor IX, protein C, protein S, and protein Z, in addition to human factor VII, revealed that it recognizes only a Ca2(+)-dependent epitope in bovine factor VII. Furthermore, this antibody VII-M31 covalently coupled with Affi-Gel allowed a simple and rapid purification of bovine factor VII. To localize the antigenic site in factor VII, various segments including a gamma-carboxyglutamic acid (Gla)-domainless protein, a Gla-domain peptide and the fragments isolated from the lysyl endopeptidase digest, were prepared. Among them, the isolated Gla-domain peptide and Gla-domainless factor VII were no longer recognized by antibody VII-M31, indicating that the sequence around the cleavage site by a-chymotrypsin is required for the interaction between the antibody and factor VII. In accordance with this result, the antibody bound specifically to a Gla-containing peptide corresponding to the NH2-terminal 23-50 residues of factor VII, which contains the chymotryptic cleavage site. These results suggest that the specific epitope of this antibody is localized in the carboxy-terminal 28 residues of the Gla-domain constituting the amino-terminal portion of bovine factor VII.

1-Carboxyglutamic Acid↗

Molecular cloning and sequence analysis of cDNA coding for rat liver hemoprotein H-450.

cDNA clones coding for hemoprotein H-450 were isolated from a rat liver cDNA library using anti-H-450 antibody. The molecular weight calculated from the deduced amino acid sequence comprising 547 amino acid residues was 60,085. The N-terminal sequence and a partial internal amino acid sequence of purified H-450, which were determined chemically, were both found in the amino acid sequence of H-450 deduced from the nucleotide sequence. H-450 mRNA is expressed in liver, kidney, and brain. A homology search of amino acid sequences indicated that H-450 shows no homology with cytochrome P-450, but shows significant homology with bacterial O-acetylserine (thiol)-lyases. However, H-450 has no O-acetylserine (thiol)-lyase activity.

Amino Acid Sequence↗