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Biomedical subjects

J Katayama

Publications and source records attributed to J Katayama.

At least 19 recordsLinked to original sources
PubMed1998

Expression of the mphB gene for macrolide 2'-phosphotransferase II from Escherichia coli in Staphylococcus aureus.

The genes mphA and mphB encode macrolide 2'-phosphotransferases I and II, respectively, and they confer resistance to macrolide antibiotics in Escherichia coli. To study the expression of these genes in Gram-positive bacteria, we constructed recombinant plasmids that consisted of an mph gene and the pUB110 vector in Bacillus subtilis. When these plasmids were introduced into Staphylococcus aureus, the mphB gene was active and macrolide 2'-phosphotransferase II was produced. The gene endowed S. aureus with high-level resistance to spiramycin, a macrolide antibiotic with a 16-membered ring. Moreover, transcription of the mphB gene in S. aureus began at the promoter that was active in E. coli.

Escherichia coli↗
PubMed1998

ERPs, semantic processing and age.

ERPs (N400, LPC and CNV) were elicited in two sets of subjects grouped according to age (young vs. elderly) using a word-pair category matching paradigm. Each prime consisted of a Japanese noun (constructed from two to four characters of the Hiragana) followed by one Chinese character (Kanji) as the target, this latter representing one of five semantic categories. There were two equally probable target conditions: match or mismatch. Each target was preceded by a prime, either belonging to, or not belonging to, the same semantic category. The subjects were required to respond with a specified button press to the given target according to the condition. We found RTs to be longer in the elderly subjects and under the mismatch condition. N400 amplitude was reduced in the elderly subjects under the mismatch condition and there was no difference between match and mismatch response, which were similar in amplitude to that under match condition for the young subjects. In addition, the CNV amplitudes were larger in the elderly subjects. These results suggested that functional changes in semantic processing through aging (larger semantic networks and diffuse semantic activation) were the cause of this N400 reduction, attributing a subsidiary role to attentional disturbance. We also discuss the importance of taking age-related changes into consideration in clinical studies.

Adult↗
PubMed1998

Expression in Pseudomonas aeruginosa of an erythromycin-resistance determinant that encodes the mphA gene for macrolide 2'-phosphotransferase I from Escherichia coli.

We studied the expression in Pseudomonas aeruginosa of an erythromycin-resistance (EMr) determinant that included the mphA gene for macrolide 2'-phosphotransferase I and originated in Escherichia coli. A recombinant plasmid, pTZ3609, that consisted of the EMr determinant and a broad-host-range vector RSF1010, endowed P. aeruginosa with high-level resistance to erythromycin. Furthermore, the EMr determinant on a self-transferable plasmid, RP1, was transferred from E. coli to P. aeruginosa by conjugation.

Anti-Bacterial Agents↗
PubMed1998

Isolation and characterization of two plasmids that mediate macrolide resistance in Escherichia coli: transferability and molecular properties.

Escherichia coli BM2506 is highly resistant to macrolide antibiotics; it produces macrolide 2'-phosphotransferase II [MPH(2')II] which inactivates such drugs. We investigated the localization and the transfer of the macrolide-resistance determinant that encoded the mphB gene for MPH(2')II in strain BM2506. Although we detected no clear band of plasmid DNA after agarose gel electrophoresis, transformation analysis using satellite DNA that corresponded to plasmid DNA after CsCl-ethidium bromide gradient centrifugation and restriction analysis of plasmid DNA in transformants showed that strain BM2506 harbored two plasmids, pTZ3721 (84 kb) and pTZ3723 (24 kb), that specified resistance to macrolides, ampicillin, streptomycin, tetracycline and sulfonamide and to macrolides and ampicillin, respectively. Southern hybridization showed that the mphB gene hybridized to both plasmids. Furthermore, pTZ3721 was transferred by conjugation to another strain of E. coli and pTZ3723 was mobilized with a self-transferable plasmid RP1 to other strains of E. coli. Therefore, it appears that the mphB gene is located on two plasmids in BM2506 and can be transferred to other strains of E. coli by conjugation or mobilization.

Anti-Bacterial Agents↗
PubMed1998

Stimulus context determines P3a and P3b.

P300 differences for target (.10), nontarget (.10), and standard tones (.80) were assessed using a three-stimulus oddball paradigm in which participants responded only to the target (n = 12). Target/standard (easy or difficult) and nontarget/standard (large or small) pitch differences were manipulated orthogonally. In all conditions, target tones elicited a parietal P300, which was affected only by the target/standard discrimination ease. Nontarget in the easy/large and difficult/small conditions elicited a parietal but smaller P300 than the target but in the easy/small condition elicited similar ERPs to the standard. However, nontarget stimuli in the difficult/large condition elicited an anterior maximum and earlier P300 (P3a) component. The findings suggest that target P300s are not influenced by the nontarget stimulus configuration, whereas the nontarget P300 outcomes are determined directly by the stimulus context. The theoretical implications are discussed.

Acoustic Stimulation↗
PubMed1997

A solid-phase assay to screen monoclonal antibodies against DNA-binding protein.

A method is described for selecting monoclonal antibodies (mAb) against DNA-binding protein. The protocol involves a non-radioactive solid-phase DNA binding assay using a 96-well plate. Because the solid-phase assay is highly specific and sensitive, partially purified antigen is sufficient for the immunization, and mAb screening can be performed with crude cell extract as the antigen. MAbs obtained by this method could supershift the DNA-protein complex in the electromobility shift assay, and were sufficient for immunoscreening of a cDNA expression library.

Animals↗
PubMed1997

Characterization of the K+ current mediated by 5-HT1A receptor in the acutely dissociated rat dorsal raphe neurons.

The action of 5-hydroxytryptamine (5-HT) via the 5-HT1A receptor on dissociated rat dorsal raphe neurons was characterized under the whole-cell mode by using the nystatin-perforated patch-clamp technique. Under voltage-clamp conditions, 5-HT induced an inwardly rectifying K+ current (I5-HT) in a concentration-dependent manner. I5-HT was mimicked by 8-OH-DPAT and buspirone, which are both 5-HT1A receptor agonists. I5-HT was reversibly blocked by such 5-HT1A receptor antagonists as (S)-UH-301 a 5-HT4 receptor antagonist. I5-HT was antagonized concentration-dependently by such K+ channel blockers as quinine, Ba2+ and 4-aminopyridine but was relatively insensitive to both CS+ and tetraethylammonium. When the neurons were loaded with guanosine 5'-O-3-thiotriphosphate through a patch pipette, the K+ current induced by 5-HT became irreversible. N-ethylmaleimide (NEM), a sulfhydryl alkylating agent, irreversibly blocked I5-HT. The intracellular perfusion with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), a Ca2+ chelator, or neomycine, a phospholipase C inhibitor, never significantly affected the 5-HT-induced response. 12-Myristate 13-acetate diester (PMA), a protein kinase C (PKC) activator, had only a weak inhibitory effect on I5-HT, and staurosporine, a PKC inhibitor, failed to significantly occlude I5-HT. Therefore, the K+ conductance activated via the 5-HT1a receptor of dorsal raphe neurons was thus characterized by the sensitivity to such K+ channel blockers as quinine, Ba2+ and 4-aminopyridine. Moreover, G protein which is NEM-sensitive and can couple to the 5-HT1A receptor, is thus considered to activate the inwardly rectifying K+ conductance without being mediated by such second messengers as Ca2+ and PKC.

Animals↗
PubMed1997

Discovery of a novel thrombopoietin mimic agonist peptide.

A random phage peptide library was constructed for the filamentous bacteriophage fuse5. The library was made by inserting a degenerate oligonucleotide which encodes 15 variable amino acids into the NH2-terminal region of the phage gene III protein. This library, containing 1x10(9) different phages, was screened with a human immunoglobulin fusion protein containing the extracellular region of human thrombopoietin receptor. Several phages were isolated following four cycles of enrichment and amplification. These phages specifically bound to the fusion protein. One phage peptide acted as an agonist of the thrombopoietin receptor, since it stimulated the proliferation of thrombopoietin-dependent cells and the differentiation of mouse bone marrow cells to megakaryocytes. The amino acid sequence of this peptide is not present in the primary amino acid sequence of thrombopoietin. This discovery may lead to the design of a small-molecular mimic of thrombopoietin.

Amino Acid Sequence↗
PubMed1996

Cloning and nucleotide sequence of the mphB gene for macrolide 2'-phosphotransferase II in Escherichia coli.

Macrolide 2'-phosphotransferase II [MPH(2')II] inactivates macrolide antibiotics. The mphB gene for MPH(2')II was cloned from Escherichia coli and sequenced. Analysis of the nucleotide sequence indicated that mphB encoded a protein of 302 amino acids with a molecular mass of 34483 Da. The carboxy terminal region of the deduced protein contained a sequence that resembled a conserved functional domain in aminoglycoside phosphotransferases.

Amino Acid Sequence↗
PubMed1996

Two components of metabotropic glutamate responses in acutely dissociated CA3 pyramidal neurons of the rat.

The excitatory and inhibitory actions of metabotropic glutamate receptor (mGluR) agonists were investigated in acutely dissociated rat hippocampal CA3 pyramidal neurons, using the conventional whole-cell and nystatin-perforated patch recording configurations under the voltage-clamp condition. With the conventional whole-cell recording, glutamate (Glu) and quisqualic acid (QA) induced only ionotropic inward currents accompanied by increased membrane conductance at a holding potential (VH) of -45 mV. The response was reversibly blocked in the presence of D-2-amino-5-phosphonopentanoic acid (D-AP5) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), the antagonists of N-methyl-D-aspartate (NMDA) receptor and non-NMDA receptor, respectively. With nystatin-perforated patch recording, mGlu responses insensitive to both D-AP5 and CNQX were observed. Fifty-five % of the cells responded by a slow inward current accompanied by conductance decrease (ImGlui) at a VH of -44 mV. One % of the neurons showed an outward current with conductance increase (ImGluo), and 34% of the neurons showed ImGluo followed by ImGlui. The onset of ImGluo occurred approximately 900 ms after the response to 30 mM K+. The time to peak of ImGluo were 32- to 79-times longer than those of ionotropic responses. ImGlui appeared at lower concentrations than ionotropic Glu responses, whereas ImGluo appeared at similar concentrations as ionotropic responses. The rank order of affinity was QA > Glu > (+/-)-1-aminocyclopentane-trans-1, 3-dicarboxylic acid (tACPD) for both ImGlui and ImGluo. Half-maximal effective concentrations (EC50) and the threshold concentrations for the three agonists were four- to tenfold lower for ImGlui than for ImGluo. The current-voltage relationship showed that the reversal potentials of ImGlui and ImGluo shifted 55 and 59 mV, respectively, for a tenfold change in extracellular K+ concentration, indicating that K+ is the charge carrier of both mGlu responses. During ImGlui, both the leakage current and muscarine-sensitive voltage-dependent K+ current (M current) were suppressed. ImGluo induced by 10(-4) M tACPD was abolished by 3.10(-7) M charybdotoxin and 10(-6) M ryanodine. These results show that there are two components of mGlu responses in CA3 pyramidal neurons and that ImGlui and ImGluo show different pharmacological properties.

Animals↗
PubMed1996

P300, probability, and the three-tone paradigm.

The effects of stimulus probability on P300 from a 3-tone paradigm were examined in two experiments. Experiment 1 manipulated the probability of the non-target tone as 0.10, 0.45, or 0.80, while the target tone probability was always 0.10. Experiment 2 manipulated the probability of 3 tones as 0.10, 0.30, or 0.60, with one of the infrequent tones assigned as the target in each condition. Subjects were required to press a button in response to the target stimulus in both experiments. The results indicated that the P300 to the target and the non-target were both affected by the probability of the eliciting stimulus, such that component amplitude was inversely related to probability; no reliable P300 latency effects were found. Target tones elicited larger P300 amplitude than the non-target tones at the same probability. The findings suggest that probability effects on P300 amplitude are independent of responding to a specific target stimulus and are discussed with reference to the clinical utility of the 3-tone paradigm.

Acoustic Stimulation↗
PubMed1996

[Functional imaging of the sensorimotor cortex using an ultra-fast MR imaging method].

UNLABELLED: The aim of this study was to assess changes in brain activity during a motor task and variable sensory stimulation using echo planar imaging, which represents the fastest clinically usefull imaging technique available. MATERIALS AND METHODS: The subjects of this study were 11 healthy-volunteers, 4 males and 11 females, with an average of 26.4 years. The subjects were instructed to tap the fingers of one hand as the motor task. Compressed air was applied 5 times a second as "simple" sensory stimulation. Simple figures were drawn on the subjects palm as "complex" sensory stimulation. In all cases, functional imaging was performed by T2*-weighted echo planar imaging (TE = 53 msec, TR = 3000 msec, flip angle = 90 degrees, matrix 64 x 64, FOV = 205 mm, slice thickness = 8 mm) alternately at rest and during the task (intervals: 30 sec). A total of 60 images was collected in 3 minutes. Images obtained by subtracting images at rest and during the task were analyzed. RESULTS AND DISCUSSION: Almost all subjects showed a transient signal increase in the contralateral paracentral region during simple sensory stimulation. Continuous signal increases in the contra- and/or ipsi-lateral para-central region were observed during complex sensory stimulation. Some exhibited signal increases in the parietal or frontal association cortex, but they disappeared when subject's attention was distracted during stimulation. All subjects displayed signal increases in the contralateral para-central region during the motor task. Some of them exhibited signal increases in the medial frontal area (supplementary motor area) and ipsilateral para-central region. These results suggest that the signal increases of functional MRI reflect not only simple reactions to stimulation but higher cerebral function as well.

Adult↗
PubMed1995

Effect of rat thyroid proliferative lesion development by intermittent treatment with sulfadimethoxine.

To determine whether production of thyroid proliferative lesions would be enhanced by intermittent rather than continuous treatment with a goitrogen, male F344 rats initiated with N-bis(2-hydroxypropyl)nitrosamine (DHPN, 2800 mg/kg body weight, single s.c. injection) were given water containing 0.1% sulfadimethoxine (SM) for 20 weeks (group 1) or 0.1% SM for the first 8 weeks followed by 2 cycles consisting of 2 weeks withdrawal and 4 weeks retreatment with 0.1% SM (group 2). Control rats (group 3) were untreated for 20 weeks after the DHPN initiation. Serum T3 and T4 levels were significantly decreased in groups 1 and 2 compared to group 3. Serum thyroid stimulating hormone level was significantly increased in all treated groups compared to group 3. The numbers of follicular cell hyperplasias were significantly increased in group 2 compared to group 1. BrdU labeling indices for follicular cells and hyperplasias were also significantly elevated in group 2 compared to group 1. Electron microscopic examination of thyrotrophs in the anterior pituitary in groups 1 and 2 revealed dilated rough ER cisternae with intracisternal dense granules. The number of intracytoplasmic secretory granules in group 2 was moderately decreased compared to group 1. Therefore, the results of the present study suggest that it may be possible to enhance production of thyroid neoplastic lesions by intermittent treatment.

Animals↗
PubMed1993

The measurement of intracellular sodium ion concentration in the horizontal cells of the stingray retina.

Membrane potentials, and intracellular concentrations of sodium ions in the horizontal cells of the stingray retina were measured directly by means of ion-selective microelectrodes. The electrodes used were of the double-barreled type and bis[(12-crown-4)methyl]methyldodecylmalonate was used as the ion-selective resin. The average intracellular concentration of Na+ ions in the horizontal cells and the extracellular concentration of Na+ ions near the horizontal cells were 123 mM and 351 mM, respectively (n = 27). The equilibrium potential was estimated to be +27.3 mV. Although each value of the equilibrium potential was scattered over a wide range, they were always positive regardless of the membrane potential. This was the first direct measurement of the sodium ion-concentration for the retinal horizontal cells. The results support the idea that the origin of the dark depolarization of the stingray horizontal cell is caused by the glutamate-induced high-sodium conductance of the membrane.

Animals↗
PubMed1992

Negative brain potentials elicited by an unexpected color patch or word.

The purpose of this study was to examine whether a physical stimulus that deviates from a semantic context can elicit the N400 component of event-related brain potentials (ERPs). ERPs were recorded while 12 students judged the veracity of a simple statement (e.g., red/is not/blue) presented with the order of subject (S), object (O), and verb (V), which is normal in Japanese grammar. In one condition, S was a color patch and O was a word representing the color, while in the other condition, S was a color name and O was a patch. In both conditions, a late additional negative potential was elicited by the O stimulus when it was mismatched with S. In addition, the negativities elicited by the incongruous color patch and word had the same morphology and scalp distribution. The results indicate that not only a word but a physical stimulus which deviates from a semantic context can elicit the N400 component.

Adult↗
PubMed1992

Intracellular and extracellular chloride ions in the horizontal cells of the stingray retina.

Intracellular and extracellular concentrations of chloride ([Cl-]i, [Cl-]o) ions in the horizontal cells of the stingray retina were studied by means of ion-selective microelectrodes. The electrodes used were of the double-barreled type and Corning's #477315 resin was used as the exchanger. The average [Cl-]o and [Cl-]i in the L-type cells were 320 and 130 mM, respectively (n = 37), and the equilibrium potential (ECl) was calculated as -23.3 mV. In some cases, dark membrane potentials were more positive than the ECl. With white light stimulus, the membrane hyperpolarized to a more negative level than ECl. Based on the experiments described above, the hypothesis that the hyperpolarizing response of horizontal cells is due to the permeability change of the membrane to chloride ion was excluded in the stingray retina.

Animals↗
PubMed1991

Negative brain potentials reflect the subjective distance between standard and deviant tones.

The purpose of this study was to examine the effects on event-related brain potentials (ERPs) of occasionally presented deviants in a sequence of tones. The sequence consisted of a series of frequent standard C5 tones from the diatonic musical scale, in which three deviants (F #5, C6, and F #6) and a target (C4) were occasionally embedded. Twenty subjects were asked to detect the target. The deviants elicited frontal N2. The amplitude of the N2 was smaller for C6 than for the other two deviants, whereas peak latencies increased as a function of the frequency distance between the deviant and the standard. The smaller N2 to C6 than that to the other deviants was considered to reflect closer subjective distance due to the octave relationship between C6 and the standard C5.

Adult↗