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Biomedical subjects

S Taguchi

Publications and source records attributed to S Taguchi.

At least 91 records · Page 5Linked to original sources
PubMed1995

Sensitization of nociceptive C-fibers in zinc-deficient rats.

A marked decrease in zinc concentration was observed in plasma (P < 0.001), hindpaw skin (P < 0.01), and dorsal skin (P < 0.01) in zinc-deficient rats (rats fed a zinc-deficient diet for 3 wk), compared with the control rats fed the same zinc-deficient diet supplemented with ZnCO3 (50 mg/kg diet). The threshold intensity needed to elicit vasodilatation in the hindpaw skin of the zinc-deficient rats on electrical stimulation of the saphenous nerve in a peripheral direction was markedly lower (P < 0.01) than that in the control rats. No difference was observed between control (n = 5) and zinc-deficient rats (n = 5) in the magnitude of the plasma extravasation evoked by either histamine or substance P. There was no difference between control and zinc-deficient rats in terms of the dose-response curve for release of histamine by substance P. Prostaglandin E2 (PGE2) concentration in the hindpaw skin of the zinc-deficient rats was nearly fourfold higher (P < 0.01) than that of the control rats, whereas no difference in the leukotriene B4 level in the hindpaw skin was observed between control and zinc-deficient rats. From the present study, it seems likely that an increased level of PGE2 in the vicinity of the nociceptive C-fiber terminals in the hindpaw skin of zinc-deficient rats may sensitize the terminals of the nociceptive C-fibers of the saphenous afferent nerve in the hindpaw and thus facilitate the production of antidromic vasodilatation.

Afferent Pathways↗
PubMed1995

Molecular cloning and sequence analysis of a gene encoding an extracellular serine protease from Streptomyces lividans 66.

A gene encoding a homolog of the chymotrypsin-like serine protease (SAM-P20), which was isolated as the target enzyme of a protease inhibitor (SSI), was cloned from Streptomyces lividans 66. This gene contained an open reading frame of 1065 nucleotides encoding 354 amino acid residues with a putative prepro portion of 157 amino acid residues. The deduced amino acid sequence of the cloned gene had significant homology to those of members of Streptomyces extracellular chymotrypsin-like protease family. By Southern blot analysis, it was suggested that protease genes of this type are found at a high frequency in Streptomyces. In this sense, we propose to categorize this protease as a member of the 'SAL' series (SAM-P20-like proteases).

Amino Acid Sequence↗
PubMed1995

[Movement-related potentials associated with motor inhibition during different types of stop signal paradigm in humans].

The motor inhibition process was examined in humans by monitoring reaction times, electromiograms, and movement-related potentials. Four subjects performed two types of visual stop signal paradigm. In one type, they were required to push a button (GO) or not to push it (NOGO), in another type, to release the button (GO) or to keep pushing it (NOGO). The results were summarized as follows; (1) The EEG waveform pattern was remarkably consistent among subjects. NO-GO-specific negative-positive potentials were observed under both tasks without significant background EMG activity changes in agonist and antagonist muscles. (2) The timing of the onset and the peak latency of the negative potential relative to the second stimulus, S 2, were constant (about 180-200 ms and 230-250 ms for each) in both tasks. The time interval between the initial onset of the negative potential and its peak latency was also constant (about 50 ms) in both tasks. The amplitude of the negative potentials was maximum at FCz (supposed to be around supplementary motor area) and their distribution was invariant among tasks. Constant features of NO-GO-specific potentials during both task performance suggested the temporally and spatially same inhibition process might specifically suppress the new motor program regardless of the existence of ongoing motor program.

Action Potentials↗
PubMed1994

Primary structure and inhibitory properties of a subtilisin-chymotrypsin inhibitor from Streptomyces virginiae.

A novel serine protease inhibitor SIL8, which was isolated from the culture medium of Streptomyces virginiae and shown to be a member of the Streptomyces subtilisin-inhibitor-like (SIL) inhibitor family by sequence analysis of its amino-terminal region [Taguchi, S., Kikuchi, H., Kojima, S., Kumagai, I., Nakase, T., Miura, K. & Momose, H. (1993) Biosci. Biotech. Biochem. 57, 522-524], is the first SIL inhibitor demonstrated to show marked inhibitory activity toward alpha-chymotrypsin, in addition to strong inhibitory activity toward subtilisin BPN', a common property of inhibitors of the Streptomyces subtilisin inhibitor (SSI) family. In this study, the complete amino acid sequence of SIL8 was determined from the sequence analysis of peptides obtained by specific cleavage at the reactive site and by enzymic digestion. SIL8 was shown to exist as a dimer protein, each subunit of which was composed of 111 amino acids, and to have less than 50% similarity with other SSI-family inhibitors, indicating its most distant relationship to other members of this family. Insertion of two residues was observed in the flexible loop region of SIL8, and amino acid replacements were found not only on the molecular surface but also in the beta-sheet and hydrophobic core, suggesting that packing rearrangements of the side chains may occur in these regions to maintain the tertiary and quaternary structures. The inhibitor constants Ki obtained using synthetic substrates are 92 pM for subtilisin BPN' and 11 nM for alpha-chymotrypsin. The P1 site was was identified as methionine, which was in good agreement with the substrate specificity of alpha-chymotrypsin. SSI, which also possesses a methionine residue at the P1 site, inhibits alpha-chymotrypsin poorly (inhibitor constant, 4.0 microM). Such a difference in the inhibitory properties of SIL8 and SSI toward alpha-chymotrypsin is discussed on the basis of the structures of the inhibitors.

Amino Acid Sequence↗
PubMed1994

Primary structure and inhibitory properties of a proteinase inhibitor produced by Streptomyces cacaoi.

Protein proteinase inhibitors showing sequence homology with Streptomyces subtilisin inhibitor (SSI) have been found to be distributed widely in Streptomyces species, and accordingly have been named SSI-like (SIL) proteins. SIL1 from S. cacaoi was the first of these proteins to be isolated and to be given a serial number. To study the structure-function relationship of SIL proteins, we determined the primary structure of SIL1 and measured its inhibitory activities. It was found to be composed of 110 amino acids and to exist in dimer form. The amino-acid sequence of SIL1 was unique among other characterized SIL proteins in having a one-residue deletion in two regions and a three-residue insertion in the flexible loop region. Sequence comparison indicated that SIL1 was distantly related to other members of the SSI family, and that amino-acid replacements had occurred not only on the surface of the SIL1 molecule but also in the beta-sheet region. The reactive site of SIL1 was considered to be Arg70-Glu71 from sequence alignment with other SSI-family inhibitors. SIL1 inhibited subtilisin BPN' strongly with an inhibitor constant (Ki) of 2.8 x 10(-11) M, like other members of the SSI family possessing an Arg residue at the P1 site. In contrast, SIL1 exhibited weak inhibition toward trypsin with a Ki value of 5.5 x 10(-8) M, possibly as a consequence of insertion of the three residues in the flexible loop region near the reactive site. This contrast seems to be due to the difference in the subsite structure of the two proteinases.

Amino Acid Sequence↗
PubMed1994

Comparative studies on the primary structures and inhibitory properties of subtilisin-trypsin inhibitors from Streptomyces.

Three novel proteinaceous inhibitors of serine proteases which had been identified as Streptomyces subtilisin inhibitor-like (SIL) inhibitors were isolated from culture supernatant of Streptomyces; SIL2 from Streptomyces parvulus, SIL3 from Streptomyces coelicolor and SIL4 from Streptomyces lavendulae. They exhibited not only strong inhibitory activity toward subtilisin BPN' but also less strong inhibition of trypsin. Their primary sequences were determined by sequence analysis of peptides obtained by specific cleavage at the reactive site and subsequent proteolytic digestion. Each inhibitor consisted of about 110 amino acids, and was considered to form a dimer. The reactive site of the inhibitors was identified as Arg-Glu for SIL2 and SIL3, and Lys-Leu for SIL4, from sequence analysis of modified forms of the inhibitors produced from the inhibitor-subtilisin complex under acidic conditions. The presence of an arginine/lysine residue at the P1 site was in agreement with their trypsin-inhibition property. Sequence comparison with other members of the Streptomyces subtilisin inhibitor family revealed that amino acid replacements in the three isolated SIL inhibitors were frequently localized on the surface region, and many of the amino acid residues in beta-sheets and the hydrophobic core were highly conserved. Values of the inhibitor constant (Ki) toward subtilisin BPN' and trypsin were also measured, and the differences were discussed on the basis of the determined structures of the inhibitors.

Amino Acid Sequence↗
PubMed1994

Improvement of a useful enzyme (subtilisin BPN') by an experimental evolution system.

In order to improve a natural enzyme so as to fit industrial purposes, we have applied experimental evolution techniques comprised of successive in vitro random mutagenesis and efficient screening systems. Subtilisin BPN', a useful alkaline serine protease, was used as the model enzyme, and the gene was cloned to an Escherichia coli host-vector system. Primary mutants with reduced activities of below 80% of that of the wild type were first derived by hydroxylamine mutagenesis directly applied to subtilisin gene DNA, followed by screening of clear-zone non-forming transformant colonies cultured at room temperature on plates containing skim-milk. Then, secondary mutants were derived from each primary mutant by the same mutagenic procedure, but screened by detecting transformant colonies incubated at 10 degrees C with clear zones that were greater in size than that of the wild type. One such secondary mutant, 12-12, derived from a primary mutant with 80% activity, was found to gain 150% activity (kcat/Km value) of the wild-type when the mutant subtilisin gene was subcloned to a Bacillus subtilis host-vector system, expressed to form secretory mutant enzyme in the medium, and the activity measured using N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide as the substrate. When N-succinyl-L-Ala-L-Ala-L-Pro-L-Leu-p-nitroanilide was used, 180% activity was gained. Genetic analysis revealed that the primary and secondary mutations corresponded to D197N and G131D, respectively. The activity variations found in these mutant subtilisins were discussed in terms of Ca(2+)-binding ability. The thermostability was also found to be related to the activity.

Amino Acid Sequence↗
PubMed1994

Reserpine-induced cell transformation without detectable genetic effects in Syrian hamster embryo cells in culture.

Reserpine, a naturally occurring rauwolfia alkaloid, used mainly as an antihypertensive drug, was examined for its ability to induce cell transformation and genetic effects in Syrian hamster embryo (SHE) cells in culture. Treatment of SHE cells with 2 micrograms/ml of reserpine had no effect on cell growth, while 4 micrograms/ml of reserpine reduced the growth rate slightly and 8 micrograms/ml resulted in a significant inhibition of cell growth. Reserpine at doses of 4-8 micrograms/ml for 48 h induced a dose-related increase in morphological transformation of the cells. Reserpine-transformed colonies were morphologically indistinguishable from colonies transformed with benzo[a]pyrene (B[a]P) or other chemical carcinogens. Over the dose range that resulted in cell transformation, treatment of SHE cells with reserpine failed to induce any detectable gene mutations at two genetic loci, chromosomal abnormalities including structural and numerical changes, or DNA adduct formation. These findings indicate that reserpine may have carcinogenic potential by unknown mechanisms that do not include direct induction of gene and/or chromosome mutations.

Aneuploidy↗
PubMed1994

Three novel subtilisin-trypsin inhibitors from Streptomyces: primary structures and inhibitory properties.

Three novel proteinaceous inhibitors, which had been identified as "Streptomyces subtilisin inhibitor-like (SIL) proteins" and exhibited trypsin inhibition in addition to strong inhibition toward subtilisin BPN', were purified from the culture broth of three Streptomyces strains: SIL10 from S. thermotolerans, SIL13 from S. galbus, and SIL14 from S. azureus. Their primary structures were determined by sequence analysis of intact SIL inhibitors and peptides obtained by enzymatic digestions of S-pyridylethylated SIL inhibitors. These inhibitors were composed of about 110 amino acids and existed as dimer proteins. The reactive site was identified as Lys-Gln for all three inhibitors by sequence analysis of their modified forms in which the reactive-site peptide bond was specifically cleaved by subtilisin BPN' under acidic conditions. Thus, their inhibition toward trypsin and subtilisin BPN' was due to the presence of a Lys residue at the P1 site. Inhibitor constants toward subtilisin BPN' and trypsin were also determined. These inhibitors showed relatively high sequence homology to other SSI-family inhibitors possessing a Lys residue at the P1 site, with amino acid replacements on their molecular surface.

Amino Acid Sequence↗
PubMed1994

The effect of stimulating voltage on time characteristics of contraction in cardiomyoplasty.

In cardiomyoplasty (CMP), the strength of a skeletal muscle increases as the stimulating voltage increases. On the other hand, timing of stimulation in CMP is a problem not yet solved. This experiment assessed the relationship between stimulating voltage and time characteristics in the contraction of skeletal muscles. In acute experiments using rabbits, the time required to reach maximal muscle contraction did not change significantly with voltage. In conclusion, stimulation voltage does not affect time characteristics. Thus, the best timing of stimulation in CMP can be assessed independently of voltage.

Animals↗
PubMed1994

A miniaturized centrifugal pump for assist circulation.

The newly developed Nikkiso HMS-15 is a miniaturized centrifugal pump. It has an impeller diameter of only 50 mm and a priming volume of only 25 ml. A totally new approach was applied to develop this very small pump. The new pump showed comparable hemolysis with pumps twice as big (e.g., the most widely used cone-shaped centrifugal pump [index of hemolysis, 0.005]). This finding refutes the belief that the pump diameter must be sufficiently large in size. Clinical application for cardiac assist was performed for 48 h without any thrombus formation despite low heparin dosage. Also, the pump showed quite favorable blood trauma when applied as a pump for cardiopulmonary bypass during open heart surgery. The compactness, the high controllability, and the system versatility proved to be very effective for clinical application. This pump is considered very reliable for its highly optimized design.

Aged↗
PubMed1994

[Synergism on the bactericidal effect of gentian violet (Gv) and acrinol (Ac) against Pseudomonas aeruginosa].

The MBCs of Ac against P. aeruginosa (7 strains) isolated from infected skin lesions of patients were more than 6400 micrograms/ml, and those of Gv were more than 1600 micrograms/ml. When either Ac or Gv was used independently, these dyes did not have the bactericidal effect of P. aeruginosa. When Gv was used in combination with Ac, predominantly synergism on the bactericidal effect of Ac and Gv against P. aeruginosa was observed. The MBCs of an Ac-Gv cocktail were between 100 micrograms/ml and 225 micrograms/ml. We have previously reported that Gv possessed significantly a bactericidal effect to MRSA isolated from clinical specimens. Therefore, these results suggested that a combination treatment by an Ac-Gv cocktail may be one of the useful drugs for the MRSA and P. aeruginosa mixed infection on the skin lesions which is frequently observed clinically.

Drug Synergism↗
PubMed1994

In vivo monitoring system for structure-function relationship analysis of the antibacterial peptide apidaecin.

A unique antibacterial peptide derivative found in immune honeybee lymph, apidaecin 1b (AP1), was randomly mutagenized and characterized by a newly established system to analyze in vivo its structure-function relationship. Initially, a high-level expression host-vector system for AP1 in Escherichia coli was constructed by creating a fusion protein with the highly stable Streptomyces subtilisin inhibitor (SSI) molecule. Expression of the SSI-AP1 fusion protein was found to depend on the concentration of the transcriptional inducer isopropyl-beta-D-thio-galactopyranoside (IPTG) and to parallel the degree of growth inhibition of the transformant cells. Subsequently, apidaecin derivatives produced by localized random mutagenesis were screened with this IPTG concentration-controlled in vivo system by monitoring the growth inhibition patterns of the transformant cells. One mutant apidaecin (P9L) that had reduced activity was purified and isolated from the periplasmic fraction of an E. coli transformant. Its antibacterial activity was reduced to one-third of that of wild-type apidaecin. When considered together with the other mutations, it was concluded that several Pro residues, including that at the ninth position, are responsible for expression of the antibacterial action of apidaecin.

Amino Acid Sequence↗
PubMed1994

[Screening for thyroid dysfunction in adults residing in Hokkaido Japan: in relation to urinary iodide concentration and thyroid autoantibodies].

The prevalence of thyroid dysfunction and its relation to thyroid autoantibodies (TAA) and urinary iodide concentration (UI) was studied in apparently healthy adults in Sapporo (n = 4110) (Sapporo group), and in five coastal areas of Hokkaido (n = 1061) (coastal group) which produce iodine-rich seaweed (kelp). The frequency of above normal UI (high UI) in the morning urinary samples of coastal group was 10.8%, significantly higher than that of Sapporo group (6.4%) (p < 0.001). Frequency of positive TAA in both groups were similar. In Sapporo group TAA was positive in 6.4% of males and 13.8% of females with an age-related increase. The overall prevalence of hyperthyroidism (TSH < 0.15 mU/L) in coastal group (0.6%) was similar to that in Sapporo group (1.1%), while that of hypothyroidism (TSH > 5.0 mU/L) in coastal group (3.8%) was significantly higher than that in Sapporo group (1.3%) (P < 0.001). The frequency of high UI correlated significantly with that of hypothyroidism with negative TAA (r = 0.829, P < 0.05), but not with positive TAA, or with that of hyperthyroidism. Hypothyroidism was more prevalent in TAA negative subjects with high UI than with normal UI. Moreover, serum TSH and thyroglobulin levels were higher and free T4 level was lower in former than in latter group. These results indicate that 1) the prevalence of TAA negative hypothyroidism in iodine sufficient areas may be associated with the amount of iodine ingested, 2) this hypothyroidism is more prevalent and marked in subjects consuming further excess amounts of iodine, and 3) excessive intake of iodine should be considered an etiology of hypothyroidism in addition to chronic thyroiditis in these areas.

Adult↗
PubMed1994

[A gastric proton pump inhibitor as preanesthetic medication].

Effect of omeprazole, a gastric proton pump inhibitor, on gastric secretion during anesthesia and surgery was evaluated in 39 elective surgical patients ranged in age from 18 to 69 years. These patients were divided into two groups according to their age either of 40 years and under or over. The patients of both groups underwent orthopedic, ophthalmic, ENT, plastic, oral or non-abdominal surgery under neuroleptanesthesia, enflurane anesthesia or total intravenous anesthesia with droperidol, fentanyl and ketamine. They all were administered omeprazole 20 mg orally at 21:00 the night before surgery and again at 7:00 on the morning of surgery. The volume and acidity of gastric juice were measured at anesthetic induction and emergence from anesthesia. The volume and pH of the gastric juice in patients of 40 years and under in age averaged to 9.8 +/- 3.2 (mean +/- SE) ml, 2.45 +/- 0.56 at the anesthetic induction and 9.3 +/- 4.1 ml, 4.66 +/- 0.60 at the emergence from anesthesia respectively. The mean volume and pH of the gastric juice in patients over 40 years of age were 5.0 +/- 1.7 ml, 4.68 +/- 0.56 at the anesthetic induction and 9.9 +/- 2.4 ml, 5.76 +/- 0.36 at the emergence from anesthesia respectively. Significant decrease in the volume and acidity of gastric juice was observed in the patients of both groups except that the average of intragastric pH of the patients under 40 years of age was below 2.50 at the induction of anesthesia.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed1994

[Inhibitory effect of running exercise on age-induced muscle atrophy].

The effects of a 10-week running exercise on the histochemical and morphological properties of muscle fibers in the soleus (SOL) and tibialis anterior (TA) muscles and also in spinal motoneurons were investigated in 4 groups of female rats. Animals of the control groups were 20, 90 and 100 weeks old. Animals of the experimental group were 100 weeks old, and were exercised on a running wheel for 10 weeks. Muscle fibers examined were classified as slow-twitch oxidative (SO), fast-twitch oxidative glycolytic (FOG) or fast-twitch glycolytic (FG) according to their contractile and metabolic profiles. Spinal motoneurons innervating the SOL and TA were identified by retrograde neuronal labelling of fluorescent dye, nuclear yellow. Oxidative enzyme activities of motoneurons in the neuron pool were examined by microspectrophotometry. When compared to 20 week old rats, a decrease in the number of FOG in the SOL and of FG in the TA was observed in 90 and 100 week old rats. Findings of atrophy were observed in SO and FOG of the SOL and in FOG and FG of the TA in 90 to 100 week old rats. Practically no changes were observed with aging in the number and oxidative capacity of motoneurons in the SOL neuron pool. However, a decrease with aging was observed in the number and oxidative capacity of motoneurons in the TA neuron pool. From these results it is indicated that exercise inhibits the age-induced atrophy of SO fibers in the SOL.

Aging↗