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Biomedical subjects

T Yakushiji

Publications and source records attributed to T Yakushiji.

At least 19 recordsLinked to original sources

Comparison of the vasodilator effects of thiopentone and pentobarbitone.

The aim of this study was to elucidate the vasodilator mechanisms of barbiturates. In helical strips of dog mesenteric artery, the effects of pretreatment with thiopentone and pentobarbitone on the contractions induced by KCl (2.0 x 10(-2) M) and norepinephrine (10(-5) M) in normal bathing fluid, and those induced by norepinephrine and caffeine (2.5 x 10(-2) M) in Ca(++)-free fluid were tested, and their effects on caffeine-induced contractions in skinned strips were also examined. Thiopentone, at concentrations over 10(-4) M, inhibited the KCl-induced contractions in normal bathing fluid and those induced by caffeine in Ca(++)-free fluid and, at concentrations over 3 x 10(-4) M, inhibited norepinephrine-induced contractions in normal and Ca(++)-free bathing fluids significantly. Pentobarbitone, at concentrations over 3 x 10(-4) M, inhibited KCl- and norepinephrine-induced contractions in normal bathing fluid significantly, whereas contractions in Ca(++)-free fluid induced by norepinephrine and caffeine were inhibited only by a high concentration (10(-3) M) of pentobarbitone. Caffeine-induced contractions of chemically skinned fibres were more susceptible to inhibition by thiopentone than by pentobarbitone. These results suggest that the vasodilator effect of thiopentone is mediated via its intracellular inhibitory effect and that, in contrast, the vasodilator effect of pentobarbitone can be attributed mainly to its Ca(++)-channel blocking action.

Animals

Serological and biological characteristics of "Streptococcus milleri" isolates from systemic purulent infections.

Ninety-one Streptococcus milleri strains isolated from various systemic purulent lesions of 68 patients were examined by physiological and serological tests. Most strains formed a smooth colony (66 strains), did not form spontaneous aggregation of cells in BHI broth culture (79), were non-beta-haemolytic (alpha-35 or non-41), and belonged to biotype Ia (49) or Ib (34) and to API taxa S. milleri I (41) or II (38). Almost all of the beta-haemolytic strains as well as two-fifths of the non-beta-haemolytic belonged to API taxon I; strains of API taxa II and III were non-beta-haemolytic and non-haemolytic, respectively. Two-fifths (38) of the isolates belonged to one of eight serotypes, a-g and k, and more than half (47) to Lancefield groups A, C, F or G, the most frequent being type b (19) and group F (33). Fifteen strains carried simultaneously type a/group A, b/C, c/C, e/G, f/F or k/G antigens. Nineteen were neither typable nor groupable. All the 38 serotypable isolates were non-beta-haemolytic and not members of API taxon III, and were serologically and physiologically similar to oral S. milleri. The isolates from various infected sites--sputum, thorax, abdomen, urogenitalia, skin, eye and dental--exhibited distinct combinations of biological and serological properties. These results suggest that serotyping, haemolytic properties and API taxon, and their combinations, would be useful methods to trace oral S. milleri in systemic infections.

Abdomen

Restricted distribution of Streptococcus milleri carbohydrate type antigens amongst other viridans streptococci.

The distribution of oral Streptococcus milleri carbohydrate type antigens in other viridans streptococcus species was examined. Rantz-Randall extracts of cells of the test strains grown in broth containing glucose were allowed to react with typing or grouping antisera for S. milleri serotypes a-k, or Lancefield groups A-G and K. Of 93 strains comprising more than 12 streptococcal species that included S. mutans and S. sanguis complexes, only 15 S. salivarius strains and one S. mitis strain were immunologically related to S. milleri serotype f. Unlike S. milleri strains, S. salivarius type f strains belonged to Lancefield group K, whereas the S. mitis strain was closely related to S. milleri serotype f but did not react with any of the Lancefield grouping antisera tested. Results suggest that oral S. milleri strains can be distinguished serologically from other oral viridans streptococci and that the typing antisera used in our researches might differentiate S. milleri isolates from the mouth from those associated with systemic purulent infections.

Antigens, Bacterial

Non-competitive inhibition of GABAA responses by a new class of quinolones and non-steroidal anti-inflammatories in dissociated frog sensory neurones.

1. The interaction of a new class of quinolone antimicrobials (new quinolones) and non-steroidal anti-inflammatory agents (NSAIDs) with the GABAA receptor-Cl- channel complex was investigated in frog sensory neurones by use of the internal perfusion and 'concentration clamp' techniques. 2. The new quinolones and the NSAIDs (both 10(-6)-10(-5) M) had little effect on the GABA-induced chloride current (ICI) when applied separately. At a concentration of 10(-4) M the new quinolones, and to a lesser degree the NSAIDs, produced some suppression of the GABA response. 3. The co-administration of new quinolones and some NSAIDs (10(-6)-10(-14) M) resulted in a marked suppression of the GABA response. The size of this inhibition was dependent on the concentration of either the new quinolone or the NSAID tested. The inhibitory potency of new quinolones in combination with 4-biphenylacetic acid (BPAA) was in rank order norfloxacin (NFLX) much greater than enoxacin (ENX) greater than ciprofloxancin (CPFX) much greater than ofloxacin (OFLX), and that of NSAIDs in combination with ENX was BPAA much greater than indomethacin = ketoprofen greater than naproxen greater than ibuprofen greater than pranoprofen. Diclofenac, piroxicam and acetaminophen did not affect GABA responses in the presence of ENX. 4. In the presence of ENX or BPAA, there was a small shift to the right of the concentration-response curve for GABA without any effect on the maximum response. However, the co-administration of these drugs suppressed the maximum of the GABA concentration-response curve, indicating a non-competitive inhibition, for which no voltage-dependency was observed.5. Simultaneous administration of ENX and BPAA also suppressed pentobarbitone (PB)-gated Icl. On the other hand, both PB and phenobarbitone reversed the inhibition of GABA-induced Ic, by coadministration of ENX and BPAA.6. The effect on GABAA responses of co-administration of new quinolones and NSAIDs was not via an interaction with benzodiazepine receptors coupled to the GABAA receptor, since this effect was not reversed by Rol5-1788 or diazepam.7. It is concluded that the co-administration of new quinolones and some of the NSAIDs inhibit GABAergic transmission, and could result in convulsions.

Animals

Blockade of 5-HT3 receptor-mediated currents in dissociated frog sensory neurones by benzoxazine derivative, Y-25130.

1. The effect of Y-25130, ((+-)-N-(1-azabicyclo[2.2.2]oct-3-yl)-6-chloro-4-methyl-3-oxo-3,4-dih ydr o- 2H-1,4-benzoxazine-8-carboxamide hydrochloride), a high affinity 5-hydroxytryptamine3 (5-HT3) receptor ligand, was examined on the 5-HT-induced response in dissociated frog dorsal root ganglion (DRG) neurones by use of the extremely rapid concentration-jump ('concentration-clamp') and the conventional whole-cell patch-clamp techniques. 2. 5-HT induced a rapid transient inward current associated with an increase in membrane conductance at a holding potential of -70 mV. The current amplitude increased sigmoidally as 5-HT concentration increased. The half-maximum value (Ka) and the Hill coefficient estimated from the concentration-response curve were 1.7 x 10(-5) M and 1.7, respectively. 3. The current-voltage (I-V) relationship of 5-HT-induced current (I5-HT) showed inward rectification at potentials more positive than -40 mV. The reversal potential (E5-HT) was -11 mV. The E5-HT value was unaffected by total replacement of intracellular K+ by Cs+, indicating that the 5-HT-gated channels might be large cation channels. 4. Both the activation and inactivation phases of I5-HT were single exponentials. The time constants of activation and inactivation (tau a and tau i) decreased with increasing 5-HT concentration. 5. The 5-HT response was mimicked by a selective 5-HT3 receptor agonist, 2-methyl-5-HT, but the maximum response induced was approximately 25% that of 5-HT. The 5-HT response was reversibly antagonized by the 5-HT3 receptor antagonists, ICS 205-930, metoclopramide and Y-25130, but not by a 5-HTIA receptor antagonist, spiperone, and a 5-HT2 receptor antagonist, ketanserin. The half-inhibition concentrations (IC50) were 4.9 x 10-10 M for Y-25130, 4.8 x 10-10 M for ICS 205-930 and 8.6 x 10-9 M for metoclopramide.6. Y-25130 (5 x 10-10 M) caused a rightward shift of the concentration-response curve for 5-HT while decreasing the maximum response.7. The results suggest that Y-25130 is a potent antagonist of the 5-HT3 receptor-channel complex.

Animals

Glycemic response and fiber content of some foods.

The proposed mechanisms for the action of dietary fiber (DF) on the glycemic response suggest a nonlinear relationship between glycemic index (GI) and DF content. This relationship was analyzed by using the newly reported total DF (TDF) values, assuming a nonlinear regression curve. The empirical equation obtained was Y = 19.9X-0.322. Similar regression curves were also obtained for soluble DF (SDF) and insoluble DF (IDF). The two regression curves indicated that the correlation coefficient between observed GI and GI calculated from the IDF regression curve (-0.781) was virtually identical to that for SDF (-0.780), but a given content (eg, 5% vs available carbohydrate) of SDF gave a lower GI (39) than did IDF (48). This stronger dependency of GI on SDF suggests a major function of SDF in the TDF hypoglycemic effect. From the regression curve of GI vs TDF, we propose a supplemental GI to predict the glycemic response to foods with no published GI.

Blood Glucose

Carbohydrate antigen of serotype g "Streptococcus milleri": immunochemical characterization.

Cell wall carbohydrate antigen of the serotype g "Streptococcus milleri" was extracted with cold trichloroacetic acid from purified cell walls of the type strain K1K. The extracts were then purified by a DEAE-Sephadex A-25 column, followed by a Sephadex G-100 column. The immunoelectrophoresis revealed that the serotype g carbohydrate antigen preparation displayed a single precipitin band against the crude anti-K1K serum. The purified type g antigen consisted of rhamnose, galactose, glucose and galactosamine in a ratio of 1.3:3.8:1.0:2.5. The quantitative precipitin inhibition test with various haptens indicated that galactosamine is a major immunodeterminant of the type g-specific antigen.

Cell Wall

Lactose-sensitive and -insensitive cell surface interactions of oral Streptococcus milleri strains and actinomyces.

Of 158 oral Streptococcus milleri strains, 46 exhibited cellular coaggregation with the reagent strains of the actinomyces coaggregation groups A, B, and/or E. All but 1 of the 33 serotype b, e, f/F, and k/G strains belonged to streptococcus coaggregation group 2, and only 14 strains of limited seroclasses (g, i, Lancefield group F, or untypeable) appeared to be members of group 5, 3, or 4 (10, 3, and 1 strain, respectively). Thus, S. milleri infrequently exhibits lactose-inhibitable coaggregation with actinomyces.

Actinomyces

Adherence of oral "Streptococcus milleri" cells to surfaces in broth cultures.

Cells of representative strains of oral "Streptococcus milleri" firmly adhered to glass surfaces when grown in glucose broths as well as in fructose and sucrose broths. Cellular adherence occurred on saliva-coated glass surfaces as well as uncoated surfaces, but the cells of only a few (6 of 69) tested adherent strains agglutinated upon the addition of human whole-saliva supernatant. Thus, the firm adherence of growing "S. milleri" cells is independent of sucrose and salivary macromolecules. Two macroscopic forms of cell adherence were observed: one was a coarse-granule type and one was a tiny-granule type covered with a thin homogeneous membranous material. Scanning electron microscopy revealed that, in both types, a stringy substance interconnected the cells and connected the cells to glass surfaces, with amorphous flocklike materials present in the intercellular space. Of the 154 tested strains, 128 were adherent, of which 90 were firmly and extensively (+2 or +3) adherent. Most strains of the serovars f/F, g, h, i, j, g-, and gj- were extensively adherent. Generally, cells of the more strongly adherent strains were more hydrophobic and spontaneously aggregating. However, there also existed a few hydrophilic and nonaggregating but extensively adherent strains. The presence of trypsin or pronase completely inhibited the adherence of some strains (mostly nonaggregating) but did not at all inhibit that of others (mostly aggregating). The adhering groups did not significantly differ in their cell surface hydrophobicities. Therefore, at least two factors, one proteinaceous and one nonproteinaceous, appear to be principally involved in the firm surface adherence of "S. milleri."

Bacterial Adhesion

Quinolones and fenbufen interact with GABAA receptor in dissociated hippocampal cells of rat.

1. Interaction of quinolone antibiotics and the anti-inflammatory agent fenbufen with the gamma-aminobutyric acid-A (GABAA) receptor-chloride channel complex in pyramidal neurons freshly dissociated from the hippocampal CA1 region of the rats was investigated in whole-cell mode, using the patch-clamp technique under voltage-clamp conditions. 2. Quinolones in clinical doses had no effects on the GABA-gated Cl- current (ICl) but slightly suppressed the response at concentrations greater than 10(-5) M. A metabolite of fenbufen, 4-biphenylacetic acid (BPA), also had little effect on the GABA response at therapeutic concentrations. 3. Coadministration of one of quinolones and BPA suppressed the GABA-gated ICl with increase in each of them in a concentration-dependent manner, and there was a parallel shift of the concentration-response curve for GABA to the right but with no effect on the maximum response, thereby indicating a competitive antagonism. The inhibitory potency of antibiotics in combination with BPA was in the order of norfloxacin much greater than enoxacin greater than cyprofloxacin greater than pipemidic acid much greater than ofloxacin greater than cinoxacin = piromidic acid = nalidixic acid = 0. 4. Norfloxacin and BPA, administered simultaneously, also strongly suppressed pentobarbital sodium (PB)-gated ICl, but they did not act on benzodiazepine (BZP) receptors. 5. Both GABA- and PB-induced ICls reversed at the Cl- equilibrium potential (ECl). In the presence of BPA, the quinolone-induced inhibition of GABA-gated ICls showed no voltage dependence. 6. It was concluded that, in the presence of an anti-inflammatory agent, the quinolone antibiotics decrease the affinity of GABAA receptors, the result being induction of epileptogenic neurotoxicities.

4-Quinolones

Studies on the synthesis of condensed pyridazine derivatives. IV. Synthesis and anxiolytic activity of 2-aryl-5,6-dihydro-(1)benzothiepino[5,4- c]pyridazin-3(2H)-ones and related compound.

A series of 2-aryl-5,6-dihydro-(1)benzothiepino[5,4-c]pyridazin-3(2H)- ones and related compounds were synthesized and evaluated for their ability to displace 3H-diazepam from rat brain membranes in vitro, and to prevent bicuculline induced convulsions in mice in vivo. Compounds with a 4'-methoxyphenyl (36) or 4'-chlorophenyl group (37, 39--42) as 2-aryl substituents showed prominent activities in both the in vitro and in vivo tests. Among them, 2-(4'-chlorophenyl)-5,6-dihydro- (37) and 2-(4'-chlorophenyl)-5,6-dihydro-10-fluoro-(1)benzothiepino[5,4-c]+ ++pyridazin- 3(2H)-one 7-oxides (41) showed activity twice as potent as diazepam in an anticonflict test (Vogel type, rats) while exhibiting less muscle relaxation (rotarod test, mice) and augmentation of gamma-aminobutyric acid-induced chloride current (Icl) in isolated frog sensory neurones than diazepam. Compound 37 (Y-23684) was selected from this series as a candidate for further development. The structure-activity relationships are discussed.

Animals

Immunochemical characterization of type i carbohydrate antigen of "Streptococcus milleri" (Streptococcus anginosus).

Type-specific carbohydrate antigen of the serotype i "Streptococcus milleri" was extracted with trichloroacetic acid from purified cell walls of the type strain K39K. The extracts were then purified by chromatography on DEAE-Sephadex A-25 and Sephadex G-100 columns. The purified serotype i carbohydrate antigen produced a single precipitin band against its homologous type-specific antiserum, which fused with the band produced by the autoclaved extract of the type strain cells. The serotype i antigen was a polysaccharide composed of rhamnose, galactose and glucose in a molar ratio of 1.6:6.8:1.0. The quantitative precipitin inhibition test with various haptenic sugars showed that galactose as well as lactose produced the greatest inhibition, which suggested that a galactose in terminal beta linkage is the immunodeterminant of the serotype i-specific antigen. Galactose was detected in the autoclaved extracts from cells of all the 15 serotype i strains tested.

Antigens, Bacterial

Comparison of the direct effects of halothane and isoflurane on large and small coronary arteries isolated from dogs.

Relaxant responses to halothane and isoflurane were compared in helical strips of dog epicardial coronary arteries of different sizes: proximal large coronary arteries with outside diameters (OD) larger than 2.5 mm and distal small arteries with 0.7-0.9 mm OD. Responses to pharmacologic vasodilators, including nitroglycerin (NTG) and adenosine, were also studied for comparison. The relaxation induced by halothane in concentrations of 0.8-2.3% and by NTG (10(-9)-10(-5) M) was greater in proximal large coronary arteries than in distal small ones contracted with 20 mM KCl. In contrast, the relaxation by isoflurane (1.2-3.5%) and by adenosine (10(-8)-10(-4) M) was greater in small coronary arteries than in large ones. These results suggest that isoflurane is, like adenosine, preferentially a small artery dilator.

Adenosine

Cellular coaggregation of oral Streptococcus milleri with actinomyces.

Oral isolates of Streptococcus milleri were examined for their ability to coaggregate with actinomyces. Of the 68 S. milleri strains tested, including 3 reference strains, 40 strains coaggregated with Actinomyces naeslundii WVU45 (actinomyces coaggregation group B) and 36 strains coaggregated with Actinomyces viscosus T14V (actinomyces coaggregation group A). All S. milleri strains of serotypes b (4 strains), e (2 strains), and f (24 strains) coaggregated with both of the actinomyces. The coaggregation reactions between the S. milleri cells and A. naeslundii WVU45 cells were optimal at about pH 7.0 and were Ca2+ or Mg2+ dependent, but they were not inhibited by the presence of simple sugars or amino sugars, including lactose (up to 0.5 M). Treatment of the S. milleri cells with heat (100 degrees C, 3 min) or proteases (trypsin, 1.0 mg/ml; pronase, 0.25 mg/ml; 37 degrees C; 3 h) and of the actinomyces cells with periodate (0.01 M, 4 degrees C, 16 h) destroyed their coaggregating abilities. The coaggregations between cells of the S. milleri strains, we well as cells of the Streptococcus sanguis H1 (reference strain for streptococcus coaggregation group 2) and the actinomyces strains (WVU45 and T14V), were inhibited by AFH1 (a carbohydrate receptor on T14V cells for a lectin on H1 cells). These interactions were also inhibited by anti-AFH1 immunoglobulin G (IgG) and by anti-b, anti-e, and anti-f S. milleri IgG or anti-f IgG Fab fragments. These results suggest that S. milleri, at least strains of serotypes b, e, and f, belongs to streptococcus coaggregation group 2.

Actinomyces

Effects of thiobarbiturates on smooth muscle reactivity in isolated aortas from spontaneously hypertensive rats.

The direct effects of thiobarbiturates on helical strips of aortas from spontaneously hypertensive (SH) rats were compared with those from Wistar-Kyoto (WKY) rats. At 5-6 wk of age, the arterial pressure of SH and WKY rats did not differ, and the effects of thiobarbiturates on aortic strips from SH and WKY rats were similar. By contrast, at the age of 10-12 or 20-21 wk arterial pressure was higher in SH than in WKY rats, and responses to thiobarbiturates differed in aortic strips from SH and WKY rats: contractile responses were greater in WKY than in SH rats, and relaxing effects were greater in SH than in WKY rats. Responses to sodium nitroprusside did not differ in the aortas of SH and WKY rats, but the effects of nifedipine were greater in strips from SH rats than from WKY rats at the age of 10-12 wk. Ca2(+)-induced contractions of strips exposed to Ca2(+)-free media and depolarized by high K+ were inhibited by treatment with thiamylal; the inhibition was greater in SH than in WKY rats. The increase in smooth-muscle relaxation induced by thiobarbiturates in strips from SH rats may be due to increased sensitivity to the Ca2(+)-channel blocking action of thiobarbiturates.

Animals

Inhibition of dental plaque formation by mouthwash containing an endo-alpha-1, 3 glucanase.

The effect of mouthrinsing with the purified endo-alpha-1, 3 glucanase (mutanase) from a Pseudomonas sp. strain on the formation of dental plaque was investigated. Twenty-two college students participated in the clinical trial. After the labial and lingual surfaces of the 12 front teeth had been subjected to a thorough prophylaxis, the participants were instructed to cease all active oral hygiene practices and to rinse their mouths with either the mutanase or placebo mouthwashes for one minute, up to four times daily, in all, eight times for three days. No restriction regarding meals was given during the experimental period. After a 5-day recess, a second experiment was performed using the same procedures, except the placebo and active mouthwashes were switched. The amount of dental plaque formed on the front teeth was scored according to the criteria of Quigley and Hein. Mean plaque scores were significantly lower for the mutanase (p less than 0.05-0.001) than for the placebo group. In the intraparticipant comparison, most plaque scores were also significantly reduced in general (p less than 0.05-0.0010) by rinsing with the mutanase mouthwash. These data indicate that mutanase is able to suppress the accumulation of dental plaque in humans.

Adult

Influences of external Ca2+ on the GABA-induced chloride current and the efficacy of diazepam in internally perfused frog sensory neurons.

The effects of extracellular Ca2+ on the gamma-aminobutyric acid (GABA)-induced Cl- current and the efficacy of diazepam in the facilitation of GABA response were studied in frog isolated sensory neurons, using a 'concentration clamp' technique which combines a suction pipette (internal perfusion and voltage clamp) and a rapid drug application system. When nominal Ca2+-free external solution was changed to the solution containing 2 mM Ca2+, the response elicited by 1 x 10(-5) M GABA was reduced by about 40% of the control obtained in nominal Ca2+-free solution. The dose-response curve for GABA was shifted to the right without affecting the maximum response. It can be suggested that the application of external Ca2+ modulates the affinity of the GABA receptor to its agonist, GABA. Diazepam at the concentration of 3 x 10(-6) M shifted the dose-response curve for GABA to the left without changing the maximum response with or without external Ca2+. However, the augmentatory action of diazepam on the GABA response was reduced in the presence of external Ca2+. Possible mechanisms for inhibitory action of external Ca2+ on the GABA-gated response and the reduced efficacy of diazepam are discussed.

Animals

Production and partial characterization of the extracellular polysaccharides from oral Streptococcus salivarius.

The production of polysaccharides from sucrose by extracellular enzymes from oral Streptococcus salivarius isolates and the physico-chemical properties of water-insoluble products (IPs) were investigated. Extracellular enzymes from all the 18 strains tested produced insoluble alpha-D-glucans (IGs) as well as soluble beta-D-fructans, and formed adhering deposits on glass. Generally, the IPs (mostly IGs) of S. salivarius strains differed from the S. sobrinus IPs by (a) containing significant proportions of alpha-D-(1----4)-, in addition to alpha-D-(1----3)- and alpha-D-(1----6)-glucosyl linkages, and much higher proportions of alpha-D-(1----3) than alpha-D-(1----6) linkages, (b) being more susceptible to hydrolysis by mutanase than by dextranase, (c) possessing low or no streptococcal cell-agglutinating ability, and (d) showing weaker adhesion to a glass surface. The degree of the polysaccharide adherence differed greatly among the S. salivarius strains and, therefore, they were divided into three groups of adherence producers; heavy, moderate, and slight. The IPs of the three groups contained, generally in descending order, a higher proportion of higher-molecular-weight fractions, and consisted of higher proportions of IG containing higher proportions of -(1----6)-alpha-D and -(1----4)-alpha-D glucosyl linkages and (1----3,6) branches, but showed higher susceptibility to hydrolysis by mutanase as well as dextranase. Thus, the production and the properties of extracellular insoluble alpha-D-glucans from sucrose differ considerably between oral S. salivarius and cariogenic S. sobrinus.

Agglutination