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Biomedical subjects

H Yoshimura

Publications and source records attributed to H Yoshimura.

At least 289 records · Page 16Linked to original sources
PubMed1991

[Enzymatic iodination of thyroglobulins obtained from patients with thyroid disease].

Iodination of the isolated thyroglobulin (Tg) by peroxidase was compared with various Tg preparations obtained from patients with thyroid diseases. For the purpose, the iodination process was observed in the incubation medium containing Tg, iodide, H2O2-generating system, and thyroid peroxidase (TPO) or lactoperoxidase (LPO). During the incubation, iodination of Tg preparations increased gradually and reached a plateau after 90 min., and 5 min. incubation with 3mIU or 14mIU of TPO, respectively. The degree of iodination level at the plateau region was different in each Tg preparation, depending on the iodine content of the original starting (native) preparation before incubation. The iodination level of cancer Tg with a very low iodine content (less than 0.1%) was low compared with the normal Tg level (obtained from normal thyroid tissue which contained about 0.4% iodine). The above findings suggest the possible existence of some structural differences of Tg in terms of the susceptibility to the iodination between the preparations of normal and diseased Tgs. As far as the immunological aspect concerned, there was no significant difference in the affinity (avidity) of Tg with polyclonal anti-Tg antibody between the native Tg and the enzymatically iodinated one. These results suggest that the changes of iodine and thyroid hormone contents of Tg by in vitro iodination, has no significant effect on the immunological property of Tg molecule.

Antigen-Antibody Reactions↗
PubMed1991

Catalytic activity of cytochrome P450 isozymes purified from rat liver in converting 11-oxo-delta 8-tetrahydrocannabinol to delta 8-tetrahydrocannabinol-11-oic acid.

Cytochrome P450 isozymes purified from rat hepatic microsomes were able to catalyse the oxidation of 11-oxo-delta 8-tetrahydrocannabinol (11-oxo-delta 8-THC) to delta 8-THC-11-oic acid in the presence of NADPH, cytochrome P450 reductase and dilauroylphosphatidylcholine. The catalytic activities (nmol/min/nmol P450) of cytochrome P450s, UT-2 (IIC11), UT-4 (IIA2), UT-5 (IIC13), PB-1, PB-2 (IIC6), PB-4 (IIB1), MC-1 (IA2), MC-5 (IA1) and IF-3 (IIA1), were 0.69, 0.08, 0.07, 0.23, 0.46, 0.02, 0.06, 0.07 and 0.34, respectively, whereas the activities of cytochrome P450s, PB-5 (IIB2) and DM (IIE1), were less than 0.02 nmol/min/nmol P450. Cytochrome P450 IIC11 showed the highest catalytic activity of the cytochromes examined. The mechanism for the oxidation of 11-oxo-delta 8-THC to delta 8-THC-11-oic acid by cytochrome P450 IIC11 was established as being an oxygenation since one atom of oxygen-18 was exclusively incorporated into the carboxylic acid formed under 18O2. The antibody raised to cytochrome P450 IIC11 inhibited by 60% the hepatic microsomal oxidation of 11-oxo-delta 8-THC to delta 8-THC-11-oic acid in male rats. These results indicate that cytochrome P450 IIC11 is a major form of the cytochrome to catalyse the oxidation of 11-oxo-delta 8-THC to delta 8-THC-11-oic acid in the hepatic microsomes of male rats and that the oxidation of aldehyde to carboxylic acid is a catalytic activity common to most isozymes of P450.

Animals↗
PubMed1991

Ethopharmacology of maternal aggression in mice: effects of diazepam and SM-3997.

The present study investigated whether there is any difference between the effects of benzodiazepine and non-benzodiazepine anxiolytics on maternal aggression in lactating mice, using an ethological technique. We used SM-3997, a 5-HT1A receptor ligand, as a non-benzodiazepine anxiolytic. Behavior towards an intruder male mouse was assessed on postpartum days 5 and 7 in female mice that had been housed alone since the end of the 4-day mating period. Acute oral administration of diazepam had a biphasic effect on the frequency of bites: 1 mg/kg diazepam significantly increased bite frequency, while 2.5 mg/kg diazepam significantly decreased it. However, 2.5 mg/kg diazepam also caused a significant decrease in locomotor activity. In contrast, SM-3997 (1, 2.5 and 5 mg/kg p.o.) significantly decreased the frequency of bites in a dose-dependent manner without causing motor dysfunction. Chronic treatment with 5 mg/kg SM-3997 significantly decreased the frequency of bites when compared with vehicle, whereas diazepam was ineffective at the doses used (0.5 and 1 mg/kg p.o.). The findings suggest that the proaggressive effect is specific to benzodiazepines, and that 5-HT1A receptors may be involved in the suppression of maternal aggression in mice.

Aggression↗
PubMed1991

Structural and functional analysis of a polyoma-related mammalian plasmid (L factor): the enhancer activity and plasmid establishment.

L factor is a unique plasmid DNA which was originally discovered in a subclone (B822) of mouse L cells at a high copy number (more than 5,000 copies/cell). The presence of L factor caused no detectable abnormalities to the plasmid-bearing cells. We determined the total DNA sequence of the L factor I (and a part of L factor II) and compared it with that of polyoma DNA. Both DNA are common to the general construction of DNA frames such as early, late and noncoding regions, suggesting the two to be closely related. On the other hand, the L factor DNA sequences differ substantially from that of polyoma in the DNA sequences corresponding to the polyoma large T antigen, capsid proteins and a portion of the enhancer region. In order to investigate the mechanism of plasmid establishment of L factor, we compared the enhancer activity, capacity of DNA replication and efficiency of plasmid establishment of L factor with those of polyoma. The results indicate that L factor enhancer activity and DNA replication capacity were considerably lower than those of polyoma, suggesting that these altered (lowered) activities associated with L factor contribute to the plasmidal establishment and stable maintenance of L factor.

Animals↗
PubMed1991

Monolayer crystallization of flagellar L-P rings by sequential addition and depletion of lipid.

The L-P ring complex is thought to be a molecular bushing that supports flagellar motor rotation at about 10,000 revolutions per minute with presumably very little friction. Structural studies of this complex have been limited because only very small amount of samples are available. Therefore devising an efficient method of crystallization was essential. The addition of a phospholipid and its subsequent slow depletion by phospholipase A2 have been used to successfully grow well-ordered monolayer crystals that extend up to about 10 micrometers. The interaction of the L-P ring complex with lipid membranes was also visualized during this process.

Cell Fractionation↗
PubMed1991

A constitutive form of guinea pig liver cytochrome P450 closely related to phenobarbital inducible P450b(e).

In order to provide evidence that a cytochrome P450 belonging to the IIB subfamily is expressed as a constitutive form in the guinea pig, we tried to purify an isozyme from liver microsomes of untreated guinea pigs by assessing its reactivity with anti-P450b antibody in the present study. One form of cytochrome P450, named P450GP-1, was obtained. The minimum molecular weight of this isozyme was estimated to be 52,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino terminal sequence up to the 33rd amino acid of P450GP-1 was determined. As expected, comparison of the amino acid sequence with those of cytochrome P450 isozymes from other species reported so far indicated that P450GP-1 was highly homologous to P450s categorized in the IIB subfamily; that is, 67% similarity to rat P450b, 82% to rabbit LM2, 76% to dog PBD-2, 70% to mouse pf 3/46, and 73% to human IIB1. On the other hand, P450GP-1 showed only low similarity, less than 41%, to other cytochrome P450s of the II subfamily and those of the I, III, and IV families. Affinity of P450GP-1 to anti-P450b immunoglobulin G was confirmed to be comparable with that of a principal antigen, P450b. Immunoblot analysis revealed that P450GP-1 in the guinea pig liver microsomes was induced by phenobarbital treatment, but the increase was not as large as in the rat. P450GP-1 efficiently catalyzed benzphetamine N-demethylation, strychnine 2-hydroxylation, and testosterone 16 beta-hydroxylation, all of which are also catalyzed by P450b. Based on these results, it was strongly suggested that the IIB-type of cytochrome P450 in guinea pigs, at least one of them, is a constitutive form which is moderately induced by phenobarbital.

Amino Acid Sequence↗
PubMed1991

Sex difference in the oxidative metabolism of delta 9-tetrahydrocannabinol in the rat.

Oxidative metabolism of delta 9-tetrahydrocannabinol (THC), one of the major components of marihuana, was studied using liver microsomes of adult male and female rats. There was no significant difference in the rates of the cannabinoid oxidation in terms of nmol per min per nmol of liver microsomal cytochrome P450 or of nmol per min per mg of microsomal protein between male and female rats. delta 9-THC was biotransformed to various metabolites including 11-hydroxy-delta 9-THC (11-OH-delta 9-THC), 8 alpha-OH-delta 9-THC, 8 alpha,11-diOH-delta 9-THC, 3'-OH-delta 9-THC by liver microsomes of male rats, while it was oxidized selectively to 11-OH-delta 9-THC by liver microsomes of female rats. After intraperitoneal administration of delta 9-THC, various metabolites were again found in the liver of the male rat, while in the female rat oxidation of the methyl group at the 9-position was a major metabolic pathway. These results demonstrate that an apparent sex-related difference exists in the oxidative metabolism of delta 9-THC in the rat.

Animals↗
PubMed1991

Oxygenation mechanism in conversion of aldehyde to carboxylic acid catalyzed by a cytochrome P-450 isozyme.

The oxygenation of an aldehyde, 11-oxo-delta 8-tetrahydrocannabinol to a carboxylic acid, delta 8-tetrahydrocannabinol-11-oic acid was catalyzed by cytochrome P-450 MUT-2 purified from hepatic microsomes of male ddN mice. The oxygenation mechanism was confirmed by the incorporation of oxygen-18 from molecular oxygen into the carboxylic acid formed. An aldehyde form but not a hydrated form of 11-oxo-delta 8-tetrahydrocannabinol may be a substrate for the cytochrome P-450. The oxygenation of aldehyde catalyzed by cytochrome P-450 might be a common metabolic reaction in biological systems, and should be considered as an additional role of cytochrome P-450 in biotransformation of endogenous compounds and xenobiotics.

Aldehydes↗
PubMed1991

Oral vaccine therapy for pneumococcal otitis media in an animal model.

We investigated whether mucosal IgA response in the middle ear cavity against Streptococcus pneumoniae type 19F is enhanced by use of enteric capsules, and whether the resulting mucosal immunity can prevent pneumococcal otitis media. Adult Hartley guinea pigs were employed. With intratympanic inoculation of 10(5) and 10(6) live S pneumoniae, the occurrence of pneumococcal otitis media significantly decreased in guinea pigs that received intraduodenal and intragastric immunization by enteric capsules. In these guinea pigs, the values of salivary IgA antibody titers against S pneumoniae were significantly greater, and histologic changes of the middle ear mucosa were also slighter than those of control guinea pigs. These findings indicate that oral vaccination by enteric capsules elicits mucosal IgA responses, as well as intraduodenal immunization, to prevent pneumococcal otitis media. These results suggest the possibility of clinical application of oral vaccination by enteric capsules for the prevention of middle ear infection.

Administration, Oral↗
PubMed1991

Fluorometric assay of microsomal aldehyde oxygenase activity: 9-anthraldehyde as a substrate.

With regard to hepatic microsomal oxidation of 9-anthraldehyde (9-AA), a fluorometric method for determination of 9-anthracene carboxylic acid (9-ACA) is described. 9-AA was incubated with hepatic microsomes prepared from male ddN mice. 9-ACA formed was fluorometrically (excitation and emission wavelengths of 255 and 458 nm, respectively) quantitated after the separation from 9-AA by an alkali extraction and ethyl acetate reextraction. Hepatic microsomes less than 0.1 mg protein were enough to assay the microsomal aldehyde oxidation. The enzyme in the microsomes that catalyzes the oxidation of 9-AA to 9-ACA has been characterized by this method.

Aldehyde Oxidoreductases↗
PubMed1991

Analysis and pharmacotoxicity of feruloyltyramine as a new constituent and p-coumaroyltyramine in Cannabis sativa L.

Feruloyltyramine (FT), a new amide compound, together with p-coumaroyltyramine (p-CT) was isolated and identified in ethanol extract of cannabis seeds. FT and p-CT were also detected in the roots, leaves and resin of Cannabis sativa L. The intracerebroventricular injection of these amides caused hypothermia and motor incoordination in mice, and the maximal effects were caused 160 to 240 min after the injection. Furthermore, p-CT also exhibited cataleptogenic effect in mice, although FT did not show any effect. These results suggest that these amide compounds may be responsible for some pharmacotoxicity of marihuana.

Animals↗
PubMed1991

Cannabielsoin as a new metabolite of cannabidiol in mammals.

Cannabielsoin (CBE) was identified as a novel metabolite of cannabidiol (CBD) in the guinea pig in vivo and in vitro. Its formation by liver microsomes of guinea pigs needed NADPH and molecular oxygen, and was inhibited with SKF 525-A, metyrapone and alpha-naphthoflavone, indicating participation of cytochrome P-450 (P-450). The CBE-forming activity was highest in guinea pigs, followed by mice, rabbits and rats. In the rat, sex difference was found in the CBE formation (male greater than female). CBD monomethylether (CBDM) was also biotransformed to CBE monomethylether (CBEM) in the guinea pig in vivo and in vitro. When CBD dimethylether (CBDD) was employed as substrate, 1S,2R-epoxy-CBDD was identified. The results suggest that CBD and CBDM are biotransformed by P-450 to CBE-type metabolites via 1S,2R-epoxides. In pharmacological studies using mice, CBDD and 1S,2R-epoxy-CBD-2',6'-diacetate produced hypothermia, and CBD, CBDM and CBEM prolonged pentobarbital-induced sleep. Moreover, 1S,2R-epoxy-CBD-2',6'-diacetate was examined in the Ames test, but had no mutagenicity.

Animals↗
PubMed1991

Ethopharmacology of copulatory disorder induced by chronic social conflict in male mice.

The present study was designed to investigate how the experience of fighting affects copulatory behavior in male mice and also to determine the effect of naloxone on the interaction between social conflict and copulatory disorder. To generate intraspecific fighting a resident-intruder paradigm was employed. Agonistic confrontations were terminated after 10 or 20 attack bites, and were repeated for 5 consecutive days. Twenty-four hours after the last confrontation test, both resident and intruder mice were tested with estrus females for 10 min. Compared to the control group without agonistic confrontation, intruder mice that had been attacked repeatedly showed a significant reduction of copulatory behavior. In contrast, attacking resident mice showed a significant increase in copulatory behavior. Pretreatment with naloxone (1 and 3 mg/kg, IP) prior to daily fighting failed to antagonize defeat-induced copulatory disorder. It would, therefore, appear that endogenous opioid mechanism may not participate in this phenomenon.

Aggression↗
PubMed1991

Evaluation of the antimicrobial activity of methylglyoxal bis(guanylhydrazone) analogues, the inhibitors for polyamine biosynthetic pathway.

Metabolic and antiproliferative effects of methylglyoxal bis(butylamidinohydrazone) (MGBB) and methylglyoxal bis(cyclopentylamidinohydrazone) (MGBCP), inhibitors for polyamine biosynthetic pathway, on Escherichia coli, Shigella sonnei, Aeromonas sobria, Aeromonas hydrophila and Vibrio cholerae were investigated. MGBB at the concentration of 100 mumol/l depleted intracellular putrescine and spermidine concentrations of E. coli to 25 and 20% of the controls, respectively, while MGBCP depressed their concentrations to 38 and 24%, respectively. In these polyamine-depleted E. coli cells the syntheses of RNA, DNA and protein decreased to 13, 54 and 29% of the control, respectively, with MGBB and to 23, 71 and 55%, respectively, with MGBCP. The minimum inhibitory concentrations (MIC) of MGBB for the growth of A. sobria, E. coli, A. hydrophila, V. cholerae and Sh. sonnei were estimated to be 50, 160, 240, 285 and 320 mumol/l, respectively, whereas those of MGBCP were slightly higher for respective bacteria.

Bacteria↗
PubMed1991

Bactericidal effect of 15-deoxyspergualin on Staphylococcus aureus.

15-Deoxyspergualin (DSG), an immunosuppressive agent used in organ transplantation, exerts metabolic and antiproliferative effects on methicillin- and gentamicin-resistant Staphylococcus aureus and other bacteria (e.g. Sarcina lutea, Bacillus subtilis, Shigella sonnei, Salmonella typhi and Citrobacter freundii). DSG, at the concentration of 20 mg/l, depleted intracellular putrescine and spermidine in S. aureus to 43 and 40% of the controls, respectively. In these polyamine-depleted S. aureus cells, the synthesis of protein, DNA and RNA was decreased to 20, 85, 78% of the controls. The minimal inhibitory concentrations (MIC) of DSG for growth of S. aureus, S. lutea, B. subtilis, S. sonnei, S. typhi and C. freundii were 17, 13, 7, 15, 4, and 29 mg/l, respectively.

Bacteria↗
PubMed1991

Site-selective oxidation of strychnine by phenobarbital inducible cytochrome P-450.

The metabolism of strychnine was studied using liver microsomes of rats treated with phenobarbital or 3-methylcholanthrene (MC). The phenobarbital-treatment resulted in 7.9-fold and 4.8-fold increases in 2-hydroxylation and N-oxidation of strychnine, respectively. The formation of 16-hydroxystrychnine, strychnine 21,22-epoxide and 22-hydroxystrychnine was induced about 2-fold. MC-treatment resulted in only 1.4-fold induction of each oxidation activity. In addition, strychnine 2-hydroxylation activity was markedly induced in liver microsomes of phenobarbital-treated mice, guinea pigs, rabbits and dogs (2.5-10.5-fold). Induction of N-oxidation activity was also higher than that of the three other oxidation activities. A reconstituted system of strychnine metabolism using cytochrome P-450 isozymes, P-450I (P450IIB1) and P-450II (P450IIB2), purified from liver microsomes of phenobarbital-treated rats showed significantly high and selective activities towards 2-hydroxylation and N-oxidation of strychnine. This characteristic metabolism of strychnine appears to occur in common with interspecies P450IIB gene subfamily. The pH Optima of 2-hydroxylation and N-oxidation of strychnine were between 8.4 and 8.6 in the microsomes of phenobarbital-treated rats, while that of N-demethylation of benzphetamine, a typical substrate of phenobarbital-inducible cytochrome P-450, was between 7.4 and 7.6. In a reconstituted system with P-450I, strychnine oxidation was little affected by pH change, while benzphetamine N-demethylation activity was decreased in the alkaline side. Among several oxidations tested, only ethylmorphine N-demethylation underwent the same pH effect as did strychnine oxidation with the microsomes and reconstituted system.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Species difference of site-selective glucuronidation of morphine.

Species difference in glucuronidation of morphine was studied using mice, rats, guinea pigs and rabbits in vivo and in vitro. Morphine-3-glucuronide (M-3-G) and morphine-6-glucuronide (M-6-G) were determined by high-performance liquid chromatography. M-3-G was the major urinary metabolite of morphine in all these animal species. However, a remarkable species difference was observed in the urinary excretion of the M-6-G. Excretion ratios of the M-3-G to M-6-G were approximately 4:1 and 50:1 in guinea pigs and rabbits, respectively. The urinary excretion of M-6-G in mice and rats was too small to be determined. On the other hand, the ratios of uridine diphosphate-glucuronyltransferase (UDPGT) activities toward 3- and 6-hydroxyl groups of morphine in liver microsomes of mice, rats, guinea pigs and rabbits were approximately 300:1, 90:1, 4:1 and 40:1, respectively. Ratios of two morphine UDPGT activities in the liver microsomes of guinea pigs and rabbits, thus, reflected those of urinary excretion of morphine glucuronides.

Animals↗