Antinociceptive activity of deltorphin analogs in the formalin test.

Two deltorphin (DLT: Tyr-D-Met-Phe-His-Leu-Met-Asp-NH2) analogs, [N alpha-n-butyl-Gly6]DLT ([nBuG6]DLT) and [N alpha-iso-butyl-Gly6]DLT ([isoBuG6]DLT), were examined for their antinociceptive activities by a formalin test in mice after subcutaneous (s.c.) injection.

[A simultaneous determination of evodiamine and rutaecarpine in oriental pharmaceutical decoctions containing Evodia fruit by ion-pair high-performance liquid chromatography].

A simple and precise method was established for the simultaneous determination of evodiamine and rutaecarpine in oriental pharmaceutical decoctions containing Evodia Fruit using high-performance liquid chromatography with sodium dodecyl sulfate (SDS) as an ion-pair reagent. Evodiamine and rutaecarpine were eluted within 60 min without interference from co-existing components using an ODS column and a mixture of water-methanol-acetonitrile-SDS-phosphoric acid (600:330:70:5:0.01, v/v/v/w/v, pH 5.

Investigation of phagocyte-inducible factor in tumor-implanted mice.

The number of spleen Mac-1-positive cells was markedly increased in methylcholanthrene-induced Meth-A fibrosarcoma (Meth-A)-implanted mice. When the sera of Meth-A-implanted mice were added to normal bone marrow cells, granulocyte and macrophage colony stimulating factor (GM-CSF)-dependent growth of bone marrow cells was significantly enhanced. Analysis of the Meth-A-implanted mice sera showed that the levels of serum transferrin were markedly elevated.

Effects of the histamine H3 agonist (R)-alpha-methylhistamine and the antagonist thioperamide in vitro on monoamine oxidase activity in the rat brain.

The effects of an H3 agonist, (R)-alpha-methylhistamine (alpha-MeHA), and an H3 antagonist, thioperamide, on monoamine oxidase (MAO) activity in rat hypothalamus were studied in vitro. Thioperamide was more potent in inhibiting MAO-B than MAO-A activity; MAO-B activity in rat hypothalamic homogenates was competitively inhibited by thioperamide with a Ki value of 175 micronM. From this in vitro experiment, the conversion of N-telemethylhistamine to N-tele-methylimidazoleacetic acid may be inhibited by thioperamide, suggesting that thioperamide may affect the regulation of histamine metabolism within histaminergic neurons.

Involvement of opioid receptors in the antinociception produced by intracerebroventricularly administered spantide in mice.

The antinociceptive effect of intracerebroventricularly (i.c.v.

Immunohistochemical determination of rat spinal cord substance P, and antinociceptive effect during development of thiamine deficiency.

During 30 days of thiamine deficiency (TD) feeding, the rat antinociceptive effect (pain threshold) to noxious heat stimulation was significantly increased in proportion to the decrease substance P (SP) fluorescent intensity in the spinal cord. Only a single injection of thiamine HCl (0.5 mg/kg, s.

Detection of beta-1,2-mannosyltransferase in Candida albicans cells.

A particulate insoluble fraction from Candida albicans J-1012 (serotype A) strain cells was obtained as the residue after extracting a 105,000 x g pellet of cell homogenate with 1% Triton X-100. Incubation of this fraction with a mannopentaose, Man beta 1-->2Man alpha 1-->(2Man alpha 1-->)(2)2Man (alpha beta Man5), in the presence of GDP-mannose followed by high performance liquid chromatography showed the formation of a mannohexaose. Analysis of the product by 1H NMR indicates that alpha beta Man5 was changed to Man beta 1-->2Man beta 1-->2Man alpha 1-->(2Man alpha 1-->)2 2Man (alpha beta Man6).

Inhibition of doxorubicin-induced cell death in vitro and in vivo by cycloheximide.

We studied the effects of the protein synthesis inhibitor, cycloheximide (CH), on cell killing by doxorubicin (DOX) in vitro and in vivo. At the concentration of CH used (1 microgram/ml) the cytotoxicity of DOX was reduced in cultured P388 leukemia cells. An analysis of the DNA histogram obtained by flow cytometry showed that DOX exerts its growth-inhibitory effect by blocking the cell cycle at the G2/M phase in P388 cells.

Novel deltorphin heptapeptide analogs with potent delta agonist, delta antagonist, or mixed mu antagonist/delta agonist properties.

A series of deltorphin (DLT: Tyr-D-Met-Phe-His-Leu-Met-Asp-NH2) analogs in which Leu5 and/or Met6 were mainly replaced by t-Leu(Tle) and/or N alpha-alkylated glycine were synthesized and examined for their receptor binding properties and in vitro bioactivities by guinea pig ileum (GPI) and mouse vas deferens (MVD) assays. [Tle5]DLT(2) showed a dramatic decrease in the MVD potency when compared to the parent peptide and was found to have a potent delta receptor antagonist activity against various delta agonists with Ke values of 16-311 nM. Interestingly, the antagonist potency of 2 against DPDPE as agonist was 20-fold weaker than that against deltorphins or Leu-enkephalin.

Effect of glutathione depletion by buthionine sulfoximine on doxorubicin toxicity in mice.

The role of the glutathione (GSH) system in vivo or in drug resistance has received much attention, since GSH is a major component of the cellular detoxification system. We Studied the effect of GSH depletion by buthionine sulfoximine (BSO), a potent inhibitor of gamma-glutamylcysteine synthetase, on doxorubicin (DOX) toxicity in mice. The administration of BSO (30 mM in drinking water for 5 days) significantly decreased the tissue GSH.

Regio- and stereo-selective oxidation of phenylbutane by rat liver.

Regio- and stereo-selective oxidation of butylbenzene (1) has been examined in vitro by rat liver supernatant fraction(S-9). When phenylbutane (1 ) was incubated at 37 degrees C for 1 hr with S-9, asymmetric oxidation occurred regioselectively at benzylic and omega-1 positions to afford preferentially (R)- and (S)-alcohol (2, 4), respectively. This enzymatic propensity was the case for the production of 1, 3-diols.

[Examination of toxin production from environmental Bacillus cereus and Bacillus thuringiensis].

Emetic toxin- and entero toxin-producing activities were examined for 524 strains of Bacillus cereus and 90 strains of Bacillus thuringiensis to determine the distribution and contamination in natural environment, including foods, vegetable, soil and etc. Emetic toxin was assayed by the production of acid from HEp-2 cells induced by the culture supernatants of Bacillus, which was developed as an improved method for the detection of B. cereus heat-stable emetic-toxin by us.

Synthesis and opioid activities of [D-Leu-8]Dynorphin(1-8) analogs containing a reduced peptide bond, psi(CH2-NH).

[D-Leu8]Dynorphin(1--8)-NH2 analogs, in which each peptide bond was systematically replaced with a psi(CH2NH) peptide bond, were synthesized by the solid-phase method. The psi(CH2NH) bond was introduced by the Boc-amino acid aldehyde/NaCNBH3 method on a solid support. In the syntheses of the analogs, undesirable double alkylation took place at the sequences of Tyr1 psi(CH2NH)Gly2 and Gly2 psi(CH2NH)Gly3, possibly due to the low steric hindrance of the glycine residue.

Role of Ca2+ on endotoxin-sensitivity by galactosamine challenge: lipid peroxide formation and hepatotoxicity in zymosan-primed mice.

This study was investigated to clarify the role of intracellular Ca2+ following endotoxin treatment (1 mg/kg, intraperitoneally) to D-galactosamine-sensitized mice (400 mg/kg, intraperitoneally), and to observe lipid peroxide levels, an index of hepatotoxicity, in endotoxin/galactosamine (Ga1N)-challenged mice under activation of macrophages, especially Kupffer cells, by zymosan. The liver lipid peroxide level and serum glutamic pyruvic transminase activity in mice 18 hr after administration of endotoxin/Ga1N were markedly higher than those in mice treated only with endotoxin. In spite of an increase in lipid peroxide formation, there was little or no effect of Ga1N administration on xanthine oxidase and superoxide dismutase activities in mice given endotoxin.

Further studies on 6-alkylandrost-4-ene-3,17-diones as aromatase inhibitors: elongation of the 6-alkyl chain.

To gain further insight on the relationship between 6-alkylandrost-4-ene-3,17-diones and their aromatase inhibition activity, a series of alkyl steroids with long alkyl chains (n-pentyl, n-hexyl, or n-octyl) at C-6 alpha and 6 beta were synthesized. All of the steroids studied inhibited human placental aromatase in a competitive manner with apparent Ki values ranging from 2.8 to 80 nM.

Competing pathway involved in allylic acetoxylation of androst-5-en-17-one, and oxidation of allylic alcohols with chromium oxides.

Allylic acetoxylation of androst-5-en-17-one (1) with bromine and silver acetate gave 6 alpha- and 6 beta-acetoxyandrost-4-en-17-ones [4 (3%) and 5 (12%)] and 5 alpha-bromo-6 beta-acetoxy steroid 8 (4%) along with the expected product 4 beta-acetoxy derivative 2 (45%). Treatment of 5 alpha,6 beta-bromide 12, an intermediate of the acetoxylation reaction, with silver acetate also produced the acetates 2, 4, 5, and 8 in relative yields similar to those above. These results indicate that the 6-acetates 4 and 5 are produced through a competing pathway involving formation of a bridged carbonium ion 13 followed by attack of an acetate anion.

[Stereoselective N-demethylation of chlorpheniramine in rat liver microsomes: studies on age and sex differences].

We have established a simple method for the chiral stationary-phase liquid chromatographic resolution of racemic chlorpheniramine (Chp) and its two N-demethylated metabolites, monodesmethylchlorpheniramine (DMChp) and didesmethylchlorpheniramine (DDMChp), using an ovomucoid-conjugated column with respective quantitation limits of 5 ng/ml. The assay was used to study the age and sex difference in stereoselective N-demethylation of Chp by rat liver microsomes. The formation rate of each DMChp from racemic Chp was about 2.

[Stereochemistry of highly selective transition metal or Lewis acid-catalyzed asymmetric reactions].

Recent progress of our studies on catalytic asymmetric synthesis is reviewed focused on transition metal-catalyzed intramolecular asymmetric reactions using chiral organo-sulfur groups as main chiral auxiliaries such as asymmetric vinyl-cyclopropane-cyclopentene rearrangements and intramolecular asymmetric metallo-ene reactions, Lewis acid-catalyzed intramolecular asymmetric ene reactions with chiral sulfinyl groups, and palladium-catalyzed intramolecular asymmetric allylations via chiral enamines.

Conformational analysis of 19-oxygenated steroids with a 4-ene or 2,4-diene structure, potential intermediates of aromatase reaction, with semiempirical molecular orbital PM3 calculations.

Conformational analysis of potent competitive inhibitors of aromatase, androst-4-enes 5, as well as 2,4-diene steroids 3, 4, and 6 was carried out, using theoretical calculations, to determine the stereochemistry of their aromatase-catalyzed oxygenation. In the steroids examined, both the 19-alcohols and the 19-aldehydes favor the above- A ring conformation among the possible three in each. The results suggest that the 3-deoxy steroid 5a as well as the 2,4-diene steroids 4a and 6a would be oxygenated at C-19 by aromatase through the same stereomechanism as that involved in the androstenedione aromatization.

Depressive effect of a traditional Chinese medicine (sho-saiko-to) on endotoxin-induced nitric oxide formation in activated murine macrophage J774A.1 cells.

The present study investigated whether or not Sho-saiko-to (crude powder extract, TJ-9) can suppress nitric oxide (NO) generation by endotoxin-activated J774A.1 cells in order to study the preventive mechanism of Sho-saiko-to against endotoxemia. In this experiment, we estimated the NO2- in the murine macrophage cell line J774A.

Effect of saiko-ka-ryukotsu-borei-to on the stress-induced increase of serum corticosterone in mice.

The effect of Saiko-ka-ryukotsu-borei-to (SRBT) on the stress-induced enhancement of serum corticosterone in various stress models was investigated in mice. The serum corticosterone was elevated significantly by immobilized stress, forced-swim stress, electric-shock stress, psychological stress and conditioned-fear stress, respectively. The concentration in the last two models was decreased in a dose-dependent manner by pre-administration of SRBT (10, 60, 100, 600 and 1000 mg/kg, p.

Biochemical studies of estr-4-ene-3,6,17-trione and 5 alpha-androstan-17-ones with or without a carbonyl function at C-3 and/or C-6 as aromatase inhibitors.

19-Nor (2) and 5 alpha-reduced (3) derivatives of androst-4-ene-3,6,17-trione (1) as well as 5 alpha-androstan-17-ones 4-6 were tested for their abilities to inhibit aromatase in human placental microsomes. All the steroids except 5 alpha-6-one 4 were fair to good competitive inhibitors of the enzyme, with apparent Ki's ranging from 50 to 820 nM in which 5 alpha-3-one 5 was the most potent among them. The inhibitory activities of the 19-nor and 5 alpha-reduced derivatives (2 and 3) were less potent than that of the parent compound 1.

Metabolism of 3-acetylcoumarin in rat 9000xg supernatant fraction (S-9)and baker's yeast.

In the incubation of 3-acetylcoumarin in rat liver supernatant fraction (S-9), four metabolites were isolated, and two of which were inseparable diastereomeric mixture as a major product. However, when 3-acetylcoumarin was fermented with baker's yeast (Saccharomyces cerevisiae), only two compounds were obtained as the same as the above mixture. The structures of those metabolites were confirmed by NMR and Mass spectra.

[Immunochemical study on mannan, the antigenic polysaccharide of pathogenic yeasts in man of genus Candida].

This article accounts for the development of immunochemical studies on the antigenic polysaccharide, mannan, a major antigen of pathogenic yeast in man, genus Candida, in order to determine the chemical structures dominating the serological specificities of the parent cells as follows. 1. The serological classification system of 7 medically important Candida species by detecting 10 antigenic factors, 1, 4, 5, 6, 8, 9, 11, 13, 13b, and 34 the corresponding antisera, established by Tsuchiya and his coworkers is documented.

Urinary and biliary metabolites of puerarin in rats.

Examination was made of the urinary and biliary excretion of the metabolites of puerarin, the major component of the roots of Pueraria lobata OHWI (Leguminosae) in rats. The urine of rats administered puerarin orally contained puerarin and four major metabolites, daidzein 4',7-di-O-sulfate (M-I), daidzein 7-O-beta-D-glucuronide (M-II), daidzein 4'-O-sulfate (M-III), daidzein (M-IV), as determined from spectroscopic and chemical data. Total cumulative amounts of the puerarin and four metabolites excreted in the urine at 48 h following the oral administration of puerarin were approximately 3.