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On the preferred rotameric conformation for dopamine agonist action: an illusory quest.

Putative dopamine agonists from the 2-aminotetrahydronaphthalene and trans-octahydrobenzo (f) and (g) quinoline series were shown to inhibit the spontaneous locomotor activity of mice. Marked potency differences were observed between the alpha-and beta-rotameric conformations, compounds having the alpha-rotameric conformation having the greater potency. Thus, 2-di-n-propylmino-5,6-dihydroxytetrahydronaphthalene was 339 times more potent than the 6, 7-dihydroxy isomer, and 2-n-propylamino-5,6-dihydroxy-compound was respectively 79 times and 179 times more than 6,7-hydroxy-and 7,8-dihydroxycompounds. trans-7,8-dihydroxy-1-n-propyl-1,2,3,4,a,9,10,10b-octahydrobenzo(f)quinoline was 11 times more potent than the beta-rotamer, the 6,7-dihydroxy compound, and within the trans-octahydrobenzo(g)quinoline series the alpha-rotameric N-propyl derivative was 467 times more potent than the beta-rotamer, and the alpha-rotameric greater than N-H analogue was 46 fold more potent than the beta-rotamer. Thus, the alpha-rotamer appears the more potent in causing the present functional dopaminergic change. The dopaminergic nature of the response was indicated by its sensitivity to spiroperidol but not to yohimbine or prazosin. The possibility that a difference in behavioural potency between the alpha- and beta-rotamers may reflect a differential metabolism by catechol-O-methyl transferase was assessed by administration of different agonists after pyrogallol pretreatment. This potentiated the activity of 2-di-n-propylamino-6,7-dihydroxytetrahydro-naphthalene but not that of the 5,6-dihydroxy analogue. However, changes in the effects of N-propyl derivatives of trans-octahydrobenzo (f) and (g)quinoline were not marked and, in all experiments, pyrogallol treatment failed by orders of magnitude to shift the dose-response curves of the beta-rotamers to indicate a comparable potency to the rotameric forms.

Animals↗

The metabolism of (3H)noradrenaline released by electrical stimulation from the isolated nictitating membrane of the cat and from the vas deferens of the rat.

1. The noradrenaline (NA) stores of the isolated medial muscle of the cat's nictitating membrane were labelled with [(3)H]NA and the tissue was set up in an isolated organ bath for field stimulation. The tritiated NA and its metabolites released spontaneously and by field stimulation were determined by scintillation counting following chromatographic separation.2. NA represented 11.8 +/- 1.0% of the total radioactivity of the spontaneously released tritiated compounds. The rest was accounted for by NA metabolites: (a) normetanephrine (NMN), 35%; (b) 4-hydroxy-3-methoxymandelic acid (VMA), 20%; (c) 3,4-dihydroxyphenylglycol (DOPEG), 10%; (d) 3,4-dihydroxymandelic acid (DOMA), 10%; (e) 4-hydroxy-3-methoxyphenylglycol (MOPEG), 14%.3. Field stimulation at 25 shocks/sec with supramaximal stimuli of 1 msec duration increased the outflow of NA six to eightfold and that of NMN, DOPEG and VMA two to threefold. The increase in outflow of DOMA and MOPEG was small. NA represented 35.8 +/- 4.1% of the total increase in radioactivity.4. After pargyline pretreatment field stimulation increased the outflow of NA and NMN. Stimulation in the presence of pyrogallol raised the release of NA, DOMA and DOPEG. Neither pargyline nor pyrogallol affected the total release induced by stimulation.5. Cocaine 0.3 mug/ml. increased the release due to stimulation at 4 shocks/sec but not at 25 shocks/sec. Cocaine did not affect the metabolism of the released transmitter.6. Phenoxybenzamine (10 mug/ml.) increased release by stimulation at 4 and 25 shocks/sec. Metabolism of the released transmitter was prevented in the presence of phenoxybenzamine.7. Phentolamine (3 mug/ml.), like phenoxybenzamine, blocked responses to field stimulation, but failed to modify release and subsequent metabolism of NA liberated by field stimulation.8. The main NA metabolites in the rat vas deferens were DOPEG, DOMA and MOPEG. Stimulation at 4 shocks/sec resulted in an increased outflow of NA and of DOPEG. Fifty per cent of the total increase of radioactive compounds was accounted for by NA metabolites.9. These experiments show that for the calculation of the actual output of transmitter it is important to include the metabolites and not to rely on the determination of [(3)H]NA alone.

Animals↗

Molecular mechanisms controlling the rate and specificity of catechol O-methylation by human soluble catechol O-methyltransferase.

Molecular mechanisms determining the turn-over rate and specificity of catechol O-methylation were studied by combining enzyme kinetic measurements, computational modeling of substrate properties and fitting ligands in a 3D model of the active site of the enzyme. Enzyme kinetic measurements were carried out for 46 compounds, including most clinically used catechol drugs, by using recombinant human soluble catechol O-methyltransferase (COMT). The most important mechanism decreasing the turnover rate and increasing affinity was the electron withdrawing effect of substituents. Several other mechanisms by which substituents affected reactivity and affinity were identified. Highest turnover rates were determined for unsubstituted catechol and pyrogallol. Pyrogallol derivatives generally seemed to be more specific substrates than catechols. Catecholestrogens were the most specific endogenous substrates, whereas catecholamines were rather poor substrates. Among the catechol drugs used in the L-DOPA treatment of Parkinson's disease, the COMT inhibitors entacapone and tolcapone were not methylated, whereas the DOPA decarboxylase inhibitor benserazide was 15 times more specific substrate than L-DOPA, the target of COMT inhibition. The structure-activity relationships found allow the prediction of reactivity, affinity, and specificity with useful accuracy for catechols with a wide range of structures and properties. The knowledge can be used in the evaluation of metabolic interactions of endogenous catechols, drugs and dietary catechols, and in the designing of drugs with the catechol pharmacophore.

Binding Sites↗

Sequential Transhydroxylations Converting Hydroxyhydroquinone to Phloroglucinol in the Strictly Anaerobic, Fermentative Bacterium Pelobacter massiliensis.

The recently isolated fermenting bacterium Pelobacter massiliensis is the only strict anaerobe known to grow on hydroxyhydroquinone (1,2,4-trihydroxybenzene) as the sole source of carbon and energy, converting it to stoichiometric amounts of acetate. In this paper, we report on the enzymatic reactions involved in the conversion of hydroxyhydroquinone and pyrogallol (1,2,3-trihydroxybenzene) to phloroglucinol (1,3,5-trihydroxybenzene). Cell extracts of P. massiliensis transhydroxylate pyrogallol to phloroglucinol after addition of 1,2,3,5-tetrahydroxybenzene (1,2,3,5-TTHB) as cosubstrate in a reaction identical to that found earlier with Pelobacter acidigallici (A. Brune and B. Schink, J. Bacteriol. 172:1070-1076, 1990). Hydroxyhydroquinone conversion to phloroglucinol is initiated in cell extracts without an external addition of cosubstrates. It involves a minimum of three consecutive transhydroxylation reactions characterized by the transient accumulation of two different TTHB isomers. Chemical synthesis of the TTHB intermediates allowed the resolution of the distinct transhydroxylation steps in this sequence. In an initial transhydroxylation, the hydroxyl group in the 1-position of a molecule of hydroxyhydroquinone is transferred to the 5-position of another molecule of hydroxyhydroquinone to give 1,2,4,5-TTHB and resorcinol (1,3-dihydroxybenzene) as products. Following this disproportionation of hydroxyhydroquinone, the 1,2,4,5-isomer is converted to 1,2,3,5-TTHB, an enzymatic activity present only in hydroxyhydroquinone-grown cells. Finally, phloroglucinol is formed from 1,2,3,5-TTHB by transfer of the 2-hydroxyl group to either hydroxyhydroquinone or resorcinol. The resulting coproducts are again cosubstrates in earlier reactions of this sequence. From the spectrum of hydroxybenzenes transhydroxylated by the cell extracts, the minimum structural prerequisites that render a hydroxybenzene a hydroxyl donor or acceptor are deduced.

Journal Article↗

Activity-guided fractionation of green tea extract with antiproliferative activity against human stomach cancer cells.

Epidemiological studies have suggested that the consumption of green tea provides protection against stomach cancer. Fractionation of green tea extract, guided by antiproliferative activity against human stomach cancer (MK-1) cells, has resulted in the isolation of six active flavan-3-ols, epicatechin (EC), epigallocatechin (EGC), epigallocatechin gallate (EGCg), gallocatechin (GC), epicatechin gallate (ECg), gallocatechin gallate (GCg), together with inactive glycosides of kaempferol and quercetin. Among the six active flavan-3-ols, EGCg and GCg showed the highest activity, EGC, GC, ECg followed next, and the activity of EC was lowest. These data suggest that the presence of the three adjacent hydroxyl groups (pyrogallol or galloyl group) in the molecule would be a key factor for enhancing the activity. Since reactive oxygen species play an important role in cell death induction, radical scavenging activity was evaluated using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical. The order of scavenging activity was ECg > or = EGCg > or = EGC > or = GC > or = EC. The compounds having a galloyl moiety showed more potent activity. The contribution of the pyrogallol moiety in the B-ring to the scavenging activity seemed to be less than that of the galloyl moiety.

Camellia sinensis↗

Antioxidant and antibacterial activities of Rumex japonicus HOUTT. Aerial parts.

We evaluated total phenolic content, antioxidant activity, reducing power and antibacterial activity of ethanol, hexane, chloroform, ethyl acetate and aqueous extracts of aerial parts of Rumex japonicus HOUTT. The ethyl acetate extract had the highest amount of phenolic compounds. It also exhibited the highest reducing power and antioxidant activity when assayed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH), beta-carotene bleaching and superoxide radical methods. The ethyl acetate extract possessed the strongest antibacterial activity against Bacillus subtilis, B. cereus and E. coli. GC-MS analysis indicated that ethyl acetate extract contained a variety of phenolic compounds. HPLC analysis showed that pyrogallol was the predominant phenolic compound in this extract. Thus, our study verified that the ethyl acetate extract has strong antioxidant and antibacterial activities which are correlated with its high level of phenolic compounds, particularly pyrogallol and pyrocatechin. This extract of R. japonicus aerial parts can be utilized as an effective and safe source of antioxidants.

Acetates↗

Biomimetic synthesis of (+/-)-galanthamine and asymmetric synthesis of (-)-galanthamine using remote asymmetric induction.

(+/-)-Galanthamine (1) was synthesized in excellent yield by applying PIFA-mediated oxidative phenol coupling of N-(4-hydroxy)phenethyl-N-(3',4',5'-trialkoxy)benzyl formamide (15b) as a key step. Because of the symmetrical characteristics of the pyrogallol moiety in the substrate (15b), the phenol coupling resulted in a sole coupling product except for volatile components from the oxidizing agent. On the basis of the successful results of the above strategy, (-)-galanthamine (1) was synthesized by employing a novel remote asymmetric induction, where conformation of the seven-membered ring in the product of the phenol coupling was restricted by forming a fused-chiral imidazolidinone ring with D-phenylalanine on the benzylic C-N bond of the tri-O-alkylated gallyl amino moiety. The conformational restriction and successive debenzylation of the protected hydroxyl groups on the pyrogallol ring caused diastereoselective cyclization to yield a cyclic ether having the desired stereochemistry for the synthesis of (-)-1.

Galantamine↗

Catalase-peroxidase from the cyanobacterium Synechocystis PCC 6803: cloning, overexpression in Escherichia coli, and kinetic characterization.

The Synechocystis PCC 6803 katG gene encodes a dual-functional catalase-peroxidase (EC 1.11.1.7). We have established a system for the high level expression of a fully active recombinant form of this enzyme. Its entire coding DNA was extended using a synthetic oligonucleotide encoding a hexa-histidine tag at the C-terminus and expressed in Escherichia coli [BL21-(DE3)pLysS] using the pET-3a vector. Hemin was added to the culture medium to ensure its proper association with KatG upon induction. The expressed protein was purified to homogeneity by two chromatography steps including a metal chelate affinity and hydrophobic interaction chromatography. The homodimeric acidic protein (pl = 5.4) had a molecular mass of 170 kDa and a Reinheitszahl (A406/A280) of 0.64. The recombinant protein contained high catalase activity (apparent Km = 4.9 +/- 0.25 mM and apparent kcat = 3500 s(-1)) and an appreciable peroxidase activity with o-dianisidine, guaiacol and pyrogallol, but not with NAD(P)H, ferrocytochrome c, ascorbate or glutathione as electron donors. By using both conventional and sequential stopped-flow spectroscopy, formation of compound I with peroxoacetic acid was calculated to be (8.74 +/- 0.26) x 10(3) M(-1) s(-1), whereas compound I reduction by o-dianisidine, pyrogallol and ascorbate was determined to be (2.71 +/- 0.03) x 10(6) M(-1) S(-1), (8.62 +/- 0.21) x 10(4) M(-1) S(-1), and (5.43 +/- 0.19) x 10(3) M(-1) S(-1), respectively. Cyanide binding studies on native and recombinant enzyme indicated that both have the same heme environment. An apparent second-order rate constant for cyanide binding of (4.8 +/- 0.1) x 10(5) M(-1) S(-1) was obtained.

Amino Acid Sequence↗

Protective effects of phenolic compounds on CCl4-induced toxicity in isolated rat hepatocytes.

The protective effects of a series of phenolic compounds, phenolic acids and flavonoids on the cytotoxicity of CCl4 in rat hepatocytes were studied. A number of flavones, 7,8-dihydroxy-flavone, luteolin and hypolaetin-8-glucoside, flavonols, morin, quercetin, robinetin and gossypin, phenolic acids, gallic, caffeic and chlorogenic acids, as well as the flavane (+)-catechin significantly inhibited alanine amine transferase (ALT) release. Catechol groups are determinant for the protective activity of flavonoids and cinnamic acid derivatives, as well as the resorcinol or pyrogallol moieties in the B ring of flavonoids. In benzoic acid derivatives a pyrogallol group is required. This feature is associated with the inhibition of ALT spontaneous release.

Animals↗

Reactivation mechanisms of thiamine with thermostable factors.

It was observed, in vitro, that the water extract of the fermented-tea customarily chewed by Thai people has a similar thermostable thiamine-inactivating factor to that found in the water extract of fern. It was also observed that the percentage of thiamine disulfide formed from thiamine with some flavones, catechol, pyrogallol, caffeic acid, dihydroxyphenylalanine, and hemin is greater at pH 7.5 than at pH 7.0. With some flavonoids, such as quercetin, rutin, and 6,7,4'-trihydroxyisoflavone, and pyrogallol, hemin, catechol and caffeic acid at pH 7.5, around 30-100% of thiamine is changed into thiamine disulfide. Water extract of shiitake, okra, coffee, black tea and fukinoto have only weak activities of thermostable thiamine-inactivating factors as a large percentage of thiamine disulfide is formed from thiamine even at pH 7.0. 2-Methyl-4-amino-5-aminomethylpyrimidine was isolated from the reaction mixture of 1 g thiamine with 20 mg catechol (1:0.5 mole) at pH 7.0, 45 degrees C, and identified with the synthesized pyrimidine.

Antimetabolites↗

[Antibacterial and anti-hemolysin activities of tea catechins and their structural relatives].

Among catechins tested, (-)epigallocatechin (EGC), (-)epicatechin gallate (ECg), (-) epigallocatechin gallate (EGCg) inhibited the growth of Staphylococcus aureus, Vibrio cholerae O1 classical Inaba 569B and El Tor Inaba V86. S. aureus was more sensitive than V. cholerae O1 to these compounds. EGCg showed also a bactericidal activity against V. cholerae O1 569B. Pyrogallol showed a stronger antibacterial activity against S. aureus and V. cholerae O1 than tannic and gallic acid. Rutin or caffein had no effect on them. ECg and EGCg showed the most potent anti-hemolysin activity against S. aureus alpha-toxin, Vibrio parahaemolyticus thermostable direct hemolysin (Vp-TDH) and cholera hemolysin. Among catechin relatives, only tannic acid had a potent anti-hemolysin activity against alpha-toxin. These results suggest that the catechol and pyrogallol groups are responsible for the antibacterial and bactericidal activities, while the conformation of catechins might play an important role in the anti-hemolysin activity.

Antitoxins↗

The action mechanism of relaxation effect of atropine on the isolated rabbit corpus cavernosum.

PURPOSE: Atropine has been used to block cholinergic neurotransmission in basic research and has received recent interest clinically in the intracavernosal pharmacotherapy of erectile dysfunction. It has been suggested that at a low dose (10(-8) M), atropine blocks muscarinic receptors, and that at a large dose (10(-3) M), it induces the release of EDRF. However, no report has supported this idea experimentally. We tried to confirm the action of atropine in cavernosal tissue and define its mechanism. MATERIALS AND METHODS: Strips of rabbit corpus cavernosum were mounted in organ chambers. On the precontracted muscle strips with phenylephrine (PHE; 5 x 10(-6) M), atropine was treated with increasing concentration from 10(-11) M. The relaxing activity of atropine was observed in deendothelialized tissue and preparation with treatment with methylene blue (10(-4) M), pyrogallol (10(-4) M), NW-nitro-L-arginine (L-NNA; 3 x 10(-4) M) and indomethacin (10(-4) M). To evaluate the relationship of atropine to Ca++, the muscle strip was incubated in Ca++ free solution, and Ca++ induced contraction by addition of CaCl2 (10(-3) M) was recorded with atropine. Depolarization by KCl was observed with atropine to investigate the relationship of atropine relaxation to K+. RESULTS: On the precontracted muscle strip with PHE, atropine induced a dose-related contraction up to 10(-8) M and began to exert a relaxing effect at the concentration of 10(-7) M and reached the 93.6% relaxation effect at the concentration of 10(-4) M, causing dose-dependent relaxation. The relaxing effect of atropine was partially inhibited by endothelial disruption, and by pretreatment with methylene blue, pyrogallol, L-NNA, and indomethacin, although they were not statistically significant. At the basal state of muscle strips in Ca++ free solution, atropine decreased basal tension as well as inhibited the contraction induced by CaCl2 dose-dependently. However, atropine did not influence depolarization by KCl. CONCLUSIONS: Atropine has both a contraction effect at lower concentrations and a relaxation effect at higher concentrations on cavernosal smooth muscle. It is presumed that the relaxation at higher concentrations is mediated via increasing intracellular calcium sequestration, not by hyperpolarization or secretion of EDRF.

Animals↗

[Biological analysis of proteinuria in the laboratory: quantitative features].

Total protein analysis is one of the most frequent laboratory analyses in urine. A proteinuria above 150 mg/L is often observed in a random way in preventive or school medicine (dipsticks) or during laboratory analysis (quantitative determination). Complete (quantitative, then qualitative) and repeated evaluation of proteinuria is of major interest for the clinician to establish a diagnosis of abnormality and for therapeutic follow-up of a nephropathy, uropathy or a non-renal disease (diabetes, multiple myeloma). Most frequent (90% of cases) and severe forms of proteinuria are of glomerular type, associated to the nephrotic syndrome, hypertension, and progressive renal failure. Attention should be paid by the biologist to the pre-analytical phase (specimen collection, treatment, and storage), to clinical data, and to prescription of drugs that could interfere with protein analysis. During the last past 10 years, significant analytical advances have been made: dipstick analysis has been dropped (false positives and most importantly false negatives) as manual precipitation techniques with turbidimetric detection (poor inter-laboratory coefficients of variation, CV), replacement of Coomassie blue by pyrogallol red (improved practicability). Urinary quality control data reflect these positive changes, as demonstrated by a dramatic reduction in reported CVs. There is, however, still no reference method for total urinary protein determination and limits of existing pyrogallol red methods should be emphasized: variable reagent composition between manufacturers (such as the presence of SDS additive), limited sensitivity, difficulty in the choice of a calibration material, underestimation of free light chains, and interference with gelatin based vascular replacement fluids.

Coloring Agents↗

[Effect of fractions isolated from Naja naja atra venom on antioxidation in animals].

OBJECTIVE: To study the antioxidation of fraction F and H isolated from Naja naja atra venom on homogenate and RBC autoxidation. And to explore the effect of two fractions on the activities of antioxidation enzymes in mice. METHOD: Samples of tissues homogenates and RBC suspension were pretreated with fraction F/H, and then their generation of malondialdehyde(MDA) was examined. The effects of fraction F/H on O2-* produced by pyrogallol autoxidation and *OH produced by Cu2+ -VitC were checked. After being administrated i.p. with fraction F/H for 15 days, superoxide dismutase and hydroperoxidase in mice were measured for their activities. RESULT: Fraction F/H inhibited RBC autohemolysis and the generation of MDA decreased. The production of lipid peroxides in normal brain, liver and heart homogenate of rat and the elevation of lipid peroxides induced by cysteine and FeSO4 in homogenate were inhibited by addition of fraction F/H. The O2-* produced by autoxidation of pyrogallol and the *OH produced by Cu2+ -VitC system could be scavenged by fraction F/H (the potency of scavenging oxygen free radical is that fraction H > fraction F). Fraction F/H all increased the activities of several antioxidation enzymes in mice to different extents. CONCLUSION: Fraction F/H have the effect of antilipid peroxidation,may scavenge active oxygen free radical and increase the activity of SOD.

Animals↗

Dioxygenase and peroxidase activities of soybean lipoxygenase: synergistic interaction between linoleic acid and hydrogen peroxide.

The interaction of H2O2 with soyabean lipoxygenase was investigated in the presence of linoleic acid. Dioxygenase activity was significantly higher at pH 9.0 than at pH 6.0. H2O2 at concentrations less than 1.0 nM stimulated linoleic acid oxidation synergistically and the magnitude of synergism was higher at pH 9.0. Linoleic acid dependent peroxidase activity towards benzidine, guaiacol, tetramethylbenzidine (TMBD) and tetramethylphenylenediamine (TMPD) was higher at pH 9.0, whereas pyrogallol and ABTS oxidation rates were higher at pH 6.0. H2O2 supported oxidation of benzidine, guaiacol, pyrogallol and ABTS was higher at pH 6.0, whereas TMPD, TMBD exhibited higher oxidation rates at pH 9.0. H2O2 in the presence of linoleic acid produced synergism in xenobotic metabolism and depending upon the substrate in question upto 11-fold increase in oxidation rate was noted.

Drug Synergism↗

High performance liquid chromatographic determination of vitamin D in fortified milks, margarine, and infant formulas.

Fortified milks were saponified overnight at room temperature with 1% ethanolic pyrogallol and KOH. The digest was extracted with hexane after adding water and ethanol, and the extract was washed consecutively with 5% KOH, water, and 55% aqueous ethanol to remove polar lipids. After evaporation, the residue was first chromatographed on a column of 5 micrometer silica. A fraction containing vitamin D was collected, evaporated, and rechromatographed on a reverse phase column for the separation and quantitation of vitamins D2 and D3. Recovery was 96-99% and the coefficient of variation was 3% (8 replicates). Infant formula was diluted and then saponified and extracted as in the analysis of milk. Margarine was saponified by shaking overnight with 1% ethanolic pyrogallol and 80% KOH. Water and ethanol were added to the digest before extraction. Extracts from formula and margarine were chromatographed as milk except, before HPLC, the extract was dissolved in isopropanol-hexane (1 + 99) and passed through 5 cm alumina in a Pasteur pipet, and the concentration of isopropanol in the first high performance liquid chromatographic (HPLC) solvent system was halved to improve the separation of vitamin D from other absorbing lipids. Usually several peaks were obtained during the final HPLC analysis, and the identification of vitamins D2 and D3 was less certain than in the analysis of milk. The coefficients of variation for formula and margarine were 6% (5 replicates) and 9% (6 replicates), respectively.

Animals↗

[Contact eczema caused by pine wood].

A 34 year-old woman, working with pine wood treated with sodium hydroxide and pyrogallol, developed dermatitis on arms, face and neck. Patch testing showed allergic reaction to colophony and pine wood, but not to sodium hydroxide or pyrogallol. Pine wood contains colophony. Since avoiding pine wood, she has had no further attack of dermatitis.

Adult↗

Triterpene antioxidants from ganoderma lucidum.

Ganoderma lucidum was studied for its antioxidative activity by bioassay guided isolation in conjunction with in vitro tests. The powdered crude drug was treated with boiling water and the aqueous extract (Ex1) was further separated to obtain terpene and polysaccharide fractions. The two fractions and Ex1 were screened for their antioxidative effect against pyrogallol induced erythrocyte membrane oxidation and Fe (II)-ascorbic acid induced lipid peroxidation. All tested samples showed antioxidative activities in a dose dependent manner and the terpene fraction was found to possess the highest effect compared with the others. Chemical isolation of the terpene fraction resulted in the detection of ganoderic acids A, B, C and D, lucidenic acid B and ganodermanontriol as major ingredients.

Animals↗