Easy DNA extraction for rapid detection of Panax ginseng C. A. Meyer in commercial ginseng products.

An investigation was carried out in order to obtain an easy and rapid detection of Panax ginseng in commercial herbal products by using molecular techniques (polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP)). Two protocols and one commercial kit for DNA extraction were used. Four forms of commercial products were considered, i.e., dried body roots, dried root tails, dried root prongs and dried extracts. The RFLP of DNA amplified products by 18df/28ccr primers, obtained using Inf I, Sau 3A1 and Taq I endonucleases, allowed the identification of P. ginseng and its differentiation from P. quinquefolium. The presence of adulterants, as Mirabilis jalapa L. and Phytolacca acinosa Roxb. was excluded in the examined commercial samples. P. ginseng was detected in 63% of the considered samples according to the declaration of the labels, whereas negative results were obtained with the dried extract form. Therefore, the Invitrogen DNA extraction kit let the easy extraction of useful amounts of DNA and a standardisation of routine work, in comparison with the other molecular protocols so far used.

Bronchodilator activity of Phymatodes scolopendria (Burm.) Ching and its bioactive constituent.

Phymatodes scolopendria (Burm.) Ching (Polypodiaceae) is widely used in the Eastern coast of Madagascar to treat respiratory disorders. Bioassay-guided fractionation using guinea pig trachea pre-contracted with histamine to monitor the activity led to the isolation of 1,2-benzopyrone (coumarin) as the main active constituent. Effectively, it induced a concentration-dependent relaxation of the histamine with a median effective concentration (EC(50)) of 35.03 microg/ml, or carbachol (EC(50) = 33.41 microg/ml) pre-contracted guinea pig trachea, and also provoked 100% relaxation at 72.10 microg/ml. It was less active either on KCl pre-contracted trachea (EC(50) = 130.78 microg/ml) or endothelium denuded trachea (153.4 +/- 22 microg/ml). It inhibited, in a non-competitive manner, the histamine and the external calcium spasm effect on the isolated trachea but it did not significantly modify the broncho-constrictive activity of KCl. When combined with theophylline, coumarin produced a significant additive relaxing effect on pre-contracted trachea. Furthermore, its bronchodilator effect was not blocked by propranolol. In vivo, pre-treated guinea pig with coumarin showed significant resistance to histamine inhalation, with an adequate dose protecting 50% of the tested animals (AD(50)) of 75 mg/kg. These results indicate that the bronchodilator effect of coumarin is partly due to the endothelium-dependent tracheal relaxation, and may be mediated through a non-specific tracheal relaxation.

Studies on vasoconstrictor activity of Curculigo pilosa extracts and of its isolated compounds.

The Curculigo pilosa total extract, its butanolic fraction (0.5 microg-100 mg/kg) and the most active in vitro compound structurally similar to adrenaline, pilosidine (10 ng-l mg/kg), caused a reversible and dose-dependent increase in blood pressure in anaesthetized rat. This hypertensive effect is partially reversed (90%) by the prior administration of phentolamine (1 mg/kg) and abolished by pre-treatment with phentolamine (1 mg/kg) and atenolol (100 microg/kg). Neither tachiphylaxis nor any toxic effects were observed. These experimental findings suggest an interaction between C. pilosa and the peripheral adrenergic system (particularly with alpha1 and beta1 receptors); the structure of the bioactive glucosides could be important in evoking this effect.

7-Epimer oxindole alkaloids of Cabucala cryptophlebia: their (13)C-NMR and CD data.

The isolation, (13)C-NMR and CD data of two 7-epimer oxindole alkaloids, 10,11-dimethoxyisomitraphylline (1) and 10,11-dimethoxymitraphylline (2) from Cabucala cryptophlebia are reported.

The co-occurrence of C(3) epimer Nb,C(21)-secocuran alkaloids in Strychnos diplotricha and Strychnos myrtoides.

From the stem bark of Strychnos diplotricha, three Nb,C(21)-secocuran alkaloids, viz., 3-epi-myrtoidine, 11-demethoxy-3-epi-myrtoidine and 11-demethoxy-12-hydroxy-3-epi-myrtoidine, were isolated together with the known myrtoidine and 11-demethoxymyrtoidine. They also occur in different parts of S. myrtoides.

Biological activities of the plant-derived bisindole voacamine with reference to malaria.

The in vivo antiplasmodial activity of voacamine was assessed in a 4-day test. It was shown to exhibit in vivo activity with 25.4% and 43.4% inhibition of parasitaemia with 2.5 and 10 mg/kg, respectively. In synchronized cultures, it was found to act on trophozoite and schizont stages of Plasmodium falciparum. Using the FMC29 strain of Plasmodium falciparum as parasite and the isobologram curve as a method to assess interaction in drug combination, it was shown to lack any chloroquine-enhancing activity and its in vitro antiplasmodial effect was not potentiated by the chemosensitizer malagashanine.

Antiplasmodial activity of the alkaloids of Peschiera fuchsiaefolia.

The tertiary and quaternary alkaloids isolated from the stem bark, root bark and seeds of Peschiera fuchsiaefolia are reported. The tertiary alkaloid crude extract from the stem bark was tested in vitro against Plasmodium falciparum on the basis of the antimalarial use of the plant. It showed good activity against both the D6 strain (IC50 = 495 ng/ml) and chloroquine-resistant W2 strain (IC50 = 817 ng/ml) and voacamine was the most active of the tested alkaloids (IC50 = 238 ng/ml for D6 and 290 ng/ml for W2). The tertiary alkaloid crude extract from the root bark of the same plant is more active than voacamine (IC50 = 179 ng/ml for D6 and 282 ng/ml for W2 strain), and is particularly rich in dimeric alkaloids (0.22% of the vegetable material).

Benzylbenzoate and norlignan glucosides from Curculigo pilosa: structural analysis and in vitro vascular activity.

From the rhizomes of Curculigo pilosa, two benzylbenzoate diglucosides, piloside A and piloside B, and a glucosyl-fused norlignan, pilosidine, previously obtained only as the tetra-O-methyl derivative, were isolated. Pilosidine showed facilitating effect on adrenaline evoked contractions in rabbit aorta isolated preparations.

Alkaloids of Hernandia voyronii: chloroquine-potentiating activity and structure elucidation of herveline D.

Further investigation of Hernandia voyronii led to the isolation of a new pavine-benzyltetrahydroisoquinoline (pavine-BTIQ) dimer, herveline D, together with herveline A, five aporphine alkaloids, two morphinane alkaloids, and their bio-synthetic precursor, i.e., the BTIQ ( S)-reticuline. Hervelines A-D have a moderate intrinsic in vitro antimalarial activity (IC (50) in the range of 1.68-3.28 microM), but displayed different effects ranging from synergism for herveline B and herveline C to simple additive effect for herveline A, and antagonism for herveline D in a chloroquine (CQ) combination evaluation and this was confirmed in vivo for hervelines A and B. Furthermore, the antiplasmoidal activity of CQ was potentiated in vitro by reticuline and its dimethyl derivative laudanosine (for the latter also in vivo), whereas herveline C moderately potentiated in vitro the anti-plasmodial activity of herveline D.

Synthesis of 1,5-diaryl-3-methyl-1H-pyrazolo[4,5-c]isoquinolines and studies of binding to specific peripheral benzodiazepine binding sites.

Some 1,5-diaryl-3-methyl-1H-pyrazolo[4,5-c]isoquinolines were synthesized and tested for their ability to displace [3H]clonazepam or [3H]Ro 5-4864 from their specific binding on the central and peripheral benzodiazepine receptors. None of the tested compounds showed any activity as central binding inhibitors, while most of them were specific as peripheral binding inhibitors, although they were not very potent.

Tricyclic heteroaromatic systems: synthesis, [3H]flunitrazepam brain membrane binding inhibition, and structure-activity relationships of 2,3-dihydro-2-aryl-4-R-[1]benzopyrano[4,3-c]pyrazole-3-ones.

We report the synthesis and binding activity to the central benzodiazepine receptors of some 2,3-dihydro-2-aryl-4-R-[1]benzopyrano[4,3-c]pyrazole-3-ones, which are isosteres of the CGS series. Although the compounds of the CGS series are potent ligands of the benzodiazepine receptors, none of the isosteres tested showed any significant inhibiting potency. This may be due to the change in electronic properties brought about by the replacement of the NH of the CGS series with an oxygen atom.

EEG spectral analysis after minor head injury in man.

A quantitative EEG analysis was performed on the posterior lead of 18 patients with their eyes closed 3-10 days after minor head injury. There was a significant increase of the mean power of slow alpha (8-10 c/sec), a reduction of fast alpha (10.5-13.5 c/sec), with a shift of mean alpha frequency towards lower values, and a reduction of fast beta (20.5-36 c/sec) in the patients with head injuries compared with the age- and sex-matched patients in the control group. Therefore, power spectral EEG analysis performed with the above-mentioned criteria may be suggested as a sensitive tool to be added to others during the neurophysiological follow-up studies of people with head injuries.

1,3-Diarylpyrazolo[4,5-c]- and -[5,4-c]quinolin-4-ones. 4. Synthesis and specific inhibition of benzodiazepine receptor binding.

A series of 1,3-diarylpyrazolo[4,5-c]- and -[5,4-c]quinolin-4-ones were prepared and tested for their ability to displace [3H]flunitrazepam from bovine brain membranes. While the 1,3-diarylpyrazolo[4,5-c]quinoline derivatives showed affinity for the receptor site, their [5,4-c] isomers were devoid of binding activity.

Synthesis and binding study on pyrazolo[4,5-c] [1,8]naphthyridines.

Following our previous reports on pyrazolo[4,5-c]quinoline derivatives some isosteric pyrazolo[4,5-c] [1,8]naphthyridines were synthesized and tested for their ability to displace [3H]flunitrazepam from rat brain membranes. The lack of activity of the synthesized compounds and structure-activity relationships for the whole series are discussed.

Pyrazolo[4,5-c]quinolines. 3. Synthesis, receptor binding, and 13C NMR study.

Some 1-aryl-3-methylpyrazolo[4,5-c]quinolin-4-ones, were prepared and tested for their ability to displace specific [3H]flunitrazepam binding from bovine brain membranes. The 1-meta-aryl derivatives were the compounds that bound with the highest affinity within this class. Our 13C NMR study suggested a correlation between the binding affinity and the chemical shift value of a carbon atom of the tricyclic system.

Pyrrolnitrin analogues. XII. Synthesis and biological activity of some pyrazole carboxylic acids.

Some 1,5-diaryl-, 1-aryl-5-(2-nitrobenzyl)-3-methylpyraloze-4-carboxylic acids and some of their 4-morpholino- and 4-N-methylpiperazino amides are prepared and tested as antibacterial agents. None of the tested compounds shows any noteworthy antimicrobial activity. The results are discussed and compared with the previously published results for other similarly structured compounds.

Pyrazolo[4,5-c]quinolines. 2. Synthesis and specific inhibition of benzodiazepine receptor binding.

A series of 1-aryl-3,5-dimethyl-4,5-dihydro-1H-pyrazolo[4,5-c]quinolin-4-ones (2a-e) and 1-aryl-3-methyl-1H-pyrazolo[4,5-c]quinolines (3-7a-e) bearing different substituents at position 4 were prepared and tested for their ability to displace specific [3H]flunitrazepam binding from bovine brain membranes. The 5-N-methyl derivatives 2a-c,e were the compounds that bound with the highest affinity within this class. The replacement of the carbonyl group with other substituents and the resulting aromatization of the pyridine moiety greatly decreased the binding affinity. From a Lineweaver-Burk analysis on the most active compound 2b, it appears that the inhibition is a competitive one.

Synthesis and binding studies of 1-arylpyrazolo[4,5-c]- and 2-arylpyrazolo[4,3-c]quinolin-4-ones. I.

The syntheses of some 2-aryl-3-methyl-4,5-dihydro-2H-pyrazolo [4,3-c] quinolin-4-ones and 1-aryl-3-methyl-4,5-dihydro-1H-pyrazolo-[4,5-c] quinolin-4-ones are reported. Some of the latter have shown a high activity in displacing specific [3H]-flunitrazepam binding from bovine brain membranes.

Pyrrolnitrin analogues. X. Synthesis and biological activity of 1-chlorophenyl-3- or 5-nitrophenyl-pyrazole-4-carboxylic acids.

The synthesis and the in vitro antimicrobial activity of all the possible 1-chlorophenyl-3-nitrophenyl-5-methylpyrazole-4-carboxylic acids and 1-chlorophenyl-3-methyl-5-nitrophenylpyrazole-4-carboxylic acids are reported. Some acids showed an interesting activity against some strains of gram-positive bacteria. The results are discussed and compared with those of other related compounds.