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PubMedDate not supplied

[Sequential isolation of superoxide dismutase and ajmaline from a tissue culture of Rauwolfia serpentina Benth].

Two biologically active compounds, the enzyme superoxide dismutase (EC 1.15.1.1) and the antiarrhythmic indole alkaloid ajmaline, were isolated from a callus culture of Rauwolfia serpentina Benth. Sequential isolation of biologically active compounds by metal-chelate affinity chromatography followed by azoadsorbent affinity chromatography allowed us to obtain highly purified products. The yields of superoxide dismutase and ajmaline were 180 mg/kg biomass and 16.5 g/kg dry weight, respectively.

Ajmaline↗
PubMedDate not supplied

[Effect of phytohormones on the protein-synthesizing ability of rauwolfia serpentina benth. tissue culture].

The accumulation of intracellular protein by the callus culture of Rauwolfia serpentina Benth. was studied in a standard or phytohormone-containing medium. Changes in the concentration of total protein in cells induced by indolylacetic and naphthylacetic acids were shown to be associated with the effects of these phytohormones on the biosynthesis and degradation of intracellular protein.

Culture Techniques↗
PubMedDate not supplied

[Formation of giant and ultramicroscopic forms of Nostoc muscorum CALU 304 during cocultivation with Rauwolfia tissues].

The study of heteromorphic Nostoc muscorum CALU 304 cells, whose formation was induced by 6- to 7-week cocultivation with the Rauwolfia callus tissues under unfavorable conditions, revealed the occurrence of giant cell forms (GCFs) with a volume which was 35-210 times greater than that of standard cyanobacterial cells. Some GCFs had an impaired structure of the murein layer of the cell wall, which resulted in the degree of impairment of the cell wall ranging from the mere loss of its rigidity to its profound degeneration with the retention of only small peptidoglycan fragments. An analysis of thin sections showed that all GCFs had enlarged nucleoids. The photosynthetic membranes of spheroplast-like GCFs formed vesicles with the contents analogous to that of nucleoids (DNA strands and ribosomes). About 60% of the vesicles had a size exceeding 300 nm. With the degradation of GCFs, the vesicles appeared in the intercellular slimy matrix. It is suggested that the vesicles are analogous to elementary bodies, which are the minimal and likely primary reproductive elements of L-forms. The data obtained in this study indicate that such L-forms may be produced in the populations of the cyanobionts of natural and model syncyanoses. Along with the other known cyanobacterial forms induced by macrosymbionts, L-forms may represent specific adaptive cell forms generated in response to the action of plant symbionts.

Adaptation, Physiological↗
PubMed1960

A new antihypertensive agent. Clinical evaluation of a Rauwolfia-flumethiazide combination (rautrax).

Twenty-eight patients were treated with Rautrax,(R) a combination of flumethiazide, rauwolfia and potassium chloride for from one to seven months. The average mean blood pressure for the group declined from 135 mm. of mercury to 107 mm. All but two of the patients had a decrease in blood pressure and 19 became normotensive. Associated symptoms of headache, dyspnea, edema and angina were completely relieved or improved in the majority of patients with these complaints. On the basis of the blood pressure response and the clinical effects seen in the patient, therapeutic results were classified as good to excellent in 22 of the 28 patients, fair in two, and poor in three. No evaluation was made in the remaining patient in the series because further adjustment in dosage was required. Three patients had side effects-moderate gastrointestinal upset in one case, headache and a sensation of the bladder's having been "wrung out" in another, and headache and paresthesia of the legs in the third. Only the third patient had persisting symptoms after the drug was discontinued. In the other two reduction of dosage sufficed.

Antihypertensive Agents↗
PubMedDate not supplied

[Change in activity of superoxide dismutase in callus culture of Rauwolfia serpentina Benth., grown in standard conditions and with temperature shock].

The rate of superoxide dismutase (SOD) accumulation in Rauwolfia serpentine Benth. cell culture under heat shock conditions (3 h, 45 degrees C) decreased insignificantly (by 4%), whereas low positive temperature (24 h, 7 degrees C) caused a drastic drop (by 48%). The observed decrease in the level of SOD activity resulted from a slowdown of the biosynthesis rate of the enzyme and decrease in its concentration in the cultivated cells. In addition, a compensatory decrease in degradation of the active protein (Kd) was observed at low positive temperatures and, consequently, an increase in its half-life (t1/2), compensating partially for a deficiency in de novo synthesized SOD molecules. The parameters studied restored after a 24-h adaptation of cells under standard temperature conditions.

Half-Life↗
PubMed1990

[Antiarrhythmic activity of ajmaline obtained from Rauwolfia serpentina biomass grown in tissue culture].

The conventional experimental techniques showed that ajmaline derived from Rouwolfia serpentina Benth. biomass grown in tissue culture was effective in arresting canine ventricular arrhythmias induced by coronary occlusion, strophanthin and epinephrine, displaying antiarrhythmic effects in aconitine-induced intoxication of rats and substantially elevating the threshold of ventricular fibrillations in cats. The antiarrhythmic activity of ajmaline produced by the biotechnological technique was identical to that of ajmaline derived from the natural root of Rauwolfia serpentina Benth, which allows one to recommend that the agent in question should be clinically tested as an antiarrhythmic.

Ajmaline↗
PubMedDate not supplied

Determination of reserpine and rescinnamine in Rauwolfia serpentina powders and tablets: collaborative study.

A liquid chromatographic (LC) method for determining reserpine and rescinnamine in Rauwolfia serpentine powders and tablets, which uses fluorescence detection, was subjected to a collaborative study. The procedure for extraction and purification is a simplified version of that used in the current official method for analysis of these products. LC separations are performed on a normal-phase column. The mobile phase is methanol to which a small volume of an aqueous solution of 1-pentanesulfonic acid sodium salt can be added to achieve desired elution characteristics. Reserpine and rescinnamine elute at approximately the same time but can be individually quantitated by appropriate settings of the fluorescence detector. Reserpine is determined at an excitation wavelength of 280 nm and an emission wavelength of 360 nm, because rescinnamine is completely non-fluorescent at these wavelengths. Rescinnamine is determined at an excitation wavelength of 330 nm and an emission wavelength of 435 nm, because reserpine is completely nonfluorescent at these wavelengths. The following materials were used for the study: one sample of United States Pharmacopeia (USP) standard R. serpentine powder, one tablet type labeled as containing 100 mg R. serpentine and 2 tablet types labeled as containing 50 mg R. serpentine. For each of the 4 materials, 2 pairs of blind duplicates were prepared. Three materials were analyzed in duplicate by 8 laboratories. One of the 2 tablets labeled to contain 50 mg R. serpentine was analyzed only by 7 of 8 participating laboratories. Average combined content of reserpine and rescinnamine was 0.144% for the USP raw material and 0.132, 0.135, and 0.137% for the 3 commercial tablets. Reproducibility relative standard deviation values were 5.72, 5.93, 8.61, and 3.48% and repeatability relative standard deviation values were 2.57, 4.87, 3.19, and 1.99% for the 4 samples. The Associate Referee conducted a study to determine recoveries of reserpine plus rescinnamine by this method from mixtures simulating sample extracts. Average recovery of 15 determinations was 100.1%, with a relative standard deviation of 1.3%. The LC method for determination of reserpine and rescinnamine in R. serpentine powders and tablets has been adopted first action by AOAC INTERNATIONAL.

Chromatography, Liquid↗
PubMed1990

Genotoxic, mutagenic and recombinogenic effects of rauwolfia alkaloids.

In the last decade, the possible correlation between the use of reserpine and rauwolfia drugs as antihypertensive agents and breast cancer incidence has been investigated. For the purpose of evaluating the mutagenic and genotoxic effects of these drugs, reserpine and ajmalicine were studied using the SOS Chromotest and the induction of gene conversion, crossing-over and reverse mutation in the yeast diploid strain XS2316. The results indicated a lack of genotoxic, mutagenic and recombinogenic effects.

Microsomes↗
PubMed1996

Combinatorial modification of natural products: preparation of unencoded and encoded libraries of Rauwolfia alkaloids.

We report the preparation of combinatorial libraries which consist of derivatives of the stereoisomeric alkaloids yohimbine and rauwolscine-members of the Rauwolfia genus. The chemistry was performed on solid support using the divide-and-pool method, and involved the derivatization of the E-ring carboxylates and hydroxyls of these alkaloids with 36 amino acids and 22 carboxylic acids, respectively, to afford 792 bifunctionalized derivatives. The rauwolscine library was prepared using an encoding strategy in which the identity of each incorporated amino acid was recorded by cosynthesizing chemically inert tags prior to the pooling step. The general strategy for library synthesis exploits existing functionality present on the natural products, and should be applicable to other families of secondary metabolites.

Chromatography, High Pressure Liquid↗
PubMed1996

Effect of the hydroalcoholic extract of rauwolfia ligustrina on smooth and cardiac muscles in-vitro.

The actions of the hydroalcoholic extract (HE) of Rauwolfia ligustrina (the whole plant) on agonist-induced contractions were analysed in the rat uterus and guinea-pig ileum and trachea, and also in rat atrium contracting spontaneously in-vitro. The HE (100-400 micrograms mL-1) caused concentration-dependent rightward shifts of the concentration-response curves and reduced the maximal contraction induced by oxytocin, bradykinin, angiotensin II, prostaglandin F2 alpha and acetylcholine in the rat uterus. The calculated mean IC50 values were: 300, 147, 158, 197 and 105 micrograms mL-1, respectively. In the guinea-pig ileum the HE also caused graded displacement to the right of the concentration-response curves for bradykinin, histamine and carbachol, associated with pronounced inhibition of the agonist-induced maximal contractions. The calculated mean IC50 values were 165, 134 and 241 micrograms mL-1, respectively. The HE (100-3000 micrograms mL-1) caused graded relaxation (IC50 of 271 micrograms mL-1) of strips of guinea-pig trachea precontracted with carbachol (0.2 microM). This effect was not influenced by propranolol (1 microM), 3-isobutyl-1-methylxanthine (1 microM) or methylene blue (10 microM), but was significantly potentiated (1.5-to 3-fold) by indomethacin (3 microM) and forskolin (1 nM). In addition, NG-monomethyl-L-arginine (L-NMMA, 100 nM) significantly reduced the HE-induced maximal relaxation, while indomethacin (3 microM) potentiated the HE response. Finally, the HE caused a concentration-dependent and long-lasting inotropic effect in the rat right atrium, contracting spontaneously with a mean EC50 value of 409 micrograms mL-1. It is suggested that the effects of the HE of R. ligustrina on smooth and cardiac muscles "in-vitro' may result from its ability to interact, at least partially, with the cAMP pathway.

Animals↗
PubMedDate not supplied

Novel glucoalkaloids from Rauwolfia cell cultures--acetylrauglucine and related glucosides.

From cell suspension cultures of Rauwolfia serpentina grown in an optimized production medium for the glucoalkaloid raucaffricine, a novel glucoalkaloid was isolated and identified as 17-O-acetyl-21-O-beta-D-glucopyranosyl-ajmaline (acetylrauglucine). This alkaloid is formed in very small amounts (less than 5 x 10(-4)%). The biogenetically related N alpha-demethylated base (acetyl-nor-rauglucine) and the deacetyl product rauglucine have also been detected in culture extracts. In addition 21(R)-(beta-D-glucopyranosyl)-hydroxy-sarpagan-17-al has been isolated and identified as an artifact which originates from raucaffricine.

Ajmaline↗
PubMed1977

[The effect of the rauwolfia-alkaloids ajmaline, rescinnamine and reserpine on the catecholamine contents of the heart (author's transl)].

The effect of three rauwolfia-alkaloids and two semi-synthetic derivatives on the catecholamine content of the hearts of guinea pigs was examined. The substances were: ajamaline, N-methyl-ajamaline, N-n-propylajmaline, rescinnamine and reserpine. The presence of catecholamines in isolated atria was tested with tyramine (10(-5) g/ml). Rescinnamine and reserpine depleted the hearts of catecholamines, ajmaline and its derivatives did not.

Ajmaline↗
PubMed2005

Characterisation of oxidation products of Rauwolfia alkaloids.

The oxidation products of three Rauwolfia alkaloids having yohimbane skeleton, namely, reserpine, rescinnamine and ajmalicine have been characterised by chromatographic and spectroscopic techniques. The Rf values of the oxidation products (3, 4-dehydro derivatives) in several solvent systems and the spectral data on UV, IR, NMR and fluorescence characteristics have been determined to confirm the structure of these compounds.

Chromatography, Thin Layer↗
PubMed1991

Acyclic monoterpene primary alcohol:NADP+ oxidoreductase of Rauwolfia serpentina cells: the key enzyme in biosynthesis of monoterpene alcohols.

Acyclic monoterpene primary alcohol:NADP+ oxidoreductase, a key enzyme in the biosynthesis of monoterpene alcohols in plants, is unstable and has been only poorly characterized. However we have established conditions which stabilize the enzyme from Rauwolfia serpentina cells, and then purified it to homogeneity. It is a monomer with a molecular weight of about 44,000 and contains zinc ions. Various branched-chain allylic primary alcohols such as nerol, geraniol, and 10-hydroxygeraniol were substrates, but ethanol was inert. The enzyme exclusively requires NADP+ or NADPH as the cofactor. Steady-state kinetic studies showed that the nerol dehydrogenation proceeds by an ordered Bi-Bi mechanism. NADP+ binds the enzyme first and then NADPH is the second product released from it. Gas chromatography-mass spectrometric analysis of the reaction products showed that 10-hydroxygeraniol undergoes a reversible dehydrogenation to produce 10-oxogeraniol or 10-hydroxygeranial, which are oxidized further to give 10-oxogeranial, the direct precursor of iridodial. The enzyme has been found to exclusively transfer the pro-R hydrogen of NADPH to neral. The N-terminal sequence of the first 21 amino acids revealed no significant homology with those of various other proteins including the NAD(P)(+)-dependent alcohol dehydrogenases registered in a protein data bank.

Alcohol Oxidoreductases↗
PubMed1985

Fluorescence histochemical studies of the effects of rauwolfia alkaloid derivatives on adrenergic vasomotor nerves.

Experiments were performed to explore the influence of two rauwolfia alkaloid derivatives, rescinnamine and syrosingopine, on noradrenaline at the sympathetic nerve-endings by delineating the localization of noradrenaline in the walls of femoral artery and abdominal aorta of rats treated with these agents compared to reserpine, using the fluorescence histochemical technique. Noradrenaline-containing nerve fibers emitting an intense yellowish-green fluorescence were demonstrated circumjacent internally to the adventitial smooth muscle layer of the femoral artery in both normal rats and spontaneously hypertensive rats (SHR). In both normal rats and SHR, treatment with rescinnamine or syrosingopine in an oral dose of 5 mg/kg caused essentially the same degree of depletion of the noradrenaline fluorescence localized subjacent to the adventitial smooth musculature of the femoral artery, as did that with 5 mg/kg of reserpine. The noradrenaline fluorescence vanished completely following treatment of rats with 15 mg/kg of rescinnamine or 30 mg/kg of syrosingopine. Noradrenaline-containing nerve fibers showing yellowish-green fluorescence were observed distributed subjacent to the entire internal circumference of the adventitial smooth muscle layer of the abdominal aorta in both the normal rats and SHR. In response to oral administration of rescinnamine or syrosingopine in a dose of 5 mg/kg, both normal and SHR rats showed a diminution of the noradrenaline fluorescence in the region of adventitial smooth musculature of abdominal aorta equivalent to that evoked by 5 mg/kg of reserpine. Treatment with 15 mg/kg of rescinnamine or 30 mg/kg of syrosingopine resulted in a complete disappearance of noradrenaline fluorescence in that region.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic Fibers↗
PubMed1986

Effects of single doses of two antihypertensive rauwolfia-diuretic combinations on urinary excretion.

In a double-blind placebo-controlled investigation, 14 healthy adult male volunteers were studied to assess and compare the urinary effects of acute single doses of two antihypertensive formulations containing: hydroflumethiazide 50 mg, Rauwolfia serpentina 50 mg and potassium chloride 625 mg (HFRK) (Rautrax-50; Squibb Laboratories); and clopamide 5 mg, reserpine 0.1 mg and dihydroergocristine 0.5 mg (CRE) (Brinerdin; Sandoz Products). Both significantly increased mean 24-hour urinary outputs of fluid, Cl-, Na+, K+ and Mg2+. CRE increased creatinine and decreased Ca2+ output. After dosing, times to maximal urinary flow of fluid, Cl-, Na+, Ca2+ and creatinine were shortened by HFRK and CRE and those corresponding to K+, Mg2+, phosphate and urate were unaffected. Both formulations thus acted mainly through their diuretic constituents.

Adolescent↗
PubMed1976

Rauwolfia derivatives and breast cancer.

During the course of a collaborative study initiated in investigate the relationship of breast cancer and the use of hormones, it was possible to study the possible relationship of rauwolfia derivatives, mainly reserpine, to breast cancer. Interviews were administered to breast cancer patients admitted to five hospitals in the Baltimore metropolitan area. The study of reserpine use was conducted for a period of approximately one and a half years and information on reserpine use was obtained on 164 breast cancer cases. Each case was matched with a control with respect to age, color, hospital of admission, and admission date. An additional set of controls was drawn from women residing in the same neighborhood as the cases. Neither reserpine, methyldopa, nor thiazides were found to be significantly associated with breast cancer. The exclusion from the analysis of hospital controls with a diagnosis of hypertensive disease and their matched cases did not significantly alter the results. Those cases who used reserpine, used more for longer periods of time than did the controls, however this was judged inconclusive due to a considerable amount of missing data. After comparison of these results with previously reported studies, it did not appear that reserpine was strongly associated with breast cancer. However, because of methodological problems and differences between the studies, a possible small overall relationship could not be definitely excluded.

Adolescent↗