Investigations into the specific effects of rosemary oil at the receptor level.

Rosemary oil is used frequently in phytotherapy. The objective of the present study was to investigate the extent to which rosemary oil shows other effects on the smooth muscles than the familiar spasmolytic effects. The effects of rosemary oil on the spontaneous contractile activity were investigated in in vitro experiments with circular smooth-muscle strips of the guinea pig stomach. Rosemary oil was found to have agonistic effects on the alpha(1) and alpha(2) adrenergic receptors. These effects can be registered at concentrations up to 25 microl/l of rosemary oil. At higher concentrations the spasmolytic effect described in other reports could be detected. At concentrations above 100 microl/l rosemary oil, the effect of 10(-5)M ACH is completely suppressed. The results permit the assumption that, besides the spasmolytic effects investigated to date, owing to its specific effects on the alpha(2) adrenergic receptors of the nerve cells, rosemary oil brings about an additional improvement of local blood circulation and alleviates pain.

Effect of Thymol on the spontaneous contractile activity of the smooth muscles.

Effects of Thymol on the spontaneous contractile activity (SCA) have been found in in vitro experiments with circular smooth-muscle strips (SMAs) from guinea pig stomach and vena portae. Thymol was found to possess an agonistic effect on the alpha(1)-, alpha(2)- and beta-adrenergic receptors. Its spasmolytic effect is registered at doses higher than 10(-6)M. Thymol in a dose of 10(-4)M inhibits 100% the SCA of the SMAs and reduces the excitatory effect of 10(-5)M ACH to 35%. It is assumed that Thymol has an analgesic effect through its action on the alpha(2)-adrenergic receptors of the nerve cells. By influencing the beta-adrenergic receptors in the adipose cells, it is possible to induce increased synthesis of fatty acids and glycerol, which is a prerequisite for increased heat release.

[The influence of the initial humidity of balneological peat on its pharmacological features]. / Der Einfluss des initialen Feuchtigkeitsgrades von Torf auf die pharmakologischen Eigenschaften von Badetorf.

The viscosity of balneological peat got special meaning according to its thermical features. Until now, there has been no answer on the question in how far balneological peat, which is prepared with different degrees of humidity, can vary the pharmacological character. Aim of this work was, on the one hand, to examine which initial degree of peat-humidity is necessary to get the wished viscosity in therapy when diluted with water. On the other hand, how far the pharmacological character of balneological is influenced by the initial degree of humidity. From peat, with initial different degrees of humidity, balneological peat is prepared, fulfilling the demands of the Quentin-Test and therefore the necessary consistence for therapy. Here in this work the in-vitro effect of aqueous peat extract (APE) on the spontaneous contractile activity of the smooth muscle fibres of guinea-pig stomachs was examined. The APE used got initial different degrees of humidity. It was found out, that the ability of peat according to a new water-uptake, with a initial humidity under 60%, was clearly reduced. Because of the reduced water content of balneological peat its thermical features were deteriorated. This goes for peat with a humidity factor < 60%. First, with the Quentin-Test, showing the optimal viscosity of peat before medical use, you can also make statements according its ability of maximum uptake of water, meaning the ability taking up water until it becomes gel-like. Second, the results show that the initial humidity of peat got no influence on the pharmacological features.

Comparative study of the thermal properties of mud and peat solutions applied in clinical practice.

Different peloids as e.g. mud and peat have been traditionally used for therapeutic purposes successfully, especially of there thermal actions. It was the aim of the experimental study to compare the thermal properties of two peloids, mud and peat, with a view to assessing their thermal effects when they are applied in clinical practice. The studies were carried out using peat of the marsh type of peats (Hochmoor), and curative Pomorie (Bulgaria) mud. As important parameters were determined the specific thermal capacity at constant pressure (Cp), the density of solutions (rho), the cooling rate (m), the coefficient of temperature transfer (a) of solutions and the coefficient of thermal conductivity (lambda) of solutions of peat and curative mud, compared to water bath. The comparative studies of the thermal properties of water and water solutions of peat and curative mud show that the thermal effect of the water bath is substantially smaller than that of the peat and mud applications. This difference is due to a greater extent to the high values of the dynamic viscosity, not allowing cooling by convection and protecting the surface of the skin upon applications of peloid solutions with a higher temperature.

Evaluation of the permeation of peat substances through human skin in vitro.

Peat and various peat extracts have been successfully applied for a variety of clinical indications. Quite apart from the physico-thermal effects, new studies point towards the so-called "chemical effects" of peat containing substances. These effects include a stimulatory response of the spontaneous contractile activity (SCA) of smooth muscle (SM) tissue. The effects are, however, dependent on the possible permeability of pharmacologically active substances as naturally occurring ingredients of peat. Since peat is a mixture of various products it is necessary to examine the various peat types based upon their biological activity on SM tissue. In order to unequivocally prove the pharmacological activity of cutaneous peat treatment, in vitro permeation measurements of these actives across excised human skin can be used.HPLC analysis revealed that aqueous peat extracts contain up to 18 fractions of water-soluble compounds of fulvic and ulmic acids. These compounds have been found to have a stimulatory response on the contractile activity of SM tissue. In vitro diffusion studies showed that the permeability of these substances across human full thickness skin (thickness: 200 um(-1)) is highly selective and the resulting stimulatory activity is dependent on the permeated fraction. Especially, the HPLC fractions 7-11 and 14 are able to permeate human skin. Fractions 7-11 show a moderate stimulatory effect of SCA on SM for more than 90 min whereas fraction 14 shows the strongest stimulatory effect which was, however, suppressed after 87 min. These results show that the cutaneous therapy with peat treatment results in transcutaneaous permeation of biologically active fulvic and ulmic acid derivatives explaining the additional "chemical" effect of peat treatment in clinical practice.

Isolation of biologically active fractions from the water soluble components of fulvic and ulmic acids from peat.

With a view to clarifying the excitatory action of aqueous peat extract (APE) on the spontaneous contractile activity (SCA) of the smooth muscles, in vitro studies were made of the influences of the water-soluble HPLC-fractions of fulvic and ulmic acids separated from peat on smooth-muscle preparations of guinea-pig stomach. The results obtained show that peat contains a large number of water-soluble components, which possess agonistic or partial agonistic actions on the alpha2-adreno- and D2-dopamine receptors. These are chemically stable substances, which retain for months their biological activity unchanged in aqueous solution. The excitatory effect of APE on the SCA of the smooth-muscle preparations was found to be more pronounced than the reactivity of alpha2-adrenoreceptors.

Metal distribution and binding in balneological peats and their aqueous extracts.

Binding of metals in typical bath peat samples ("Grosses Gifhomer Moor", Sassenburg/North Germany) and their aqueous extracts was characterized by means of a multi-method approach. For that purpose a sequential extraction procedure based on peat-filled chromatography columns was developed. Water-soluble metal and DOM (dissolved organic matter) fractions were subdivided by use of a stepwise increased pH gradient (pH 3.8-5), finally by the chelator EDTA and 0.1 mol L(-1) hydrochloric acid. Metal fractions very strongly bound to peat were assessed by an aqua regia extraction. Metal determinations required were performed by atomic spectrometry methods (AAS, ICP-OES, and TXRF). The metal and DOM concentrations in the peat extracts varied significantly, depending on the natural variety of the peat matter under study (e.g., Al: 25-674, Cd: 0.05-0.2, Cu: 5-15.4, Fe: 77-1785, Mn: 21-505, Ni: 2-33, Pb: < 1, Zn: 9-715 (microg L(-1)); Na: 8-45, K: 1.3-14.9, Ca: 2-51, Mg: 1.1-7.9 (mg L(-1)); 26-73 mg L(-1) DOC). An increase of the pH increased the DOC (dissolved organic carbon) of the peat extracts, but hardly the concentration of heavy metals. The latter could only be re-mobilized by EDTA and dilute hydrochloric acid. Additional investigations of the peat extracts using tangential-flow ultrafiltration revealed that the heavy metals extracted at pH < 4 were predominantly dissociated. At higher pH (pH > 4.5) they were preferentially bound to macromolecular DOM. Moreover, using multistage ultrafiltration the size distribution of the DOM and their metal species was assessed.

The effect of peat components on endocrine and immunological parameters and on trace elements--results of two pilot studies.

Peat and different peat preparations are successfully used in clinical therapies for different indications (as, for instance, in the field of gynecology). New studies show the biochemical effects of peat components which they have aside from their physical-thermal effects. This is of extraordinary interest with regard to the medical use of peat, because considerable concentrations of trace elements and heavy metals have been found in different kinds of peat. By means of atomic spectrometry it was investigated in 17 female patients with irritable bladder whether and how variations of the concentration of special trace elements and heavy metals (lead, cadmium, copper, manganese) could be measured within 24-hour urine after vaginal peat-mush treatments had been applied serially. Additionally, the effect of peat-mush baths compared to the effect of water baths (n=6) - both of which were applied to 17 female patients with degenerative diseases - was examined with regard to their special endocrinological parameters. The results concerning safety did not show any changes of the concentration of the trace-elements or heavy metals within the 24-hour urine. These results can be explained by the chelating features of the peat components, which are the reason for the absorption of the trace elements. Examinations done to compare the effects of peat-mush baths and water baths have shown that peat components - independent from their thermal effects - are the reason for the occurrence of special effects. This applies in particular to the parameter soluble interleukin-2-receptor. As regards estradiol, a significant increase could be measured after peat-mush baths had been applied to 17 postmenopausal female patients (n=11). Comparing these results with those of the group of patients treated with water baths, we noticed that the increase of estradiol was remarkably lower and not significant. The effect of the peat components is thought to be the reason for this.

[Mechanism of the effect of aqueous peat extract on spontaneous contractile activity of smooth muscles]. / Der Wirkungsmechanismus von wässrigem Torfextrakt auf die spontane kontraktile Aktivität der glatten Muskulatur.

BACKGROUND: There are reports in the literature about the effects of peat and its products on spontaneous contractile activity (SCA) of smooth muscles. In our earlier work we showed that the aqueous peat extract is a stimulus for SCA of smooth muscle fibres. After longstanding (4-5 h) influence from aqueous peat extract, a self-blocking effect of these influences can be observed. The mechanism of action could not be determined up to now. PURPOSE: The purpose of this study is to elucidate the peat mechanism of action. MATERIAL AND METHODS: In organ baths, the effects of aqueous peat extract on the SCA from smooth muscles have been studied under normal conditions and with presence of different receptor blocking substances. RESULTS: Our investigations show that only with simultaneous blocking of alpha(2) and D(2) receptors the stimulating activity of aqueous peat extract on the SCA of smooth muscle fibres can be suppressed. The self-blocking actions of the stimulating activity of aqueous peat extract on the SCA of smooth muscles are interconnected with blocking of alpha(2 ) adreno and D(2) receptors. CONCLUSION: The aqueous peat extract contains biologically active substances, which have a partial agonistic effect on alpha(2 ) and D(2) dopamine receptors. Corresponding to the results we start out at pharmacological effects as they are known from the mentioned receptors. The clinical efficacy have to be shown by clinical trials.

The influence of fulvic and ulmic acids from peat, on the spontaneous contractile activity of smooth muscles.

The aim of the presented studies was to evaluate which classes of compounds of peat ingredients could be responsible for the partial agonistic effect of aqueous peat extract on the alpha2 adreno and D2 dopamine receptors of smooth muscles, which we have reported from former investigations. Based on the different solubility of peat ingredients, water-soluble components of fulvic and ulmic acids were separated according to the pH-value and chemical structure of the solvent. The biological activity of these acids was examined in peat baths using smooth muscle fibers of guinea pig stomach. The results demonstrate that the water-soluble components of fulvic and ulmic acids have a partially agonistic effect on the alpha2 adreno and D2 dopamine receptors, but at the same time quite different effects in terms of their influence on the spontaneous contractile activity (SCA) of smooth muscles have to be noted. From these investigations, we can conclude that aqueous peat extract and the water-soluble components of fulvic acid exhibit similar partial agonistic effects on the alpha2 adreno and D2 dopamine receptors. Therefore it is likely that the mentioned effects of the peat extract derive from the fulvic- and ulmic acids only. The water-soluble component of ulmic acid also showed partial agonistic effects on alpha2 adreno and D2 dopamine receptors. In this case, another substance group is involved, which has a faster blocking effect on these receptors, but was barely soluble in water at a normal pH-value (pH 7).

Effect of fractions of water extract from peat with a specific molecular weight on the spontaneous contractile activity of guinea pig stomach smooth muscles.

OBJECTIVE: Peat and water extracts from peat have been reported in the literature to have an effect on the spontaneous contractile activity of certain smooth muscles. The purpose of the present study was to investigate the effect of organic substances of various molecular weight contained in the water peat extracts on the spontaneous contractile activity of smooth muscles. METHODS: Using Amicon Centriprep molecular filters the substances contained in the water extracts from peat were separated into fractions of various molecular mass. Their effect on spontaneous contractile activity was studied on strips of guinea pig stomach. RESULTS: All fractions containing the low-molecular weight component of the water peat extract (under 3000 Da) showed an excitatory effect on smooth muscle spontaneous contractile activity. The spontaneous contractile activity of smooth muscles was not influenced by the microelements Ba, Mn, and Cu applied in doses equal to the original water peat extract. CONCLUSION: The excitatory effect of water peat extracts on the spontaneous contractile activity of smooth muscles is due to the organic compounds with molecular weight under 3000 Da.

The ethosuximide-induced hyperpolarization of smooth muscle tissues--a cause of functional changes in the gastrointestinal tract of rats--is provoked by CA(2+)-dependent K(+)-efflux.

Ethosuximide is an antiepileptic drug successfully used in the treatment of petit mal especially in childhood. Clinical investigations reveal that ethosuximide has a number of adverse side effects on the gastrointestinal tract (GIT) of patients which may include heaviness, anorexia, pains in the region of the stomach, accompanied sometimes with nausea and vomiting. In the present study we attempt to explain the mechanisms of some of the drug's adverse side effects using an experimental animal model. White rats were employed in the experiments during which they were treated daily with ethosuximide (100 mg/kg) for 100 days. We made use of the following methods in the study: 1. Contrast X-rays study of the gastrointestinal tract. 2. Recording of the bioelectric activity of isolated smooth muscle strips using the sucrose-gap method. 3. Recording of the contractile activity of isolated smooth muscle. Characteristic changes occurring after treatment of rats with ethosuximide include atony of the stomach and the intestines, hypertonus in separate duodenal segments, diminished peristaltic activity, and delayed passage at the 24th hour. The drug inhibits the spontaneous contractile activity of isolated smooth muscle strips from rat gastrointestinal tract. In some duodenal preparations is minimised in the presence of apamin, an inhibitor of Ca2+(-dependent) K+ channels (Ka+(Ca)), while in duodenal preparations this effect may undergo a reversion. In the presence of coffeine the action of ethosuximide on the smooth musculature of rat gastrointestinal tract is reduced significantly. We hypothesis that the observed functional changes in the gastrointestinal tract occur as a result of the hyperpolarizing effect of ethosuximide on the gastrointestinal musculature caused by the outcoming K+(CA)-efflux.

Effect of sodium valproate on the spontaneous contractile and bioelectric activity of smooth muscle fibres isolated from experimental animals.

Experiments were carried out with smooth muscles preparations of 24 rats (gastric corpus) and 36 guinea pigs (gastric corpus and taenia coli). Their contractile responses were recorded isometrically using tensile transducers. The bioelectric activity was determined by the single sucrose-gap method. Depending on the concentration sodium valproate elicits two types of responses from the smooth muscle of gastrointestinal tract of rats and guinea pigs: a contractile response, at concentrations less than 10(-4) g/ml, and a relaxant response, at concentrations higher than 10(-4) g/ml. At concentrations below 10(-4) g/ml sodium valproate induces contractions of the smooth muscle taken from the rat and guinea pig gastrointestinal tract. These contractions can be abolished by indomethacin. It may be concluded, therefore, that the contractile effect of sodium valproate has a prostaglandin basis. At concentrations higher than 10(-4) g/ml, sodium valproate hyperpolarizes the smooth muscle tissues, reduces the frequency and amplitude of the spontaneously generated spike potentials and relaxes the preparations. These effects are blocked by picrotoxin. Sodium valproate in high concentrations increases the endogenous level of the gamma-aminobutyric acid stimulating at the same time the prostaglandin synthesis which is inadequate to compensate the relaxant effect induced by elevated level of the endogenous gamma-aminobutyric acid.

Effects of the medium chain octanoic fatty acid on the contractile activity of vascular smooth muscle tissues.

Multiple elevation of the octanoic fatty acid level in human sera is often observed in some hepatic diseases which are accompanied by cardiovascular disorders, hypotension, and increased cardiac stroke volume. Different experiments reveal the hypotensive effect of octanoate. The present study investigates the octanoate action on the bioelectric and contractile activity of vascular smooth muscle tissues. Octanoate is shown to cause hyperpolarization of smooth muscle cells, reduction of spike potential frequency, and relaxation of vascular muscles. These effects are inhibited by indomethacin, which proves their prostaglandin nature. Octanoate action on stomach smooth muscle strips is inhibited by SC 19220, a specific competitive inhibitor of the contractile action of prostaglandins E2 and F2 alpha on the same tissues in vitro. Using thin-layer chromatography two PG fractions were isolated from arterial blood of octanoate treated rats and from a control group of rats. These fractions have identical chromatographic characteristics with those of PGE2 and PGF2 alpha. The level of the PG fraction with Rf value similar to that of PGE2 is significantly increased in octanoate treated animals while the other fraction tends to decrease.

Influence of short-chain fatty acids on vascular tone.

Results from research on the influence of short-chain fatty acids (SCFA) (C3-C6) on vascular tone are reported. Isolated vascular smooth muscle strips were studied in vitro and the arterial blood pressure of guinea-pigs injected with SCFA was taken. On the basis of results from these first two methods, the level of PG F2 alpha was determined by radioimmunoassay. The experiments indicate that in vitro the SCFA have spasmogenic effects which are blocked by indomethacin and aspirin and which are reduced by the inhibitor of the activating effect of PG F2 alpha, PG E2, and PG I2-SC19220. The effects of SCFA on smooth muscle strips used as a bioassay are analogous to the effects of PG F2 alpha on the same tissue. The injection of butyric acid into guinea-pigs causes hypertension which is not manifest if indomethacin pretreatment is carried out. Radioimmunoassay results indicate that the level of PG F2 alpha in the blood of animals treated with butyric acid is significantly increased.

Determination of certain physical and chemical characteristics of the activating serum factor from preparturient women and women with habitual abortions.

Follow-up investigation of the blood sera from preparturient women and women with habitual abortions showed the presence of a factor which has an activating effect on smooth muscle preparations because it causes the release of prostaglandins. Gel-chromatographic counter flow separation and microelectrophoresis of the blood sera have shown that the isolated serum factor is a water soluble glycopeptide with a molecular weight of about 2000.