Valerian extract characterized by high valerenic acid and low acetoxy valerenic acid contents demonstrates anxiolytic activity.

Valerian is one of the most commonly used herbal remedies for the treatment of insomnia and anxiety. Valerian extracts allosterically modulate GABAA receptors, an action related to valerenic acid, which is one of the active compounds determined from pharmacological studies. Derivatives of valerenic acid, i.e. acetoxy valerenic acid or hydroxy valerenic acid, do not allosterically modulate GABAA receptors, but they bind to identical binding sites. Therefore, the question arises whether they might interfere with the effects of valerenic acid. Two valerian extracts were tested in the elevated plus maze test and the tail suspension test for anxiolytic and antidepressive activity, respectively. Reference substances were diazepam (1.0mg/kg) and imipramine (30mg/kg). The extracts were standardized to the identical total amounts of the acids (0.1; 0.5; 1.0 and 2.0mg/kg), i.e. valerenic and acetoxy valerenic acid, but the ratio between the acids was different (12:1 and 1:1.5). The extract with the ratio 12:1 prolonged the time spent on the open arm significantly when 0.5mg/kg was applied. Of the other extract, with the ratio 1:1.5, four times that amount was required (2.0mg/kg). Both of the tested extracts did not show any antidepressive effect, rather the other way around, the extract with the ratio 1:1.5 prolonged the immobility phase. However, since the core body temperature was reduced by the 1.0 and 2.0mg/kg extract dose, the prolongation may be related to the temperature phenomenon and is not indicative of a specific depressive action. In conclusion, the anxiolytic activity of the valerian extract seems rather related to valerenic acid and, moreover, standardization with respect to the total amount of valerenic acids, i.e. valerenic acid together with acetoxy valerenic acid, is misleading.

The effect of genotype and cultivation method on the total salicylate yield of dark-leaved willows (Salix myrsinifolia).

Ten clones of dark-leaved willows ( Salix myrsinifolia Salisb.) were grown for two years on two different sites, Luikonlahti and Punkaharju, using various combinations of soil cultivation, fertilization and mulch treatments, with the objective of comparing the effect of different cultivation methods on the growth and total salicylate yield. In cultivated soil, the total salicylate yield ranged from 20 to 220 kg/ha, depending on the clone and cultivation method. The use of black polythene mulch and the addition of mineral fertilizer reduced the total salicylate concentrations of several clones. On the other hand, polythene mulch noticeably increased the hectare yield of total salicylates by promoting biomass accumulation.

[Efficacy of tea blends in the treatment of dyspeptic disorders--an application observation]. / Wirksamkeit von Teemischungen bei dyspeptischen Beschwerden - eine Anwendungsbeobachtung.

BACKGROUND: Medicinal teas present one of the oldest galenic preparations of herbal remedies. Their use is primarily determined by tradition and empirics. One of their traditional domains are gastrointestinal disorders, which belong to the most frequent wellness disorders. While the effectiveness and compatibility of essential oils, modern drug extracts, and alcoholic extracts from bitters and etheric-oil drugs in the treatment of dyspeptic disorders have been documented in placebo-controlled clinical trials, little attention has so far been given to aqueous extracts from bitters and etheric-oil drugs, which are equivalent to the standard method of preparing herbal teas. PURPOSE: The presented application observation in clinical practice tried to give evidence for effectiveness and tolerability of medicinal teas in the treatment of dyspeptic disorders and to quantify their extent. MATERIAL AND METHODS: We collected information about effectiveness, compatibility, and side effects of herbal teas from 89 patients (w = 56; m = 33) suffering from dyspeptic disorders. The data were reported with a questionnaire that was sent to physicians and pharmacists experienced in phytotherapy. RESULTS: It could be shown that complaints in patients with primary dyspeptic symptoms (n = 79) decreased by an average of 74%. Final overall assessment revealed that the physicians as well as the patients estimated an effectiveness of 2.9 points as good (3 = good). Compatibility was considered as good to very good (4 = very good), with an average rating of 3.3 points. Two patients stopped therapy because of an extreme aversion to the bitter taste of the teas. No other serious side effects were reported. CONCLUSION: The herbal teas can be considered effective, very well tolerable and to a large extent free from serious side effects. However, due to the limited observation time, no final conclusion could be given concerning long-term compatibility.

[From medicinal plant to phytotherapeutic drug]. / Von der Arzneipflanze zum Phytotherapeutikum.

The effects of phytotherapy are generally attributed to a specific plant as a whole. However, plants only provide the raw material from which the active ingredients are collected by special procedures. Most of the herbal medicinal preparations as the active ingredients in herbal medicinal products are obtained by extraction using water or organic solvents such as ethanol or methanol. This yields a large variety of preparations with diverse properties. Particular to phytotherapeutic preparations in contrast to synthesized drugs is that they contain a complex mixture of active principles that is difficult to characterize chemically and biologically. Nevertheless, the quality of medicinal preparations from plants can be guaranteed if the raw material, the mode of preparation and the end product are well defined by the manufacturer.

Comparing phytopharmaceuticals: the example of St. John's Wort.

At present, dosage in phytotherapy often falls victim to an undifferentiated point of view. In the case of St. John's Wort, for example, 900 mg of extract has become the standard dosage; however, no studies have investigated the dose-effect ratio, even though there are numerous indications that smaller amounts, particularly of ethanolic extracts, are sufficient to achieve the desired effect. Moreover, listing extract dosages is of no use unless other identifying characteristics are also specified. If phytopharmaceuticals are to be compared with one another, the scope of consideration cannot be limited to single components. The comparison must be carried out comprehensively.

Absorbance data of hypericin and pseudohypericin used as reference compounds for medicinal plant analysis.

The evaluation of the absorbance data of hypericin and pseudohypericin revealed the molar/specific coefficients of absorbance in methanol-pyridine (99:1, v/v) at the maximum of the longest wavelength to be 51936/1030 and 43486/836, respectively. The absorbance data of hypericin were also determined in methanol. They were not significantly different from those in the presence of pyridine. The decrease of the coefficients by water addition was found to be the same for hypericin and pseudohypericin. It was concluded that hypericin and pseudohypericin reveal the same homoassociation behavior.

Efficacy of Vitex agnus castus L. extract Ze 440 in patients with pre-menstrual syndrome (PMS).

In a prospective, multicentre trial the efficacy of an Vitex agnus castus L extract Ze 440 was investigated in 50 patients with pre-menstrual syndrome (PMS). The patients were treated daily with one tablet (20 mg native extract) during three menstrual cycles. 43 patients completed the study protocol which encompassed 8 menstrual cycles (2 baseline, 3 treatment and 3 post-treatment). 13/43 patients were receiving concomitant oral contraceptives. 6 patients did not complete the study for reasons not related to study medication, and one patient complained of fatigue possibly related to study medication. All evaluated patients took at least 85% of the prescribed medication. The main effect parameter was the validated Moos' menstrual distress questionnaire (MMDQ), and secondary parameters were a visual analogue scale (VAS; self-assessment) and a global impression scale (GI, self-assessment). The study population was homogenous in age (31.3+/-7.7 years) weight (58.9+/-6.9 kg) and cycle length (28.4+/-0.3 d). The diagnosis was according to DMS-III. At the end of the study, PMS-related symptoms were reduced by treatment. There was a significant score reduction (42.5%) of the MMDQ as the main effect parameter (p<0.001). Symptoms gradually returned after treatment cessation. However, a difference from baseline remained (20%; p<0.001) up to 3 cycles thereafter. 20/43 patients were considered "responders", with a reduction in MMDQ score by at least 50% relative to baseline. At baseline, the VAS score was elevated in the late luteal phase and low at the follicular phase, as expected. During treatment, VAS score decreased in the late luteal phase (47.2%; p<0.01) and remained 21.7% (p<0.001) below baseline after 3 cycles post-cessation of treatment. The low VAS score within the follicular phase remained unchanged over the whole observation period. 38 patients judged the global efficacy moderate to excellent, 5 patients indicated no global efficacy. The number of days patients sustained PMS symptoms was reduced slightly from 7.5 to 6. Resting levels of blood prolactin remained within the physiological range throughout. No differences were seen between patients on or off oral contraceptives. 20 patients reported 37 adverse events (AE). No serious AE were reported. One patint withdrew after four days of treatment due to fatigue and headache. Laboratory safety control parameters were not affected. In conclusion, patients with PMS can be treated successfully with Vitex agnus-castus extract Ze 440, as indicated by clear improvement in the main effect parameter during treatment and the gradual return after cessation of treatment. The main response to treatment seems related to symptomatic relief rather than to the duration of the syndrome.

Pharmacological activities of Vitex agnus-castus extracts in vitro.

The pharmacological effects of ethanolic Vitex agnus-castus fruit-extracts (especially Ze 440) and various extract fractions of different polarities were evaluated both by radioligand binding studies and by superfusion experiments. A relative potent binding inhibition was observed for dopamine D2 and opioid (micro and kappa subtype) receptors with IC50 values of the native extract between 20 and 70 mg/mL. Binding, neither to the histamine H1, benzodiazepine and OFQ receptor, nor to the binding-site of the serotonin (5-HT) transporter, was significantly inhibited. The lipophilic fractions contained the diterpenes rotun-difuran and 6beta,7beta-diacetoxy-13-hydroxy-labda-8,14-dien . They exhibited inhibitory actions on dopamine D2 receptor binding. While binding inhibition to mu and kappa opioid receptors was most pronounced in lipophilic fractions, binding to delta opioid receptors was inhibited mainly by a aqueous fraction. Standardised Ze 440 extracts of different batches were of constant pharmacological quality according to their potential to inhibit the binding to D2 receptors. In superfusion experiments, the aqueous fraction of a methanolic extract inhibited the release of acetylcholine in a concentration-dependent manner. In addition, the potent D2 receptor antagonist spiperone antagonised the effect of the extract suggesting a dopaminergic action mediated by D2 receptor activation. Our results indicate a dopaminergic effect of Vitex agnus-castus extracts and suggest additional pharmacological actions via opioid receptors.

Quantitative high performance liquid chromatographic analysis of diterpenoids in agni-casti fructus.

Pharmacological data have indicated that part of the dopaminergic activity of Vitex agnus-castus is attributed to the labdan diterpenoids found in the fruits. Therefore an analytical method for the standardization of rotundifuran (1), vitexilactone (2) and 6 beta,7 beta-diacetoxy-13-hydroxy-labda-8,14-diene (3) was developed. Because of the time-consuming and expensive isolation of the diterpenoids, p-cymene was chosen as an internal standard. The concentration of rotundifuran in different extracts and trade samples of the drug varies between 0.04 and 0.30% in the drug and between 1.04 and 2.23% in the extract. The concentration of vitexilactone was generally lower between 0.016 and 0.167% for the drug and between 0.34 and 1.01% for the extract. The determined concentration of 6 beta,7 beta-diacetoxy-13-hydroxy-labda-8,14-diene in the drug was in the range of 0.02 and 0.10% and in the extract in the range of 0.18 and 0.80%. Determination of the factors of correction of p-cymene gave 5.63 for rotundifuran, 2.73 for vitexilactone and 3.74 for 6 beta,7 beta-diacetoxy-13-hydroxy-labda-8,14-diene.

Bisanthraquinone glycosides of Hypericum perforatum with binding inhibition to CRH-1 receptors.

Four new bisanthraquinone glycosides, S-(+)-skyrin-6-O-beta-glucopyranoside (1), R-(-)-skyrin-6-O-beta-glucopyranoside (2), S-(+)-skyrin-6-O-beta-xylopyranoside (3) and S-(+)-skyrin-6-O-beta-alpha-arabinofuranoside (4), have been isolated from an ethanol-water (1:1, v/v) dry extract of the aerial parts of Hypericum perforatum L. The structures were elucidated by spectroscopic methods, mainly NMR and mass spectrometry. Circular dichroism was used to determine their axial stereochemistry revealing 1 and 2 to be atropisomers. 1 and 2 inhibited [125I]sauvagine binding to corticotropin releasing hormone (CRH-1) receptors.

Petasites hybridus: a tool for interdisciplinary research in phytotherapy.

The 3rd Petasites gathering took place in Romanshorn, Switzerland on March 29, 1996 and gave 16 European scientists the opportunity to transmit their latest considerable discoveries to interested researchers working in different scientific disciplines such as pharmacognosy, botany, chemistry, pharmacology, medicine or clinical pharmacy. The newest findings on Petasites hybridus as a significant plant drug showed very promising aspects of therapeutic utility. Great progress has been made in chemical analytical methods and the determination of pharmacological activities. Substantial advances have also occurred in the production of bioassay procedures and plant materials, particularly utilizing cell- and tissue-culture techniques.

Cardioprotection in patients undergoing chemo- and/or radiotherapy for neoplastic disease. A pilot study.

OBJECTIVES: To assess the cardioprotective efficiency of an antioxidant regimen (vitamins E, C and N-acetylcysteine) in patients receiving high dose chemo- and/or radiotherapy for malignant disease. METHODS: Prospective, placebo controlled, randomized and double blinded pilot study involving 13 patients receiving chemotherapy and 12 patients receiving radiotherapy. RESULTS: In patients receiving antioxidants, left ventricular ejection fraction did not change (63 +/- 4% to 63 +/- 4%). In the placebo group, ejection fraction changed from 67 +/- 6% to 61 +/- 4% (p = 0.03). No patient in the antioxidant group and 6/13 (46%) patients in the placebo group showed a fall of > 10% in the left ventricular ejection fraction. In the chemotherapy group, the left ventricular ejection fraction changed from 62% (+/- 2) to 63% (+/- 2) in the patients treated with antioxidants (ns) and from 63% (+/- 5) to 61% (+/- 5) in patients treated with placebo (ns). No patient showed a significant fall in ejection fraction in the antioxidant group, whereas 2/7 (29%) in the placebo group showed a reduction > or = 10%. In the radiotherapy group, left ventricular ejection fraction did not change ¿64% (+/- 6) to 64% (+/- 5)¿ in patients treated with antioxidants (ns) and changed from 70% (+/- 8) to 60% (+/- 4) in patients treated with placebo (p = 0.008). No patient in the antioxidant group, but 4/6 (66%) patients in the placebo group showed a fall of > or = 10% in ejection fraction. CONCLUSION: The small number of patients in the study precludes a definitive statement. The preliminary results however suggest efficient cardioprotection by this nontoxic and inexpensive antioxidant combination, so larger studies are warranted for confirmation.

Metal uptake of recombinant cambialistic superoxide dismutase from Propionibacterium shermanii is affected by growth conditions of host Escherichia coli cells.

We constructed the complete nucleotide sequence coding for the cambialistic superoxide dismutase from Propionibacterium shermanii by ligation of a synthetic linker to a polymerase chain reaction amplification product obtained using degenerate primers. We set up an expression system yielding large amounts of recombinant superoxide dismutase in the cytoplasm of Escherichia coli and purified the enzyme from cells grown in a complex medium. The physicochemical properties of the recombinant enzyme were identical to those of the natural protein. Under anaerobic conditions the enzyme produced in an iron-supplemented medium incorporated iron as metal cofactor, while the enzyme purified from cells grown under aerobic conditions contained a variable amount of iron and manganese depending on metal availability. Functional equivalence of the two metals in this superoxide dismutase variant was indicated by independence of enzyme activity from Fe/Mn ratio.

[Cardioprotection in chemo- and radiotherapy for malignant diseases--an echocardiographic pilot study]. / Kardioprotektion bei Chemo- und Radiotherapie für maligne Erkrankungen--eine echokardiographische Pilotstudie.

AIM: Pilot study, examining the cardioprotective effect of an antioxidant regimen in patients with malignancies receiving high dose chemo- or radiotherapy. PATIENTS AND METHODS: 14 patients with chemotherapy and 10 patients with radiotherapy were randomized in a double-blind fashion (placebo versus vitamin E and C and N-acetylcysteine). Systolic and diastolic echocardiographic parameters were determined before and within three weeks of treatment completion. RESULTS: Left ventricular ejection fraction fell significantly in patients receiving placebo (radiotherapy: 67 +/- 6 to 56 +/- 2%, p = 0.008, chemotherapy: 67 +/- 7 to 60 +/- 8%, p = 0.05). Patients on antioxidants showed no significant fall in EF (radiotherapy: 63 +/- 8 to 61 +/- 7%, chemotherapy: 67 +/- 5 to 64 +/- 6%). CONCLUSION: The small number of patients in the study precludes a definitive statement. The preliminary results, however, suggest efficient cardioprotection by this cheap and safe antioxidant combination, so that larger studies are warranted for confirmation.

Spontaneous conscious covert cognition states and brain electric spectral states in canonical correlations.

Correlations between subjective, conscious, spontaneous cognitions and EEG power spectral profiles were investigated in 20 normal volunteers (2 sessions each) during relaxation-drowsiness-sleep onset. Four-channel EEG (temporal-parietal and parietal-central, left and right) was continuously recorded. The subjects were prompted 15 times per session to give brief reports of their ongoing thoughts. The reports were rated on 23 scales, and the 16 seconds of EEG recording preceding the prompts were spectral analyzed. Canonical correlation analysis was applied to the data (23 cognition ratings and 124 EEG spectral values for each of the 538 prompts). Four of the 23 pairs of canonical EEG variables and cognition variables were significant (p < 0.016) with correlation coefficients ranging from 0.78 to 0.62. The four pairs of canonical variables showed distinctive features in EEG spectra and cognition styles. The results demonstrate ruleful correspondences between EEG states and spontaneous, conscious, covert, cognitive-emotional states in a no-input, no-task, no-response paradigm.

Regulation of an in vivo metal-exchangeable superoxide dismutase from Propionibacterium shermanii exhibiting activity with different metal cofactors.

The anaerobic, but aerotolerant Propionibacterium freudenreichii sp. shermanii contains a single superoxide dismutase [EC 1.15.1.1.] exhibiting comparable activity with iron or manganese as metal cofactor. The formation of superoxide dismutase is not depending on the supplementation of iron or manganese to the culture medium. Even in the absence of these metals the protein is built in comparable amounts. Bacteria grown in the absence of iron and manganese synthesize a superoxide dismutase with very low activity which had incorporated copper. If the medium was also depleted of copper, cobalt was incorporated, leading to an enzymically inactive form. In the absence of cobalt an enzymically inactive superoxide dismutase was built with unknown metal contents. Upon aeration the amount of superoxide dismutase activity increased continuously up to 9 h, due to a de novo synthesis of the protein. This superoxide dismutase had incorporated iron into the active centre. The superoxide dismutase of Propionibacterium shermanii is able to form a much wider variety of complexes with trace metal ions in vivo than previously recognized, leading to the hypothesis that the original function of these proteins was the binding of cytoplasmic trace metals present in excess.

In vivo incorporation of cobalt into Propionibacterium shermanii superoxide dismutase.

Propionibacterium shermanii, an aerotolerant anaerobic bacterium, has already been shown to incorporate, depending on the metal supplementation to the medium, either iron or manganese or copper into the same superoxide dismutase protein. The in vivo incorporation of cobalt in the same superoxide dismutase was obtained in an iron-, manganese- and copper-depleted medium. The protein was isolated and characterized by NMR which offers the possibility to identify the amino acid residues at the active site exploiting isotropically shifted proton resonance.

Spatial and temporal accumulation of defense gene transcripts in bean (Phaseolus vulgaris) leaves in relation to bacteria-induced hypersensitive cell death.

Primary leaves of 7- to 9-day-old Red Mexican bean plants were inoculated with virulent or avirulent isolates of Pseudomonas syringae pv. phaseolicola, or saprophytic P. fluorescens either by vacuum infiltration of the whole leaf lamina, or by syringe-inoculation of selected leaf panels. In the incompatible combination, resistance was associated with a hypersensitive response (HR). Syringe-inoculated leaves were sampled in three zones: zone 1, the inoculated leaf area; zone 2, the surrounding 0.5-0.7 cm of leaf tissue; and zone 3, the remainder of the leaf. Northern blots of RNA from zones 1, 2, and 3 were probed with bean cDNAs for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chitinase (CHT), and lipoxygenase (LOX). Accumulation of PAL, CHS, and CHT transcripts was more rapid and generally of greater magnitude in the incompatible than in the compatible interaction and, in both cases, was observed essentially only in zone 1 tissues. Similarly, antibacterial phytoalexins were only detected in zone 1 from the incompatible interaction. Young primary leaves have a background level of LOX transcripts, which declines as leaves age. This decline was accelerated over the first 12 hr postinoculation (hpi) with avirulent bacteria, whereas a weak transient induction, peaking at 5-6 hpi, was observed in the compatible interaction. A subsequent, strong accumulation of LOX transcripts was seen in both the compatible and incompatible interactions outside the inoculation site starting about 14 hpi. LOX transcripts did not accumulate at the inoculation site itself in the incompatible interaction compared to a relatively strong induction in the compatible interaction. Interestingly, inoculation of leaves with cells of the saprophyte P. fluorescens also induced the accumulation of transcripts for CHS, CHT, and LOX, but generally to a lesser degree than in the incompatible interaction. No HR occurred and no macroscopic cell damage was apparent in leaves inoculated with P. fluorescens. However, at the microscopic level individual, trypan blue-stained, necrotic plant cells were visible. In spite of this and the accumulation of CHS transcripts, no phytoalexin accumulation was found up to 48 hr after inoculation. The spatial and temporal relationship of the hypersensitive reaction to defense gene transcript and phytoalexin accumulation is discussed.

The enzymatic decomposition of salicin and its derivatives obtained from Salicaceae species.

The enzymatic catalysis of the decomposition of Salicaceae phenolic glucosides was tested using almond beta-glucosidase and rabbit and porcine liver esterases. The beta-glucosidase catalyzed the complete hydrolysis of salicin and salicortin, yielding saligenin and glucose. Salicortin also produced (+)-6-hydroxycyclohexen-2-one (6-HCH). The acylglucosides were not decomposed by the beta-glucosidase. Both esterases catalyzed the decomposition of tremulacin, salicortin, and 2'-O-acetylsalicortin, releasing tremuloidin, salicin, and 2'-O-acetylsalicin as the main products, accompanied by 6-HCH and catechol. Tremuloidin and 2'-O-acetylsalicin were quite stable under the esterase hydrolysis, and salicin was not decomposed at all.