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PubMed1999

Induction of mitochondrial Apo2.7 molecules and generation of reactive oxygen-intermediates in cultured lymphocytes by the toxic proteins from Viscum album L.

We analysed mitochondrial alterations in human lymphocytes incubated with toxins exerting RNA and/or protein synthesis/transport inhibitory activity. We found that all toxins known to affect macromolecule synthesis, such as ricin from Ricinus communis, mistletoe lectin I (ML I) from Viscum album, cycloheximide, actinomycin D, and brefeldin A but also the thionins from Viscum album (viscotoxins; VT) generated reactive oxygen intermediates (ROI) and induced expression of newly described mitochondrial membrane proteins Apo2.7, however, with different kinetics. Apart from a rapid permeabilisation of cell membranes by the VT with swelling of mitochondria, loss of their cristae and ROI generation within 2-4 h, the majority of the cells may have received a distinct 'death signal' resulting in an induction of Apo2.7 molecules within 24 h. In contrast, protein synthesis/transport inhibition may signal for apoptosis within 24 h by decreasing distinct 'survival promotors' which remain to be characterised.

Annexin A5↗
PubMed2003

Some medicinal plants as immunostimulant for fish.

Immunostimulant effects of the dietary intake of various medicinal plant extracts on fish, rainbow trout (Oncorhynchus mykiss), were investigated. For this purpose fish were fed with diets containing aqueous extracts of mistletoe (Viscum album), nettle (Urtica dioica), and ginger (Zingiber officinale). Food containing lyophilized extracts of these plants as 0.1 and 1% was used at a rate of 2% of body weight per day for three weeks. At the end of the experimental period, various parameters of non-specific defence mechanisms, including extracellular and intracellular respiratory burst activities, phagocytosis in blood leukocytes and total plasma protein level were examined. Specific growth rates (SGRs) and condition factors (CFs) of the fish were also measured. Plant materials tested for immunostimulatory food additives caused an enhanced extracellular respiratory burst activity (P<0.001) compared to the control group. Especially the rainbow trout fed with a diet containing 1% aqueous extract of powdered ginger roots for three weeks exhibited a significant non-specific immune response. Phagocytosis and extracellular burst activity of blood leukocytes were significantly higher in this group than those in the control group. All plant extracts added to fish diet increased the total protein level in plasma except 0.1% ginger. The highest level of plasma proteins was observed in the group fed with 1% ginger extract containing feed.

Adjuvants, Immunologic↗
PubMed1998

Effects of the Turkish subspecies of Viscum album on macrophage-derived cytokines.

Viscum album L. herbs are used for the treatment of various diseases. Apart from their immunostimulatory, antitumor, and hypotension inducing activities, anti-inflammatory effects has also been recorded in the literature. Since the immunostimulatory activity of mistletoe is mainly attributed to a stimulation of the mononuclear phagocytic system and to an induction of inflammation by macrophage-derived cytokines, utilization based on anti-inflammatory activity seems in contrast with its use as immunostimulant. The inhibitory effects of 80% ethanol extracts and subfractions obtained by petroleum ether, diethylether, ethylacetate and n-butanol from three Turkish subspecies of V. album L. (ssp. album, ssp. abietis, ssp. austriacum) against interleukins (IL-1alpha and IL-1beta) and tumor necrosis factor-alpha were studied. Ethanolic (80%) extracts of all three subspecies exhibited almost no significant inhibitory activity on the proinflammatory cytokines, at least in the concentrations applied in this study. In contrast, they induce a concentration dependent stimulation of these cytokines. On the other hand, moderate to weak, dose dependent inhibitory effects were observed on fractionation, which may be attributed to the existence of less polar components with anti-inflammatory activity. Based on our results, it was concluded that the utilization of ethanol extracts for anti-inflammatory purposes would not be beneficial probably due to the partial extraction of the low-molecular-weight polypeptides, i.e. viscotoxins, which induce an inflammatory response. Moreover, for anti-inflammatory effects, extracts of Viscum album ssp. austriacum may be preferred, since higher inhibitory ratios were obtained against inflammatory cytokines than with the other two subspecies.

Dose-Response Relationship, Drug↗
PubMed1998

High-level expression of a viscotoxin in Arabidopsis thaliana gives enhanced resistance against Plasmodiophora brassicae.

Viscotoxins are a group of toxic thionins found in several mistletoe species. The constitutive CaMV-omega promoter was used to drive the expression of the viscotoxin A3 cDNA from Viscum album in transgenic Arabidopsis thaliana C24. Lines with high viscotoxin A3 levels in all parts of the plant were selected and tested for resistance against the clubroot pathogen Plasmodiophora brassicae. The transgenic lines were more resistant to infection by this pathogen than the parental line.

Animals↗
PubMed1998

Selective killing of CD8+ cells with a 'memory' phenotype (CD62Llo) by the N-acetyl-D-galactosamine-specific lectin from Viscum album L.

As reported previously by our group, among the toxic proteins from Viscum album L. only the mistletoe lectins (MLs) induce the apoptotic killing pathway in human lymphocytes. Although one may expect a homogenous distribution of carbohydrate domains on cell surface receptors for the carbohydrate binding B chains of the toxic protein, the sensitivity of cells to these B chains obviously differ. Here we report a selective killing of CD8+ CD62Llo cells from healthy individuals by the galNAc-specific ML III (and RCA60, which binds to gal and galNAc), while the gal-specific ML I was less effective. This selective killing is not sufficiently explained by protein synthesis inhibition alone, since this subset was not affected by other ribosome inhibiting proteins such as the lectin from Ricinus communis (RCA120), lectin from Abrus precatorus (APA), abrin A, and inhibitors of RNA, DNA and/or protein synthesis such as actinomycin D, mitomycin C, and cycloheximide. We conclude that CD8+ cells with 'memory' phenotype (CD62Llo) are more sensitive to the ML III-mediated killing than their CD8+ CD62Lhi counterparts, CD4+ T cells, and CD19+ B cells. These cells probably express a distinct receptor with galNAc domains that is missing or not active on CD8+ cells with a 'naive' phenotype.

Acetylgalactosamine↗
PubMed2000

Genetic evidence for host specificity in the hemi-parasitic Viscum album L. (Viscaceae).

Nuclear ribosomal DNA (nrDNA) ITS sequences and partial sequences of three non-coding chloroplast DNA (cpDNA) introns and spacers were used to assess genetic variation within and among three presumed host races of the hemi-parasite Viscum album L. Currently, identification of host races occurs via the host trees, and morphological differences are minute at best. cpDNA and nrDNA ITS sequences revealed little sequence variation, but the variation found consistently supported the distinction of three host races. cpDNA and ITS sequences were not incongruent, as assessed by the incongruence length difference test. A combined analysis supported the sister group relationship between mistletoes from deciduous trees and fir.

Cycadopsida↗
PubMed1998

[Use of alternative medicine in oncology patients].

BACKGROUND AND OBJECTIVE: Up to 60% of all oncological patients use methods of alternative medicine in the course of their illness. In earlier blinded studies demographic characteristics and the patients' motives of using alternative medicine had been recorded, but a correlation with individual illnesses had not been possible. PATIENTS AND METHODS: 142 patients of a oncological outpatient clinic gave their experience with alternative medicine in interviews and non-blinded questionnaires, 103 of them (72.5%; 46 men and 57 women; median age 58 years, range 18-91 years) returning questionnaires that could be evaluated. RESULTS: 46 patients stated that they had used alternative medicine. There was no difference between users and non-users regarding sex; age, profession, education, family status or religion. 58% of all patients with advanced disease used alternative medicine, compared with only 31% with partial remission or stable disease and 41% in complete remission (P = 0.042). Vitamins and mistletoe preparations were the most commonly used substances (50 and 45%, respectively). The predominant purpose was to stimulate the immune system (77%) and strengthen general physical capacity (64.5%). As main stimulus for using alternative medicine the patients came from their family doctor (56%), followed by family and friends (41%). Alternative medicine was used largely as complementary and not an alternative to conventional medicine. Health insurance met all or some of the costs of alternative treatment in 59% of patients. CONCLUSION: A large number of patients treated with conventional oncological regimens also use alternative medicine, most of them because of a polypragmatic attitude to tumor treatment. Family doctors and health insurance companies are playing a more important role than had hitherto been assumed in spreading the use of treatment options without providing scientifically based evidence of their efficacy.

Adult↗
PubMed1986

[The use of paramedical treatment methods by cancer patients. A inquiry on 101 ambulatory patients].

In an anonymous inquiry 101 consecutive tumour patients and their doctors were questioned on the use of treatment methods and preparations without proven efficacy. 32 patients stated they were using such preparations and methods. Three-quarters of the patients had adopted the following dietetic measures, in descending order of frequency: beetroot juice (34%), vitamins (22%), lactic acid or herbal products (9% each). Iscador (mistletoe extract) was used by 21%, and homoeopathic preparations by 6%. Patients' estimate of the total cost, in 69% of cases, was below 300 Swiss francs, but in one case it exceeded 1200. There was no difference in the use or non-use of the products in regard to the doctor's assessment of the prognosis, age, the therapeutic success according to established medical criteria, or the diagnosis and its correct appreciation by the patient. Male patients employed these methods or preparations significantly more often (43%) than female patients (22%); such patients also showed a higher incidence of severe suffering from side effects of chemotherapy (19% vs. 7%), especially nausea. The use of Iscador was known to the doctor in 71% of the cases, whereas the use of the other methods and preparations was known in only 25%.

Antineoplastic Agents, Phytogenic↗
PubMed1997

Metabolic inhibitors as tools to delineate participation of distinct intracellular pathways in enhancement of lactose-induced dissociation of neutrophil and thymocyte aggregates formed by mediation of a plant lectin.

Signaling processes in the course of the formation of the lectin-mediated aggregates may partake in conveying enhanced stability to the cell clusters. To prove the validity of this reasoning in a model, we have studied the impact of addition of three metabolic inhibitors (N-ethylmaleimide, nordihydroguaiaretic acid, and trifluoperazine) on lactose-dependent dissociation of cell aggregates, formed in the presence of the galactoside-binding mistletoe lectin. Using both human neutrophils and rat thymocytes to avoid measurement of responses restricted to a single cell type, an enhanced dissociation of lectin-formed cell aggregates was observed, when lactose and an inhibitor were present. Among the tested inhibitors, nordihydroguaiaretic acid and N-ethylmaleimide were more potent enhancers of cell dissociation than trifluoperazine. These results suggest that biosignalling pathways connected with lipoxygenase activity as well as the level of intracellular sulfhydryl groups confer further stability to lectin-dependent cell aggregates. The systematic evaluation of inhibitors for defined activities is thus suggested as a tool to disclose the nature and the contribution of individual signaling mechanisms to post-binding effects following lectin-initiated cell contact formation.

Animals↗
PubMed2002

The evolution of parasites from their hosts: intra- and interspecific parasitism and Emery's rule.

In some taxa of Hymenoptera, fungi, red algae and mistletoe, parasites and their hosts are either sibling species or at least closely related (Emery's rule). Three evolutionary mechanisms have been proposed for this phenomenon: (i) intraspecific parasitism is followed by sympatric speciation; (ii) allopatric speciation is followed by secondary sympatry and the subsequent parasitism of one sibling species by the other; and (iii) allopatric speciation of a species with intraspecific parasitism is followed by secondary sympatry, in which one species becomes an obligate parasite of the other. Mechanisms (i) and (ii) are problematic, while mechanism (iii) has not, to our knowledge, been analysed quantitatively. In this paper, we develop a model for single- and two-species evolutionary stable strategies (ESSs) to examine the basis for Emery's rule and to determine whether mechanism (iii) is consistent with ESS reasoning. In secondary sympatry after allopatric speciation, the system's evolution depends on the relative abundances of the two sibling species and on the proportional damage wrought by parasites of each species on non-parasitic members of the other. Depending on these interspecific effects, either the rarer or the commoner species may become the parasite and the levels of within-species parasitism need not determine which evolves to obligate parasitism.

Animals↗
PubMed2006

Antifungal effects and mechanism of action of viscotoxin A3.

Viscotoxins are cationic proteins, isolated from different mistletoe species, that belong to the group of thionins, a group of basic cysteine-rich peptides of approximately 5 kDa. They have been shown to be cytotoxic to different types of cell, including animal, bacterial and fungal. The aim of this study was to obtain information on the cell targets and the mechanism of action of viscotoxin isoform A3 (VtA3). We describe a detailed study of viscotoxin interaction with fungal-derived model membranes, its location inside spores of Fusarium solani, as well as their induced spore death. We show that VtA3 induces the appearance of ion-channel-like activity, the generation of H2O2, and an increase in cytoplasmic free Ca2+. Moreover, we show that Ca2+ is involved in VtA3-induced spore death and increased H2O2 concentration. The data presented here strongly support the notion that the antifungal activity of VtA3 is due to membrane binding and channel formation, leading to destabilization and disruption of the plasma membrane, thereby supporting a direct role for viscotoxins in the plant defence mechanism.

Amino Acid Sequence↗
PubMed1979

Plants and hypotensive, antiatheromatous and coronarodilatating action.

However great the success in the therapy of hypertension, atherosclerosis and ischemic heart disease has been gained today by recent efficient drugs, the definite healing of patients is not yet attained. The late discovery of reserpine, such an efficient drug of plant origin against hypertension, convinced so far reluctant scientists to consider the chemical compounds of the plant world. With respect to this traditional medical knowledge, it seems necessary to define more accurately the specificity of these healings-sometimes recommended unspecifically for a whole branch of medicine. This experimental verification should not use inconsiderately the present-day classification of diseases; there should be an awareness that conventional experimental methods in pharmacology are often unsuitable for revealing the real biological activity of one or another medicinal plant. The interest in the millennial empirical field of health care is acknowledged by the World Health Organization which promotes research and development of traditional medicine, along with investigations into its psychosocial and ethnographic aspects. These studies cover a number of plants growing in Bulgaria that have a healing effect in hypertension, atherosclerosis and ischemic heart disease according to the data of traditional medicine. Using screening methods, extracts and chemically pure substances were investigated; extraction was done with solvents such as water, ether, chloroform, dichloretan, ethanol, methanol, and acetone. Most of the experiments were carried out on anesthetized cats, rabbits and dogs. The substances tested were applied mainly intravenously, and in some experiments orally. Chronic experiments were also carried out on wakeful dogs with induced hypertension, on animals fed on an atherogenic diet, and on animals with induced arrhythmia and coronary spasm. Data are presented of clinical examination of some plants or of active substances isolated from them. Major results of these studies are presented for the following plants: Garlic, Geranium; Hellebore; Mistletoe; Olive; Valerian; Hawthorn; Pseucedanum arenarium; Periwinkle; Fumitory. For another 50 plants growing in Bulgaria and in other countries the author presents his and other investigators' experimental and clinical data about hypotensive, antiatheromatous and coronarodilatating action.

Animals↗
PubMed1986

Natural killer and antibody-dependent cell-mediated cytotoxicity activities and large granular lymphocyte frequencies in Viscum album-treated breast cancer patients.

20 breast cancer patients received a single intravenous infusion of Iscador, a mistletoe (Viscum album L.) extract. Natural killer (NK) and antibody-dependent cell-mediated cytotoxicity (ADCC) activities as well as the number of large granular lymphocytes (LGL) were investigated in their peripheral blood at various times. Six hours after the start of the infusion, significant decreases and 24 h later, significant increases in NK/ADCC activities and LGL frequencies were observed. These responses followed a kinetic pattern similar to that which has been described by others to occur after treatment with interferon.

Adult↗
PubMed1986

Structure and properties of polysaccharides from Viscum album (L.).

Polysaccharides are possibly involved in the pharmacological effects of Viscum album (mistletoe) extracts, which are used in cancer therapy. Therefore the water-soluble polysaccharides of the fresh plant and the fermented proprietary preparation Iscador were isolated and characterized inter alia by methylation analysis, partial hydrolysis and C-13-NMR spectroscopy. The main polysaccharide of the green parts of Viscum is a highly esterified galacturonan whereas in Viscum 'berries' a complex arabinogalactan is predominant. Both types of these constituents were found in Iscador but with definite changes in molecular weight and structure. An interaction between the arabinogalactan and the galactose-specific lectin (ML I) in Viscum could be demonstrated. In three immunological tests (granulocyte, chemiluminescence, carbon clearance test) the polysaccharides failed to increase phagocytic activity of granulocytes and macrophages.

Carbon↗
PubMed1986

Immunomodulatory effects of iscador: a Viscum album preparation.

The immunomodulatory effects of Iscador (a mistletoe, Viscum album extract) were investigated. After a single intravenous infusion of Iscador several immunological parameters in the peripheral blood of breast cancer patients were examined. Parallel with neutrophilia, and with an increase of juvenile neutrophils, a significant enhancement of phagocytic activity of granulocytes was shown. After short decreases during the first 24 h, significant increases in natural killer (NK) and antibody-dependent cell-mediated cytotoxicity (ADCC) activities as well as augmented levels of large granular lymphocytes were observed. These NK/ADCC responses followed a kinetic pattern similar to that after treatment with alpha-interferon described by others. Further significant increases in mitogenic responses to phytohemagglutinin and concanavalin A were observed.

Adjuvants, Immunologic↗
PubMed2001

Involvement of caspase-3 in apoptosis induced by Viscum album var. coloratum agglutinin in HL-60 cells.

A cytotoxic lectin (Viscum album L. coloratum agglutinin, VCA) from Korean mistletoe was isolated by affinity chromatography on Sepharose 4B immobilized with asialofetuin. In HL-60 cells, addition of VCA resulted in a dose- and time-dependent growth suppression, morphological changes of apoptotic nuclei, and DNA fragmentation characteristics of apoptosis. To investigate how caspase-3 activation during VCA-induced apoptosis induces cleavages of PARP, the expression of PARP and the pattern of caspase-3 activation in HL-60 cells were investigated. The native and processed PARP forms typically seen in apoptotic cells were observed, and a decrease in expression of the 32-kDa form of caspase-3 in a dose-dependent manner was observed. The VCA-induced apoptosis was significantly inhibited by a caspase-3 specific inhibitor, z-DEVD-FMK, and the PARP processing and caspase-3 activation were also inhibited by the inhibitor. A possible involvement of cell cycle arrest in VCA-induced apoptosis was investigated by flow cytometry and the results suggested that the apoptotic effect of VCA is not involved in the induction of cell cycle arrest.

Apoptosis↗
PubMed1997

Amino acid sequence, S-S bridge arrangement and distribution in plant tissues of thionins from Viscum album.

The complete primary structure of a cytotoxic 5 kDa polypeptide, viscotoxin A1, isolated from Viscum album L., has been determined by combining classical Edman degradation methodology with advanced mass spectrometric procedures. The same integrated approach allowed correction of the sequence of viscotoxin A2 and definition of the pattern of the disulfide bridges. The arrangement of the cysteine pairing was determined as Cys3-Cys40, Cys4-Cys32 and Cys16-Cys26. The primary structure of viscotoxin A1 shares a high degree of similarity with the known viscotoxins and more generally with the plant alpha- and beta-thionins. The pattern of S-S bridges determined for viscotoxin A2 and A1 is similar to that inferred by X-ray and NMR analysis in crambin and related to that present in alpha-purothionin and beta-hordothionin, thus indicating a highly conserved organization of the S-S pairings within the entire family. This arrangement of S-S bridges describes a peculiar structural motif, indicated as 'concentric motif', which is suggested to stabilize a common structure occurring in various small proteins able to interact with cell membranes. The distribution of the new variant toxin in different mistletoe subspecies was investigated. Viscotoxin A1 is abundant in the seeds of the three European subspecies of V. album whereas it represents a minor component in the shoots.

Amino Acid Sequence↗
PubMed2004

Botanical medicine and cancer: a review of the safety and efficacy.

It is currently estimated that > 50% of all patients diagnosed with cancer explore complementary and alternative medicine - especially herbal medicine. We conducted a comprehensive review to assess the safety and efficacy of herbal medicines commonly used by patients in an attempt to: prevent cancer; treat cancer; and treat adverse effects associated with conventional cancer treatments. Current evidence suggests that Asian ginseng, garlic, green tea, tomatoes and soy intake as part of the diet may be useful in preventing various cancers; additional research is needed in order to determine the efficacy of essiac, evening primrose oil, mistletoe, reishi, shiitake and turmeric as cancer treatments; and ginger may be effective in treating chemotherapy-induced nausea and vomiting.

Beverages↗