Mistletoe Extract Research Articles

Korean mistletoe (Viscum album var. coloratum) extends the lifespan via FOXO activation induced by dSir2 in Drosophila melanogaster.

We examined the underlying mechanisms associated with the longevity effects of Korean mistletoe extract (KME) in Drosophila melanogaster. We measured the lifespan of sirtuin, chico and foxo mutant flies fed KME, the expression of the forkhead box O (FOXO) target genes and insulin-like peptide genes, and the localization of FOXO in flies fed the KME. The longevity effect of KME was abolished in sirtuin, chico and foxo null mutant flies. In addition, the expression of FOXO target genes and the localization of FOXO into nuclei were increased in flies fed KME, but the expression of the insulin-like peptide genes was decreased by KME supplementation. The results show that KME extends the fly lifespan through sirtuin-induced FOXO activation. We suggest that KME has potential use as a beneficial anti-aging and longevity supplement. Geriatr Gerontol Int 2021; 21: 725-731.

The Effect of Mango Mistletoes (Dendrophthoe pentandra) Leaves Extract on Percentage of CD4+CD28+, CD8+CD28+, and interleukin-2 Levels of Aged Balb/c Mice

BACKGROUND: Population aging is considered to be a global phenomenon today. Age-associated immune system dysfunction or “immunosenescence” is indicated by increased susceptibility to infections and chronic diseases, such as hypertension, diabetes mellitus, autoimmune diseases, heart disease, and atherosclerosis. One of the immunosenescence markers is a significant drop in CD28 and reduced proinflammatory cytokine interleukin-2 (IL-2). The mango mistletoes are deemed to have a better affinity for docking the CD28 and IL-2R receptors of α and β subunits than other plants. AIM: This study aims to determine the effect of ethanol extract of mango mistletoes on IL-2 levels, the percentage of CD4+CD28+, and the percentage of CD8+CD28+ in aged female mice. METHODS: The leaves of mango mistletoes were extracted using 96% ethanol solvent, and the extract was administered to aged female mice (18–20 months) orally with different doses for each group, namely 150, 300, and 600 mg/kg. Mango mistletoe leaves extract was administered once a day for 14 days. Then, the IL-2 levels of the mice were checked from their heart blood samples using Enzyme-Linked Immunosorbent Assay, while the percentages of CD4+CD28+ and CD8+CD28+ were examined from the spleen samples using flow cytometry. RESULTS: The ethanol extract of mango mistletoe leaves was able to increase the percentage of CD4+CD28+ significantly (p < 0.05) at doses of 300 and 600 mg/kg and increase the percentage of CD8+CD28+ significantly (p < 0.05) at a dose of 600 mg/kgBW, while other various doses had a strong enough correlation (r = 0.48) to increase IL-2 levels. CONCLUSION: The ethanol extract of mango mistletoe leaves has the good potential to inhibit the aging process in the immune system, as characterized by an increase in IL-2 levels and the percentage of CD4+CD28+ and CD8+CD28+.

Complementary and Alternative Medicine for the Treatment of Gliomas: Scoping Review of Clinical Studies, Patient Outcomes, and Toxicity Profiles

Complementary and alternative medicine (CAM) are highly used among those diagnosed with glioma. Further research is warranted, however, as it remains important to clearly delineate CAM practices that are unproven, disproven, or promising for future research and implementation. A systematic review was conducted to identify all articles that investigated the effect of any CAM therapy on survival of patients with newly diagnosed or recurrent glioma. Eighteen papers and 4 abstracts pertaining to the effects of ketogenic diet (4), antioxidants (3), hyperbaric oxygen (4), cannabinoids (2), carbogen and nicotinamide (3), mistletoe extract (2), hypocupremia and penicillamine (1), and overall CAM use (3) on overall and progression-free survival in patients with low- and high-grade glioma were identified (Levels of Evidence I-IV). Ketogenic diets, hyperbaric oxygen therapy, and cannabinoids appear to be safe and well tolerated by patients; preliminary studies demonstrate tumor response and increased progression-free survival and overall survival when combined with standard of care therapies. Antioxidant usage exhibit mixed results perhaps associated with glioma grade with greater effect on low-grade gliomas; vitamin D intake was associated with prolonged survival. Conversely, carbogen breathing and hypocupremia were found to have no effect on the survival of patients with glioma, with associated significant toxicity. Most modalities under the CAM umbrella have not been appropriately studied and require further investigation. Despite widespread use, Level I or II evidence for CAM for the treatment of glioma is lacking, representing future research directions to optimally counsel and treat glioma patients.

Effects of Mango Mistletoe (Dendrophthoe pentandra L. Miq) Extracts on Brain in Hypertensive Rats Treated with Deoxycorticosterone Acetate (DOCA)-Salt

Mango mistletoe (Dendrophthoe pentandra) is a plant that has the potential as a medicinal plant. Mango mistletoe is reported to contain the flavonoids quercetin, mesoinositol, rutin, and tannins. Hypertension as a non-communicable disease is a very serious health problem experienced by people in developing countries. Hypertension can be caused by high ROS (reactive oxygen species) which can inhibit oxygen flow to the heart and brain. Antioxidants can reduce the risk of hypertension. Damage to the brain due to hypertension is manifest in the white matter area. This study aimed to see how the histopathological profile of the brain in hypertensive rats (DOCA-Salt) which were given mango mistletoe methanolic extract (MMME) by experimental method. The number of tested animals was 25 male Wistar rats divided into 5 groups, each group contained 5 rats, they were negative control (Control-) as normotension, positive control (Control+) as inducer, and the treatment group with the provision of MMME at a dose of 50 mg / kgBW, 100 mg / kgBW, and 200 mg / kgBW. Data were analyzed using JAMOVI 1.1.9.0. Based on the results of the study showed that the difference in significant values between all groups was p<0.05. So, that giving EMBM to male rat significantly can decrease the number of cell necrosis and affect the width of the white matter area of the brain. The three dose variations did not show a significant difference, so this effect was controlled by the MMME dose of 50 mg / KgBW which was the optimum dose in reducing the amount of cell necrosis in the white matter area of the male Wistar rat brain.

Präklinische Untersuchungen von Wechselwirkungen zwischen Mistel und Radio- oder Chemotherapie auf pädiatrische Tumorzellen

Hintergrund: Mistelanwendungen werden als komplementäre Therapien häufig in der pädiatrischen Onkologie zusammen mit einer Radio- oder Chemotherapie verabreicht. Wechselwirkungen bei simultaner Applikation sind gerade in der Pädiatrie von großer Bedeutung, sie sind allerdings nach wie vor unzureichend untersucht. Material und Methoden: Zytotoxische Effekte des Mistelextraktes abnobaVISCUM Fraxini (aVF) auf LAN-1 Neuroblastomzellen und deren Etoposid- bzw. Cisplatin-resistente Subzelllinien wurden mittels Viabilitätstest untersucht, sowie mögliche Synergieeffekte zwischen aVF und den Chemotherapeutika durch die Softwareprodukte Combenefit und CompuSyn analysiert. Effekte einer Kombinationstherapie aus aVF und Bestrahlung auf SH-SY5Y Zellen wurden mittels Koloniebildungstest untersucht und Auswirkungen auf die Reparatureffizienz strahleninduzierter Doppelstrangbrüche mit Hilfe durchflusszytometrischer Quantifizierungen von γ-H2AX-Foci nach PI/FITC Doppelfärbung analysiert. Ergebnisse: Die Chemotherapie-resistenten LAN-1 Subzelllinien erwiesen sich als resistenter gegenüber der Mistelbehandlung als die Ursprungszelllinie. Auf Basis vier verschiedener Referenz­modelle konnten vor allem synergistisch/additive Effekte zwischen aVF und den Zytostatika Etoposid und Cisplatin berechnet werden. Die Kombination aus Mistelbehandlung und Bestrahlung führte zu einer Verringerung der Koloniebildung und zu einer Verzögerung der Reparaturgeschwindigkeit von strahleninduzierten Doppelstrangbrüchen. Schlussfolgerung: Die präklinischen Daten könnten darauf hinweisen, dass die Verwendung des Mistelextraktes, aVF, eine unterstützende Wirkung auf Radio- und Chemotherapien hat.

The effects of dietary supplementation of mistletoe (Viscum album) extract on the growth performance, antioxidant, and innate, immune responses of rainbow trout (Oncorhynchus mykiss)

In the present study, rainbow trout fingerlings, Oncorhynchus mykiss, were fed diets supplemented with 0% (C), 0.5 (T1), 1.5 (T2), 2.5 (T3), and 4% (T4) mistletoe extract (ME), Viscum album over eight weeks. At the end of feeding trial, growth performance, serum antioxidant and innate immune responses, and serum bactericidal activity against Aeromonas hydrophila were evaluated. According to results, the highest final wight and wight gain (%), and the lowest feed conversion ratio (FCR) were observed in T2 and T3 treatments. Regression analysis showed that fish growth parameters were polynomially-related to dietary ME levels. The optimum dietary ME level, based upon the varied growth parameters, was 1.78 to 1.94%. The highest moisture content was observed in the control group followed by the T1 treatment, whereas the highest protein contents were observed in the T2 and T3 treatments. The lipid and ash contents were not affected by ME supplementation. Serum lysozyme, ACH50, total Ig, SOD, and bactericidal activity significantly increased in the ME-fed fish, with the highest values observed within the T3 treatment. The highest CAT and GPx values were recorded in the T2 and T3 treatments. ME treatment significantly decreased serum MDA levels, which was lowest in the T3 treatment. Serum ALT activities were similar in the T2, T3, and T4 treatments, and were significantly lower than those of T1 and control (C) treatments. Serum AST and ALP activities exhibited declines along with an increase in dietary ME levels; in which the lowest activities were occurred in the T4 treatment. In conclusion, dietary ME supplementation proved beneficial to rainbow trout, as it stimulated its growth performance, innate immune and antioxidant systems, and promoted bactericidal activity. Moreover, serum hepatic health indicators suggest that dietary ME may improve the fishes' hepatic health.

Mistletoe Extract Targets the STAT3-FOXM1 Pathway to Induce Apoptosis and Inhibits Metastasis in Breast Cancer Cells.

Mistletoe extracts (Viscum album L.) have been widely used as complementary and alternative medicines for the treatment of cancer, and their cytotoxic effects have been reported on various types of cancer. However, the molecular targets of mistletoe extracts have not been well studied. Herein, we investigated molecules associated with the in vitro and in vivo anticancer effects of mistletoe extract using 4T1 murine breast cancer cells. Mistletoe extract induced apoptosis and inhibited the signal transducer and activator of transcription3 (STAT3) phosphorylation. This inhibition was accompanied by the downregulations of forkhead box M1 (FOXM1) and the DNA repair proteins, RAD51 and survivin. Mistletoe extract simultaneously increased the expression of the DNA damage marker proteins, phosphorylated H2A histone family member X (H2A.X), and phosphorylated p38. Furthermore, mistletoe extract effectively suppressed tumor growth in 4T1 tumor-bearing BALB/c mice. In addition to tumor growth inhibition, mistletoe extract inhibited lung metastasis in the tumor-bearing mice and cell invasiveness by downregulating the expressions of matrix metalloproteinases (MMPs), urokinase-type plasminogen activator (uPA), uPA receptor, and markers of epithelial-mesenchymal transition (snail and fibronectin). Taken together, our results suggest that mistletoe extract targets the STAT3-FOXM1 pathway for its cytotoxic effects, and that mistletoe extracts might be useful for the treatment of patients with cancers highly expressing the STAT3-FOXM1 pathway.

Some aspects of the adaptogenic potential of European mistletoe (Viscum album L.) extracts under variable physical performance

Some aspects of the adaptogenic potential of European mistletoe (Viscum album L.) extracts under variable physical performance

Farklı Konukçu Bitkilerden Toplanan Ökse Otu (Viscum album L.) Ekstraktlarının Bazı Bitki Patojeni Bakteriler Üzerine Etkisi

Bu çalışma ile farklı ağaçlarda yarı parazitik olarak bulunan ökse otu (Viscum album L.)‘ndan elde edilen ekstraktların domates bakteriyel kanser ve solgunluk hastalık etmeni Clavibacter michiganensis subsp. michiganensis, biber ve domates bakteriyel yaprak lekesi hastalık etmeni Xanthomonas axonopodis pv. vesicatoria ve domates bakteriyel benek hastalık etmeni Pseudomonas syringae pv. tomato‘ya olan etkileri belirlenmiştir. Çalışmada çam, armut, akasya ve mahlep ağaçlarından toplanan ökse otunun yaprak ve saplarından elde edilen su ekstraktları son konsantrasyonları %1, %2,5, %5 ve %10 olacak şekilde otoklav edilmeden önce Nutrinet agar besi yerine ilave edilmiştir. Spektrofotometrede 600 nm dalga boyunda 0,2 absorbans değerinde 108 hücre/ml yoğunluğunda hazırlanan bakteri süspansiyonları ekstraktlı besi yerlerine ekilmiştir. Kontrol grubu olarak ekstraktların ilave edilmediği besi yerlerine patojenlerin ekimi yapılmıştır. Çalışma 3 tekerrürlü olarak kurulmuş olup, 2 kez tekrarlanmıştır. İnkübasyon süresi sonunda uygulama yapılan tüm petrilerde gelişen bakteriler toplanarak spektrofotometrede yoğunlukları ölçülmüştür. Yapılan çalışmada, farklı ağaçlardan toplanan ökse otunun yaprak ve saplarından elde edilen ekstraktların %1 ve %2,5‘luk dozları testlenen üç patojen üzerinde etkili olmazken, %5 ve %10‘luk dozları patojen bakterilerin gelişimini tamamen (%100) durdurmuştur. Ayrıca patojenler üzerinde ökse otunun toplandığı konukçu bazında bir farklılığın olmadığı da görülmüştür. In vitro koşullar altında yürütülen bu çalışma sonucuna göre, patojenler üzerinde etkili bulunan ökse otu ekstraktı ile in vivo çalışmaların yürütülmesi gerekmektedir.

Efficacy of mistletoe extract as a complement to standard treatment in advanced pancreatic cancer: study protocol for a multicentre, parallel group, double-blind, randomised, placebo-controlled clinical trial (MISTRAL)

BackgroundMost pancreatic cancer patients present with advanced stage at diagnosis with extremely short expected survival and few treatment options. A multimodal palliative approach is necessary for symptom relief and optimisation of health-related quality of life. In a recent open-label trial of mistletoe extract for advanced pancreatic cancer patients not eligible for chemotherapy, promising results on improved overall survival and better health-related quality of life were reported.The objective of the present study is to assess the value of mistletoe extract as a complement to standard treatment (palliative chemotherapy or best supportive care) in advanced pancreatic cancer patients with regard to overall survival and health-related quality of life.MethodsThe trial is prospective, randomised, double-blind, multicentre, parallel group and placebo-controlled. In total, 290 participants are randomly assigned to placebo or mistletoe extract given subcutaneously in increasing dosage from 0.01 to 20 mg three times per week for 9 months. Stratification is performed for site and palliative chemotherapy. Main inclusion criteria are advanced pancreatic cancer and Eastern Cooperative Oncology Group performance status 0 to 2; main exclusion criteria are life expectancy less than 4 weeks and neuroendocrine tumour of the pancreas. Two ancillary studies on sub-sets of participants are nested in the trial: a biomarker study collecting blood samples and a cross-sectional qualitative study with semi-structured face-to-face interviews.DiscussionTo our knowledge, this is the first placebo-controlled randomised trial assessing the impact of mistletoe extract as a complement to standard treatment on overall survival and health-related quality of life in patients with advanced pancreatic cancer. The presented trial with its two nested ancillary studies exploring biomarkers and patient experiences is expected to give new insights into the treatment of advanced pancreatic cancer.Trial registrationEU Clinical Trial Register, EudraCT Number 2014-004552-64. Registered on 19 January 2016.ClinicalTrials.gov NCT02948309. Registered on 28 October 2016.

Abstract 1831: Mistletoe lectin induces apoptosis in MYC-driven cancers through post-translational downregulation of c-MYC protein

Abstract c-Myc is part of the Myc family proteins associated with cellular proliferation, differentiation, and apoptosis. Dysregulated c-Myc expression is associated with tumorigenesis, and cancers with c-Myc overexpression are often associated with poor prognosis, partly due to difficulty in targeting c-Myc. Mistletoe lectin (ML), derived from the European mistletoe plant, has shown variable efficacy in cancer patients. One case series following 12 patients with advanced hepatocellular carcinoma (HCC) showed 2 patients with &gt30% reduction in the tumor marker alpha-fetoprotein and 1 patient who had stable disease for 9 months. These cases indicate promising results. ML consists of a heteroprotein complex of an A- and B-chain connected by a disulfide bond. The A-chain was previously described to inhibit translation by 28s rRNA cleavage, while the B-chain binds cell surface glycoproteins to trigger internalization of the lectin. We found that purified mistletoe extract, consisting primarily of ML, downregulates c-Myc protein and induces apoptosis in in vitro and in vivo models of human-derived cancer, and that the efficacy of ML treatment is strongly correlated with MYC gene expression and c-Myc protein abundance in in vitro human models of HCC, ovarian cancer, small cell lung cancer, and lymphoma. MTS assay data demonstrated that exogenous MYC expression can sensitize low-MYC cell lines to ML treatment, suggesting that anti-cancer effects of ML are dependent on its ability to downregulate c-Myc. While ML may be able to downregulate many proteins via general translational inhibition, we found that of 296 proteins profiled by RPPA, c-Myc was one of 11 proteins decreased by ML with at least a 2-fold change (p&lt0.01). We also found that ML promotes c-Myc ubiquitylation, suggesting that ML regulates c-Myc protein stability, rather than synthesis. In summary, here we describe a novel mechanism of action for ML in downregulating c-Myc and promoting apoptotic cell death in various models of MYC-driven cancer, and provide evidence for MYC gene expression as a predictive marker for response. This research supports the development of ML-based therapies that may provide a c-Myc-targeting treatment in multiple cancer types. Citation Format: Gabrielle Kennelley, Eric Yuan, Mohammad Shatat, Betsy Gauthier, Peiying Yang, Analisa Difeo, Richard Lee. Mistletoe lectin induces apoptosis in MYC-driven cancers through post-translational downregulation of c-MYC protein [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1831.

Quality of life in cancer patients treated with mistletoe: a systematic review and meta-analysis

BackgroundMistletoe extracts are used as an adjunct therapy for cancer patients, but there is dissent as to whether this therapy has a positive impact on quality of life (QoL).MethodsWe conducted a systematic review searching in several databases (Medline, Embase, CENTRAL, CINAHL, PsycInfo, Science Citation Index, clinicaltrials.gov, opengrey.org) by combining terms that cover the fields of “neoplasm”, “quality of life” and “mistletoe”. We included prospective controlled trials that compared mistletoe extracts with a control in cancer patients and reported QoL or related dimensions. The quality of the studies was assessed with the Cochrane Risk of Bias tool version 2. We conducted a quantitative meta-analysis.ResultsWe included 26 publications with 30 data sets. The studies were heterogeneous. The pooled standardized mean difference (random effects model) for global QoL after treatment with mistletoe extracts vs. control was d = 0.61 (95% CI 0.41–0.81, p < 0,00001). The effect was stronger for younger patients, with longer treatment, in studies with lower risk of bias, in randomized and blinded studies. Sensitivity analyses support the validity of the finding. 50% of the QoL subdomains (e.g. pain, nausea) show a significant improvement after mistletoe treatment. Most studies have a high risk of bias or at least raise some concern.ConclusionMistletoe extracts produce a significant, medium-sized effect on QoL in cancer. Risk of bias in the analyzed studies is likely due to the specific type of treatment, which is difficult to blind; yet this risk is unlikely to affect the outcome.PROSPERO registrationCRD42019137704.

Anticancer Effects of Viscum album Fraxini Extract on Medulloblastoma Cells in vitro

Background: Mistletoe therapy is frequently administered as a supportive treatment in diverse pediatric cancer entities including brain tumors. Medulloblastoma is the most common brain tumor in childhood. Its high risk to metastasize and its long-term sequelae caused by aggressive chemo- or radiotherapies are still challenging. Material and Methods: Effects of a lectin-rich mistletoe extract, abnobaVISCUM Fraxini, were investigated in two medulloblastoma cell lines (Daoy and ONS-76). Responsiveness of tumor cells was assessed by cell viability assays and xCELLigence real-time analyses. Moreover, impacts on proliferation, cell cycle and apoptosis were investigated. Apoptosis was studied by staining of vital mitochondria and assessing the involvement of caspases. In addition, effects on migration and invasion were analyzed. Results: Both medulloblastoma cell lines were more susceptible to treatment with the mistletoe extract than a nontumorigenic fibroblast cell line. In mistletoe-sensitive Daoy cells, reduction of proliferation and induction of caspase-mediated apoptosis were observed upon administration of 0.05 and 0.5 mg/mL abnobaVISCUM Fraxini treatment, respectively. Furthermore, mistletoe extract inhibited migration and invasion properties in Daoy and significantly impaired invasive capabilities of ONS-76 cells. Conclusion: AbnobaVISCUM Fraxini has cell line dependent antitumoral effects in medulloblastoma models. These results call for further investigations, to reveal mechanistic insights into antitumorigenic properties of mistletoe extracts.

Transiently Expressed Mistletoe Lectin II in Nicotiana benthamiana Demonstrates Anticancer Activity In Vitro.

Mistletoe (Viscum album) extracts have been used as alternative and complementary therapeutic preparations in multiple cancers for decades. Mistletoe lectins (ML-I, ML-II, and ML-III) are considered to be the main anticancer components of such preparations. In the present study, ML-II was transiently expressed in Nicotiana benthamiana using the pEAQ-HT expression system. Expression levels of up to 60 mg/kg of the infiltrated plant tissue were obtained, and a three-fold increase was achieved by adding the endoplasmic reticulum (ER) retention signal KDEL to the native ML-II sequence. The native protein containing His-tag and KDEL was purified by immobilized metal affinity chromatography (IMAC) and gel filtration. We found that the recombinant ML-II lectin was glycosylated and retained its carbohydrate-binding activity. In addition, we demonstrated that plant produced ML-II displayed anticancer activity in vitro, inhibiting non-small cell lung cancer H460 and A549 cells with EC50 values of 4 and 3.5 µg/mL, respectively. Annexin V-448A and PI double staining revealed that cell cytotoxicity occurred via apoptosis induction. These results indicate that ML-II transiently expressed in N. benthamiana plants is a promising candidate as an anticancer agent, although further optimization of production and purification methods is required to enable further in vitro testing, as well as in vivo assays.

Possibility of using lichen and mistletoe extracts as potential natural wood preservative

Increasing environmental pressures on toxic chemical wood preservatives lead to the development of natural and environmentally friendly wood preservatives. In this study, using possibilities of lichen (Usnea filipendula) and leaves of mistletoe (Viscum album) as potential natural wood preservative were researched. Impregnation procedure was applied at four different concentration levels and with two different extraction methods (hot water and methanol). The concentration levels were arranged as 3%, 5%, 10%, 15% for hot water and as 3.75%, 6.25%, 12.5%, 18.75% for methanol. The treatment procedure has been applied according to the ASTM D 1413 (1988) standard test method. The fungal decay test has been done according to the EN 113 (1996) standard test method using a brown rot fungus, Coniophora puteana for both treated test and untreated control samples. The best results were obtained at the highest concentration level of the solutions. However, the weight losses in treated test specimen have not met the standard requirements. Nevertheless, it can be assumed that both natural extracts provide promising protection performance.

Anticancer and biochemical effects of Viscum album L. protein extracts on HeLa cells

European mistletoe (Viscum album) is a medicinal plant with significant anticancer properties. In vitro callus production provides an essential alternative for biomass production and medicinal compounds propagation. In this study, we investigated (a) the biotechnological production of somaclonal proteins from mistletoe callus propagation and (b) the in vitro cytotoxic properties of these mistletoe protein extracts against cervical cancer. Mistletoe proteins (MPEs) were extracted from natural resources (plant leaves and stems) and in vitro propagated variant calluses. Potential cytotoxic effects of the extracted proteins on HeLa cells were assessed by the MTT proliferation assay, whereas oxidative damage and induction of apoptosis were measured fluorometrically by the reactive oxygen species (ROS) sensitive carboxy-H2-DCFDA method and caspase-3 activation assays, respectively. Electrophoretic results indicated differences in the protein bands between callus and donor plant tissues. Leaf and stem protein extracts (PEs) demonstrated a strong impact on HeLa cells viability by promoting oxidative damage and inducing apoptosis in contrast to callus PEs. This study shows that MPEs application in HeLa cells causes oxidative alterations and apoptosis through caspase-3 activation and increased intracellular ROS levels. These findings may be useful for uncovering novel bioactive products of pharmaceutical interest as well as for gaining an understanding of the biochemical pathways of mistletoe proteins in cancer treatment. In spite of the low cytotoxicity observed for biotechnologically propagated somaclonal variant calluses, towards cervical cancer, the ability to establish biotechnological production of said mistletoe extracts is important because it allows for standardization of protein production.

Time and Dose-Dependent Effects of Viscum Album Quercus on Rabbit Spermatozoa Motility and Viability in Vitro.

The target of this study was to evaluate the effect of extract of the European mistletoe - Viscum album quercus L. on spermatozoa motility and viability in vitro. The CASA system was used to determine the spermatozoa motility parameters at different time intervals (0, 1, 2 and 3 h) and spermatozoa viability was determined in five different doses of Viscum album quercus L [10 (QA), 6.6 (QB), 3.3 (QC), 2.5 (QD) and 2 (QE) mg/ml]. Results in experimental groups detected a significant deterioration on rabbit spermatozoa after 1, 2 and 3 hours, compared to the control. The initial total spermatozoa motility showed increased value for all doses of Viscum album quercus in comparison to control. After in vitro culture a dose-dependent decrease (QA: reduction of 69.7 %, QB: reduction of 40.9 %) was found. For the progressive spermatozoa most significant decrease (86.8 % for QA vs. 48.5 % for QB) was detected compared to the control after 3 hours of culture. Spermatozoa viability (MTT test) was decreased in all experiment groups at the end of experiment, but the differences were not significant. Significant alterations of membrane integrity were found in groups with the highest Viscum album quercus concentration (QA, QB), but acrosome integrity showed no significant changes. Results suggest negative dose- and time-dependent effect of Viscum album quercus at higher doses on spermatozoa motility and viability parameters in vitro.

3246 Long-Term Survival in a Patient With Recurrent Gastric Cancer With Peritoneal Seeding Undergoing Chemotherapy and Mistletoe Extract Treatment

INTRODUCTION: Recurrent gastric cancer with peritoneal seeding is associated with poor overall survival. Mistletoe extract has been widely used for complementary cancer therapy such as pancreatic cancer, ovarian cancer, and melanoma which support the quality of life and prolong overall survival. However, the effect of treatment for recurrent gastric cancer remains controversial. CASE DESCRIPTION/METHODS: A 71-year-old man with advanced gastric cancer underwent radical total gastrectomy (stage IIIb, T4b N1 M0). Abdominopelvic computed tomography performed 7 months after surgery revealed the increased ascites with an enhanced peritoneal thickening. He was diagnosed with peritoneal seeding and received 2nd line chemotherapy (Irinotecan). However, he was refused the chemotherapy due to side effects after 1 cycle. He starts receiving alternative therapy with Viscum album extract (mistletoe) instead of chemotherapy. Malignant ascites had decreased gradually after mistletoe extract therapy. So we start 3rd line chemotherapy (docetaxel) with mistletoe extract. The patient achieved good health with complete response postoperative 32 months after combination treatment. The patient has survived for 51 months after surgery without disease progression. We report a case of recurrent gastric cancer with peritoneal seeding that was successfully treated by docetaxel monotherapy with long-term mistletoe extract treatment. DISCUSSION: Mistletoe extract contributed to the reduction of malignant ascites and improved quality of life during chemotherapy. This case suggests that the synergistic effect of mistletoe extract treatment could be expected when used together with chemotherapy for recurrent gastric cancer.

Preclinical Evaluation of Antitumoral and Cytotoxic Properties of Viscum album Fraxini Extract on Pediatric Tumor Cells.

In Europe, especially in German-speaking countries, administration of mistletoe extracts is the most common and popular complementary and alternative therapy approach reported in oncology. Mistletoe therapy is applied to children with cancer for curative and palliative therapeutic regimes with increasing frequency, but at the same time, there are only a few studies on the effectiveness of this therapy. Therefore, we have investigated the response of various pediatric cell lines (acute myeloid leukemia, Ewing's sarcoma, hepatocellular carcinoma, medulloblastoma, neuroblastoma, and osteosarcoma) to mistletoe extract, abnobaVISCUM Fraxini. Effects on cell proliferation, cell cycle distribution as well as on mitochondrial integrity and caspase-mediated apoptosis were investigated in neuroblastoma cell lines, SH-SY5Y and Kelly. Additionally, in vitro tumor cell migration and invasion were studied. In vivo effects of the mistletoe extract were investigated in a syngeneic neuroblastoma mouse model. We could show that tumor cell lines were from 5- to 640-fold more sensitive to abnobaVISCUM Fraxini treatment than non-tumorigenic fibroblasts, whereby neuroblastoma cell lines were the most sensitive. For two neuroblastoma cell lines, SH-SY5Y and Kelly, induction of caspase-9-mediated apoptosis, a decrease of mitochondrial integrity as well as attenuation of migration and invasion were observed. In vivo experiments revealed a reduction of tumor growth and a prolonged survival of tumor-bearing animals. In summary, we can state that these results provide the first preclinical data for cytotoxic activities of abnobaVISCUM Fraxini for a broad panel of pediatric tumor cell lines, in particular, neuroblastoma cells. Thus, it might be a potential remedy for the supportive treatment of neuroblastoma.

Velutin, an Aglycone Extracted from Korean Mistletoe, with Improved Inhibitory Activity against Melanin Biosynthesis.

The abnormal regulation of melanin synthesis leads to a wide range of pigmentary disorders. Although various melanin biosynthesis inhibitors have been developed, their efficacy and long-term safety needs to be further improved, and thus the goal of this study is to develop promising natural compound inhibitors of melanin biosynthesis. Here, we obtained aglycone flavonoid extract through the microwave-assisted hydrolysis of glycoside extract from Korean mistletoe in acidic condition. The aglycone extract inhibited tyrosinase activity more efficiently with better antioxidant activity than glycoside extract in vitro. The microwave-assisted aglycone extract of mistletoe was further analyzed for in vivo activity, and the results showed the aglycone extract inhibited both early melanocyte development and melanin synthesis more efficiently in zebrafish embryo in a dose-dependent manner. Our in vivo toxicity assay quantitatively measured cell death in zebrafish embryos and showed that the microwave-assisted aglycone extract of mistletoe had no significant effect on cell death (p < 0.001), indicating that aglycone extract is more biocompatible than glycoside extract. Furthermore, our in vitro and in vivo analyses successfully identified and characterized velutin, an aglycone of a homoflavoyadorinin B glycoside, as a major inhibitory component in the microwave-assisted mistletoe extract. Ultimately, this study showed that the novel natural compound inhibitor velutin, which was generated through microwave-assisted extraction from mistletoe, improved the efficacy of melanin biosynthesis inhibition with little toxicity.