In vitro evaluation of the effects of Viscum album homeopathic preparations on murine melanoma cells

Viscum album extract (VA) is a complementary treatment in cancer, with in vitro and in vivo cytotoxic effects on several tumor types when applied in phytochemical doses. However, highly diluted ethanolic homeopathic preparations' effects and mechanisms need further study. Aims:To assess the in vitro effects of highly diluted VA from the subspecies V. album abietis and V. album album at different potency levels in different dilution ratios on murine melanoma cells. Methodology:The VA mother tinctures (MT)from Abies alba (MTA) and Quercus robur (MTQ) were prepared with summer and winter samples, harvested in Switzerland. They were submitted to homeopathic ethanolic maceration and a subsequent dynamization process. MTA, MTQ and the following respective potencies were tested in B16F10 murine cells: 3x, 12x, 30x, 6cH, 12cH, 200cH, 2LM, 3LM, and 5LM. Dynamized water, dynamized and non-dynamized ethanol, and carboplatin were used as control groups. The mitochondrial activity and cell viability analysis were performed at 1, 24, 48, and 72 hours by in vitro incubation. MTA and MTQ harvested in summer, as well as 12x, 200cH and 5LM potencies were also tested to cell apoptosis and necrosis markers, reactive oxygens species (ROS) production, inflammatory cytokines profile, cell morphology, and migratory capacity. Results and discussion: MTA and MTQ induced a decrease in cell metabolism and higher cytotoxicity within 1 hour, with significant morphological changes and increased production of ROS and inflammatory cytokines. Both homeopathic dilutions 12x and 5LM showed an influence on cell metabolism, cell replication, and oxidative stress modulation with inflammatory cytokines, mitosis, and migration pattern changes. On the other hand, Quercus robur and Abies alba 200cH showed increased on cytotoxicity and ROS levels, respectively. Conclusion:The in vitro effects of Viscum album homeopathic solutions in melanoma cells highlight the promising antitumoral potential and reinforce the need for further research to better understanding their mechanisms of action.

Variación de los perfiles de polifenoles en Ligaria cuneifolia (R. et. P) Tiegh (Loranthaceae), su estudio mediante electroforesis capilar y su relación con la actividad farmacológica. / Polyphenolic profile variation in Ligaria cuneifolia (R. et P.) Tiegh (Loranthaceae), capillary electrophoretic analysis and its relationship with the pharmacological activity

Ligaria cuneifolia (R. et P.) Tiegh. ­Loranthaceae­ es una especie hemiparásita que se desarrolla sobre diferentes hospedantes. Es conocida con el nombre vulgar de "liga" o "liguilla". Debido a su similitud morfológica, constituye el sustituto natural del "muérdago europeo", por lo cual es denominado "muérdago criollo". Las drogas vegetales son matrices complejas en las cuales múltiples componentes actúan en forma sinérgica y son responsables de la acción farmacológica. Con el fin de dar sustento científico al uso folclórico de L. cuneifolia se estudiaron distintas formas de obtención de los extractos, se evaluaron diferentes hospedantes y regiones fitogeográficas. Se desarrolló y validó un método de electroforesis capilar para construir fingerprints o perfiles cromatográficos característicos que permitan evaluar los distintos componentes con el fin de estandarizar los extractos. Se efectuó la comparación con otras técnicas cromatográficas, tales como en cromatografía en capa delgada (TLC) y líquida de alta resolución (HPLC). A su vez, se procedió al aislamiento, purificación y análisis estructural de los compuestos de interés por técnicas espectroscópicas y cromatográficas. Se identificaron diez compuestos, de los cuales cuatro son reportados por primera vez en esta especie. La electroforesis capilar probó ser una técnica adecuada para el control de calidad de los extractos y una alternativa atractiva a las técnicas cromatográficas tradicionales.

Ligaria cuneifolia (R. et P.) Tiegh. ­Loranthaceae­ is a hemiparasite plant which grows on different host trees. It is popularly referred to as "liga" or "liguilla". Due to its morphological similarity, it is considered as the natural substitute for the European mistletoe, for which is known as the "Argentine mistletoe". Herbal drugs are complex matrices in which multiple components acting synergistically are responsible for the pharmacological activity. In order to provide scientific support to the popular use of L. cuneifolia, a capillary electrophoretic method was developed and validated to build a chromatographic profile or fingerprint that allows the evaluation of different components for extract standardization. A comparison was made with other chromatographic techniques such as TLC and HPLC. Isolation, purification and structural analysis of compounds were performed by chromatographic and spectroscopic methods. Ten analytes were identified, four of which are reported for the first time in L. cuneifolia. Capillary electrophoresis proved to be an appropriate tool for the quality control of herbal drugs, as well as an attractive alternative to traditional chromatographic techniques.

Conocimiento local del género Arceuthobium en un área natural protegida del centro de México / Local knowledge of Arceuthobium genus in a natural protected area of the centre of Mexico

El género Arceuthobium (muérdago enano) es una de las principales plagas en los bosques templados de México, con antecedentes de uso medicinal y forrajero. El objetivo de este trabajo fue documentar el conocimiento local sobre los usos del muérdago enano en el Área Natural Protegida Nevado de Toluca, México, bajo las perspectivas emic y etic, a través de la etnobotánica. Mediante entrevistas se identificó que ambas especies (A. vaginatum y A. globosum) son conocidas como muérdago y forman parte de la flora del bosque; sin embargo, los entrevistados desconocen los efectos negativos dentro de los ecosistemas forestales de esta plaga. Destacan los usos lúdicos, como juguete y tinta, y los medicinales para el tratamiento de afecciones respiratorias y del sistema nervioso, siendo éste el primer registro para dicho género localmente. Se requieren estudios que validen el conocimiento local para su inclusión en programas locales de manejo de plagas forestales.

Arceuthobium (dwarf mistletoe) genus is one of the main pests in temperate forests of Mexico, with records of medicinal and forage uses. The objective of this work was to document local knowledge regarding uses of dwarf mistletoe in the Natural Protected Area Nevado de Toluca, Mexico, under emic and etic perspectives, through ethnobotany. Throughout interviews it was identified that both species (A. vaginatum and A. globosum) are known as mistletoe and they are part of flora in forest, however, interviewees are not aware of negative effects within forest ecosystems of this pest. It stands out ludic uses as a toy and ink, and medicinal ones for treatment of respiratory diseases and nervous system, being this the first record for such genus locally. Studies are required to validate the local knowledge for its inclusion in local programmes for management of forest pests.

Anti-Inflammatory Properties of Flavone di-C-Glycosides as Active Principles of Camellia Mistletoe, Korthalsella japonica

The chemical components and biological activity of Camellia mistletoe, Korthalsella japonica (Loranthaceae) are relatively unknown compared to other mistletoe species. Therefore, we investigated the phytochemical properties and biological activity of this parasitic plant to provide essential preliminary scientific evidence to support and encourage its further pharmaceutical research and development. The major plant components were chromatographically isolated using high-performance liquid chromatography and their structures were elucidated using tandem mass spectrometry and nuclear magnetic resonance anlysis. Furthermore, the anti-inflammatory activity of the 70% ethanol extract of K. japonica (KJ) and its isolated components was evaluated using a nitric oxide (NO) assay and western blot analysis for inducible NO synthase (iNOS) and cyclooxygenase (COX)-2. Three flavone di-C-glycosides, lucenin-2, vicenin-2, and stellarin-2 were identified as major components of KJ, for the first time. KJ significantly inhibited NO production and reduced iNOS and COX-2 expression in lipopolysaccharide-stimulated RAW 264.7 cells at 100 μg/mL while similar activity were observed with isolated flavone C-glycosides. In conclusion, KJ has a simple secondary metabolite profiles including flavone di-C-glycosides as major components and has a strong potential for further research and development as a source of therapeutic anti-inflammatory agents.

Alteraciones anatómicas en epidermis de hojas infectadas de Phoradendron bathyoryctum Eichler (Viscaceae) / Anatomical alterations in epidermis of infected leaves of Phoradendron bathyoryctum Eichler (Viscaceae)

Phoradendron bathyoryctum Eichler (Viscaceae) es una planta hemiparásita que crece en la República Argentina sobre varias especies arbóreas. Se utiliza en medicina popular de la misma manera que el muérdago europeo (Viscum album L. ­Viscaceae­), para disminuir la presión arterial. Los muérdagos, a veces, pueden ser susceptibles de epiparasitismo, o también sufrir el ataque de diferentes agentes fitopatógenos. El objetivo de este estudio es analizar los cambios anatómicos que se producen en las hojas de Ph. bathyoryctum cuando son afectadas por algún agente infeccioso. Se colectaron ejemplares sanos e infectados, los últimos, con lesiones foliares superficiales. Se realizaron técnicas de raspado, diafanizado y obtención de epidermis en las hojas de los ejemplares. También se practicaron reacciones histoquímicas. En las hojas infectadas se observó una destrucción progresiva de los estomas y cambios en el resto de los tejidos epidérmico y subepidérmico, que dieron como resultado la formación de estructuras visibles macroscópicamente, y semejantes a lenticelas foliares. De acuerdo con las observaciones realizadas y por comparación con la bibliografía consultada, se supone que el daño foliar se inicia en el estoma donde se producen cambios estructurales hasta su destrucción, la modificación de las células adyacentes y la formación de lesiones superficiales en las hojas infectadas. En cuanto al agente, si bien no se pudo determinar en esta oportunidad, podría asociarse a algún tipo de virus. Se sugiere realizar futuros estudios para confirmar si los virus son los causantes de las lesiones. (AU)

Korean Mistletoe (Viscum album var. coloratum) Inhibits Amyloid beta Protein (25-35)-induced Cultured Neuronal Cell Damage and Memory Impairment

The present study aims to investigate the effect of methanol extract of Korean mistletoe (KM; Viscum album var. coloratum), on amyloid beta protein (Abeta) (25-35), a synthetic 25-35 amyloid peptide, -induced neurotoxicity in cultured rat cerebral cortical neurons and memory impairment in mice. Exposure of cultured neurons to 10 microM Abeta (25-35) for 24 h induced a neuronal cell death, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. KM (10, 30 and 50 microg/ml) significantly inhibited the Abeta (25-35)-induced apoptotic neuronal death. KM (50 microg/ml) inhibited 10 microM Abeta (25-35)-induced elevation of intracellular calcium concentration ([Ca2+]i), which was measured by a fluorescent dye, Fluo-4 AM. Glutamate release into medium and generation of reactive oxygen species (ROS) induced by 10 microM Abeta (25-35) were also inhibited by KM (10, 30 and 50 microg/ml). These results suggest that KM may mitigate the Abeta (25-35)-induced neurotoxicity by interfering with the increase of [Ca2+]i and then inhibiting glutamate release and generation of ROS in cultured neurons. In addition, orally administered KM (25 and 50 mg/kg, 7 days) significantly prevented memory impairment induced by intracerebroventricular injection of Abeta (25-35) (8 nmol). Taken together, it is suggested that anti-dementia effect of KM is due to its neuroprotective effect against Abeta (25-35)-induced neurotoxicity and that KM may have therapeutic role in prevention of the progression of Alzheimer's disease.

Immunologic Response to Mistletoe Extract (Viscum album L.) after Conventional Treatment in Patients with Operable Breast Cancer / 한국유방암학회지

PURPOSE: To reduce the side effects and improve the effectiveness of standard chemoradiation therapy, many complementary or alternative medicines have been tried. However, little is known about its immunologic effects in breast cancer patients. The aim of this study was to assess the immunologic effects of mistletoe extract (Viscum album L., VAE) in patients with early breast cancer after surgery followed by standard adjuvant chemoradiation therapy. METHODS: A total 20 patients with early breast cancer treated with breast conserving surgery followed conventional chemoradiation therapy. Ten of these patients received subcutaneous injections of VAE for 7 weeks. IL-2, IL-4, IL-6, IL-10, TGF-beta, and IFN-gamma levels in serum samples were measured in all patients. RESULTS: The concentrations of IL-2, IL-4, IL-10, and TGF-beta were not significantly changed between before and after VAE treatment in both test and control group. The concentration of IL-6 in the test group was increased from 8.19+/-1.75 pg/mL to 9.86+/-1.46 pg/mL after treatment (p=0.013). The concentration of IFN-gamma in the test group was remarkably increased from 91.76+/-17.16 pg/mL to 167.42+/-66.61 pg/mL after treatment (p=0.009). CONCLUSION: Significant increases in the concentration of IL-6 and IFN-gamma were observed after VAE treatment. These results suggest that VAE treatment can stimulate immune responses, especially cell-mediated immunity in immune-compromised patients received the chemoradiation for breast cancer.

Frugivoria de larvas de Neosilba McAlpine (Diptera, Lonchaeidae) sobre Psittacanthus plagiophyllus Eichler (Santalales, Loranthaceae) no sudoeste de Mato Grosso do Sul, Brasil / Larval frugivory of Neosilba McAlpine (Diptera, Lonchaeidae) on Psittacanthus plagiophyllus Eichler (Santalales, Loranthaceae) in southwestern Mato Grosso do Sul State, Brazil

Loranthaceae são plantas hemiparasitas com distribuição geográfica mundial e representadas no Brasil por seis gêneros. Os mais importantes são Phthirusa, Psittacanthus e Struthanthus, os quais parasitam uma grande diversidade de plantas hospedeiras. Este trabalho avaliou a occurrência e a flutuação sazonal de moscas infestando os frutos de Psittacanthus plagiophyllus nos municípios de Anastácio, Aquidauana e Miranda, Mato Grosso do Sul, Brasil, onde ervas-de-passarinho são disseminadas. As coletas foram realizadas de junho de 1998 a julho de 2000 para obtenção de frutos maduros de P. plagiophyllus, seus insetos associados e catalogação dos seus hospedeiros. Os insetos foram criados para identificação, sendo obtidos 1.522 adultos de Neosilba spp., destes, 612 machos foram identificados a nível específico: Neosilba bifida Strikis & Prado (6 indivíduos), N. certa (Walker) (26 indivíduos), N. pendula (Bezzi) (16 indivíduos), N. zadolicha McAlpine & Steyskal (4 indivíduos) e duas diferentes espécies: morfotipo MSP1 (478 indivíduos) e morfotipo 4 (82 indivíduos). O período de mais alta infestação por Neosilba spp. ocorreu durante agosto de 1998 e de 1999 e, a espécie morfotipo MSP1 foi significativamente mais abundante que todas as outras. A morfotipo 4 foi a segunda mais abundante, diferindo significativamente de Neosilba zadolicha. Neosilba foi o único gênero de moscas frugívoras (Tephritoidea) infestante dos frutos de P. plagiophyllus e comportou-se como um invasor primário neste hospedeiro.

Loranthaceae are hemiparasite plants with worldwide distribuition, represented in Brazil by six genera. The most important are Phthirusa, Psittacanthus and Struthanthus that parasitize a great diversity of host plants. This paper evaluated the occurrence and the seasonal fluctuation of flies infesting the fruits of Psittacanthus plagiophyllus in the municipalities of Anastácio, Aquidauana and Miranda, Mato Grosso do Sul, Brazil, where mistletoes are widespread. The samples were taken from June 1998 to July 2000 to obtain the hosts and its associated insects. All insects were reared until adult stage, and it were obtained 1,522 adults of Neosilba spp., of which 612 males were identified at species level: Neosilba bifida Strikis & Prado (6 specimens), N. certa (Walker) (26 specimens), N. pendula (Bezzi) (16 specimens), N. zadolicha McAlpine & Steyskal (4 specimens) and two different species, morphotype MSP1 (478 specimens) and morphotype 4 (82 specimens). The period of highest infestation by Neosilba spp. occurred during August 1998 and 1999, and morphotype MSP1 was significantly more abundant than all other species. The species morphotype 4 was the second most abundant, differing significantly from Neosilba zadolicha. Neosilba was the only genus of frugivorous fly infesting fruits of P. plagiophyllus and behaves as primary invader in this host.

The Effects of Mistletoe Extract and Anti-cancer Drugs on the Apoptosis of Gastric Cancer Cells

PURPOSE: Mistletoe extract was widely used for cancer treatment as complementary or alternative therapy in European area from early twenty century. It is currently used as alternative anti-cancer remedy by piecemeal in domestic medical group, however, the anti-cancer mechanism of mistletoe extract was not known precisely until now. In this study the effect of mistletoe extract on gastric cancer was studied vis cell line experiments. MATERIALS AND METHODS: The SNU719 gastric cancer cell line was used, and ABNOBAviscum-Q and ABNOBAviscum-F were treated to cells as mistletoe extract, or 5-FU and cisplatin were used with mistletoe extract. The cell viability and cell death rate were estimated by CCK-8 assay kit and lactate dehydrogenase (LDH) assay kit in each. Caspase 3 assay kit was used to measure caspase 3 activity. The protein expression amounts of Bcl2, p53, and PTEN were estimated through Western blot analysis. RESULTS: The co-treatments of mistletoe extract Q/F and 5-FU/cisplatin decreased lesser cell viability than only mistletoe treat. Caspase 3 activity was increased 4~6 times in co-treatment of mistletoe extracts and 5-FU than control. Bcl2 protein expression was reduced by mistletoe extracts or anti-cancer drugs, further more, the co-treatment of mistletoe extracts and 5-FU/cisplatin diminished more the expression than only mistletoe treatment. Mistletoe extracts did not affect the protein expressions of p53 and PTEN. CONCLUSION: It was concluded that the anti-cancer mechanism of mistletoe extracts was made by caspase 3 activation and lowered Bcl2 expression, and this apoptosis inducing mechanism was independent to p53.

Complementary Therapies and Cancer Treatment

Many patients with cancer use complementary medicine that is used together with conventional medicine. The widespread use of complementary therapies in cancer patients has major implications for research and clinical practices. Some complementary therapies, such as polysaccharide K, Astragalus membranaceus and A. mistletoe, may offer therapeutic benefits for patients with cancer. However, others did not show significant clinical effects for cancer treatment. Therefore, it is important for physicians to communicate openly with cancer patients about the use of complementary therapy. In order to encourage open communications with their patients, physicians should be knowledgeable about the most commonly used remedies, or at least they should be able to direct patients to reliable sources of information.

Complementary Therapy for Improvement of Quality of Life in Cancer Patients

Complementary Therapy for Improvement of Quality of Life in Cancer Patients. As a results of reviews of database for effects of complementary therapies for improvement of Quality of Life on cancer patients, there were 8 mind-body therapies, 2 manual based therapies, 17 biologic based therapies(3 vitamin or minerals, 6 health nutrient products, 8 herbs). There were relative much more proportional studies of mind-body therapies and manual based therapies for quality of life of cancer. It were reported much more positive results in cognitive behavioral therapy, meditation, art therapy, music therapy, yoga included to mind-body therapies also it were reported much more positive result in massage & acupressure included to manual based therapies. And then these findings can be considered to us clinical application of these therapies by additional studies. On the other hand, the amount of study for quality of life of cancer in biologic based therapies was relatively more smaller than mind-body therapies and the results were contrary to each other. The mistletoe, one of the biological therapeutic products, can be considered to relatively useful material as a results of this review and its seems like to have positive value for further study, materials such as honey, ginseng or heat-treated ginseng, AHCCR, selenium. The others of biologic based therapies were inconclusive due to relatively small amounts of studies or revealed negative conclusion or negative tendency by their studies. The high usual rate of complementary therapy in cancer patients in Korea is applying the screw to us study for complementary medicine and is producing needs of complementary closed co-works or cooperation between the citizens, the government, medical doctors for high quality study.

Korean Mistletoe (Viscum album Coloratum) Extract Induces Eel (Anguilla japonica) Non-specific Immunity

BACKGROUND: The immunomodulatory effects of Korean mistletoe (Viscum album Coloratum) on the innate immune responses of eel (Anguilla japonica) were studied. METHODS: Mistletoe, Freund's complete adjuvant (FCA), or phosphate-buffered saline (PBS) as a control was injected into eel peritoneal cavities. RESULTS: Nitroblue tetrazolium (NBT)-positive cells in the head kidney of eel were significantly augmented by the second day post-injection of mistletoe. Reactive oxygen intermediates (ROI) were more produced in mistletoe-injected fish kidney leucocytes than in FCA-injected ones. The level of lysozyme activity in the serum of fish 2 days after injection with mistletoe was also significantly higher than that in the serum of the control fish. The optimal concentration of mistletoe in inducing the highest serum lysozyme activity was revealed to 500microgram/200 g of fish. In phagocytic activity assay, mistletoe-sensitized eel kidney phagocytes captured more zymosan than did the control fish. CONCLUSION: Korean mistletoe appeared to be a good activator of the non-specific immune responses of eel.

The Immunological Effect of Mistletoe Extract on Gastric Cancer Patients

PURPOSE: Mistletoe (Viscum album L.) extract is one of the most widely used agents in alternative cancer therapeutic regimens in Europe. This study was conducted to determine the effect of mistletoe extract on immune function in gastric cancer patients. MATERIALS AND METHODS: Ten patients that had undergone a curative gastrectomy were enrolled in the prospective study. ABNOBAviscum Q(R) was injected subcutaneously three times a week from postoperative-day 7 to week 16 with an increasing dose. All of the patients simultaneously received chemotheraphy with mitomycin, oral 5-FU and a cisplatin regimen. The WBC count, differential count, lymphocyte/WBC ratio and the level of cytokines (IL-1beta, IL-2, IL-6, IFN-gamma, TNF-alpha) were checked in the peripheral blood preoperatively, at postoperative week 8 and at postoperative week 16. RESULTS: The WBC and neutrophil counts significantly decreased after treatment on week 8 and week 16 (P=0.001), but the total eosinophil count was slightly increased (P=0.15). The total lymphocyte count also decreased during treatment but the lymphocyte/WBC ratio was slightly increased without statistical significance (P=0.91). The cytokine levels did not significantly change during treatment. CONCLUSION: It is somewhat difficult to determine the direct effect of mistletoe therapy on immune function as the effect may be compromised by the concurrent chemotherapy. It can be assumed that the slightly increased lymphocyte/WBC ratio and eosinophil count may be a result of the immunomodulatory effect of the mistletoe extract.

Radiofrequency thermal ablation versus trans-catheter hepatic arterial chemo embolization for hepatocellular carcinoma: follow up with and without giving mistletoe [fraxini 2]

Hepatocellular carcinoma [HCC] represents a major health problem world wide. It accounts for 90% of all primary liver cancers and is the fifth most common malignancy [1]. Evaluation of radiofrequency thermal ablation versus transarterial hepatic chemoembolization with the effect of Viscum [Fraxini 2] on tumour recurrence. 60 patients with HCC were enrolled in the study. Group 1 include 30 patients and were treated with radiofrequency thermal ablation [15 patients of them received viscum 2 by subcutaneous route for 2 years]. Group 11 included 30 patients with HCC and were treated by TACE [15 patients of them received viscum 2 subcutaneously for 2 years]. Group 1 patients showed total ablation in 70% with persistant inactivity during 2 years follow up. Group II did not show significant difference from Group 1 as regards relapse rate nor the performance status. Complications as nausea, vomiting, fever, jaundice, and elevation of transaminases were significantly more encountered with TA CE. Viscum 2 did not significantly arrest tumour recurrence. Non surgical patients with HCC can achieve curative treatment with radiofrequency with minimal side effects. TACE is a palliative treatment option for large HCC

Treatment strategy for postoperative persistent pain of endometriosis / 대한산부인과학회지

OBJECTIVE: This study was designed to establish the treatment strategy on postoperative persistent pain of endometriosis. METHODS: During a 12-month period from March 2000 to February 2001, 62 patients had postoperative persistent pelvic pain. 62 patients of them were treated with GnRH analogue for 6 months. If they were not responsive to those treatment, It were recommended high dose progesterone treatment for 6 months after trigger point injection to differentiate from trigger point pain. and then for remained pain, Nonsteroidal antiinflammatory drugs (NSAIDs) and tricyclic antidepressants (TCAs) were supplied for 6 months. Finally, contineously remained pelvic pain was treated by mistletoe extract. RESULTS: 62 patients had a postoperative persistent pelvic pain. GnRH analogue were supplied to 62 patients, and then 23 patients had persistent pain. These 23 patients undertook lidocaine injection on trigger point to differentiate from trigger point pain. Two patients were responsive to injection. Remained 21 patients were treated with high dose progesterone for 6 months. Thereafter unresponded 14 patients were supplied with NSAIDs and TCAs. Finally, remained 5 patients that were uncontrolled with previous treatments were injected with mistletoe extract. Three patients were controlled with mistletoe extract. CONCLUSION: Most of persistent pain after laparoscopic operation of endometriosis were successfully controlled by sequential treatment of GnRH analogue, trigger point injection, high dose progesterone, NSAIDs and TCAs, and mistletoe extract injection.

An experimental study on mistletoe extract-induced apoptosis in oral squamous cell carcinoma

This study was performed to investigate mistletoe extract-induced apoptosis in oral squamous cell carcinoma. In vivo study, HN22 cells were xenografted in nude mice. After tumor was experimentally induced, mistletoe extract was directly injected on the tumor mass. The specimens were evaluated using light and transmission electron microscopes. In vitro study, HN22 cells were cultured and exposed to mistletoe extract. The cells were evaluated using transmissin electron microscope. To evaluate apoptotic cells, flow cytometric analysis was done. The results were obtained as follows: 1. Light microscopic view of tumor mass showed necrosis at 2-4 weeks. 2. Transmission electron micrographs of tumor mass showed apoptosis and necrosis. 3. In TEM view of cell lines, necrosis and apoptosis were shown with mistletoe extract at 300microgram/ml, apoptosis was shown with mistletoe extract at 100microgram/ml. 4. In flow cytometric analysis, early and late apoptosis was shown when using caspase-3Ab and annexin-V, but no significant change was noted when using mebstain and Apo2.7 Ab. In this study, mistletoe extract induced necrosis and apoptosis in the tumor mass was induced by HN22 cells, early and late apoptosis in vitro study. Mistletoe extract was likely to induce cell death in oral squamous cell carcinoma through apoptosis.

The Effect of subcutaneous injection of Mistletoe extract for the treatment of chronic pelvic pain / 대한산부인과학회지

OBJECTIVE: Chronic pelvic pain is pelvic pain which is not completely controlled by classical treatments. In many reports, immune alternation is related to original diseases that induce the chronic pelvic pain. Mistletoe extract has been known that it has immune modulation effect. Therefore the purpose of this study is to determine the pain-reducing effects of mistletoe extract for chronic pelvic pain that is not controlled by classical treatments. METHODS: The fitty-eight patients who visited to outpatient department of Department of Obstetrics and Gynecology, Chonnam National University Hospital due to chronic pelvic pain. Mistletoe extract (Helixor-A(R)) 5-10 mg was injected subcutaneously 3 times per week for 5 weeks, and the change of pain and short-term side-effects after injections were observed. In patients after 6 month of follow-up, the change of pain and long-term side-effects were also observed. RESULTS: In 2 of 58 patients (0.03%), treatments were discontinued doe to severe general side effect. In 48 of 56 patients (85.7%), chronic pelvic pains were improved. In 40 of 56 patients (69%), mild side effects as like mild itching were appeared and they were easily controlled by anti-histamines and conservative treatments. In 6 months after injection, general itching sensation and easily catched common cold were complaint in 4 of 10 patients. CONCLUSION: The subcutaneous injection of mistletoe extract may effective treatment method for the chronic pelvic pain that is not controlled by classical treatment. However more studies about log-term effect of mistletoe therapy are needed.

The Effect of intraperitoneal instillation of Mistletoe extract during the diagnostic laparoscopy for pain of endometriosis / 대한산부인과학회지

OBJECTIVE: Immune alternation is related to the pathogenesis and progression of endometriosis. Helixor-A, mistletoe extract, has been known that it has immune modulation effect. Therefore the purpose of this study is to determine the effects of Helixor-A for pain of endometriosis by intraperitoneal instillation of mistletoe extract during the diagnostic laparoscopy. METHODS: Among the paitients who visited to outpatient clinic due to chronic pelvic pain, 30 patients who had laparoscopically-confirmed endometriosis were included in this study. Helixor-A 5 mg was instilled into the posterior cul-de-sac during the diagnostic laparoscopy. Before diagnostic laparoscopy and one month after laparoscopy, the patients recorded pain scores. RESULTS: In 11 of 30 patients (36.7%), pelvic pain of endometriosis improved after laparoscopy. In 2 of 30 patients, the side-effect of Helixor- A appeared and they were easily controlled by antihistamines. CONCLUSION: The instillation of Helixor-A during diagnostic laparoscopy, that is essential for confirmation of endometriosis, may be helpful as a supportive treatment for treatment of pelvic pain due to endometriosis with classical treatment such as GnRH agonist therapy.

Establishment and Characteristics of the B Cell Line (KEB1) from the Bone Marrow Cells of the Patient Infected with Epstein-Barr Virus / 대한혈액학회지

BACKGROUND: Cell lines can be established when the cells are clonally selected and propagated continuously in vitro culture system. Recently we established a B cell line (KEB1) from the bone marrow cells of the patient infected with Epstein-Barr virus (EBV). METHODS: The patient's initial platelet count was 1,000/microliter and peripheral blood smear showed atypical lymphocytes accounting 20% of the differentials of WBC. Antibodies to EBV and PCR for EBV were positive but heterophil antibody was negative. Mononuclear cells were obtained by Ficoll-paque separation and suspended in RPMI media with 10% FCS. After incubation in 37degrees C, 5% CO2 incubator, cells grew continuously and finally immortalized to B cell line. RESULTS: Cells showed abundant, clear basophilic cytoplasms and a few vacuoles. Cells had granular reaction in PAS stain and were positive to B cell antibodies. Immunohistochemical stain showed positive expression for EBV antibody. Electron microscopic finding of cultured cells showed several viral particles, and immunoelectron microscopic finding showed electron dense expression. Immunophenotyping of cultured cells was positive for B lymphoid lineage, and karyotypings had hypotetraploidy. Cells expressed MAGE and SSX gene. Cytotoxicity showed relative resistance to mistletoe and several chemotherapeutic agents compared to leukemic cell line. CONCLUSION: KEB1 cell line was established from the bone marrow cells of the patient with infectious mononucleosis. The characteristics of the cell lines including morphology, immunophenotype, karyotype, gene analysis (MAGE, SSX) and chemosensitivity were analyzed. There should be further studies of these cell lines including gene analysis, telomerase activity and cytokine production. This cell line might be helpful to establish another normal lymphocyte cell line and to predict the toxicity of chemotherapy.

Effect of the aqueous extract of African mistletoe, Tapinanthus sessilifolius (P. Beauv) van Tiegh leaf on gastrointestinal muscle activity.

Effects of the aqueous extract of T. sessilifolius on the gastrointestinal muscle were investigated on smooth muscle preparations isolated from rabbit jejunum, guinea pig ileum and on gastrointestinal transit in mice. Elemental analysis of the extract was also carried out. The aqueous extract of T. sessilifolius evoked a concentration dependent contraction of the rabbit jejunum and guinea pig ileum. The contractions evoked by the extract were not attenuated either by atropine or mepyramine, but they were completely blocked by verapamil. The elemental analysis revealed the presence of Mg, Zn, Fe, Cu, and very high concentration of Ca. The intraperitoneal LD50 in mice was found to be 1500 mg/kg. The aqueous extract of T. sessilifoliius possesses active components that may be mediating the observed biological activity through calcium mobilization.