In-Vitro Growth Kinetics of Mesenchymal Stem Cells in Cytotoxicity Tests Using Low-Diluted Viscum Album.

INTRODUCTION: The use of mesenchymal stem cells (MSC) in cytotoxicity tests is an in-vitro alternative model for predicting initial doses. Homeopathic medicines may stimulate the immune system to combat a pathology effectively and have been used for over two centuries. Viscum album (VA) extracts are widely used in the treatment of cancer, due to their immunomodulatory, cytotoxic and pro-apoptotic properties. OBJECTIVE: This study aimed to evaluate the in-vitro growth kinetics of canine MSC in relation to cytotoxicity, cell differentiation and expression of pluripotentiality markers, using a VA preparation at the D1D2 (1×10-1, 1×10-2 potency (VAD1D2). METHODS: MSC were obtained from adipose tissue sampled from a healthy dog that was undergoing an elective veterinary procedure and with its owner's permission. The experiments were performed in three groups: MSC treated with VAD1D2 or diluent or untreated (control). The cytotoxicity was evaluated by MTT assay. The differentiation was induced in three lineages, and apoptotic cell labeling was performed by an Annexin-V test. RESULTS: At the concentration of 10 µL/mL of VA, the number of cells after in-vitro culture was maintained when compared with the control (untreated) group. A significant and gradual decrease in cell viability was recorded as VA concentrations increased. The apoptosis analysis showed that VA at 20 µL/mL presented absolute percentages of initial apoptosis twice as high as at 10 µL/mL, which was similar to the control (untreated group). CONCLUSION: The results suggest that the use of efficient methods to assess the in-vitro cytotoxicity of VA-based homeopathic medicines using MSC lineages may predict the potential action at different concentrations. These findings demonstrated that VAD1D2 interferes with canine MSC growth kinetics.

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.

Homeopathic mistletoe adverse reaction mimics nodal involvement in 18F-FDG PET/CT performed for evaluation of response to chemotherapy in lymphoma.

Some patients use complementary medicine. We present a patient with Hodgkin's lymphoma, scanned with 18F-FDG PET/CT for evaluation of response after chemotherapy, who was self-administering mistletoe as a homeopathic medicine product. The careful review of the images of the entire scan and patient collaboration in anamnesis were crucial to avoid a false positive result. A review of the published scientific data on the effects of mistletoe is also presented.

Viscumalbum L. homeopathic mother tinctures: metabolome and antitumor activity

Viscum album L. is a semi-parasitic plant with antitumor activity attributed to theaqueous extracts. However, European V. album ethanolic extracts (VAE) have also demonstrated invitro activity in tumor models. Aims: Evaluate the metabolic profiles of fifty VAE harvested duringsummer and winter seasons and their antitumor activity through 2D and 3D models. Methodology:VAEwerepreparedbymacerationfrom:V.albumsubsp.albumgrowingonMalus domestica,Quercus sp.and Ulmus sp.; V. album subsp. austriacum from Pinus sylvestris; V. album subsp. abietis from Abies alba.Chemical analyses were performed through liquid chromatography coupled with high resolutionmass spectrometry and Partial Least Squares Discriminant Analysis (PLS-DA) was performed in theMetaboanalyst 4.0. The antitumor potential of the selected VAE was evaluated in 2D and 3D models(MDA-MB-231 cancer cells) by MTT, crystal violet and glycolytic pathway analysis. Results anddiscussion:Thefirst3principalcomponentsinPLS-DAexplained60%and40%ofdatavariationin positive andnegativemodesrespectively.Threegroupswereformedandshowedchemicalsimilarityamong V. album subspecies. The compounds responsible for group separation were tentativelyidentifiedas:pinobankasinornaringenin hexoside;isorhamnetin-3-hexoside,meglutolanddifferent aminoacids.ThesummerVAEat0.5%v/vinducedhighercytotoxicdamagethanthewinterpreparations, and Abies alba and Quercus sp. VAE promoted 49% and 42% reduction of tumorviability in 3D model (72h incubation), respectively. MDA-MB-231 glycolytic pathway in 2D modelshowed a decrease in the glucose consumption and extracellular lactate production. Also, PFK (6-phosphofructo-1-kinase)andPK(Pyruvatekinase)activitieswereinhibitedbyAbiesalbaandQuercus sp. VAE at 48h of incubation. Conclusion: VAE extracts showed different metabolomes andthe glycolytic pathway should be an important target involved in the inhibition of tumor growth bytheseextracts

Homoeopathic Viscum album (10-3) is More Cytotoxic to in vitro Culture of Human Breast Cancer Cell Line than to Human Mesenchymal Stem Cells

BackgroundBreast cancer has been considered a public health problem and homeopathic treatments are becoming increasingly recommended due to its ways of action and absence of adverse effects. MCF-7 is an adenocarcinoma of human breast cell line useful as preclinicalmodel to screen therapeutic agents such as ultra-diluted Viscum album, an European plant which extract is commonly used in cancer therapy. AIMS MCF-7 and mesenchymal stem cells (MSC) were used to evaluate the in vitrocytotoxicity of homoeopathic Viscum album 1x10-3(VA3X). Methodscells were cultured for 24 hours in controlled environment (37.5oC and 5% CO2) in 96-well plates. After this time, VA3X was added to the culture medium in concentrations varying from 10 to 100 L/mL.A control group was maintained with culture medium only. Cells were cultivated for 48 hours in these conditions for evaluation of cell viability by MTT assay. ResultsHigher cytotoxicity was observed in MCF-7 when compared to MSC, as the lower concentration of VA3X was capable of inducing tumor cell death and not healthy cell death. The MTT assay results were that 42 L/mL of VA3X reduced MCF-7 cells viability to 50% and 62 L/mL reduced MSC cells to the same percentage, what means that tumor cells are more sensible to VA3X than heathy cells. ConclusionViscum albumpresented higher cytotoxic action on human breast cancer cell line culture than on mesenchymal stem cells. This medicine is extensively used against cancer, and the use of the homoeopathic form of it brings new possibilities as no or fewer adverse effects would be present.(AU)

Clinical evaluation of Viscum album mother tincture as an antihypertensive: a pilot study.

Viscum album, an infrequently used antihypertensive in homeopathy, is evaluated by 1-group pretest-posttest model in primary hypertension. The drug was administered for 12 weeks at a dosage of 10 drops 3 times a day. Using paired t test, a significant drop in blood pressure (P < .0001) and serum triglyceride (P < .0001) was observed in the treatment group. This dual effect of Viscum album shows promise in optimizing therapy for primary hypertension.

Dissertation abstract: Viscum album citotoxicity in Caco-2 cells (in vitro) analyzed by Dispersive Raman Spectroscopy

In the last decades, it has been observed a gradual increase in the diagnosis of neoplasm in both human beings and animals. This can be attributed to the advances in research and technology, which facilitated detection, promoting, thus, survival. On the other hand, the current incidence of cancer may also be attributed to an excess of chemicals in food products, radiation arising from antennae, environmental pollutants, indiscriminate use of hormone therapies, besides genetic alterations. Conventional oncologic treatments traditionally employ chemotherapic drugs. Later on, vegetal immunomodulators were also introduced, such as Viscum album. In the present study, the effectiveness of injectable extract Viscum album was tested in vitro in Caco-2 cells, in dilutions D3 (0.1ìg/ml ), D6 (0.1 ng/ml), D9 (0.1x10-³ ng/ml), D12 (0.1x10-6 ng/ml) and D30 (0.1x10-6 ng/ml). The optical method used was the Dispersive Raman Spectroscopy at 830nm by comparison to MTT Assay. The results showed that Viscum album has an action on mitochondrial activity, reducing cellular viability in Caco-2 cells in dilutions D3, D6 and D9. Correlation of Raman spectrum to the values obtained in the MTT test was statistically analyzed by PLS, revealing a sensitive tool for the detection of changes in the mitochondrial activity of Caco-2 cells. Raman spectroscopy is a non invasive diagnostic procedure, allowing for real-time diagnosis and reduction of costs, besides indicating the best therapy for each type of tumor as well as modifications in the therapeutic strategy.

Dissertation Abstract: Viscum album citotoxicity in Caco-2 cells (in vitro) analyzed by Dispersive Raman Spectroscopy

In the last decades, it has been observed a gradual increase in the diagnosis of neoplasm in both human beings and animals. This can be attributed to the advances in research and technology, which facilitated detection, promoting, thus, survival. On the other hand, the current incidence of cancer may also be attributed to an excess of chemicals in food products, radiation arising from antennae, environmental pollutants, indiscriminate use of hormone therapies, besides genetic alterations. Conventional oncologic treatments traditionally employ chemotherapic drugs. Later on, vegetal immunomodulators were also introduced, such as Viscum album. In the present study, the effectiveness of injectable extract Viscum album was tested in vitro in Caco-2 cells, in dilutions D3 (0.1ìg/ml ), D6 (0.1 ng/ml), D9 (0.1x10-³ ng/ml), D12 (0.1x10-6 ng/ml) and D30 (0.1x10-6 ng/ml). The optical method used was the Dispersive Raman Spectroscopy at 830nm by comparison to MTT Assay. The results showed that Viscum album has an action on mitochondrial activity, reducing cellular viability in Caco-2 cells in dilutions D3, D6 and D9. Correlation of Raman spectrum to the values obtained in the MTT test was statistically analyzed by PLS, revealing a sensitive tool for the detection of changes in the mitochondrial activity of Caco-2 cells. Raman spectroscopy is a non invasive diagnostic procedure, allowing for real-time diagnosis and reduction of costs, besides indicating the best therapy for each type of tumor as well as modifications in the therapeutic strategy.(AU)

A medicina dos druidas e o Viscum album / The medicine of the druids and the Viscum album

Os autores apresentam uma revisão histórica sobre o Viscum album, planta utilizada na Farmacopeia homeopática e sua relação com a cultura Druida.

A medicina dos druidas e o Viscum album / The medicine of the druids and the Viscum album

Os autores apresentam uma revisão histórica sobre o Viscum album, planta utilizada na Farmacopeia homeopática e sua relação com a cultura Druida.(AU)

Estudos de casos clínicos oncológicos em caninos e felinos tratados com homeopatia injetável (2005-2007) / Clinical case studies cancer in dogs and cats treated with homeopathy injectable (2005-2007)

Este experimento clínico relata o resultado do tratamento de oito animais com tipos variados de neoplasias que foram tratados com uma combinação de medicamentos homeopáticos injetáveis, Arnica montana, Lachesis muta, thuya occidentalis, Echinacea e o Viscum album - um antineoplásico que estimula o sistema imune - e possuem efeitos citotóxicos, sendo uma alternativa ao tratamento convencional empregado nos animais domestícos. O resultado obtido no decorrer de 12 meses de acompanhamento indica que a medicação estuda auxilia na qualidade de vida demostrando ser uma possibilidade terapêutica auxiliando no controle da evolução da doença neoplásica em cães e gatos.

Estudos de casos clínicos oncológicos em caninos e felinos tratados com homeopatia injetável (2005-2007) / Clinical case studies cancer in dogs and cats treated with homeopathy injectable (2005-2007)

Este experimento clínico relata o resultado do tratamento de oito animais com tipos variados de neoplasias que foram tratados com uma combinação de medicamentos homeopáticos injetáveis, Arnica montana, Lachesis muta, thuya occidentalis, Echinacea e o Viscum album - um antineoplásico que estimula o sistema imune - e possuem efeitos citotóxicos, sendo uma alternativa ao tratamento convencional empregado nos animais domestícos. O resultado obtido no decorrer de 12 meses de acompanhamento indica que a medicação estuda auxilia na qualidade de vida demostrando ser uma possibilidade terapêutica auxiliando no controle da evolução da doença neoplásica em cães e gatos.(AU)

Viscum album preparations inhibit DNA methylation by preventing DNA methyltransferase-induced methyl group transfer

Interactions between viscum album (Iscador) and DNA were studies by amplification of specific human pepsinogen A gene promoter sequences by polymerase chain reaction (PCR). Gel retardation assays showed no binding of Viscum album to these promoter sequences. Incubation of plasmid constructs consisting of human pepsigen A promoter fragments coupled to thechloramphenicol acetyl-transferase (CAT) reporter gene with Iscador had no effect on transcriptional activity. Promoter sequences incubated with Iscador became insensitive to methylation by specific DNA methyltransferases by destruction of DNA methyltransferase activity

Hipertensión arterial / Arterial hypertension

La hipertensión arterial (HTA) afecta aproximadamente al 25 por ciento de la población de edad igual o superior a los 18 años. La prevalencia de la hipertensión aumenta con la edad, la padecen más de la mitad de todas las personas mayores de 65 años. La HTA sistólica-diastólica del adulto se define arbitrariamente como presión arterial igual o superior a 140/90 mm Hg. Más del 90 por ciento de los hipertensos sufren HTA esencial (causa desconocida). Menos del 10 por ciento de los hipertensos presentan HTA secundaria, es decir HTA asociada al uso de anticonceptivos orales, corticoides, patología renovascular, feocromocitoma, síndrome de Cushing, hiperaldosteronismo primario, hiperparatiroidismo, coartación de aorta, etc. El objetivo del tratamiento antihipertensivo es reducir la morbi-mortalidad cardiovascular. Es importante tener en cuenta modificaciones en el estilo de vida como p.e.: reducción del peso, moderación en la ingestión de bebidas alcohólicas, actividad física regular, mantener un aporte adecuado de calcio y potasio y suspender el tabaquismo. La medicación homeopática es de suma utilidad en: 1) hipertensión limítrofe, sin riesgos de afección cardiovascular o daño de órgano blanco, 2) asociada al tratamiento farmacológico

In vitro effects of Viscum album preparations on human fibroblasts and tumor cell lines

The effects of several preparations of Iscador, a commercially obtainable extract of mistletoe, Viscum album L., on cell proliferation and cell viability was studied. Iscador Quercus was shown to inhibit cell proliferation of: human fibroblast cell lines, mouse tumour cell lines, human carcinoma tumour cell lines and a human lymphoblastic tumour cell line. Iscador Quercus, Iscador Ulnus and Iscador Malus were about equally cytostatic to mouse 3T3 fibroblasts. Iscador Pinus was somewhat less cytostatic. Iscador Quercus had a very direct cytotoxic effect on MOLT4 tumour cells as assessed by trypan blue dye exclusion. The cytostatic and cytotoxic properties of Iscador could be annulled by heating Iscador form 30 min. at 90 "graus"C. Iscador reduced the cloning efficiency of ASG experiments it can be concluded that a small percentage of CHO cells is insensitive to high concentrations of Iscador at short incubation times. The importance of these findings in relation to the cytostatic and cytotoxic compounds possibly present in Iscador is discussed

Effect of Viscum album preparations on gastric chief cells and DNA methylation by N-methyl-N-nitro-N-nitrosoguanidine

The protective effect of extracts of mistletoe Viscum album (Iscador) with reference to carcinogenesis was tested on in vitro cultured porcine gastric chief cells. Putative protection against in vitro methylation of lambda DNA by N-methyl-N-nitro-N-nitrosoguanidine (MNNG) was studied with restriction endonucleases. Preparations of Iscador from mistletoe grown on oak as host tree had a high cytotoxic effect on gastric chief cells, with an LC50 after 24hrs that ranged between 0.01 and 0.03 mg/ml. At 0.01 mg/ml, Iscador was also found to protect lambda DNA from methylation or destruction by MNNG. During the course of this study a high variability was found both in cytotoxicity and protection rate against methylation which we attribute to batch to batch differences. Thus the LC50 for Iscador MH86L12 was 0.005 mg/ml, while for MH87 D24 it was 0.05 mg/ml after 24hrs. The Iscador batch W frf 50 mg/ml, which was made from a fresh plant extract at the Hiscia Institute, showed less variability. Results are therefore given for this batch of Iscador only