Gene expression signatures of pathways alterations in tumor cells caused by plant extracts

Extracts of Viscum album from different host trees (Iscador®Qu, M, P and A) and isolated mistletoe lectin showed distinct gene-expression profiles using cDNA microarrays. We were able to confirm that genes of apoptosis, the cell cycle and the immune system were affected by mistletoe preparations and we also found that signal transduction, cell communication and cancer related pathways were overregulated in the tested breast cancer (HCC-1937, MCF-7, KPL-1, Mfm-223) and lymphoma cell lines (WSZ-NHL). The effect of a specific mistletoe extract on a particular cell line was individual, so that the distinction between the reaction of the cell type and/or the influence of the mistletoe preparation could be used in targeted mistletoe therapy

Mistletoe lectin is not the only cytotoxic component in fermented preparations of Viscum album from white fir (Abies pectinata)

<p>Abstract</p> <p>Background</p> <p>Preparations of mistletoe (<it>Viscum album</it>) are the form of cancer treatment that is most frequently used in the complementary medicine. Previous work has shown that these preparations are able to exert cytotoxic effects on carcinoma cells, the extent of which might be influenced by the host tree species and by the content of mistletoe lectin.</p> <p>Methods</p> <p>Using colorimetric assays, we have now compared the cytotoxic effects of <it>Viscum album </it>preparations (VAPs) obtained from mistletoe growing on oak (<it>Quercus robur </it>and <it>Q. petraea</it>, VAP-Qu), apple tree (<it>Malus domestica</it>,, VAP-M), pine (<it>Pinus sylvestris</it>, VAP-P) or white fir (<it>Abies pectinata</it>, VAP-A), on the <it>in vitro </it>growth of breast and bladder carcinoma cell lines. While MFM-223, KPL-1, MCF-7 and HCC-1937 were the breast carcinoma cell lines chosen, the panel of tested bladder carcinoma cells comprised the T-24, TCC-SUP, UM-UC-3 and J-82 cell lines.</p> <p>Results</p> <p>Each of the VAPs inhibited cell growth, but the extent of this inhibition differed with the preparation and with the cell line. The concentrations of VAP-Qu, VAP-M and VAP-A which led to a 50 % reduction of cell growth (IC₅₀) varied between 0.6 and 0.03 mg/ml. Higher concentrations of VAP-P were required to obtain a comparable effect. Purified mistletoe lectin I (MLI) led to an inhibition of breast carcinoma cell growth at concentrations lower than those of VAPs, but the sensitivity towards purified MLI did not parallel that towards VAPs. Bladder carcinoma cells were in most cases more sensitive to VAPs treatment than breast carcinoma cells. The total mistletoe lectin content was very high in VAP-Qu (54 ng/mg extract), intermediate in VAP-M (25 ng/mg extract), and very low in VAP-P (1.3 ng/mg extract) and in VAP-A (1 ng/mg extract). As to be expected from the low content of mistletoe lectin, VAP-P led to relatively weak cytotoxic effects. Most remarkably, however, the lectin-poor VAP-A revealed a cytotoxic effect comparable to, or even stronger than, that of the lectin-rich VAP-Qu, on all tested bladder and breast carcinoma cell lines.</p> <p>Conclusion</p> <p>The results suggest the existence of cytotoxic components other than mistletoe lectin in VAP-A and reveal an unexpected potential of this preparation for the treatment of breast and bladder cancer.</p

Heat transfer analysis of high pressure hydrogen tank fillings

Fast fillings of hydrogen vehicles require proper control of the temperature to ensure the integrity of the storage tanks. This study presents an analysis of heat transfer during filling of a hydrogen tank. A conjugate heat transfer based on energy balance is introduced. The numerical model is validated against fast filling experiments of hydrogen in a Type IV tank by comparing the gas temperature evolution. The impact of filling parameters, such as initial temperature, inlet nozzle diameter and filling time is then assessed. For the considered Type IV tank, the results show that both a higher and lower tank shell thermal conductivity results in lower inner wall peak temperatures. The presented model provides an analytical description of the temperature evolution in the gas and in the tank shell, and is thus a useful tool to explore a broad range of parameters, e.g., to determine new hydrogen filling protocols.ISSN:1879-3487ISSN:0360-319

URANS Simulations of Vehicle Exhaust Plumes with Insight on Remote Emission Sensing

Remote Emission Sensing (RES) is a measurement method based on absorption spectroscopy for the determination of pollutant concentrations. The absorption of the exhaust plume of a vehicle is measured from the roadside without intervention by means of a light/laser barrier during a short measurement (∼0.5 s) and concentration ratios of pollutants (e.g., NO (Formula presented.) to CO (Formula presented.)) are estimated. Unsteady Reynolds-Averaged Navier-Stokes (URANS) simulations of exhaust plumes in vehicle wakes are performed using the k- (Formula presented.) SST turbulence model with focus on pollutant dispersion. The simulation setup has been validated by a comparison with experimentally obtained drag coefficients. The resulting concentration fields represent the pollutants available for measurements by a RES device. The influence of the characteristics of the RES device on the measurement is assessed. In addition, investigations involve several environmental and vehicle related parameters. The results demonstrate that due to strong turbulence, mixing is enhanced and the exhaust plumes rapidly disperse in the near vehicle wakes. Results show that emission characteristics of a vehicle are contained downstream for approximately half the vehicle length, regardless of different vehicle configurations, driving and ambient parameters. Further downstream dispersion of pollutants results in concentrations that are less than (Formula presented.) of the pollutant concentration in the vehicle’s exhaust tailpipe implying that RES devices have to measure at a high sampling frequency. Therefore, reliable determination of the concentration ratios of pollutant at high vehicle velocities requires the RES device to operate in the order of 1000 Hz sampling frequency. Ultimately, the numerical simulations not only help to understand exhaust plume dispersion, but provide a very useful tool to minimize RES uncertainties.ISSN:2073-443

Direct conversion of porcine primary fibroblasts into hepatocyte-like cells.

The pig is an important model organism for biomedical research, mainly due to its extensive genetic, physiological and anatomical similarities with humans. Until date, direct conversion of somatic cells into hepatocyte-like cells (iHeps) has only been achieved in rodents and human cells. Here, we employed lentiviral vectors to screen a panel of 12 hepatic transcription factors (TF) for their potential to convert porcine fibroblasts into hepatocyte-like cells. We demonstrate for the first time, hepatic conversion of porcine somatic cells by over-expression of CEBPα, FOXA1 and HNF4α2 (3TF-piHeps). Reprogrammed 3TF-piHeps display a hepatocyte-like morphology and show functional characteristics of hepatic cells, including albumin secretion, Dil-AcLDL uptake, storage of lipids and glycogen and activity of cytochrome P450 enzymes CYP1A2 and CYP2C33 (CYP2C9 in humans). Moreover, we show that markers of mature hepatocytes are highly expressed in 3TF-piHeps, while fibroblastic markers are reduced. We envision piHeps as useful cell sources for future studies on drug metabolism and toxicity as well as in vitro models for investigation of pig-to-human infectious diseases

Prolonged cytotoxic effect of aqueous extracts from dired Viscum album on bladder cancer cells

Aqueous extracts from whole dried mistletoe (Viscum album L., Iscucin®) are often used in anti-cancer treatment. We studied the effect of extracts obtained from mistletoe bushes that grew on different host trees on bladder cancer cells by means of MTT-colorimetric cell proliferation/survival assays. The extracts possessed concentration-dependent cytotoxic properties whose extent varied with the host tree, but did not always correlate with the corresponding mistletoe lectin content. A 2-hours treatment of bladder cancer cells triggered a later, strong cytotoxic effect. This prolonged effect suggests that instillation with Iscucin® has therapeutic potential for bladder cancer patients

Gene expression profiles of different breast cancer cells compared with their responsiveness to fermented mistletoe (Viscum album L.) extracts Iscador from oak (Quercus), pine (Pinus), white fir (Abies) and apple tree (Malus) in vitro

Cytotoxicity assays in vitro (MTT test) showed that the different breast cancer cell lines Kpl-1, MCF-7 and Mfm-223 respond differently to the mistletoe (Viscum album L.) preparations Iscador. Quercus (Qu), Abies (A), Malus (M) and Pinus (P). In order to determine the differences in the responsiveness of the cells more exactly, the gene expression profiles were determined by cells, which were treated with Mistletoe extracts, compared with untreated control cells. Such differences can be analysed in more detail by looking at the gene expression using Human Whole Genome microarray chips (41,000 genes). The results of the transcriptome analyses suggested that Iscador preparations influenced the overregulation of genes regarding immune defense, stress response, apoptosis and cell-cell adhesion pathways. Within the Mfm-223-Zellen was the Genexpression in MCF-7 and Kpl-1. The MCF-7 cells were affected on the genes which are involved in cell-cell contacts whereas Kpl-1 responded to the mistletoe extracts by changing the mRNA levels of the immune and stress response pathways. Concerning the effects of the mistletoe extract, we conclude that Iscador Qu and M have a greater influence on the immune defense and stress response genes whereas Iscador A tends to affect the cell-cell adhesion and cytoskeleton pathways. In summary, cDNA microarray analyses give us information on whether a cancer cell is sensitive to mistletoe extracts in relation to how many genes are significantly overrepresented after mistletoe treatment, and whether a particular mistletoe extract is more effective on a specific cancer cell than the other preparation