Piezo1-induced durotaxis of pancreatic stellate cells depends on TRPC1 and TRPV4 channels.

Pancreatic stellate cells (PSCs) are primarily responsible for producing the stiff tumor tissue in pancreatic ductal adenocarcinoma (PDAC). Thereby, PSCs generate a stiffness gradient between the healthy pancreas and the tumor. This gradient induces durotaxis, a form of directional cell migration driven by differential stiffness. The molecular sensors behind durotaxis are still unclear. To investigate the role of mechanosensitive ion channels in PSC durotaxis, we established a two-dimensional stiffness gradient mimicking PDAC. Using pharmacological and genetic methods, we investigated the role of the ion channels Piezo1, TRPC1, and TRPV4 in PSC durotaxis. We found that PSC migration towards a stiffer substrate is diminished by altering Piezo1 activity. Moreover, disrupting TRPC1 along with TRPV4 abolishes PSC durotaxis even when Piezo1 is functional. Hence, PSC durotaxis is optimal with an intermediary level of mechanosensitive channel activity, which we simulated using a numerically discretized mathematical model. Our findings suggest that mechanosensitive ion channels, particularly Piezo1, detect the mechanical microenvironment to guide PSC migration.

Poincaré and Log-Sobolev Inequalities for Mixtures.

This work studies mixtures of probability measures on R n and gives bounds on the Poincaré and the log-Sobolev constants of two-component mixtures provided that each component satisfies the functional inequality, and both components are close in the χ 2 -distance. The estimation of those constants for a mixture can be far more subtle than it is for its parts. Even mixing Gaussian measures may produce a measure with a Hamiltonian potential possessing multiple wells leading to metastability and large constants in Sobolev type inequalities. In particular, the Poincaré constant stays bounded in the mixture parameter, whereas the log-Sobolev may blow up as the mixture ratio goes to 0 or 1. This observation generalizes the one by Chafaï and Malrieu to the multidimensional case. The behavior is shown for a class of examples to be not only a mere artifact of the method.

Reprint of "Fast and sensitive in vivo studies under controlled environmental conditions to substitute long-term field trials with genetically modified plants".

We introduce an easy, fast and effective method to analyze the influence of genetically modified (GM) plants on soil and model organisms in the laboratory to substitute laborious and time consuming field trials. For the studies described here we focused on two GM plants of the so-called 3rd generation: GM plants producing pharmaceuticals (PMP) and plant made industrials (PMI). Cyanophycin synthetase (cphA) was chosen as model for PMI and Choleratoxin B (CTB) as model for PMP. The model genes are expressed in transgenic roots of composite Vicia hirsuta plants grown in petri dishes for semi-sterile growth or small containers filled with non-sterile soil. No significant influence of the model gene expression on root induction, growth, biomass, interaction with symbionts such as rhizobia (number, size and functionality of nodules, selection of nodulating strains) or arbuscular mycorrhizal fungi could be detected. In vitro, but not in situ under field conditions, structural diversity of the bulk soil microbial community between transgenic and non-transgenic cultivars was determined by PLFA pattern-derived ratios of bacteria: fungi and of gram+: gram- bacteria. Significant differences in PLFA ratios were associated with dissimilarities in the quantity and molecular composition of rhizodeposits as revealed by Py-FIMS analyses. Contrary to field trials, where small effects based on the transgene expression might be hidden by the immense influence of various environmental factors, our in vitro system can detect even minor effects and correlates them to transgene expression with less space, time and labour.

Fast and sensitive in vivo studies under controlled environmental conditions to substitute long-term field trials with genetically modified plants.

We introduce an easy, fast and effective method to analyze the influence of genetically modified (GM) plants on soil and model organisms in the laboratory to substitute laborious and time consuming field trials. For the studies described here we focused on two GM plants of the so-called 3rd generation: GM plants producing pharmaceuticals (PMP) and plant made industrials (PMI). Cyanophycin synthetase (cphA) was chosen as model for PMI and Choleratoxin B (CTB) as model for PMP. The model genes are expressed in transgenic roots of composite Vicia hirsuta plants grown in petri dishes for semi-sterile growth or small containers filled with non-sterile soil. No significant influence of the model gene expression on root induction, growth, biomass, interaction with symbionts such as rhizobia (number, size and functionality of nodules, selection of nodulating strains) or arbuscular mycorrhizal fungi could be detected. In vitro, but not in situ under field conditions, structural diversity of the bulk soil microbial community between transgenic and non-transgenic cultivars was determined by PLFA pattern-derived ratios of bacteria: fungi and of gram+: gram- bacteria. Significant differences in PLFA ratios were associated with dissimilarities in the quantity and molecular composition of rhizodeposits as revealed by Py-FIMS analyses. Contrary to field trials, where small effects based on the transgene expression might be hidden by the immense influence of various environmental factors, our in vitro system can detect even minor effects and correlates them to transgene expression with less space, time and labour.

Effects of elm bark extracts from Ulmus laevis on human chorion carcinoma cell lines.

PURPOSE: The potential of substances from elm bark extracts to affect cancer has been described in several studies. In this study, the anticancer effects of extracts from Ulmus laevis bark were tested in hormone-dependent gynecological tumours using human chorion carcinoma cell lines. METHODS: The molecular-chemical composition of the bark extract was analysed by pyrolysis-field ionisation mass spectrometry. The influence of the extracts was determined on cell vitality and cytotoxicity in the human chorion carcinoma cell lines Jeg3 and BeWo in comparison with primary trophoblast cells. RESULTS: The elm bark extract was mainly composed of triterpenes, phytosterols, free fatty acids and suberins with lower amounts of dilignols and lipids. The elm bark extract significantly inhibited the vitality of Jeg3 and BeWo cells but increased the vitality of primary trophoblast cells. CONCLUSIONS: Substances extracted from elm bark might have beneficial effects for the prevention of hormone-dependent tumours.

Pyrolysis-field ionization mass spectrometry and nitrogen K-edge XANES spectroscopy applied to bulk soil leachates--a case study.

Although dissolved organic matter (DOM) is an important component of C- and N-fluxes in the environment, its structural composition is still poorly understood due to methodological challenges. We explored the potential of combining pyrolysis-field ionization mass spectrometry (Py-FIMS) and N X-ray absorption near edge structure (N-XANES) spectroscopy to study the molecular-chemical composition of lyophilized bulk soil solution samples that were not subjected to pretreatment like dialysis. Soil leachates were collected at 90 cm and 220 cm depth from an arable and a fallow site. Py-FIMS spectra reflected differences in DOM composition related to land use and sampling depth. Land use effects were expressed in higher abundances of carbohydrates and peptides at the arable than at the fallow site. The relative proportions of carbohydrates decreased and the proportions of lignin-derived compounds increased with depth, indicating a relative enrichment to more stabilized DOM along the flow path. Nitrogen XANES spectra were dominated by the signal of NO(3)-salts but also indicated the presence of organic, non-amidic N as found in imidazoles, pyrazoles, purines and/or nitrile-N, whereas N-compounds like pyridines, pyrroles, quinoline and indole were detected by Py-FIMS. Thus, the combined application of Py-FIMS and N-XANES yielded complementary information regarding the molecular-chemical composition of DOM. Future applications of these techniques may benefit from selectively analyzing soil solution samples with lower nitrate concentrations collected in early spring.

Inhibitory effects of bark extracts from Ulmus laevis on endometrial carcinoma: an in-vitro study.

Anti-inflammatory effects of elm tree have been shown in several studies. Besides this, protective effects of components of elm bark on damaged tissue have also been described. This study was carried out to investigate the antitumour potential of an ethanolic extract isolated from Ulmus laevis in the hormone-dependent endometrial carcinoma cell line RL95-2. A range of 2.5-500 microg/ml of elm bark extract was used as standard concentrations. The molecular-chemical composition of the bark extract was analysed by pyrolysis-field ionization mass spectrometry. The influence of the bark extracts was determined on cell vitality [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test], cell proliferation (5-bromo-2-deoxyuridine test) and cytotoxicity (lactate dehydrogenase test) in the human endometrial carcinoma cell line RL 95-2. By pyrolysis-field ionization mass spectrometry, the main substance classes of the extract as a composition of sterols/triterpenes, free fatty acids and a group of phenols, lignin monomers and flavonoids was identified. Our study showed a significant inhibition of cell vitality and proliferation measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test up to 5 microg/ml extract and up to 100 microg/ml according to the 5-bromo-2-deoxyuridine test. Concentrations of 500 microg/ml induced a significant inhibition of cell vitality up to 80% and cell proliferation up to 81.5%. A significant cytotoxity was not observed. The results lead to the assumption that the bark extract from Ulmus laevis has antiproliferation and anticancer potential in hormone-dependent endometrial carcinoma cells.

New evidence for the molecular-chemical diversity of potato plant rhizodeposits obtained by pyrolysis-field Ionisation mass spectrometry.

INTRODUCTION: Detailed descriptions of the molecular-chemical diversity in plant rhizodeposits are scarce. The vast majority of our knowledge is derived from a priori methods of analysis, such as GC-MS and HPLC. OBJECTIVE: To analyse the composition of rhizodeposits from the potato cultivar Solanum tuberosum L. cv. Albatros by pyrolysis -field ionisation mass spectrometry (Py-FIMS) and to explain differences in relation to plant growth stage and photoperiod. METHODOLOGY: Potato (Solanum tuberosum L.) plants were grown in non-sterile, native soil under controlled environmental conditions (plant chamber). Rhizodeposit samples were collected by leaching during two different growth stages and after the physiological day- and night-cycle. All leachate samples were investigated by Py-FIMS. Mass spectrometric data were evaluated by multivariate statistics. RESULTS: Screening of the rhizodeposits by Py-FIMS revealed a broad range of m/z signals. Low-molecular-weight substances of m/z 15-56 (8.1-18.6%), alkylaromatics (12.0-15.9%), phenols and lignin monomers (8.8-13.1%) and carbohydrates (6.0-11.2%) comprised the largest proportions of total ion intensity (TII). Mass signals with significantly different abundance at the various sampling dates were assigned to compound classes of carbohydrates, phenols and lignin monomers, lignin dimers, lipids, N-containing compounds, sterols, peptides and free fatty acids; these were supplemented by marker signals for N-acetylmuramic acid from bacterial cell walls and signal molecules for the regulation of secondary pathways such as 4-hydroxycinnamic acid and linolenic acid. CONCLUSION: Py-FIMS was well suited to detect the molecular-chemical diversity of potato plant rhizodeposits and, compared with traditional a priori analytical methods, provided detailed evidence for significant differences in the composition of rhizodeposits depending on growth stage and diurnal period.