Oligomerization and Nitration of the Grass Pollen Allergen Phl p 5 by Ozone,Nitrogen Dioxide,and Peroxynitrite: Reaction Products,Kinetics, and Health Effects

The allergenic and inflammatory potential of proteins can be enhanced by chemical modification upon exposure to atmospheric or physiological oxidants. The molecular mechanisms and kinetics of such modifications, however, have not yet been fully resolved. We investigated the oligomerization and nitration of the grass pollen allergen Phl p 5 by ozone (O₃), nitrogen dioxide (NO₂), and peroxynitrite (ONOO^–). Within several hours of exposure to atmospherically relevant concentration levels of O₃ and NO₂, up to 50% of Phl p 5 were converted into protein oligomers, likely by formation of dityrosine cross-links. Assuming that tyrosine residues are the preferential site of nitration, up to 10% of the 12 tyrosine residues per protein monomer were nitrated. For the reaction with peroxynitrite, the largest oligomer mass fractions (up to 50%) were found for equimolar concentrations of peroxynitrite over tyrosine residues. With excess peroxynitrite, the nitration degrees increased up to 40% whereas the oligomer mass fractions decreased to 20%. Our results suggest that protein oligomerization and nitration are competing processes, which is consistent with a two-step mechanism involving a reactive oxygen intermediate (ROI), as observed for other proteins. The modified proteins can promote pro-inflammatory cellular signaling that may contribute to chronic inflammation and allergies in response to air pollution.     

Expression of Connexin43 Stimulates Endothelial Angiogenesis Independently of Gap Junctional Communication In Vitro

Connexins (Cx) form gap junctions (GJ) and allow for intercellular communication. However, these proteins also modulate gene expression, growth, and cell migration. The downregulation of Cx43 impairs endothelial cell migration and angiogenetic potential. Conversely, endothelial Cx43 expression is upregulated in an in vivo angiogenesis model relying on hemodynamic forces. We studied the effects of Cx43 expression on tube formation and proliferation in HUVECs and examined its dependency on GJ communication. Expectedly, intercellular communication assessed by dye transfer was linked to Cx43 expression levels in HUVECs and was sensitive to a GJ blockade by the Cx43 mimetic peptide Gap27. The proliferation of HUVECs was not affected by Cx43 overexpression using Cx43 cDNA transfection, siRNA-mediated knockdown of Cx43, or the inhibition of GJ compared to the controls (transfection of an empty vector, scrambled siRNA, and the solvent). In contrast, endothelial tube and sprout formation in HUVECs was minimized after Cx43 knockdown and significantly enhanced after Cx43 overexpression. This was not affected by a GJ blockade (Gap27). We conclude that Cx43 expression positively modulates the angiogenic potential of endothelial cells independent of GJ communication. Since proliferation remained unaffected, we suggest that Cx43 protein may modulate endothelial cell migration, thereby supporting angiogenesis. The modulation of Cx43 expression may represent an exploitable principle for angiogenesis induction in clinical therapy.     

„Die kommen alle her und gaffen.“ Hohes Körpergewicht als medizinische,ethische und gesellschaftliche Herausforderung

Ethik in der Medizin -     

A Gel‐Based Model of Selective Cell Motility: Implications for Cell Sorting,Diagnostics, and Screening

The ability to precisely control cell‐loaded material systems is essential for in vitro testing of novel therapeutics poised to advance to clinic. In this report, unique patterns of cell migration are devised into an in vitro gel‐in‐gel model for the purpose of obtaining cell response data to potentially therapeutic chemical agonists. The model consists of co‐cultures in a cell‐loaded microgel invading an acellular “sorting” gel. Material properties including biophysical and chemical compositions of the sorting gel are carefully controlled to guide a desired cell‐specific behavior, leading to massive tumor cell invasion by amoeboid migration mechanisms. Optical transparency enables straightforward and high‐throughput measurements of outgrowth response in the presence of either chemical and photoradiation therapy. Important dosing and drug sensitivity information are obtained with the gel‐in‐gel model using no more than a light microscope, without further need for arduous genomic or proteomic screening of the tissue samples.     

Field assessment of lead immobilization in a contaminated soil after phosphate application

A pilot-scale field demonstration was conducted at a Pb-contaminated site to assess the effectiveness of Pb immobilization using P amendments. The test site was contaminated by past battery recycling activities, with average soil Pb concentration of 1.16%. Phosphate amendments were applied at a 4.0 molar ratio of P/Pb with three treatments: T1, 100% P from H(3)PO(4); T2, 50% from H(3)PO(4)+50% from Ca(H(2)PO(4))(2); and T3, 50% from H(3)PO(4)+5% phosphate rock. Soil samples were collected and characterized 220 days after P application. Surface soil pH was reduced from 6.45 to 5.05 in T1, to 5.22 in T2, and to 5.71 in T3. Phosphate treatments effectively transformed up to 60% of total soil Pb from the non-residual fraction (sum of water soluble and exchangeable, carbonate, Fe-Mn oxide, and organic fractions) to the residual fraction relative to the control. In addition, P treatments reduced Toxicity Characteristic Leaching Procedure (TCLP) Pb from 82 mg l(-1) to below EPA's regulatory level of 5 mg l(-1) in the surface soil. Scanning electron microscopy-energy dispersive X-ray elemental analysis and X-ray diffraction analysis indicated formation of insoluble chloropyromorphite [Pb(5)(PO(4))(3)Cl] mineral in the P-treated soils. Although H(3)PO(4) is necessary to dissolve meta-stable Pb in soil for further lead immobilization, it should be used with caution due to its potential secondary contamination. A mixture of H(3)PO(4) and Ca(H(2)PO(4))(2) or phosphate rock was effective in immobilizing Pb with minimum adverse impacts associated with pH reduction.     

Growth responses of three ornamental plants to Cd and Cd-Pb stress and their metal accumulation characteristics

Up to now, there was no document on ornamental plants that had been applied to phytoremediation, which can remedy contaminated environment and beautify it at the same time. Thus, the growth responses and possible phytoremediation ability of three ornamental plants selected from the previous preliminary experiments were further examined under single Cd or combined Cd-Pb stress. The results showed that these tested plants had higher tolerance to Cd and Pb contamination and could effectively accumulate the metals, especially for Calendula officinalis and Althaea rosea. For C. officinalis, it grew normally in soils containing 100 mg kg(-1) Cd without suffering phytotoxicity, and the Cd concentration in the roots was up to 1084 mg kg(-1) while the Cd concentration in the shoots was 284 mg kg(-1). For A. rosea, the Cd accumulation in the shoots was higher than that in the roots when the Cd concentration in soils was <100 mg kg(-1), and reached 100 mg kg(-1) as the criteria of a Cd hyperaccumulator when the Cd concentration in soils was 100 mg kg(-1). Their accumulation and tolerance to Cd and Pb were further demonstrated through the hydroponic-culture method. And A. rosea had a great potential as a possible Cd hyperaccumulator under favorable or induced conditions. Furthermore, the interactive effects of Cd and Pb in the three ornamentals were complicated, not only additive, antagonistic or synergistic, but also related to many factors including concentration combinations of heavy metals, plant species and various parts of plants. Thus, it can be forecasted that this work will provide a new way for phytoremediation of contaminated soils.     

Antibacterial activity of fullerene water suspensions (nC60) is not due to ROS-mediated damage

The cytotoxic and antibacterial properties of nC 60, a buckminsterfullerene water suspension, have been attributed to photocatalytically generated reactive oxygen species (ROS). However, in this work, neither ROS production nor ROS-mediated damage is found in nC 60-exposed bacteria. Furthermore, the colorimetric methods used to evaluate ROS production and damage are confounded by interactions between nC 60 and the reagents, yielding false positives. Instead, we propose that nC 60 exerts ROS-independent oxidative stress, thus reconciling conflicting results in the literature.