Consensus guidelines for the use and interpretation of angiogenesis assays

The formation of new blood vessels, or angiogenesis, is a complex process that plays important roles in growth and development, tissue and organ regeneration, as well as numerous pathological conditions. Angiogenesis undergoes multiple discrete steps that can be individually evaluated and quantified by a large number of bioassays. These independent assessments hold advantages but also have limitations. This article describes in vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis and highlights critical aspects that are relevant for their execution and proper interpretation. As such, this collaborative work is the first edition of consensus guidelines on angiogenesis bioassays to serve for current and future reference

Non-variant phenomena in heterogeneous systems. New type of solubility diagrams points

The article gives a general classification of non-invariant points in phase equilibrium diagrams of all possible types. The complete topological isomorphism of the diagrams of fusibility, solubility, and liquid-vapor equilibria in various sets of variables is demonstrated. The stability of mono-variant equilibria near the non-variant points is investigated. Recurrent formulas for calculating the number of topological elements of phase diagrams are given. A previously undescribed type of non-invariant points and phase processes in the solubility diagrams is described and characterized. The last ones have no topological analogs in other types of diagrams. Thus, we have carried out, as far as is available to the authors, a complete classification of invariant points and invariant processes in phase equilibrium diagrams of an arbitrary type and with an arbitrary number of components

Multiphase Open Phase Processes Differential Equations

The thermodynamic approach for the description of multiphase open phase processes is developed based on van der Waals equation in the metrics of Gibbs and incomplete Gibbs potentials. Examples of thermodynamic modeling of the multiphase and multicomponent A3B5 systems (In-Ga-As-Sb and In-P-As-Sb) and Na+, K+, Mg2+, Ca2+//Cl−, SO42−-H2O water–salt system are presented. Topological isomorphism of different type phase diagrams is demonstrated

Fullerenol‑<i>d</i> Solubility in Fullerenol‑<i>d</i>–Inorganic Salt–Water Ternary Systems at 25 °C

In this work, solubility in ternary systems fullerenol-<i>d</i>–NaCl–H₂O, fullerenol-<i>d</i>–Pr (NO₃) ₃–H₂O, fullerenol-<i>d</i>–YCl₃–H₂O, fullerenol-<i>d</i>–uranyl sulfate–water, and fullerenol-<i>d</i>–CuCl₂–water at 25 °C by the method of isothermal saturation in ampules was studied; a description of the results is presented

Pre-existence of DRG neurons within the Matrigel increased B16 melanoma growth in the Matrigel in mice.

<p>C57BL/6 mice (5 mice/group) received s.c. injection of control Matrigel (500μl ECM gel diluted 1:1 in RPMI 1640) and Matrigel with cultured DRG neurons (1x10⁴ cells). B16 cells (5x10⁴/300μl PBS) were administered into the Matrigel plaques two weeks later. Animals were sacrificed two weeks post tumor inoculation, and Matrigel plaques were weighted to characterize tumor growth (A). 50x10³ tumor cells alone or mixed with cultured DRG neurons (1:5 cell:cell ratio). (A). The presence of live DRG neurons in the Matrigel harvested two weeks after administration, i.e., right before the tumor cell injection, was determined by immunohistochemistry with anti-Neurofilament H antibody as described in Materials and Methods (B). Left panel show control Matrigel, right panel show Matrigel with added DRG neurons two weeks after injections. Matrigel weight is expressed as the mean ± SEM. *, p<0.05 (ANOVA, n = 3).</p

B16 cells stimulated the growth of DRG neurons <i>in vitro</i>.

<p>DRG cultures were treated with B16-conditioned medium (10% v/v, 72 h), stained for Neuronal class III ß-tubulin (A, upper panels) and subjected to 3D neuron and dendrite reconstruction (A, lower panels).Quantitative analysis was performed to determine DRGdendritic length, area, volume, and the number of segments and t branches per slide using the FilamentTracer module of Imaris (Bitplane) software package as described in Materials and Methods (B). Bars represent morphometric parameters of neurons treated with B16 conditioned medium as the percentage of control non-treated neurons. Representative images and automated data from one experiment are shown. Data from four independent cultures were combined to determine means±SEM.</p

DRG neurons do not affect proliferation of B16 cells in vitro.

<p>B16 cells were cultured in the presence of DRG culture medium (10% v/v), and DRG conditioned medium (10% v/v) and cell proliferation were determined by the ³H-thymidine incorporation assay as described in Materials and Methods (A). The results are expressed as counts per minute (cpm) and shown as the mean±SEM (n = 3). Analysis of cell cycle was done by flow cytometry using propidium iodide (PI) to label DNA content in B16 cells cultured with and w/o DRG conditioned medium (B). The results of a representative experiment are shown (n = 2).</p

Co-administration of DRG neurons with B16 melanoma cells up-regulated tumor growth in mice.

<p>C57BL/6 mice (5 mice/group) received s.c. injection of 50x10³ tumor cells alone or mixed with cultured DRG neurons (1:5 cell:cell ratio). Tumor size was assessed using a caliper and expressed in mm² (A). Administration of DRG cells alone, as expected, did not induce any tumor growth. Tumor tissues were harvested two weeks later, and the presence of live DRG neurons in the tumor was determined by DAB staining after 14 days using an antibody against the anti-Neurofilament H epitope (B). Left panels show control tumors (administration of B16 cell alone), right panels show tumors growing in mice receiving B16+DRG neurons injections. Tumor size is expressed as the mean ± SEM. *, p<0.05 (ANOVA, n = 3).</p

Accelerated growth of melanoma in the presence of DRG neurons was accompanied by an increased tumor infiltration by MDSC <i>in vivo</i>.

<p>C57BL/6 mice received s.c. injection of control Matrigel and Matrigel with DRG neurons as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156095#pone.0156095.g002" target="_blank">Fig 2</a> legend. B16 cells were administered into the Matrigel plaques two weeks later. Animals were sacrificed two weeks post tumor inoculation, and Matrigel plaques were harvested, dissolved and subject for the presence of granulocytic CD11b⁺Ly6G⁺ and monocytic CD11b⁺Ly6C⁺ MDSC among CD45⁺ leukocytes by flow cytometry. The results of a representative experiment from six mice analyzed in two independent studies are shown.</p

Approach for the Description of Chemical Equilibrium Shifts in the Systems with Free and Connected Chemical Reactions

Approach for the description of chemical equilibrium shifts in the systems with free and connected chemical reactions was elaborated. Driving forces of chemical equilibrium shifts were temperature change (at P = const), pressure change (at T = const), and input or output of reagents or products (at T, P = const). It was demonstrated how the conditions for passing through the extremes of the state parameters (T, P, and components molar numbers) in one of the reactions transmitted to other reactions, connected with the first one by reagents or products