Differentiation in Neuroblastoma: Diffusion-Limited Hypoxia Induces Neuro-Endocrine Secretory Protein 55 and Other Markers of a Chromaffin Phenotype

Background: Neuroblastoma is a childhood malignancy of sympathetic embryonal origin. A high potential for differentiation is a hallmark of neuroblastoma cells. We have previously presented data to suggest that in situ differentiation in tumors frequently proceeds along the chromaffin lineage and that decreased oxygen ( hypoxia) plays a role in this. Here we explore the utility of Neuro-Endocrine Secretory Protein 55 ( NESP55), a novel member of the chromogranin family, as a marker for this process.Methodology/Principal Findings: Immunohistochemical analyses and in situ hybridizations were performed on human fetal tissues, mouse xenografts of human neuroblastoma cell lines, and on specimens of human neuroblastoma/ganglioneuroma. Effects of anaerobic exposure on gene expression by cultured neuroblastoma cells was analyzed with quantitative real-time PCR. Fetal sympathetic nervous system expression of NESP55 was shown to be specific for chromaffin cell types. In experimental and clinical neuroblastoma NESP55 immunoreactivity was specific for regions of chronic hypoxia. NESP55 expression also correlated strikingly with morphological evidence of differentiation and with other chromaffin-specific patterns of gene expression, including IGF2 and HIF2 alpha. Anaerobic culture of five neuroblastoma cell lines resulted in an 18.9-fold mean up-regulation of NESP55.Conclusions/Significance: The data confirms that chronic tumor hypoxia is a key microenvironmental factor for neuroblastoma cell differentiation, causing induction of chromaffin features and NESP55 provides a reliable marker for this neuronal to neuroendocrine transition. The hypoxia-induced phenotype is the predominant form of differentiation in stroma-poor tumors, while in stroma-rich tumors the chromaffin phenotype coexists with ganglion cell-like differentiation. The findings provide new insights into the biological diversity which is a striking feature of this group of tumors

Dielectric Properties of Liquid Crystalline Isothiocyanato-tolane Derivatives with Fluorine Atom at Various Lateral Positions

Dielectric studies were performed for several isothiocyanato-tolane compounds having the F atom attached to the benzene rings at different lateral positions and the alkoxy or alkyl chain at the p-position. They form the nematic phase in broad temperature intervals. The static as well as dynamic properties of the compounds in the nematic and isotropic phases were studied. Tensor permittivity components: ε$\text{}_{ǁ}$, ε$\text{}_{⊥}$, Δε, as functions of temperature were measured. The complex dielectric permittivity, ε$\text{}^{*}$(f)=ε'(f)-iε''(f), was measured in the frequency range of 1 kHz - 3 GHz from which the relaxation times τ$\text{}_{ǁ}$ and τ$\text{}_{⊥}$ were calculated. The order parameter S(T) was determined from the dielectric data. It was found that the close vicinity of the F atom and the alkoxy group results in blocking of the internal rotation of the wing around the O-phenyl bond

Dynamics of 4,4'-di-n-heptylazoxybenzene (HAB) studied using dielectric and 2H NMR relaxation measurements

Results of studies of 4,4'-di-n-heptylazoxybenzene (HAB) in the isotropic, nematic and smectic A phases are presented. Two experimental methods were employed: broad band dielectric spectroscopy and nuclear magnetic resonance spectroscopy. The complex dielectric permittivity, epsilon*(omega)= epsilon'(omega) iepsilon"(omega), was measured in the frequency range 1 kHz - 4 GHz. This allowed two main relaxation processes to be separated in all the phases studied: the low frequency (l.f.) process connected with molecular reorientations around the short axes, and the high frequency ( h.f.) process connected with the rotations around the long axes. The corresponding relaxation times and activation enthalpies were obtained. The l.f. relaxation time changes step-wise at the phase transitions, whereas the h.f. relaxation time passes smoothly through all the phases. The measurement of H-2 spin-lattice relaxation times was carried out throughout the mesophase range at 61.38 MHz. These data were analysed together with the relaxation times measured at 10.00 and 46.04 MHz, available from previous studies. Using suitable theoretical models the principal components of the diffusional tensor, D-parallel to and D-perpendicular to, as well as the diffusion coefficients D-R relative to the internal rotation of the phenyl rings, were determined. The results of both studies are compared and discussed

Dielectric behavior of aqueous solutions of plasmid DNA at microwave frequencies.

The relative permittivity and dielectric loss of aqueous solutions of plasmid (pUC8.c1 and pUC8.c2) DNA have been measured at 20 degrees C over the frequency range 100 MHz-10 GHz. The solutions had a concentration of 0.1% DNA, and were studied both in the relaxed and the supercoiled form. The dielectric measurements were made using a variety of techniques including frequency domain and time domain methods of operation. No evidence of any resonance absorption, nor of any other kind of enhanced absorption, was observed