Investigation of PAHs, nitrated PAHs and oxygenated PAHs in PM10 urban aerosols. A comprehensive data analysis.

Polycyclic aromatic compounds (PACs) in particulate matter contribute considerably to the health risk of air pollution. As such, we have optimized a method to determine the levels of polycyclic aromatic hydrocarbons, especially nitrated and oxygenated polycyclic aromatic hydrocarbons, in samples of PM10 particulate matter using microwave-assisted extraction (MAE) and gas chromatography coupled to a triple quadrupole mass spectrometer (GC-MS/MS). The proposed method was applied to the analysis of real samples collected in the urban area of Ciudad Real (Spain) during one year. The median total concentrations of eighteen PAHs (∑PAHs) and seven OPAHs (∑OPAHs) were 0.54 and 0.23 ng m-3, respectively, with the corresponding value for NPAH (∑NPAHs) being 0.03 ng m-3 (only detected in 40% of samples). A clear seasonal trend was observed, with higher levels in the cold season and lower in the warm season for ∑PAHs. The same effect was observed for ∑OPAHs, which exhibited a median concentration of 0.72 ng m-3 in the cold season and 0.10 ng m-3 in the warm season, and for ∑NPAH, which exhibited a median of 0.04 ng m-3 in the cold season but were not detected in the warm season. Molecular diagnostic ratios and PCA (principal component analysis) showed a predominantly traffic origin for PACs. The sources of PAHs also depend on meteorological conditions and/or atmospheric reactions, as confirmed by means of statistical analysis. The ∑OPAH/∑PAH and ∑NPAH/∑PAH ratios were higher in the cold season than the warm season, thus suggesting that PAH derivatives originated from primary combustion emission sources together with their parent PAHs. The concentration range found for benzo(a)pyrene was 0.006-0.542 ng m-3, which is below the threshold value of 1 ng m-3 established in European legislation as the annual average value. The lifetime lung risk from inhalation of PM10-bound PACs was estimated to be six cancer cases per million people using the World Health Organization method.

Cofilin activation mediates Bax translocation to mitochondria during excitotoxic neuronal death.

During excitotoxic neuronal death, Bax translocates to the mitochondria where it plays an important role by contributing to the release of proapoptotic factors. However, how Bax translocates to the mitochondria during excitotoxicity remains poorly understood. Herein, our data suggest the presence of a novel signalling mechanism by which NMDA receptor stimulation promotes Bax translocation. This signalling pathway is triggered by dephosphorylation of cofilin. Once dephosphorylated, cofilin might interact physically with Bax acting as a carrier for it, translocating it to the mitochondria, where it contributes to mitochondrial membrane despolarization, permeabilization and to the release of apoptotic factors, thus leading to neuronal death. Lack-of-function studies indicate that only the Slingshot family of phosphatases, more specifically the enzyme Slingshot 1L phosphatase, but not cronophin participates in the cofilin activation process during excitotoxicity. Indeed, cofilin-mediated Bax translocation seems to be a key event in excitotoxic neuronal death as knock down of either cofilin or Slingshot 1L phosphatase has a marked neuroprotective effect on NMDA-mediated neuronal death. This novel biochemical pathway may therefore be a good target to develop future therapeutic molecules for neurodegenerative diseases.

Dendrimer-mediated siRNA delivery knocks down Beclin 1 and potentiates NMDA-mediated toxicity in rat cortical neurons.

Autophagy is an important process which plays a key role in cellular homeostasis by degrading cytoplasmic components in the lysosomes, which facilitates recycling. Alterations to normal autophagy have been linked to excitotoxicity, but the mechanisms governing its signal transduction remain unclear. The aim of this study was to explore the role of autophagy in neuronal excitotoxic death by delivering small interfering RNA (siRNA) to rat cortical neurons, using a dendrimer to silence the autophagy-related gene 6 (beclin 1) and to determine the role of autophagy in excitotoxicity. We have found that the dendrimer is very efficient to deliver siRNA to rat cortical neurons, leading to almost complete removal of the target protein Beclin 1. In addition, NMDA increases autophagy markers, such as the protein levels of Beclin 1, the microtubule-associated light chain 3 (LC3) B-II/LC3B-I ratio, and monodansylcadaverine (MDC) labeling in rat cortical neurons. Moreover, NMDA also increases the formation of autophagosomes observed under a transmission electron microscope. Silencing beclin 1 expression blocked NMDA-induced autophagy. Moreover, Beclin 1 removal potentiated NMDA-induced neuronal death indicating that autophagy plays a protective role during excitotoxicity and suggesting that targeting autophagy might be a helpful therapeutic strategy in neurodegenerative diseases.

Efficient, non-toxic hybrid PPV-PAMAM dendrimer as a gene carrier for neuronal cells.

A novel hybrid dendrimer (TRANSGEDEN) that combines a conjugated rigid polyphenylenevinylene (PPV) core with flexible polyamidoamine (PAMAM) branches at the surface was synthesized and characterized. The potential of this material as a nonviral gene delivery system was also examined, and it was observed that dendriplexes formed by TRANSGEDEN and small interfering ribonucleic acids (siRNAs) can be incorporated into >90% of neuronal cells without any toxicity up to a dendrimer concentration of 3 µM. TRANSGEDEN was used to deliver a specific siRNA to rat cerebellar granular neurons (CGNs) to knock down the cofilin-1 protein. Cofilin-1 removal partially protects CGNs from N-methyl D-aspartate (NMDA)-mediated neuronal death.

Control of surface functionality in poly(phenylenevinylene) dendritic architectures.

The efficient synthesis of new asymmetric poly(phenylenevinylene) dendritic macromolecules using a stepwise convergent-growth approach is described. By an iterative methodology that made use of the Horner-Wadsworth-Emmons (HWE) reaction, dendrons and dendrimers up to the third generation, with eight different functional groups located at the periphery, were prepared in good yields. Both the number and placement of functionalities can be accurately controlled to afford a large variety of dendritic architectures.