Aggregation is a critical cause of poor transfer into the brain tissue of intravenously administered cationic PAMAM dendrimer nanoparticles.

Dendrimers have been expected as excellent nanodevices for brain medication. An amine-terminated polyamidoamine dendrimer (PD), an unmodified plain type of PD, has the obvious disadvantage of cytotoxicity, but still serves as an attractive molecule because it easily adheres to the cell surface, facilitating easy cellular uptake. Single-photon emission computed tomographic imaging of a mouse following intravenous injection of a radiolabeled PD failed to reveal any signal in the intracranial region. Furthermore, examination of the permeability of PD particles across the blood-brain barrier (BBB) in vitro using a commercially available kit revealed poor permeability of the nanoparticles, which was suppressed by an inhibitor of caveolae-mediated endocytosis, but not by an inhibitor of macropinocytosis. Physicochemical analysis of the PD revealed that cationic PDs are likely to aggregate promptly upon mixing with body fluids and that this prompt aggregation is probably driven by non-Derjaguin-Landau- Verwey-Overbeek attractive forces originating from the surrounding divalent ions. Atomic force microscopy observation of a freshly cleaved mica plate soaked in dendrimer suspension (culture media) confirmed prompt aggregation. Our study revealed poor transfer of intravenously administered cationic PDs into the intracranial nervous tissue, and the results of our analysis suggested that this was largely attributable to the reduced BBB permeability arising from the propensity of the particles to promptly aggregate upon mixing with body fluids.

Biodistribution of gold nanoparticles in mice and investigation of their possible translocation by nerve uptake around the alveolus.

The effect of nanoparticles in the environment on our health is a cause of concern. The greatest concern with respect to the biological effect of nanoparticles is that they remain in the body and invade tissues, overcoming the protective mechanisms of the body. It is generally believed that nanoparticles invading a living body move into the blood and are carried by the bloodstream to all organs. However, some studies have shown that the inhaled nanoparticles directly translocate to the central nervous system by nerve uptake. Here quantification of the amount of migration of nanoparticles to organs in short time spans (1, 3, and 6 hr) was attempted by animal experiments. Furthermore, the possibility of migration of nanoparticles through the nerves that project around the alveolus, including the nodose ganglion and dorsal root ganglion (DRG), was investigated. Gold (Au) nanoparticles (15 nm) were administered to mice by intratracheal instillation and tail vein injection. After tail vein injection, most nanoparticles were distributed in the liver. After intratracheal instillation, approximately 80% of detected nanoparticles remained in the lungs at 1 hr and were believed to be translocated to digestive organs, including the stomach and intestine, at 3 and 6 hr. With respect to quantification in ganglia, the levels in most samples were lower than the limit of quantification of inductively coupled plasma mass spectrometry (ICP-MS). However, Au nanoparticles were detected in DRG in only some samples of intratracheal instillation. Therefore, this suggests the possibility of translocation of nanoparticles to DRG via nerves.

Expression and localization of two SecA homologs in the unicellular red alga Cyanidioschyzon merolae.

SecA is an ATP-driven motor for Sec translocase that participates in bacterial protein export and thylakoidal import in plants. We have reported that Cyanidioschyzon merolae, a unicellular red alga, possesses a nuclear-encoded secA(nuc) and a plastid-encoded secA(pt) gene. In this study we found that the amount of SecA(nuc) protein almost quadrupled at high temperature, whereas that of the SecA(pt) protein increased far less. We were also able to determine the localization of both SecAs to the chloroplast by immunofluorescence and immunoelectron microscopy. We suggest that SecA(nuc) has an important role in the chloroplast at high temperatures.

Characterization of the nuclear- and plastid-encoded secA-homologous genes in the unicellular red alga Cyanidioschyzon merolae.

SecA is an ATP-driven motor for protein translocation in bacteria and plants. Mycobacteria and listeria were recently found to possess two functionally distinct secA genes. In this study, we found that Cyanidioschyzon merolae, a unicellular red alga, possessed two distinct secA-homologous genes; one encoded in the cell nucleus and the other in the plastid genome. We found that the plastid-encoded SecA homolog showed significant ATPase activity at low temperature, and that the ATPase activity of the nuclear-encoded SecA homolog showed significant activity at high temperature. We propose that the two SecA homologs play different roles in protein translocation.