Light triggering goldsomes enable local NO-generation and alleviate pathological vasoconstriction.

While nitric oxide (NO) can remedy vasoconstriction, inhalation of NO may cause systematic toxicity. We report a goldsome, which comprises a hollowed poly(lactic-co-glycolic acid) (PLGA) polymersome with S-nitrosoglutathione (GSNO, a NO donor) molecules and gold nanoparticles (Au NPs) incorporated in its hydrophilic core and hydrophobic membrane, respectively. Photothermal heating caused breakdown of polymersomes and enabled NO generation through reaction between GSNO and Au NPs. Photo-illumination at the zebrafish head led to local NO generation and selective cerebral vasodilation while it had little effects in regions away from the illumination site, and effectively mitigated hypoxia induced cerebral vasoconstriction. We demonstrate a translational potential by showing photo-stimulated NO generation with a clinical intravascular optical catheter. In conclusion, the goldsome, which enables light stimulated local NO generation and can be delivered with clinical intravascular optical catheters, should extend applications of NO therapies while surmounting limitations associated with systemic administration.

A Novel Metallo-ß-Lactamase Involved in the Ampicillin Resistance of Streptococcus pneumoniae ATCC 49136 Strain.

Streptococcus pneumoniae, a penicillin-sensitive bacterium, is recognized as a major cause of pneumonia and is treated clinically with penicillin-based antibiotics. The rapid increase in resistance to penicillin and other antibiotics affects 450 million people globally and results in 4 million deaths every year. To unveil the mechanism of resistance of S. pneumoniae is thus an important issue to treat streptococcal disease that might consequently save millions of lives around the world. In this work, we isolated a streptococci-conserved L-ascorbate 6-phosphate lactonase, from S. pneumoniae ATCC 49136. This protein reveals a metallo-ß-lactamase activity in vitro, which is able to deactivate an ampicillin-based antibiotic by hydrolyzing the amide bond of the ß-lactam ring. The Michaelis parameter (Km) = 25 µM and turnover number (kcat) = 2 s(-1) were obtained when nitrocefin was utilized as an optically measurable substrate. Through confocal images and western blot analyses with a specific antibody, the indigenous protein was recognized in S. pneumoniae ATCC 49136. The protein-overexpressed S. pneumonia exhibits a high ampicillin-tolerance ability in vivo. In contrast, the protein-knockout S. pneumonia reveals the ampicillin-sensitive feature relative to the wild type strain. Based on these results, we propose that this protein is a membrane-associated metallo-ß-lactamase (MBL) involved in the antibiotic-resistant property of S. pneumoniae.

Toward live-cell imaging of dopamine neurotransmission with fluorescent neurotransmitter analogues.

We report a novel 'fluorescent dopamine' that possesses essential features of natural dopamine. Our method is simple and is readily extended to monoamine neurotransmitters such as L-norepinephrine, serotonin and GABA, providing a more practical approach. Because of its compatibility with sensitive fluorescent measurements, we envisage that our approach will have a broad range of applications in neural research.

Interplay between structure and fluidity of model lipid membranes under oxidative attack.

A proper regulation of membrane fluidity is critical for cellular activities such as communication between cells, mitosis, and endocytosis. Unsaturated lipids, a main component of biological membranes, are particularly susceptible to oxidative attack of reactive oxygen species. The oxidation of lipids can produce structural derangement of membranes and eventually alter the membrane fluidity. We have applied fluorescence correlation spectroscopy (FCS) and Raman spectroscopy to investigate the fluidity and structure of model membranes subject to oxidative attack. Hydrogen peroxide has little effect on the lateral fluidity of membranes, whereas hydroxyl radical causes a significantly increased fluidity. The latter is rationalized with the cleavage of the acyl chains of lipids caused by hydroxyl radical; this interpretation is founded on the diminished intensities of lines in Raman spectra associated with -CH(2) and C═C moieties in lipids and supported by mass-spectral measurements. The same approach provides a mechanistic account of the inhibitory capability of vitamins C and E against the increased membrane fluidity resulting from an oxidative attack. Membranes with much cholesterol exhibit a novel resistance against altered membrane fluidity induced with oxidative attack; this finding has biological implications. Our approach combining FCS and Raman measurements reveals the interplay between the structure and fluidity of membranes and provides insight into the pathophysiology of cellular oxidative injury.