Single-cell mapping of focused ultrasound-transfected brain.

Gene delivery via focused ultrasound (FUS) mediated blood-brain barrier (BBB) opening is a disruptive therapeutic modality. Unlocking its full potential will require an understanding of how FUS parameters (e.g., peak-negative pressure (PNP)) affect transfected cell populations. Following plasmid (mRuby) delivery across the BBB with 1 MHz FUS, we used single-cell RNA-sequencing to ascertain that distributions of transfected cell types were highly dependent on PNP. Cells of the BBB (i.e., endothelial cells, pericytes, and astrocytes) were enriched at 0.2 MPa PNP, while transfection of cells distal to the BBB (i.e., neurons, oligodendrocytes, and microglia) was augmented at 0.4 MPa PNP. PNP-dependent differential gene expression was observed for multiple cell types. Cell stress genes were upregulated proportional to PNP, independent of cell type. Our results underscore how FUS may be tuned to bias transfection toward specific brain cell types in vivo and predict how those cells will respond to transfection.

Analysis of diterpenoid compounds from the latex of two Euphorbiaceae by liquid chromatography‒electrospray ionisation mass spectrometry.

The latex of Euphorbia papillosa A.St.-Hil. and Sapium glandulosum (L.) Morong was phytochemically investigated by liquid chromatography/mass spectrometry. The results reveal the presence of tigliane-type diterpenoid compounds, particularly 12-deoxyphorbol esters, in both species. In addition, ingenane-type diterpene esters were found in the latex of E. papillosa. The latex of S. glandulosum showed only tigliane-type diterpenes, confirming the literature data on this genus. According to mass data, we are proposing the presence of 10 compounds derived from 12-deoxyphorbol monoesters and diesters, from one 12-deoxyphorbaldehyde, from two ingenol derivatives and from one nitrogen-containing phorbol ester in the latex of the analysed species. Considering that 12-deoxy tigliane diterpenes are described as antitumour and antiviral agents, these results indicate a pharmacological potential for these two Euphorbiaceae species.

Gniditrin is the main diterpenoid constituent in the bark of Daphne mezereum L.

The bark of Daphne mezereum L. is known as toxic drug due to the presence of diterpene esters. The phytochemical analysis of the bark used for preparation of homeopathic mother tinctures showed that gniditrin was the main diterpene constituent, only in the fruits of D. mezereum mezerein could be detected. The complete NMR data of gniditrin are published for the first time.

Proteolytic activity in latex of the genus Euphorbia--a chemotaxonomic marker?

Eighteen species of the genus Euphorbia are known to have proteolytic enzymes in their latices, 9 of them are characterized by the type of endopeptidases (Cysteine-, Serine-, Metallo- or Aspartatic-endopeptidase) which are responsible for the activity, and all nine are serine endopeptidases. In our study we examined the latices of 64 different species of the genus Euphorbia concerning proteolytic activity and serine protease activity, five of them are mentioned in the literature to be proteolytic active and four are known to contain at least one serine endopeptidase. All tested samples were able to degrade labelled casein, the activity of six latices were completely inhibited by specific serine protease inhibitors, 15 samples were not influenced, and in 43 latices a remaining activity was measured, indicating that other types of endopeptidases seem to be involved.

Enhancement of cytotoxicity of ribosome-inactivating-protein type I by saponinum album is not based on stimulation of phagocytosis.

Saponinum album, a mixture of triterpenoic saponins derived from Gypsophila species, led to an increased internalization of agrostin, a ribosome-inactivating-protein (RIP) type I in U-937 cells differentiated with interferon-gamma or phorbol myristate acetate. Treatment with agrostin only showed no cytotoxicity. It was hypothesized that saponinum album stimulated phagocytosis and by that the uptake of agrostin. For this purpose phagocytosis experiments with Alexa-Fluor-488-labelled 1-microm amino-latex beads and FITC-labelled Escherichia coli (K-12 strain) were performed. The results indicated no stimulation of phagocytosis by treatment with saponinum album.