Archaea, bacteria, and eukarya secrete membrane microvesicles (MVs) as a mechanism for intercellular communication. We report the isolation and characterization of MVs from the probiotic strain Lactobacillus casei BL23. MVs were characterized using analytical high performance techniques, DLS, AFM and TEM. Similar to what has been described for other Gram-positive bacteria, MVs were on the nanometric size range (30–50 nm). MVs carried cytoplasmic components such as DNA, RNA and proteins. Using a proteomic approach (LC-MS), we identified a total of 103 proteins; 13 exclusively present in the MVs. The MVs content included cell envelope associated and secretory proteins, heat and cold shock proteins, several metabolic enzymes, proteases, structural components of the ribosome, membrane transporters, cell wall-associated hydrolases and phage related proteins. In particular, we identified proteins described as mediators of Lactobacillus’ probiotic effects such as p40, p75 and the product of LCABL_31160, annotated as an adhesion protein. The presence of these proteins suggests a role for the MVs in the bacteria-gastrointestinal cells interface. The expression and further encapsulation of proteins into MVs of GRAS (Generally Recognized as Safe) bacteria could represent a scientific novelty, with applications in food, nutraceuticals and clinical therapies.
The
diffusion of alkaline chlorides (LiCl, KCl, and CsCl) and water
in mesoporous silica samples with pore sizes covering the range from
micropores (2 nm) up to mesopores larger than 30 nm have been measured
by resorting to a simple diffusional technique in the case of electrolytes
and 1H NMR in the case of water. The morphology of the
silica samples varies from a microporous structure, an interconnected
network of pores, and typical mesoporous materials with ink-bottle
pores, with increasing pore size. The release of electrolytes from
the silica as a function of time exhibits two differentiated regimes,
at short and long times, which correlates quite well with the size
of the pores and that of necks of the pores, respectively. The diffusion
of water inside the pores follows the same trend with pore size that
the diffusion of electrolytes, indicating a coupling between the ions
and water diffusional mobilities. The tortuosity effect on the diffusion
of all studied electrolytes and water shows a monotonic slight increase
with decreasing diameter for pores larger than 5 nm, while the tortuosity
factor increases markedly for smaller pores. In microporous and mesoporous
silica with pore sizes below 10 nm, the tortuosity factor of Li+ ion is much larger than those for K+ and Cs+ ions, since its diffusion is hindered by a stronger electrostatic
interaction with the ionizable silanol groups on the pore wall; and
also larger than that for water diffusion which it is retarded by
a weaker hydrogen bond interaction with the silanol groups. The differences
in tortuosity factors among alkaline chlorides and water become negligible
for pore sizes larger than 10 nm. The spin–lattice relaxation
time measurements of 1H-water and Li+ ions confirm
this behavior.
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