A solid-phase dot assay using silica/gold nanoshells

We report on the first application of silica-gold nanoshells to a solid-phase dot immunoassay. The assay principle is based on staining of a drop (1 µl) analyte on a nitrocellulose membrane strip by using silica/gold nanoshells conjugated with biospecific probing molecules. Experimental example is human IgG (hIgG, target molecules) and protein A (probing molecules). For usual 15-nm colloidal gold conjugates, the minimal detectable amount of hIgG is about 4 ng. By contrast, for nanoshell conjugates (silica core diameter of 70 nm and gold outer diameter of 100 nm) we have found significant increase in detection sensitivity and the minimal detectable amount of hIgG is about 0.5 ng. This finding is explained by the difference in the monolayer particle extinction

Synthesis of silymarin−selenium nanoparticle conjugate and examination of its biological activity in vitro

Silymarin (Sil) was conjugated to selenium nanoparticles (SeNPs) to increase Sil bioavailability. The conjugates were monodisperse; the average diameter of the native SeNPs was ~ 20-50 ± 1.5 nm, whereas that of the conjugates was 30-50 ± 0.5 nm. The use of SeNPs to increase the bioavailability of Sil was examined with the MH-22a, EPNT-5, HeLa, Hep-2, and SPEV-2 cell lines. The EPNT-5 (glioblastoma) cells were the most sensitive to the conjugates compared to the conjugate-free control. The conjugates increased the activity of cellular dehydrogenases and promoted the penetration of Sil into the intracellular space. Possibly, SeNPs play the main part in Sil penetration of cells and Sil penetration is not associated with phagocytosis. Thus, SeNPs are promising for use as a Sil carrier and as protective antigens

Biodynamic parameters of micellar diminazene in sheep erythrocytes and blood plasma

In this work, we used a preparation of diminazene, which belongs to the group of aromatic diamidines. This compound acts on the causative agents of blood protozoan diseases produced by both flagellated protozoa (Trypanosoma) and members of the class Piroplasmida (Babesia, Theileria, and Cytauxzoon) in various domestic and wild animals, and it is widely used in veterinary medicine. We examined the behavior of water-disperse diminazene (immobilized in Tween 80 micelles) at the cellular and organismal levels. We assessed the interaction of an aqueous and a water-disperse preparation with cells of the reticuloendothelial system. We compared the kinetic parameters of aqueous and water-disperse diminazene in sheep erythrocytes and plasma. The therapeutic properties of these two preparations were also compared. We found that the surface-active substances improved intracellular penetration of the active substance through interaction with the cell membrane. In sheep blood erythrocytes, micellar diminazene accumulated more than its aqueous analog. This form was also more effective therapeutically than the aqueous analog. Our findings demonstrate that use of micellar diminazene allows the injection dose to be reduced by 30%

Immunization of Mice with Gold Nanoparticles Conjugated to Thermostable Cancer Antigens Prevents the Development of Xenografted Tumors

Gold nanoparticles as part of vaccines greatly increase antigen stability, antigen accumulation in the lymph nodes, and antigen uptake by antigen-presenting cells. The use of such particles as part of anticancer vaccines based on heat shock proteins to increase vaccine effectiveness is timely. We prepared and characterized nanoconjugates based on 15-nm gold nanoparticles and thermostable tumor antigens isolated from MH22a murine hepatoma cells. The whole-cell lysate of MH22a cells contained the main heat shock proteins. BALB/c mice were injected with the conjugates and then received transplants of MH22a cells. The highest titer was produced in mice immunized with the complex of gold nanoparticles + antigen with complete Freund’s adjuvant. The immunized mice showed no signs of tumor growth for 24 days. They also showed a decreased production of the INF-γ, IL-6, and IL-1 proinflammatory cytokines compared to the mice immunized through other schemes. This study is the first to show that it is possible in principle to use gold nanoparticles in combination with thermostable tumor antigens for antitumor vaccination. Antitumor vaccines based on thermostable tumor antigens can be largely improved by including gold nanoparticles as additional adjuvants

Identification and Characterization of Rothia amarae sp. nov. in a Suspension Culture of Arabidopsis thaliana (Heynh.) Cells

We report, for the first time, that a nonpathogenic bacterial microflora has been found in a suspension culture of Arabidopsis thaliana cells. The 16S rRNA gene sequencing showed that the isolate belonged to Rothia amarae. Identification was confirmed by microbiological, microscopic, and immunochemical methods. The growth of the isolate on blood agar preserves the morphological and immunochemical properties of the isolate from the plant cell suspension culture. Whether the isolated strain is a contaminant or a true symbiont remains an open question. It is known that Rothia bacteria live mostly in oceanic and waste water and in benthos. Members of Rothia are part of the normal microflora of the oral cavity, respiratory tract, and stomach of humans. Endophytic Rothia are inhibitory against several pathogenic fungi and bacteria. In addition, some actinobacteria, including members of Rothia, are nitrogen fixers. It cannot be ruled out that the R. amarae strain isolated in this work can be endosymbiotic with a suspension culture of A. thaliana. The bacterial “inclusions” found by us in a suspension culture of A. thaliana merit further investigation to identify them more deeply and clarify their symbiotic properties