A novel IRQ ligand-modified nano-carrier targeted to a unique pathway of caveolar endocytic pathway.

In the present study, the cellular uptake and subsequent intracellular trafficking of liposomes was investigated, in which a novel peptide (IRQ), identified with in vivo phage display, was modified on the surface. Since the novel peptide IRQRRRR is rich in arginine, the cellular uptake mechanism was compared with octaarginine (R8)-modified liposomes, which are known to be taken up by cells via macropinocytosis. The uptake mechanism and intracellular trafficking of peptide-modified liposomes was determined by confocal laser scanning microscopy and flow cytometry analysis. Modification of the liposomal surface with the IRQ peptide (IRQ-Lip), induced internalization via a novel pathway-caveolar endocytosis-in parallel with clathrin-mediated endocytosis. Furthermore, the IRQ peptide stimulated escape from endocytic vesicles, leading to efficient gene silencing. When siRNA was condensed and encapsulated in an IRQ-modified multifunctional envelope-type nano-device (IRQ-MEND), transgene expression was reduced 52% with the fusogenic lipid, DOPE/CHEMS. This result shows that the novel IRQ can be utilized for cytoplasmic delivery of macromolecules. Moreover, the IRQ has the potential to be useful for delivery therapeutic agents to parenchymal cells via caveolar endocytosis, as this uptake pathway also plays an important role in transcytosis.

Evaluation of modified CMC and CMC-PVA as miscible polymer blend membranes for hepatocytes.

CMC and CMC-PVA were blended either with type I collagen, BSA or CS to obtain biocompatible membranes for evaluation as potential hepatocyte culture substrates. Pure and modified forms of CMC showed distinct surface, mechanical, and cell attachment properties. While the hydrophilicity decreased, the mechanical stability and the porosity of CMC membranes increased after blending. Serum proteins were adsorbed by all types of membranes. Among eight membranes tested, collagen-modified CMC was found to be a suitable membrane material for hepatocyte culture, in terms of mechanical and cell interaction properties.

Determination of a translocation chromosome by atomic force microscopy.

Atomic force microscopy (AFM) has been used to study the translocation involving chromosomes 11 and 13. An amniocentesis procedure was performed at 18 weeks of pregnancy on a familial balanced translocation carrier mother whose karyotype was 46,XX,t(11;13) (q23;q34). After harvesting the tissue cultures, light microscopy studies (LM) have indicated that the fetus had the same translocation. A 0.3 microm gap region on the derivative chromosome 13 was determined by AFM; it was equivalent to a mid-sized G-band. The enhanced resolution of AFM with respect to its line measure analysis and three-dimensional image capture capability has allowed an extension and reconsideration of conclusions about chromosomal aberrations based on the study of LM preparations. In this manner, chromosomal disorders will be studied at nanoscale to help in the planning of new therapy strategies.