Targeting Design to the Lung and Pulmonary Intracellular Structure of Endogenous Gene by Irq Modified Nano Carrier

Inhibition of angiogenesis is a novel strategy for the treatment of lung cancer. For efficient therapy, vectors must firstly reach the target tissue and subsequently demonstrate an efficient intracellular targeting. In this study, we attempted to design a vector for in vivo pulmonary targeting which was able to deliver small interfering Ribonucleic Acid (siRNA) for endogenous gene of angiogenesis in pulmonary endothelial cells. siRNA was condensed with polycation agent and encapsulated in lipidous nano carrier. To obtain high level of lung accumulation, we controlled the surface of nano-carrier by changing the length of Polyethylene glycol (PEG) moiety. These nano carriers showed prominent Ribonucleic acid interference (RNAi) effect, when luciferase gene was used as a target. In addition, an efficient transgene knockdown ofVascular Endothelial Growth Factor Receptor 1 (VEGFR1), a responsible gene of angiogenesis, can be obtained by the Instantaneous Respiratory Exchange Ratio (IRQ) modified nano carrier with the use of Stearyl-R8 (STR-R8) peptide, known as an endosomal membrane inducer. In conclusion, pulmonary targeting of nano carrier by encapsulating siRNA can be developed by controlling the PEG length and the structure of nano carrier for efficient intracellular targeting

Molecular Pathogenesis of Preeclampsia: MicroRNA Hypothesis

The discovery of micro RNA (miRNA) in 1993 by Ambros and colleagues has a huge influence in pathogenesis theory, diagnosis and treatment approach of some diseases. Some studies have conducted to seek the association alterations of miRNA expression to incidences and severity of preeclampsia (PE). We have reviewed some studies that conducted to seek the association of miRNA and PE and we discussed the role of various miRNAs in PE pathogenesis. In summary, we have shown that many researchers have given evident that the different placental and plasma miRNA expression is associated with PE. Some studies also identified the novel candidate of miRNAs (and their pathways) that may be of etiologic relevance in the pathogenesis of PE. Base on review, specific miRNA have a role to down regulate of anti apoptosis genes, regulate angiogenics growth factors such as angiogenin, vascular endothelial growth factor (VEGF) B (VEGF-β), cysteine-rich 61 (CYR61), Placental growth factor (PlGF) and VEGF-A that have a role in angiogenesis. miRNA also have a role in survival, migration, and capillary tube formation of HUVEC by targeted of c-kit. Some miRNAs target genes that participate in immunologic dysfunction, cell adhesion, cell cycle, and signaling. miRNA also have a roles in endothelial cell response to hypoxia, cell differentiation, and survival. A miRNA influence calcium signaling through negative regulations of the calmodulin-coding mRNAs, Mef2a and Gata4, mainly in smooth muscle cells that contribute to PE pathogenesis. These investigations provide novel targets for further investigation of the pathogenesis of PE and these differential miRNAs may be potential markers for the diagnosis and provide a potential therapeutic target for PE. Further investigations on posttranscriptional regulation in PE to evaluate biologic effects of identified miRNAs (including confirmations of miRNA and target gene interactions) are neede

Genome variations associated with viral susceptibility and calcification in Emiliania huxleyi

Emiliania huxleyi, a key player in the global carbon cycle is one of the best studied coccolithophores with respect to biogeochemical cycles, climatology, and host-virus interactions. Strains of E. huxleyi show phenotypic plasticity regarding growth behaviour, light-response, calcification, acidification, and virus susceptibility. This phenomenon is likely a consequence of genomic differences, or transcriptomic responses, to environmental conditions or threats such as viral infections. We used an E. huxleyi genome microarray based on the sequenced strain CCMP1516 (reference strain) to perform comparative genomic hybridizations (CGH) of 16 E. huxleyi strains of different geographic origin. We investigated the genomic diversity and plasticity and focused on the identification of genes related to virus susceptibility and coccolith production (calcification). Among the tested 31940 gene models a core genome of 14628 genes was identified by hybridization among 16 E. huxleyi strains. 224 probes were characterized as specific for the reference strain CCMP1516. Compared to the sequenced E. huxleyi strain CCMP1516 variation in gene content of up to 30 percent among strains was observed. Comparison of core and non-core transcripts sets in terms of annotated functions reveals a broad, almost equal functional coverage over all KOG-categories of both transcript sets within the whole annotated genome. Within the variable (non-core) genome we identified genes associated with virus susceptibility and calcification. Genes associated with virus susceptibility include a Bax inhibitor-1 protein, three LRR receptor-like protein kinases, and mitogen-activated protein kinase. Our list of transcripts associated with coccolith production will stimulate further research, e.g. by genetic manipulation. In particular, the V-type proton ATPase 16 kDa proteolipid subunit is proposed to be a plausible target gene for further calcification studies

Clathrin-Mediated Endocytosis in Living Host Cells Visualized through Quantum Dot Labeling of Infectious Hematopoietic Necrosis Virus▿†

Infectious hematopoietic necrosis virus (IHNV) is an important fish pathogen that infects both wild and cultured salmonids. As a species of the genus Novirhabdovirus, IHNV is a valuable model system for exploring the host entry mechanisms of rhabdoviruses. In this study, quantum dots (QDs) were used as fluorescent labels for sensitive, long-term tracking of IHNV entry. Using live-cell fluorescence microscopy, we found that IHNV is internalized through clathrin-coated pits after the virus binds to host cell membranes. Pretreatment of host cells with chlorpromazine, a drug that blocks clathrin-mediated endocytosis, and clathrin light chain (LCa) depletion using RNA interference both resulted in a marked reduction in viral entry. We also visualized transport of the virus via the cytoskeleton (i.e., actin filaments and microtubules) in real time. Actin polymerization is involved in the transport of endocytic vesicles into the cytosol, whereas microtubules are required for the trafficking of clathrin-coated vesicles to early endosomes, late endosomes, and lysosomes. Disrupting the host cell cytoskeleton with cytochalasin D or nocodazole significantly impaired IHNV infectivity. Furthermore, infection was significantly affected by pretreating the host cells with bafilomycin A1, a compound that inhibits the acidification of endosomes and lysosomes. Strong colocalizations of IHNV with endosomes indicated that the virus is internalized into these membrane-bound compartments. This is the first report in which QD labeling is used to visualize the dynamic interactions between viruses and endocytic structures; the results presented demonstrate that IHNV enters host cells via clathrin-mediated endocytic, cytoskeleton-dependent, and low-pH-dependent pathways

A Highly Efficient Synthetic Vector: Nonhydrodynamic Delivery of DNA to Hepatocyte Nuclei in Vivo

Multifunctional membrane-core nanoparticles, composed of calcium phosphate cores, arginine-rich peptides, cationic and PEGylated lipid membranes, and galactose targeting ligands, have been developed as synthetic vectors for efficient nuclear delivery of plasmid DNA and subsequent gene expression in hepatocytes in vivo. Targeted particles exhibited rapid and extensive hepatic accumulation and were predominantly internalized by hepatocytes, while the inclusion of such peptides in LCP was sufficient to elicit high degrees of nuclear translocation of plasmid DNA. Monocyclic CR8C significantly enhanced in vivo gene expression over ten-fold more than linear CR8C, likely due to a release-favoring mechanism of the DNA/peptide complex. Though 100-fold lower in activity than that achieved via hydrodynamic injection, this formulation presents as a much less invasive alternative. To our knowledge, this is the most effective synthetic vector for liver gene transfer