Nanoparticle delivery of a peptide targeting EGFR signaling

EGFR serves as an important therapeutic target because of its over-expression in many cancers. In this study, we investigated a peptide-based therapy aimed at blocking intracellular protein-protein interactions during EGFR signaling and evaluated a targetable lipid carrier system that can deliver peptides to intracellular targets in human cancer cells. EEEEpYFELV (EV), a nonapeptide mimicking the Y845 site of EGFR which is responsible for STAT5b phosphorylation, was designed to block EGFR downstream signaling. EV was loaded onto LPH nanoparticles that are comprised of a membrane/core structure including a surface-grafted polyethylene glycol (PEG) used to evade the reticuloendothelial system (RES) and anisamide (AA) for targeting the sigma receptor over-expressed in H460 human lung cancer cells. EV formulated with PEGylated and targeted LPH (LPH-PEG-AA) was taken up by the tumor cells and trafficked to the cytoplasm with high efficiency. Using this approach, EV acted as a dominant negative inhibitor of STAT5b phosphorylation, arrested cell proliferation, and induced massive apoptosis. Intravenous administration of EV loaded in LPH-PEG-AA led to efficient EV peptide delivery to the tumor in a xenograft mouse model, and multiple injections inhibited tumor growth in a dose-dependent manner. Our findings offer proof-of-concept for an intracellular peptide-mediated cancer therapy that is delivered by carefully designed nanoparticles

The targeted intracellular delivery of cytochrome C protein to tumors using lipid-apolipoprotein nanoparticles

Intracellular-acting therapeutic proteins offer a promising clinical alternative to extracellular-acting agents, but are limited in efficacy by their low permeability into the cell cytoplasm. We have developed a nanoparticle (NP) composed of lipid (DOTAP/DOPE) and apolipoprotein (APO A-I) to mediate the targeted delivery of intracellular-acting protein drugs to non-small cell lung tumors. NPs were produced with either GFP, a fluorescent model protein, or cytochrome C (cytC), an inducer of apoptosis in cancer cells. GFP and cytC were separately conjugated with a membrane permeable sequence (MPS) peptide and were admixed with DOPE/DOTAP nanoparticle formulations (NPs) to enable successful protein loading. Protein-loaded NPs were modified with DSPE-PEG-Anisamide to enable specific NP targeting to the tumor site in a xenograft model. The resulting particle was 20–30 nm in size and exhibited a 64–75% loading efficiency. H460 cells treated with the PEGylated MPS-cytC-NPs exhibited massive apoptosis. When MPS-GFP-NPs or MPS-cytC-NPs were intravenously administered in H460 tumor bearing mice, a specific tumor targeting effect with low NP accumulation in the liver was observed. In addition, MPS-cytC-NP treatment provoked a tumor growth retardation effect in H460 xenograft mice. We conclude that our NP enables targeted, efficacious therapeutic protein delivery for the treatment of lung cancer

Targeted delivery of EV peptide to tumor cell cytoplasm using lipid coated calcium carbonate nanoparticles

Intracellular-acting peptide drugs are effective for inhibiting cytoplasmic protein targets, yet face challenges with penetrating the cancer cell membrane. We have developed a lipid nanoparticle formulation that utilizes a pH-sensitive calcium carbonate complexation mechanism to enable the targeted delivery of the intracellular-acting therapeutic peptide EEEEpYFELV (EV) into lung cancer cells. Lipid-calcium-carbonate (LCC) nanoparticles were conjugated with anisamide, a targeting ligand for the sigma receptor which is expressed on lung cancer cells. LCC EV nanoparticle treatment provoked severe apoptotic effects in H460 non-small cell lung cancer cells in vitro. LCC NPs also mediated the specific delivery of Alexa- 488-EV peptide to tumor tissue in vivo, provoking a high tumor growth retardation effect with minimal uptake by external organs and no toxic effects

COMPARISON OF ANGULAR KINEMATIC PATTERNS BETWEEN CARVING TURN AND SKIDDING TURN DURING ALPINE SKIING

The purpose of this study was to investigate the movement patterns between segments (lower spine, pelvis, thigh, shank) and ski using the relative angular displacement on anteroposterior and vertical axis. Fourteen alpine ski instructors were participated in this study. Eight inertial measurement units were used to measure kinematic variables. Each skier was asked to perform ten carving turns and ten skidding turns on the groomed 15°slope, respectively. On the vertical axis, relative angular displacement of lower spine-ski was significantly increased during carving turn, whereas relative angular displacement of shank-ski was significantly increased during skidding turn. On the anteroposterior axis, relative angular displacement of lower spine-ski, pelvis-ski and thigh-ski were significantly increased during carving turn

SUCCESSFUL FACTORS OF 540° DWIHURYEOCHAGI IN TAEKWONDO

The purpose of our study was to provide fundamental information about success factors of 540° Dwihuryeochagi in Taekwondo. Twenty Taekwondo athletes who participated in the 2012 Taekwondo Kyukpa Wang (breaking king) championship: ten successful athletes (S, age: 23.1±1.6 yrs, height: 171.0±3.5 cm, body mass: 66.4±7.1 kg) and ten failed athletes (F, age: 22.3±1.8 yrs, height: 172.1±5.4 cm, body mass: 64.4±4.2 kg) were selected. Three-dimensional motion analysis using a system of 3 video cameras with a sampling of 60 fields/s was performed during the competition of 540 ° Dwihuryeochagi. Based on the findings, it is concluded that success factors of 540° Dwihuryeochagi were horizontal velocity of COM during P1, vertical velocity of COM during P2, and the time, kick distance, velocity and angle of lower extremities of P3-P4

Genetic and Metabolic Characterization of Insomnia

Insomnia is reported to chronically affect 10∼15% of the adult population. However, very little is known about the genetics and metabolism of insomnia. Here we surveyed 10,038 Korean subjects whose genotypes have been previously profiled on a genome-wide scale. About 16.5% reported insomnia and displayed distinct metabolic changes reflecting an increase in insulin secretion, a higher risk of diabetes, and disrupted calcium signaling. Insomnia-associated genotypic differences were highly concentrated within genes involved in neural function. The most significant SNPs resided in ROR1 and PLCB1, genes known to be involved in bipolar disorder and schizophrenia, respectively. Putative enhancers, as indicated by the histone mark H3K4me1, were discovered within both genes near the significant SNPs. In neuronal cells, the enhancers were bound by PAX6, a neural transcription factor that is essential for central nervous system development. Open chromatin signatures were found on the enhancers in human pancreas, a tissue where PAX6 is known to play a role in insulin secretion. In PLCB1, CTCF was found to bind downstream of the enhancer and interact with PAX6, suggesting that it can probably inhibit gene activation by PAX6. PLCB4, a circadian gene that is closely located downstream of PLCB1, was identified as a candidate target gene. Hence, dysregulation of ROR1, PLCB1, or PLCB4 by PAX6 and CTCF may be one mechanism that links neural and pancreatic dysfunction not only in insomnia but also in the relevant psychiatric disorders that are accompanied with circadian rhythm disruption and metabolic syndrome

SPRINTING SPEED OF ELITE SPRINTERS AT THE WORLD CHAMPIONSHIPS

The purpose of this study was to analyze the sprinting speed of the sprinters during the men's 100 m events at the IAAF World Championships (WC) in order to provide important information to track and field coaches and athletes. Sprinting speed of the sprinters was collected by using laser guns (Laveg Sport, Jenoptik, Germany) during the World Championships in Daegu 2011. Then, data from Osaka 2007 WC and Berline 2009 WC were included in the analysis. The findings indicated that a reduction of a sprinter’s maximum speed is correlated with their performance time (

SPRINTING CHARACTERISTICS OF WOMEN’S 100 METER FINALS AT THE IAAF WORLD CHAMPIONSHOPS DAEGU 2011

This study analyzed the sprinting characteristics of the finalists during the women's 100 m event in the IAAF World Championships Daegu 2011 in order to provide important information to track and field coaches and athletes. Five high speed cameras (Casio, Japan) with a sampling frequency of 300 Hz were used to calculate the number of steps, step length, and stride frequency of the eight sprinters in the women’s final event. There was a tendency to show a better performance time with a high number of steps (p=0.13) and shorter stride length (p=0.14) among the eight sprinters. Furthermore, stride frequency and performance time were negatively correlated as a higher stride frequency had a positive impact on performance time (p=0.02). Based on the relationship between COM velocity and lower extremity joint angles, the 4 top ranked sprinters showed the different strategies to maintain a high COM velocity during the mid portion of the race