WorldWideScience

Sample records for activatable cell penetrating

  1. Angiopep-2 and activatable cell penetrating peptide dual modified nanoparticles for enhanced tumor targeting and penetrating.

    Science.gov (United States)

    Mei, Ling; Zhang, Qianyu; Yang, Yuting; He, Qin; Gao, Huile

    2014-10-20

    Delivering chemotherapeutics by nanoparticles into tumor was influenced by at least two factors: specific targeting and highly efficient penetrating of the nanoparticles. In this study, two targeting ligands, angiopep-2 and activatable cell penetrating peptide (ACP), were functionalized onto nanoparticles for tumor targeting delivery. In this system, angiopep-2 is a ligand of low-density lipoprotein receptor-related protein-1 (LRP1) which was highly expressed on tumor cells, and the ACP was constructed by the conjugation of RRRRRRRR (R8) with EEEEEEEE through a matrix metalloproteinase-2 (MMP-2) sensitive linker, enabling the ACP with tumor microenvironment-responsive cell penetrating property. 4h incubation of ACP with MMP-2 leads to over 80% cleavage of ACP, demonstrating ACP indeed possessed MMP-2 responsive property. The constructed dual targeting nanoparticles (AnACNPs) were approximately 110 nm with a polydispersity index of 0.231. In vitro, ACP modification and angiopep-2 modification could both enhance the U-87 MG cell uptake because of the high expression of MMP-2 and LRP-1 on C6 cells. AnACNPs showed higher uptake level than the single ligand modified nanoparticles. The uptake of all particles was time- and concentration-dependent and endosomes were involved. In vivo, AnACNPs showed best tumor targeting efficiency. The distribution of AnACNPs in tumor was higher than all the other particles. After microvessel staining with anti-CD31 antibody, the fluorescent distribution demonstrated AnACNPs could distribute in the whole tumor with the highest intensity. In conclusion, a novel drug delivery system was developed for enhanced tumor dual targeting and elevated cell internalization.

  2. Single-cell resolution imaging of retinal ganglion cell apoptosis in vivo using a cell-penetrating caspase-activatable peptide probe.

    Directory of Open Access Journals (Sweden)

    Xudong Qiu

    Full Text Available Peptide probes for imaging retinal ganglion cell (RGC apoptosis consist of a cell-penetrating peptide targeting moiety and a fluorophore-quencher pair flanking an effector caspase consensus sequence. Using ex vivo fluorescence imaging, we previously validated the capacity of these probes to identify apoptotic RGCs in cell culture and in an in vivo rat model of N-methyl- D-aspartate (NMDA-induced neurotoxicity. Herein, using TcapQ488, a new probe designed and synthesized for compatibility with clinically-relevant imaging instruments, and real time imaging of a live rat RGC degeneration model, we fully characterized time- and dose-dependent probe activation, signal-to-noise ratios, and probe safety profiles in vivo. Adult rats received intravitreal injections of four NMDA concentrations followed by varying TcapQ488 doses. Fluorescence fundus imaging was performed sequentially in vivo using a confocal scanning laser ophthalmoscope and individual RGCs displaying activated probe were counted and analyzed. Rats also underwent electroretinography following intravitreal injection of probe. In vivo fluorescence fundus imaging revealed distinct single-cell probe activation as an indicator of RGC apoptosis induced by intravitreal NMDA injection that corresponded to the identical cells observed in retinal flat mounts of the same eye. Peak activation of probe in vivo was detected 12 hours post probe injection. Detectable fluorescent RGCs increased with increasing NMDA concentration; sensitivity of detection generally increased with increasing TcapQ488 dose until saturating at 0.387 nmol. Electroretinography following intravitreal injections of TcapQ488 showed no significant difference compared with control injections. We optimized the signal-to-noise ratio of a caspase-activatable cell penetrating peptide probe for quantitative non-invasive detection of RGC apoptosis in vivo. Full characterization of probe performance in this setting creates an important in

  3. Challenges associated with the targeted delivery of gelonin to claudin-expressing cancer cells with the use of activatable cell penetrating peptides to enhance potency

    Directory of Open Access Journals (Sweden)

    Howell Stephen B

    2011-02-01

    Full Text Available Abstract Background Treatment of tumors with macromolecular toxins directed to cytoplasmic targets requires selective endocytosis followed by release of intact toxin from the endosomal/lysosomal compartment. The latter step remains a particular challenge. Claudins 3 and 4 are tight junction proteins that are over-expressed in many types of tumors. This study utilized the C-terminal 30 amino acid fragment of C. perfringens enterotoxin (CPE, which binds to claudins 3 and 4, to deliver a toxin in the form of recombinant gelonin (rGel to the cytoplasm of the human ovarian carcinoma cell line 2008. Results CPE was fused to rGel at its N-terminal end via a flexible G4S linker. This CPE-G4S-rGel molecule was internalized into vesicles from which location it produced little cytotoxicity. To enhance release from the endosomal/lysosomal compartment a poly-arginine sequence (R9 was introduced between the CPE and the rGel. CPE-R9-rGel was 10-fold more cytotoxic but selectivity for claudin-expressing cells was lost. The addition of a poly-glutamic acid sequence (E9 through a G4S linker to R9-rGel (E9-G4S-R9-rGel largely neutralized the non-selective cell membrane penetrating activity of the R9 motif. However, introduction of CPE to the E9-G4S-R9-rGel fusion protein (CPE-E9-G4S-R9-rGel further reduced its cytotoxic effect. Treatment with the endosomolytic reagent chloroquine increased the cytotoxicity of CPE-E9-G4S-R9-rGel. Several types of linkers susceptible to cleavage by furin and endosomal cathepsin B were tested for their ability to enhance R9-rGel release but none of these modifications further enhanced the cytotoxicity of CPE-E9-G4S-R9-rGel. Conclusion We conclude that while a claudin-3 and -4 ligand serves to deliver rGel into 2008 cells the delivered molecules were entrapped in intracellular vesicles. Incorporation of R9 non-specifically increased rGel cytotoxicity and this effect could be masked by inclusion of an E9 sequence. However, the putative

  4. Polypeptide micelles with dual pH activatable dyes for sensing cells and cancer imaging.

    Science.gov (United States)

    Gong, Ping; Yang, Yueting; Yi, Huqiang; Fang, Shengtao; Zhang, Pengfei; Sheng, Zonghai; Gao, Guanhui; Gao, Duyang; Cai, Lintao

    2014-05-21

    pH is an important control parameter for maintenance of cell viability and tissue functions. pH monitoring provides valuable information on cell metabolic processes and the living environment. In this study, we prepared dual pH-sensitive, fluorescent dye-loaded polypeptide nanoparticles (DPNs) for ratiometric sensing of pH changes in living cells. DPNs contain two types of dyes: N-(rhodamine B) lactam cystamine (RBLC), an acid activatable fluorescent dye with increased fluorescence in an acidic environment, and fluorescein isothiocyanate (FITC), a base activatable fluorescent dye with enhanced fluorescence in an alkaline environment. Hence, DPNs exhibited a dual response signal with strong red fluorescence and weak green fluorescence under acidic conditions; in contrast, they showed strong green fluorescence and almost no red fluorescence under alkaline and neutral conditions. The favorable inverse pH responses of the two fluorescent dyes resulted in ratiometric pH determination for DPNs with an optimized pH-sensitive range of pH 4.5-7.5. Quantitative analysis of the intracellular pH of intact MCF-7 cells has been successfully demonstrated with our nanosensor. Moreover, single acid activatable fluorescent dye doped polypeptide nanoparticles that only contained RBLC can distinguish tumor tissue from normal tissue by monitoring the acidic extracellular environment.

  5. Activatable photodynamic destruction of cancer cells by NIR dye/photosensitizer loaded liposomes.

    Science.gov (United States)

    Yuan, Ahu; Tang, Xiaolei; Qiu, Xuefeng; Jiang, Ke; Wu, Jinhui; Hu, Yiqiao

    2015-02-25

    The phototoxicity of Chlorin e6 (Ce6) for photodynamic therapy (PDT) was found to be effectively suppressed by indocyanine green (ICG), a near infrared (NIR) dye. Upon NIR laser irradiation at 808 nm, ICG in the liposomes containing ICG and Ce6 could be degraded, while the phototoxicity of Ce6 could be recovered. In addition, we demonstrate that this newly developed liposomal component can be successfully used for activatable PDT to destroy cancer cells in vitro.

  6. Fluorescence in vivo imaging of live tumor cells with pH-activatable targeted probes via receptor-mediated endocytosis

    Science.gov (United States)

    Asanuma, Daisuke; Urano, Yasuteru; Nagano, Tetsuo; Hama, Yukihiro; Koyama, Yoshinori; Kobayashi, Hisataka

    2009-02-01

    One goal of molecular imaging is to establish a widely applicable technique for specific detection of tumors with minimal background. Here, we achieve specific in vivo tumor visualization with a newly-designed "activatable" targeted fluorescence probe. This agent is activated after cellular internalization by sensing the pH change in the lysosome. Novel acidic pH-activatable probes based on the BODIPY fluorophore were synthesized, and then conjugated to a cancer-targeting monoclonal antibody, Trastuzumab, or galactosyl serum albumin (GSA). As proof of concept, ex and in vivo imaging of two different tumor mouse models was performed: HER2-overexpressed lung metastasis tumor with Trastuzumab-pH probe conjugates and lectin-overexpressed i.p. disseminated tumor with GSA-pH probe conjugates. These pH-activatable targeted probes were highly specific for tumors with minimal background signal. Because the acidic pH in lysosomes is maintained by the energy-consuming proton pump, only viable cancer cells were successfully visualized. Furthermore, this strategy was also applied to fluorescence endoscopy in tumor mouse models, resulting in specific visualization of tumors as small as submillimeter in size that could hardly detected by naked eyes because of their poor contrast against normal tissues. The design concept can be widely adapted to cancer-specific cell-surface-targeting molecules that result in cellular internalization.

  7. Activated thrombin-activatable fibrinolysis inhibitor (TAFIa) attenuates breast cancer cell metastatic behaviors through inhibition of plasminogen activation and extracellular proteolysis

    OpenAIRE

    Bazzi, Zainab A.; Lanoue, Danielle; El-Youssef, Mouhanned; Romagnuolo, Rocco; Tubman, Janice; Cavallo-Medved, Dora; Porter, Lisa A.; Boffa, Michael B.

    2016-01-01

    Background Thrombin activatable fibrinolysis inhibitor (TAFI) is a plasma zymogen, which can be converted to activated TAFI (TAFIa) through proteolytic cleavage by thrombin, plasmin, and most effectively thrombin in complex with the endothelial cofactor thrombomodulin (TM). TAFIa is a carboxypeptidase that cleaves carboxyl terminal lysine and arginine residues from protein and peptide substrates, including plasminogen-binding sites on cell surface receptors. Carboxyl terminal lysine residues ...

  8. Protease-activatable organometal-Peptide bioconjugates with enhanced cytotoxicity on cancer cells.

    Science.gov (United States)

    Splith, Katrin; Hu, Wanning; Schatzschneider, Ulrich; Gust, Ronald; Ott, Ingo; Onambele, Liliane A; Prokop, Aram; Neundorf, Ines

    2010-07-21

    Over the past years, numerous promising new metalorganic lead structures have been developed exhibiting highly active cytostatic properties. However, the efficiency of such chemotherapeutics in the treatment of tumors is often limited by their low therapeutic index due to their short half-life, lack of tumor selectivity, and associated side effects. Furthermore, the membrane barrier often restricts their cellular uptake by passive diffusion. In this contribution, we describe the synthesis, cellular uptake, and biologic activity of a series of cymantrene-peptide conjugates. Cymantrene CpMn(CO)(3) is a robust organometallic group, which is stable in air and water and easy to functionalize. In this work, some new cymantrene derivatives with different linkers between the half-sandwich complex and the carboxylate group were attached to the cell-penetrating peptide sC18 that should act as a transporter for the metal moiety. All conjugates were characterized for their cytotoxic activity on human breast adenocarcinoma cells (MCF-7) and human colon carcinoma cells (HT-29). We found that bioconjugates bearing two cymantrene groups were more active than the monofunctionalized ones. By the introduction of a cathepsin B cleavage site next to the organometallic group, the biologic activity could be in increased even further. Fluorescence microscopy studies and apoptosis assays gave preliminary hints on the mode of action of these systems. PMID:20586419

  9. A Cell-Based Internalization and Degradation Assay with an Activatable Fluorescence-Quencher Probe as a Tool for Functional Antibody Screening.

    Science.gov (United States)

    Li, Yan; Liu, Peter Corbett; Shen, Yang; Snavely, Marshall D; Hiraga, Kaori

    2015-08-01

    For the development of therapeutically potent anti-cancer antibody drugs, it is often important to identify antibodies that internalize into cells efficiently, rather than just binding to antigens on the cell surface. Such antibodies can mediate receptor endocytosis, resulting in receptor downregulation on the cell surface and potentially inhibiting receptor function and tumor growth. Also, efficient antibody internalization is a prerequisite for the delivery of cytotoxic drugs into target cells and is critical for the development of antibody-drug conjugates. Here we describe a novel activatable fluorescence-quencher pair to quantify the extent of antibody internalization and degradation in the target cells. In this assay, candidate antibodies were labeled with a fluorescent dye and a quencher. Fluorescence is inhibited outside and on the surface of cells, but activated upon endocytosis and degradation of the antibody. This assay enabled the development of a process for rapid characterization of candidate antibodies potentially in a high-throughput format. By employing an activatable secondary antibody, primary antibodies in purified form or in culture supernatants can be screened for internalization and degradation. Because purification of candidate antibodies is not required, this method represents a direct functional screen to identify antibodies that internalize efficiently early in the discovery process. PMID:26024945

  10. Molecular Imaging with Activatable Reporter Systems

    Directory of Open Access Journals (Sweden)

    Gang Niu, Xiaoyuan Chen

    2012-01-01

    Full Text Available Molecular imaging is a newly emerged multiple disciplinary field that aims to visualize, characterize and quantitatively measure biological processes at cellular and molecular levels in humans and other living systems. A reporter gene is a piece of DNA encoding reporter protein, which presents as a readily measurable phenotype that can be distinguished easily from the background of endogenous protein. After being transferred into cells of organ systems (transgenes, the reporter gene can be utilized to visualize transcriptional and posttranscriptional regulation of gene expression, protein-protein interactions, or trafficking of proteins or cells in living subjects. Herein, we review previous classification of reporter genes and regroup the reporter gene based imaging as basic, inducible and activatable, based on the regulation of reporter gene transcription and post-translational modification of reporter proteins. We then focus on activatable reporters, in which the signal can be activated at the posttranslational level for visualizing protein-protein interactions, protein phosphorylation or tertiary structure changes. The applications of several types of activatable reporters will also be summarized. We conclude that activatable reporter imaging can benefit both basic biomedical research and drug development.

  11. Magnetic resonance imaging of stem cell apoptosis in arthritic joints with a caspase activatable contrast agent.

    Science.gov (United States)

    Nejadnik, Hossein; Ye, Deju; Lenkov, Olga D; Donig, Jessica S; Martin, John E; Castillo, Rostislav; Derugin, Nikita; Sennino, Barbara; Rao, Jianghong; Daldrup-Link, Heike

    2015-02-24

    About 43 million individuals in the U.S. encounter cartilage injuries due to trauma or osteoarthritis, leading to joint pain and functional disability. Matrix-associated stem cell implants (MASI) represent a promising approach for repair of cartilage defects. However, limited survival of MASI creates a significant bottleneck for successful cartilage regeneration outcomes and functional reconstitution. We report an approach for noninvasive detection of stem cell apoptosis with magnetic resonance imaging (MRI), based on a caspase-3-sensitive nanoaggregation MRI probe (C-SNAM). C-SNAM self-assembles into nanoparticles after hydrolysis by caspase-3, leading to 90% amplification of (1)H MR signal and prolonged in vivo retention. Following intra-articular injection, C-SNAM causes significant MR signal enhancement in apoptotic MASI compared to viable MASI. Our results indicate that C-SNAM functions as an imaging probe for stem cell apoptosis in MASI. This concept could be applied to a broad range of cell transplants and target sites.

  12. Market penetration scenarios for fuel cell vehicles

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, C.E.; James, B.D.; Lomax, F.D. Jr. [Directed Technologies, Inc., Arlington, VA (United States)

    1997-12-31

    Fuel cell vehicles may create the first mass market for hydrogen as an energy carrier. Directed Technologies, Inc., working with the US Department of Energy hydrogen systems analysis team, has developed a time-dependent computer market penetration model. This model estimates the number of fuel cell vehicles that would be purchased over time as a function of their cost and the cost of hydrogen relative to the costs of competing vehicles and fuels. The model then calculates the return on investment for fuel cell vehicle manufacturers and hydrogen fuel suppliers. The model also projects the benefit/cost ratio for government--the ratio of societal benefits such as reduced oil consumption, reduced urban air pollution and reduced greenhouse gas emissions to the government cost for assisting the development of hydrogen energy and fuel cell vehicle technologies. The purpose of this model is to assist industry and government in choosing the best investment strategies to achieve significant return on investment and to maximize benefit/cost ratios. The model can illustrate trends and highlight the sensitivity of market penetration to various parameters such as fuel cell efficiency, cost, weight, and hydrogen cost. It can also illustrate the potential benefits of successful R and D and early demonstration projects. Results will be shown comparing the market penetration and return on investment estimates for direct hydrogen fuel cell vehicles compared to fuel cell vehicles with onboard fuel processors including methanol steam reformers and gasoline partial oxidation systems. Other alternative fueled vehicles including natural gas hybrids, direct injection diesels and hydrogen-powered internal combustion hybrid vehicles will also be analyzed.

  13. Cell Penetration Properties of a Highly Efficient Mini Maurocalcine Peptide

    Directory of Open Access Journals (Sweden)

    Michel De Waard

    2013-03-01

    Full Text Available Maurocalcine is a highly potent cell-penetrating peptide isolated from the Tunisian scorpion Maurus palmatus. Many cell-penetrating peptide analogues have been derived from the full-length maurocalcine by internal cysteine substitutions and sequence truncation. Herein we have further characterized the cell-penetrating properties of one such peptide, MCaUF1-9, whose sequence matches that of the hydrophobic face of maurocalcine. This peptide shows very favorable cell-penetration efficacy compared to Tat, penetratin or polyarginine. The peptide appears so specialized in cell penetration that it seems hard to improve by site directed mutagenesis. A comparative analysis of the efficacies of similar peptides isolated from other toxin members of the same family leads to the identification of hadrucalcin’s hydrophobic face as an even better CPP. Protonation of the histidine residue at position 6 renders the cell penetration of MCaUF1-9 pH-sensitive. Greater cell penetration at acidic pH suggests that MCaUF1-9 can be used to specifically target cancer cells in vivo where tumor masses grow in more acidic environments.

  14. Prediction of cell penetrating peptides by support vector machines.

    Directory of Open Access Journals (Sweden)

    William S Sanders

    2011-07-01

    Full Text Available Cell penetrating peptides (CPPs are those peptides that can transverse cell membranes to enter cells. Once inside the cell, different CPPs can localize to different cellular components and perform different roles. Some generate pore-forming complexes resulting in the destruction of cells while others localize to various organelles. Use of machine learning methods to predict potential new CPPs will enable more rapid screening for applications such as drug delivery. We have investigated the influence of the composition of training datasets on the ability to classify peptides as cell penetrating using support vector machines (SVMs. We identified 111 known CPPs and 34 known non-penetrating peptides from the literature and commercial vendors and used several approaches to build training data sets for the classifiers. Features were calculated from the datasets using a set of basic biochemical properties combined with features from the literature determined to be relevant in the prediction of CPPs. Our results using different training datasets confirm the importance of a balanced training set with approximately equal number of positive and negative examples. The SVM based classifiers have greater classification accuracy than previously reported methods for the prediction of CPPs, and because they use primary biochemical properties of the peptides as features, these classifiers provide insight into the properties needed for cell-penetration. To confirm our SVM classifications, a subset of peptides classified as either penetrating or non-penetrating was selected for synthesis and experimental validation. Of the synthesized peptides predicted to be CPPs, 100% of these peptides were shown to be penetrating.

  15. Prediction of cell-penetrating peptides with feature selection techniques.

    Science.gov (United States)

    Tang, Hua; Su, Zhen-Dong; Wei, Huan-Huan; Chen, Wei; Lin, Hao

    2016-08-12

    Cell-penetrating peptides are a group of peptides which can transport different types of cargo molecules such as drugs across plasma membrane and have been applied in the treatment of various diseases. Thus, the accurate prediction of cell-penetrating peptides with bioinformatics methods will accelerate the development of drug delivery systems. The study aims to develop a powerful model to accurately identify cell-penetrating peptides. At first, the peptides were translated into a set of vectors with the same dimension by using dipeptide compositions. Secondly, the Analysis of Variance-based technique was used to reduce the dimension of the vector and explore the optimized features. Finally, the support vector machine was utilized to discriminate cell-penetrating peptides from non-cell-penetrating peptides. The five-fold cross-validated results showed that our proposed method could achieve an overall prediction accuracy of 83.6%. Based on the proposed model, we constructed a free webserver called C2Pred (http://lin.uestc.edu.cn/server/C2Pred). PMID:27291150

  16. Antimicrobial and cell-penetrating properties of penetratin analogs

    DEFF Research Database (Denmark)

    Bahnsen, Jesper Søborg; Franzyk, Henrik; Sandberg-Schaal, Anne;

    2013-01-01

    Cell-penetrating peptides (CPPs) and antimicrobial peptides (AMPs) show great potential as drug delivery vectors and new antibiotic drug entities, respectively. The current study deals with the properties of a variety of peptide analogs derived from the well-known CPP penetratin as well as...

  17. Cell-penetrating peptides: breaking through to the other side.

    Science.gov (United States)

    Koren, Erez; Torchilin, Vladimir P

    2012-07-01

    Cell-penetrating peptides (CPPs) have been previously shown to be powerful transport vector tools for the intracellular delivery of a large variety of cargoes through the cell membrane. Intracellular delivery of plasmid DNA (pDNA), oligonucleotides, small interfering RNAs (siRNAs), proteins and peptides, contrast agents, drugs, as well as various nanoparticulate pharmaceutical carriers (e.g., liposomes, micelles) has been demonstrated both in vitro and in vivo. This review focuses on the peptide-based strategy for intracellular delivery of CPP-modified nanocarriers to deliver small molecule drugs or DNA. In addition, we discuss the rationales for the design of 'smart' pharmaceutical nanocarriers in which the cell-penetrating properties are hidden until triggered by exposure to appropriate environmental conditions (e.g., a particular pH, temperature, or enzyme level), applied local microwave, ultrasound, or radiofrequency radiation. PMID:22682515

  18. Discovery and characterization of a new cell-penetrating protein.

    Science.gov (United States)

    Simeon, Rudo L; Chamoun, Ana Maria; McMillin, Thomas; Chen, Zhilei

    2013-12-20

    We describe a new cell-penetrating protein, B1, capable of delivering conjugated proteins and nucleic acids into mammalian cells. B1 is a 244-amino-acid product of a single-base frameshift in the gene encoding enhanced green fluorescent protein (eGFP). The molecule has a net positive charge of 43 and a very high charge-to-mass ratio of 1.5. eGFP-fused B1 potently penetrates both adherent and suspension cells with >80% of cells taking up the protein when exposed to concentrations as low as 1 μM. The protein was found to cluster in the paranuclear region of TZM-bl cells. Most importantly, we show that B1 not only facilitates cellular uptake but allows biomolecular cargo to reach sites of biological relevance. For example, baby hamster kidney cells underwent DNA recombination when exposed to B1-tagged Cre recombinase at protein concentrations as low as 2.5 μM, indicating potent nuclear delivery of functional protein cargos. Additionally, B1 delivers noncovalently conjugated RNA and DNA across the cell membrane to cytosolic and nuclear sites accessible to the cellular translation and transcription machinery, as gauged by detection of encoded reporter functions, with efficiency comparable to commercially available cationic lipid reagents. B1 appears to utilize cell-surface glycans and multiple competing endocytic pathways to enter and traffic through cells. These studies provide both a new tool for intracellular delivery of biomolecules and insights that could aid in the design of more effective cell penetrating proteins.

  19. Strategies to stabilize cell penetrating peptides for in vivo applications.

    Science.gov (United States)

    Fominaya, Jesús; Bravo, Jerónimo; Rebollo, Angelita

    2015-10-01

    In the era of biomedicines and engineered carrier systems, cell penetrating peptides (CPPs) have been established as a promising tool for therapeutic application. Likewise, other therapeutic peptides, successful in vivo application of CPPs will strongly depend on peptide stability, the bottleneck for this type of biodegradable molecules. In this review, the authors describe the current knowledge of the in vivo degradation for known CPPs and the different strategies available to provide a higher resistance to metabolic degradation while preserving cell penetration efficiency. Peptide stability can be improved by different means, either modifying the structure to make it unrecognizable to proteases, or preventing access of proteolytic enzymes by applying conformation restriction or shielding strategies. PMID:26448473

  20. Investigation of penetration force of living cell using an atomic force microscope

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Eun Young; Kim, Young Tae; Kim, Dae Eun [Yonsei University, Seoul (Korea, Republic of)

    2009-07-15

    Recently, the manipulation of a single cell has been receiving much attention in transgenesis, in-vitro fertilization, individual cell based diagnosis, and pharmaceutical applications. As these techniques require precise injection and manipulation of cells, issues related to penetration force arise. In this work the penetration force of living cell was studied using an atomic force microscope (AFM). L929, HeLa, 4T1, and TA3 HA II cells were used for the experiments. The results showed that the penetration force was in the range of 2{approx}22 nN. It was also found that location of cell penetration and stiffness of the AFM cantilever affected the penetration force significantly. Furthermore, double penetration events could be detected, due to the multi-membrane layers of the cell. The findings of this work are expected to aid in the development of precision micro-medical instruments for cell manipulation and treatment

  1. Intracellular Delivery of Nanoparticles with Cell Penetrating Peptides.

    Science.gov (United States)

    Salzano, Giuseppina; Torchilin, Vladimir P

    2015-01-01

    The functionalization of nanoparticles (NPs) with cell penetrating peptides (CPPs) constitutes a breakthrough for the intracellular delivery of therapeutic and diagnostic payloads. In late 1998, a significant cellular uptake of a small protein from the HIV-1 virus, namely TAT peptide (TATp), was observed. Thereafter, research began on design of similarly acting peptides, and the coupling of NPs with these novel CPPs. Here, we describe recent methods used to modify the surface of NPs with CPPs and the in vitro and in vivo effects of such functionalization on the intracellular delivery of various cargos. In particular, we highlight recent advances aimed at reducing the non-selectivity of CPPs and the prevention of their enzymatic cleavage en route to target tissues. PMID:26202282

  2. Enthalpy-driven interactions with sulfated glycosaminoglycans promote cell membrane penetration of arginine peptides.

    Science.gov (United States)

    Takechi-Haraya, Yuki; Nadai, Ryo; Kimura, Hitoshi; Nishitsuji, Kazuchika; Uchimura, Kenji; Sakai-Kato, Kumiko; Kawakami, Kohsaku; Shigenaga, Akira; Kawakami, Toru; Otaka, Akira; Hojo, Hironobu; Sakashita, Naomi; Saito, Hiroyuki

    2016-06-01

    The first step of cell membrane penetration of arginine peptides is thought to occur via electrostatic interactions between positive charges of arginine residues and negative charges of sulfated glycosaminoglycans (GAGs) on the cell surface. However, the molecular interaction of arginine peptides with GAG still remains unclear. Here, we compared the interactions of several arginine peptides of Tat, R8, and Rev and their analogues with heparin in relation to the cell membrane penetration efficiency. The high-affinity binding of arginine peptides to heparin was shown to be driven by large favorable enthalpy contributions, possibly reflecting multidentate hydrogen bondings of arginine residues with sulfate groups of heparin. Interestingly, the lysine peptides in which all arginine residues are substituted with lysine residues exhibited negligible binding enthalpy despite of their considerable binding to heparin. In CHO-K1 cells, arginine peptides exhibited a great cell-penetrating ability whereas their corresponding lysine peptides did not penetrate into cells. The degree of cell penetration of arginine peptides markedly decreased by the chlorate treatment of cells which prevents the sulfation of GAG chains. Significantly, the cell penetration efficiency of arginine peptides was found to be correlated with the favorable enthalpy of binding to heparin. These results suggest that the enthalpy-driven strong interaction with sulfated GAGs such as heparan sulfate plays a critical role in the efficient cell membrane penetration of arginine peptides.

  3. Infectious rotavirus enters cells by direct cell membrane penetration, not by endocytosis

    Energy Technology Data Exchange (ETDEWEB)

    Kaljot, K.T.; Shaw, R.D.; Greenberg, H.B. (Stanford Univ. School of Medicine, CA (USA) Palo Alto Veterans Administration Medical Center, CA (USA)); Rubin, D.H. (Univ. of Pennsylvania, Philadelphia (USA))

    1988-04-01

    Rotaviruses are icosahedral viruses with a segmented, double-stranded RNA genome. They are the major cause of severe infantile infectious diarrhea. Rotavirus growth in tissue culture is markedly enhanced by pretreatment of virus with trypsin. Trypsin activation is associated with cleavage of the viral hemagglutinin (viral protein 3 (VP3); 88 kilodaltons) into two fragments (60 and 28 kilodaltons). The mechanism by which proteolytic cleavage leads to enhanced growth is unknown. To determine whether trypsin treatment affected rotavirus internalization, the authors studied the kinetics of entry of infectious rhesus rotavirus (RRV) into MA104 cells. Trypsin-activated RRV was internalized with a half-time of 3 to 5 min, while nonactivated virus disappeared from the cell surface with a half-time of 30 to 50 min. In contrast to trypsin-activated RRV, loss of nonactivated RRV from the cell surface did not result in the appearance of infection, as measured by plaque formation. Purified trypsin-activated RRV added to cell monolayers at pH 7.4 mediated {sup 51}Cr, ({sup 14}C)choline, and ({sup 3}H)inositol released from prelabeled MA104 cells. This release could be specifically blocked by neutralizing antibodies to VP3. These results suggest that MA104 cell infection follows the rapid entry of trypsin-activated RRV by direct cell membrane penetration. Cell membrane penetration of infectious RRV is initiated by trypsin cleavage of VP3. Neutralizing antibodies can inhibit this direct membrane penetration.

  4. Membrane adsorption and binding, cellular uptake and cytotoxicity of cell-penetrating peptidomimetics with α-peptide/β-peptoid backbone

    DEFF Research Database (Denmark)

    Jing, Xiaona; Yang, Mingjun; Kasimova, Marina Robertovna;

    2012-01-01

    Cell-penetrating peptides (CPPs) provide a promising approach for enhancing intracellular delivery of therapeutic biomacromolecules by increasing transport through membrane barriers. Here, proteolytically stable cell-penetrating peptidomimetics with α-peptide/β-peptoid backbone were studied...

  5. Visualization of the Nucleolus in Living Cells with Cell-Penetrating Fluorescent Peptides.

    Science.gov (United States)

    Martin, Robert M; Herce, Henry D; Ludwig, Anne K; Cardoso, M Cristina

    2016-01-01

    The nucleolus is the hallmark of nuclear compartmentalization and has been shown to exert multiple roles in cellular metabolism besides its main function as the place of ribosomal RNA synthesis and assembly of ribosomes. The nucleolus plays also a major role in nuclear organization as the largest compartment within the nucleus. The prominent structure of the nucleolus can be detected using contrast light microscopy providing an approximate localization of the nucleolus, but this approach does not allow to determine accurately the three-dimensional structure of the nucleolus in cells and tissues. Immunofluorescence staining with antibodies specific to nucleolar proteins albeit very useful is time consuming, normally antibodies recognize their epitopes only within a small range of species and is applicable only in fixed cells. Here, we present a simple method to selectively and accurately label this ubiquitous subnuclear compartment in living cells of a large range of species using a fluorescently labeled cell-penetrating peptide.

  6. Visualization of the Nucleolus in Living Cells with Cell-Penetrating Fluorescent Peptides.

    Science.gov (United States)

    Martin, Robert M; Herce, Henry D; Ludwig, Anne K; Cardoso, M Cristina

    2016-01-01

    The nucleolus is the hallmark of nuclear compartmentalization and has been shown to exert multiple roles in cellular metabolism besides its main function as the place of ribosomal RNA synthesis and assembly of ribosomes. The nucleolus plays also a major role in nuclear organization as the largest compartment within the nucleus. The prominent structure of the nucleolus can be detected using contrast light microscopy providing an approximate localization of the nucleolus, but this approach does not allow to determine accurately the three-dimensional structure of the nucleolus in cells and tissues. Immunofluorescence staining with antibodies specific to nucleolar proteins albeit very useful is time consuming, normally antibodies recognize their epitopes only within a small range of species and is applicable only in fixed cells. Here, we present a simple method to selectively and accurately label this ubiquitous subnuclear compartment in living cells of a large range of species using a fluorescently labeled cell-penetrating peptide. PMID:27576711

  7. Mitochondrial transit peptide exhibits cell penetration ability and efficiently delivers macromolecules to mitochondria.

    Science.gov (United States)

    Jain, Aastha; Chugh, Archana

    2016-09-01

    Mitochondrial malfunction under various circumstances can lead to a variety of disorders. Effective targeting of macromolecules (drugs) is important for restoration of mitochondrial function and treatment of related disorders. We have designed a novel cell-penetrating mitochondrial transit peptide (CpMTP) for delivery of macromolecules to mitochondria. Comparison between properties of cell-penetrating peptides (CPPs) and mitochondrial signal sequences enabled prediction of peptides with dual ability for cellular translocation and mitochondrial localization. Among the predicted peptides, CpMTP translocates across HeLa cells and shows successful delivery of noncovalently conjugated cargo molecules to mitochondria. CpMTP may have applications in transduction and transfection of mitochondria for therapeutics.

  8. Recent Developments in Thrombin-Activatable Fibrinolysis Inhibitor Research

    NARCIS (Netherlands)

    P.F. Marx; C.J.N. Verkleij; M.V. Seron; J.C.M. Meijers

    2009-01-01

    Thrombin-activatable fibrinolysis inhibitor (TAFI) provides an important molecular link between the coagulation and fibrinolytic systems. In this review, recent major advances in TAFI research, including the elucidation of crystal structures, the development of small inhibitors and the role of TAFI

  9. Oligonucleotide delivery with cell surface binding and cell penetrating Peptide amphiphile nanospheres.

    Science.gov (United States)

    Mumcuoglu, Didem; Sardan, Melis; Tekinay, Turgay; Guler, Mustafa O; Tekinay, Ayse B

    2015-05-01

    A drug delivery system designed specifically for oligonucleotide therapeutics can ameliorate the problems associated with the in vivo delivery of these molecules. The internalization of free oligonucleotides is challenging, and cytotoxicity is the main obstacle for current transfection vehicles. To develop nontoxic delivery vehicles for efficient transfection of oligonucleotides, we designed a self-assembling peptide amphiphile (PA) nanosphere delivery system decorated with cell penetrating peptides (CPPs) containing multiple arginine residues (R4 and R8), and a cell surface binding peptide (KRSR), and report the efficiency of this system in delivering G-3129, a Bcl-2 antisense oligonucleotide (AON). PA/AON (peptide amphiphile/antisense oligonucleotide) complexes were characterized with regards to their size and secondary structure, and their cellular internalization efficiencies were evaluated. The effect of the number of arginine residues on the cellular internalization was investigated by both flow cytometry and confocal imaging, and the results revealed that uptake efficiency improved as the number of arginines in the sequence increased. The combined effect of cell penetration and surface binding property on the cellular internalization and its uptake mechanism was also evaluated by mixing R8-PA and KRSR-PA. R8 and R8/KRSR decorated PAs were found to drastically increase the internalization of AONs compared to nonbioactive PA control. Overall, the KRSR-decorated self-assembled PA nanospheres were demonstrated to be noncytotoxic delivery vectors with high transfection rates and may serve as a promising delivery system for AONs. PMID:25828697

  10. Methods to Study the Role of the Glycocalyx in the Uptake of Cell-Penetrating Peptides

    NARCIS (Netherlands)

    Schmidt, S.; Wallbrecher, R.; Kuppevelt, T.H. van; Brock, R.E.

    2015-01-01

    Cells are covered by a layer of negatively charged oligo- and polysaccharides, the glycocalyx. Cell-penetrating peptides and other drug delivery vehicles first encounter these polyanions before contacting the lipid bilayer of the plasma membrane. While a large body of data supports the notion that i

  11. SKOV-3 cell imaging by paramagnetic particles labeled with hairpin cell-penetrating peptides

    Institute of Scientific and Technical Information of China (English)

    ZHAI Xiao-hui; LIU Min; GUO Xiao-juan; WANG Si-cen; ZHANG Hong-xia; GUO You-min

    2011-01-01

    Background The hairpin cell-penetrating peptides (hCPPs) demonstrate an interesting characteristic of conditioned activation by molecules. We hypothesized that hCPPs have the potential to selectively deliver a paramagnetic gadolinium probe into the matrix metalloproteinase 2 (MMP-2) positive human ovary adenocarcinoma cell lines,SKOV-3.Methods hCPPs were synthesized and labeled with 1,4,7,10-tetraazacyclododecane-N,N',N",N'"-tetraacetic acid gadolinium (Ⅲ) (Gd-DOTA) and fluorescein isothiocyanate (FITC) by f-moc strategy using a standard solid phase peptide synthesis protocol. MMP-2 expression and activity were demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Internalization and location of hCPPs in SKOV-3 cells were observed by fluorescein imaging and flow cytometery. Selective delivery of Gd-DOTA in SKOV-3 cells was observed by magnetic resonance imaging (MRI) and transmission electron microscopy (TEM).Results The uptake of hCPPs by SKOV-3 cells depended on the activity of MMP-2. T1WI signals of SKOV-3 cells treated with Gd-DOTA-hCPPs suggested the uptake of Gd-DOTA-hCPPs increased in a time- (r=0.990, P <0.01) and concentration-dependent manner (r=0.964, P <0.001), but was inhibited by a MMP-2 inhibitor. Electron-dense particles observed in the cytoplasm and nucleus by transmission electron microscopy proved the intracellular penetration of gadolinium.Conclusions hCPPs can be used as an effective vector for an MRI molecular probe to assess the activity of MMP-2.

  12. Cell-penetrating antimicrobial peptides - prospectives for targeting intracellular infections

    DEFF Research Database (Denmark)

    Bahnsen, Jesper S; Franzyk, Henrik; Sayers, Edward J;

    2015-01-01

    La WT cells and analyzed by flow cytometry and confocal microscopy. Furthermore, the effects of the peptides on eukaryotic cell viability as well as their antimicrobial effect were tested. In addition, the disrupting ability of the peptides in the presence of bilayer membranes of different composition...

  13. Liposome-based engineering of cells to package hydrophobic compounds in membrane vesicles for tumor penetration.

    Science.gov (United States)

    Lee, Junsung; Kim, Jiyoung; Jeong, Moonkyoung; Lee, Hyoungjin; Goh, Unbyeol; Kim, Hyaeyeong; Kim, Byungji; Park, Ji-Ho

    2015-05-13

    Natural membrane vesicles (MVs) derived from various types of cells play an essential role in transporting biological materials between cells. Here, we show that exogenous compounds are packaged in the MVs by engineering the parental cells via liposomes, and the MVs mediate autonomous intercellular migration of the compounds through multiple cancer cell layers. Hydrophobic compounds delivered selectively to the plasma membrane of cancer cells using synthetic membrane fusogenic liposomes were efficiently incorporated into the membrane of MVs secreted from the cells and then transferred to neighboring cells via the MVs. This liposome-mediated MV engineering strategy allowed hydrophobic photosensitizers to significantly penetrate both spheroids and in vivo tumors, thereby enhancing the therapeutic efficacy. These results suggest that innate biological transport systems can be in situ engineered via synthetic liposomes to guide the penetration of chemotherapeutics across challenging tissue barriers in solid tumors.

  14. Potential of activatable FAP-targeting immunoliposomes in intraoperative imaging of spontaneous metastases.

    Science.gov (United States)

    Tansi, Felista L; Rüger, Ronny; Böhm, Claudia; Kontermann, Roland E; Teichgraeber, Ulf K; Fahr, Alfred; Hilger, Ingrid

    2016-05-01

    Despite intensive research and medical advances met, metastatic disease remains the most common cause of death in cancer patients. This results from late diagnosis, poor therapeutic response and undetected micrometastases and tumor margins during surgery. One approach to overcome these challenges involves fluorescence imaging, which exploits the properties of fluorescent probes for diagnostic detection of molecular structures at the onset of transformation and for intraoperative detection of metastases and tumor margins in real time. Considering these benefits, many contrast agents suitable for fluorescence imaging have been reported. However, most reports only demonstrate the detection of primary tumors and not the detection of metastases or their application in models of image-guided surgery. In this work, we demonstrate the influence of fibroblast activation protein (FAP) on the metastatic potential of fibrosarcoma cells and elucidate the efficacy of activatable FAP-targeting immunoliposomes (FAP-IL) for image-guided detection of the spontaneous metastases in mice models. Furthermore, we characterized the biodistribution and cellular localization of the liposomal fluorescent components in mice organs and traced their excretion over time in urine and feces. Taken together, activatable FAP-IL enhances intraoperative imaging of metastases. Their high accumulation in metastases, subsequent localization in the bile canaliculi and liver kupffer cells and suitable excretion in feces substantiates their potency as contrast agents for intraoperative imaging.

  15. Effects of antiphagocytic agents on penetration of Eimeria magna sporozoites into cultured cells.

    Science.gov (United States)

    Jensen, J B; Edgar, S A

    1976-04-01

    Madin-Darby Bovine Kidney cells were treated with sodium flouride, iodoacetate, and 2-deosyglucose, reagents that block glycolysis, and thus reduce phagocytosis. Sporozoites readily entered cells whose ATP stores were largely depleted. They also entered cells treated with colchicine, colcemid, and vinblastine. These latter agents did not inhibit sporozite motility after 6 hr incubation. Cytochalasin B prevented penetration of cells by inhibiting the motility of sporozoites. This effect was reversible. Warm sporozoites entered cold cells 4 times more radily than cold sporozoites into warm cells. The above findings suggest that phagocytosis is not the mechanism for entry of E. magna sporozoites into cultured cells, but that sporozoite motility is of primary importance.

  16. Nanoformulated cell-penetrating survivin mutant and its dual actions

    Directory of Open Access Journals (Sweden)

    Sriramoju B

    2014-07-01

    Full Text Available Bhasker Sriramoju, Rupinder K Kanwar, Jagat R Kanwar Nanomedicine Laboratory of Immunology and Molecular Biomedical Research (NLIMBR, School of Medicine, Faculty of Health, Deakin University, Geelong, Australia Abstract: In this study, we investigated the differential actions of a dominant-negative survivin mutant (SurR9-C84A against cancerous SK-N-SH neuroblastoma cell lines and differentiated SK-N-SH neurons. In both the cases, the mutant protein displayed dual actions, where its effects were cytotoxic toward cancerous cells and proliferative toward the differentiated neurons. This can be explained by the fact that tumorous (undifferentiated SK-N-SH cells have a high endogenous survivin pool and upon treatment with mutant SuR9-C84A causes forceful survivin expression. These events significantly lowered the microtubule dynamics and stability, eventually leading to apoptosis. In the case of differentiated SK-N-SH neurons that express negligible levels of wild-type survivin, the mutant indistinguishably behaved in a wild-type fashion. It also favored cell-cycle progression, forming the chromosome-passenger complex, and stabilized the microtubule-organizing center. Therefore, mutant SurR9-C84A represents a novel therapeutic with its dual actions (cytotoxic toward tumor cells and protective and proliferative toward neuronal cells, and hence finds potential applications against a variety of neurological disorders. In this study, we also developed a novel poly(lactic-co-glycolic acid nanoparticulate formulation to surmount the hurdles associated with the delivery of SurR9-C84A, thus enhancing its effective therapeutic outcome. Keywords: survivin mutant, neurological disorders, protein therapeutics, inhibitor of apoptosis protein family, poly(lactic-co-glycolic acid

  17. Cell-penetrating peptides for drug delivery across membrane barriers

    DEFF Research Database (Denmark)

    Foged, Camilla; Nielsen, Hanne Moerck

    2008-01-01

    , proteins and colloidal carriers such as liposomes and polymeric nanoparticles. Their ability to cross biological membranes in a non-disruptive way without apparent toxicity is highly desired for increasing drug bioavailability. This review provides an overview of the application of cell...

  18. Binding and Clustering of Glycosaminoglycans: A Common Property of Mono- and Multivalent Cell-Penetrating Compounds

    OpenAIRE

    Ziegler, André; Seelig, Joachim

    2007-01-01

    Recent observations in cell culture provide evidence that negatively charged glycosaminoglycans (GAGs) at the surface of biological cells bind cationic cell-penetrating compounds (CPCs) and cluster during CPC binding, thereby contributing to their endocytotic uptake. The GAG binding and clustering occur in the low-micromolar concentration range and suggest a tight interaction between GAGs and CPCs, although the relation between binding affinity and specificity of this interaction remains to b...

  19. Application of Cell Penetrating Peptide in Magnetic Resonance Imaging of Bone Marrow Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Min LIU; You-Min GUO; Jun-Le YANG; Peng WANG; Lin-Yu ZHAO; Nian SHEN; Si-Cen WANG; Xiao-Juan GUO; Qi-Fei WU

    2006-01-01

    Tracking the distribution and differentiation of stem cells by high-resolution imaging techniques would have significant clinical and research implications. In this study, a model cell-penetrating peptide was used to carry gadolinium particles for magnetic resonance imaging (MRI) of mesenchymal stem cells (MSCs).MSCs were isolated from rat bone marrow and identified by osteogenic differentiation in vitro. The cellpenetrating peptide labeled with fluorescein-5-isothiocyanate (FITC) and gadolinium was synthesized by a solid-phase peptide synthesis method. Fluorescein imaging analysis confirmed that this new peptide could internalize into the cytoplasm and nucleus at room temperature, 4℃ and 37℃. Gadolinium were efficiently internalized into mesenchymal stem cells by the peptide in a time or concentration-dependent manner,resulting in intercellular shortening of longitudinal relaxation enhancements, which were obviously detected by 1.5 Tesla Magnetic Resonance Imaging. Cytotoxicity assay and flow cytometric analysis showed that the intercellular contrast medium incorporation did not affect cell viability at the tested concentrations. The in vitro experiment results suggested that the new constructed peptides could be a vector for tracking MSCs.

  20. Polymeric pH nanosensor with extended measurement range bearing octaarginine as cell penetrating peptide

    DEFF Research Database (Denmark)

    Ke, Peng; Sun, Honghao; Liu, Mingxing;

    2016-01-01

    A synthetic peptide octaarginine which mimics human immunodeficiency virus-1, Tat protein is used as cell penetrating moiety for new pH nanosensors which demonstrate enhanced cellular uptake and expanded measurement range from pH 3.9 to pH 7.3 by simultaneously incorporating two complemental pH-s...

  1. Cationic cell-penetrating peptides induce ceramide formation via acid sphingomyelinase: implications for uptake.

    NARCIS (Netherlands)

    Verdurmen, W.P.R.; Thanos, M.; Ruttekolk, I.R.R.; Gulbins, E.; Brock, R.E.

    2010-01-01

    Cationic cell-penetrating peptides (CPP) are receiving increasing attention as molecular transporters of membrane-impermeable molecules. Import of cationic CPP occurs both via endocytosis and - at higher peptide concentrations - in an endocytosis-independent manner via localized regions of the plasm

  2. A cell-penetrating peptide derived from human lactoferrin with conformation-dependent uptake efficiency.

    NARCIS (Netherlands)

    Duchardt, F.; Ruttekolk, I.R.R.; Verdurmen, W.P.R.; Lortat-Jacob, H.; Burck, J.; Hufnagel, H.; Fischer, R.; Heuvel, M. van den; Lowik, D.W.; Vuister, G.W.; Ulrich, A.; Waard, M. de; Brock, R.E.

    2009-01-01

    The molecular events that contribute to the cellular uptake of cell-penetrating peptides (CPP) are still a matter of intense research. Here, we report on the identification and characterization of a 22-amino acid CPP derived from the human milk protein, lactoferrin. The peptide exhibits a conformati

  3. Efficient Cargo Delivery into Adult Brain Tissue Using Short Cell-Penetrating Peptides.

    Directory of Open Access Journals (Sweden)

    Caghan Kizil

    Full Text Available Zebrafish brains can regenerate lost neurons upon neurogenic activity of the radial glial progenitor cells (RGCs that reside at the ventricular region. Understanding the molecular events underlying this ability is of great interest for translational studies of regenerative medicine. Therefore, functional analyses of gene function in RGCs and neurons are essential. Using cerebroventricular microinjection (CVMI, RGCs can be targeted efficiently but the penetration capacity of the injected molecules reduces dramatically in deeper parts of the brain tissue, such as the parenchymal regions that contain the neurons. In this report, we tested the penetration efficiency of five known cell-penetrating peptides (CPPs and identified two- polyR and Trans - that efficiently penetrate the brain tissue without overt toxicity in a dose-dependent manner as determined by TUNEL staining and L-Plastin immunohistochemistry. We also found that polyR peptide can help carry plasmid DNA several cell diameters into the brain tissue after a series of coupling reactions using DBCO-PEG4-maleimide-based Michael's addition and azide-mediated copper-free click reaction. Combined with the advantages of CVMI, such as rapidness, reproducibility, and ability to be used in adult animals, CPPs improve the applicability of the CVMI technique to deeper parts of the central nervous system tissues.

  4. A cell penetrating peptide-integrated and enediyne-energized fusion protein shows potent antitumor activity.

    Science.gov (United States)

    Ru, Qin; Shang, Bo-Yang; Miao, Qing-Fang; Li, Liang; Wu, Shu-Ying; Gao, Rui-Juan; Zhen, Yong-Su

    2012-11-20

    Arginine-rich peptides belong to a subclass of cell penetrating peptides that are taken up by living cells and can be detected freely diffusing inside the cytoplasm and nucleoplasm. This phenomenon has been attributed to either an endocytotic mode of uptake and a subsequent release from vesicles or a direct membrane penetration. Lidamycin is an antitumor antibiotic, which consists of an active enediyne chromophore (AE) and a noncovalently bound apoprotein (LDP). In the present study, a fusion protein (Arg)(9)-LDP composed of cell penetrating peptide (Arg)(9) and LDP was prepared by DNA recombination, and the enediyne-energized fusion protein (Arg)(9)-LDP-AE was prepared by molecular reconstitution. The data in fixed cells demonstrated that (Arg)(9)-LDP could rapidly enter cells, and the results based on fluorescence activated cell sorting indicated that the major route for (Arg)(9)-mediated cellular uptake of protein molecules was endocytosis. (Arg)(9)-LDP-AE demonstrated more potent cytotoxicity against different carcinoma cell lines than lidamycin in vitro. In the mouse hepatoma 22 model, (Arg)(9)-LDP-AE (0.3mg/kg) suppressed the tumor growth by 89.2%, whereas lidamycin (0.05 mg/kg) by 74.6%. Furthermore, in the glioma U87 xenograft model in nude mice, (Arg)(9)-LDP-AE at 0.2mg/kg suppressed tumor growth by 88.8%, compared with that of lidamycin by 62.9% at 0.05 mg/kg. No obvious toxic effects were observed in all groups during treatments. The results showed that energized fusion protein (Arg)(9)-LDP-AE was more effective than lidamycin and would be a promising candidate for glioma therapy. In addition, this approach to manufacturing fusion proteins might serve as a technology platform for the development of new cell penetrating peptides-based drugs. PMID:22982402

  5. Alternative Mechanisms for the Interaction of the Cell-Penetrating Peptides Penetratin and the TAT Peptide with Lipid Bilayers

    NARCIS (Netherlands)

    Yesylevskyy, Semen; Marrink, Siewert-Jan; Mark, Alan E.

    2009-01-01

    Cell-penetrating peptides (CPPs) have recently attracted much interest due to their apparent ability to penetrate cell membranes in an energy-independent manner. Here molecular-dynamics simulation techniques were used to study the interaction of two CPPs: penetratin and the TAT peptide with 1,2-Dipa

  6. Topical Delivery of Protein and Peptide Using Novel Cell Penetrating Peptide IMT-P8

    Science.gov (United States)

    Gautam, Ankur; Nanda, Jagpreet Singh; Samuel, Jesse S.; Kumari, Manisha; Priyanka, Priyanka; Bedi, Gursimran; Nath, Samir K.; Mittal, Garima; Khatri, Neeraj; Raghava, Gajendra Pal Singh

    2016-01-01

    Skin, being the largest organ of the body, is an important site for drug administration. However, most of the drugs have poor permeability and thus drug delivery through the skin is very challenging. In this study, we examined the transdermal delivery capability of IMT-P8, a novel cell-penetrating peptide. We generated IMT-P8-GFP and IMT-P8-KLA fusion constructs and evaluated their internalization into mouse skin after topical application. Our results demonstrate that IMT-P8 is capable of transporting green fluorescent protein (GFP) and proapoptotic peptide, KLA into the skin and also in different cell lines. Interestingly, uptake of IMT-P8-GFP was considerably higher than TAT-GFP in HeLa cells. After internalization, IMT-P8-KLA got localized to the mitochondria and caused significant cell death in HeLa cells signifying an intact biological activity. Further in vivo skin penetration experiments revealed that after topical application, IMT-P8 penetrated the stratum corneum, entered into the viable epidermis and accumulated inside the hair follicles. In addition, both IMT-P8-KLA and IMT-P8-GFP internalized into the hair follicles and dermal tissue of the skin following topical application. These results suggested that IMT-P8 could be a potential candidate to be used as a topical delivery vehicle for various cosmetic and skin disease applications. PMID:27189051

  7. Conformational analysis of Infectious bursal disease virus (IBDV derived cell penetrating peptide (CPP analogs

    Directory of Open Access Journals (Sweden)

    Vinay G. Joshi

    2013-12-01

    Full Text Available Aim: This study was designed to develop peptide analogs of Infectious Bursal Disease (IBD virus VP5 protein segment having cell penetrating ability to improve their interaction with cargo molecule (Nucleic acid without affecting the backbone conformation. Materials and Methods: IBDV VP5 protein segment designated as RATH peptide were synthesized using solid phase peptide synthesis and their solution conformation was elucidated using CD spectroscopy in polar (water and apolar (TFE solvents. Cell penetrating ability of RATH-CONH2 was observed using FITC labeled peptide internalization in to HeLa cells under fluorescent microscopy. The efficacy of RATH analog interactions with nucleic acids was evaluated using FITC labeled oligonucleotides by fluorescence spectroscopy and plasmid constructs in gel retardation assay. Results: CD spectra of RATH analogs in water and apolar trifluroethanol (TFE helped to compare their secondary structures which were almost similar with dominant beta conformations suggesting successful induction of positive charge in the analogs without affecting back bone conformation of CPP designed. Cell penetrating ability of RATH CONH2 in HeLa cell was more than 90%. The fluorescence spectroscopy and plasmid constructs in gel retardation assay demonstrated successful interaction of amide analogs with nucleic acid. Conclusion: Intentional changes made in IBDV derived peptide RATH COOH to RATH CONH2 did not showed major changes in backbone conformation and such modifications may help to improve the cationic charge in most CPPs to interact with nucleic acid. [Vet World 2013; 6(6.000: 307-312

  8. Topical Delivery of Protein and Peptide Using Novel Cell Penetrating Peptide IMT-P8.

    Science.gov (United States)

    Gautam, Ankur; Nanda, Jagpreet Singh; Samuel, Jesse S; Kumari, Manisha; Priyanka, Priyanka; Bedi, Gursimran; Nath, Samir K; Mittal, Garima; Khatri, Neeraj; Raghava, Gajendra Pal Singh

    2016-01-01

    Skin, being the largest organ of the body, is an important site for drug administration. However, most of the drugs have poor permeability and thus drug delivery through the skin is very challenging. In this study, we examined the transdermal delivery capability of IMT-P8, a novel cell-penetrating peptide. We generated IMT-P8-GFP and IMT-P8-KLA fusion constructs and evaluated their internalization into mouse skin after topical application. Our results demonstrate that IMT-P8 is capable of transporting green fluorescent protein (GFP) and proapoptotic peptide, KLA into the skin and also in different cell lines. Interestingly, uptake of IMT-P8-GFP was considerably higher than TAT-GFP in HeLa cells. After internalization, IMT-P8-KLA got localized to the mitochondria and caused significant cell death in HeLa cells signifying an intact biological activity. Further in vivo skin penetration experiments revealed that after topical application, IMT-P8 penetrated the stratum corneum, entered into the viable epidermis and accumulated inside the hair follicles. In addition, both IMT-P8-KLA and IMT-P8-GFP internalized into the hair follicles and dermal tissue of the skin following topical application. These results suggested that IMT-P8 could be a potential candidate to be used as a topical delivery vehicle for various cosmetic and skin disease applications. PMID:27189051

  9. Dimerization of a cell-penetrating peptide leads to enhanced cellular uptake and drug delivery

    Directory of Open Access Journals (Sweden)

    Jan Hoyer

    2012-10-01

    Full Text Available Over the past 20 years, cell-penetrating peptides (CPPs have gained tremendous interest due to their ability to deliver a variety of therapeutically active molecules that would otherwise be unable to cross the cellular membrane due to their size or hydrophilicity. Recently, we reported on the identification of a novel CPP, sC18, which is derived from the C-terminus of the 18 kDa cationic antimicrobial protein. Furthermore, we demonstrated successful application of sC18 for the delivery of functionalized cyclopentadienyl manganese tricarbonyl (cymantrene complexes to tumor cell lines, inducing high cellular toxicity. In order to increase the potential of the organometallic complexes to kill tumor cells, we were looking for a way to enhance cellular uptake. Therefore, we designed a branched dimeric variant of sC18, (sC182, which was shown to have a dramatically improved capacity to internalize into various cell lines, even primary cells, using flow cytometry and fluorescence microscopy. Cell viability assays indicated increased cytotoxicity of the dimer presumably caused by membrane leakage; however, this effect turned out to be dependent on the specific cell type. Finally, we could show that conjugation of a functionalized cymantrene with (sC182 leads to significant reduction of its IC50 value in tumor cells compared to the respective sC18 conjugate, proving that dimerization is a useful method to increase the drug-delivery potential of a cell-penetrating peptide.

  10. The bacteriophage ϕ29 tail possesses a pore-forming loop for cell membrane penetration.

    Science.gov (United States)

    Xu, Jingwei; Gui, Miao; Wang, Dianhong; Xiang, Ye

    2016-06-23

    Most bacteriophages are tailed bacteriophages with an isometric or a prolate head attached to a long contractile, long non-contractile, or short non-contractile tail. The tail is a complex machine that plays a central role in host cell recognition and attachment, cell wall and membrane penetration, and viral genome ejection. The mechanisms involved in the penetration of the inner host cell membrane by bacteriophage tails are not well understood. Here we describe structural and functional studies of the bacteriophage ϕ29 tail knob protein gene product 9 (gp9). The 2.0 Å crystal structure of gp9 shows that six gp9 molecules form a hexameric tube structure with six flexible hydrophobic loops blocking one end of the tube before DNA ejection. Sequence and structural analyses suggest that the loops in the tube could be membrane active. Further biochemical assays and electron microscopy structural analyses show that the six hydrophobic loops in the tube exit upon DNA ejection and form a channel that spans the lipid bilayer of the membrane and allows the release of the bacteriophage genomic DNA, suggesting that cell membrane penetration involves a pore-forming mechanism similar to that of certain non-enveloped eukaryotic viruses. A search of other phage tail proteins identified similar hydrophobic loops, which indicates that a common mechanism might be used for membrane penetration by prokaryotic viruses. These findings suggest that although prokaryotic and eukaryotic viruses use apparently very different mechanisms for infection, they have evolved similar mechanisms for breaching the cell membrane. PMID:27309813

  11. Cell-Penetrating Peptides—Mechanisms of Cellular Uptake and Generation of Delivery Systems

    Directory of Open Access Journals (Sweden)

    Sara Trabulo

    2010-03-01

    Full Text Available The successful clinical application of nucleic acid-based therapeutic strategies has been limited by the poor delivery efficiency achieved by existing vectors. The development of alternative delivery systems for improved biological activity is, therefore, mandatory. Since the seminal observations two decades ago that the Tat protein, and derived peptides, can translocate across biological membranes, cell-penetrating peptides (CPPs have been considered one of the most promising tools to improve non-invasive cellular delivery of therapeutic molecules. Despite extensive research on the use of CPPs for this purpose, the exact mechanisms underlying their cellular uptake and that of peptide conjugates remain controversial. Over the last years, our research group has been focused on the S413-PV cell-penetrating peptide, a prototype of this class of peptides that results from the combination of 13-amino-acid cell penetrating sequence derived from the Dermaseptin S4 peptide with the SV40 large T antigen nuclear localization signal. By performing an extensive biophysical and biochemical characterization of this peptide and its analogs, we have gained important insights into the mechanisms governing the interaction of CPPs with cells and their translocation across biological membranes. More recently, we have started to explore this peptide for the intracellular delivery of nucleic acids (plasmid DNA, siRNA and oligonucleotides. In this review we discuss the current knowledge of the mechanisms responsible for the cellular uptake of cell-penetrating peptides, including the S413-PV peptide, and the potential of peptide-based formulations to mediate nucleic acid delivery.

  12. Paramagnetic particles carried by cell-penetrating peptide tracking of bone marrow mesenchymal stem cells, a research in vitro

    International Nuclear Information System (INIS)

    The ability to track the distribution and differentiation of stem cells by high-resolution imaging techniques would have significant clinical and research implications. In this study, a model cell-penetrating peptide was used to carry gadolinium particles for magnetic resonance imaging of the mesenchymal stem cells. The mesenchymal stem cells were isolated from rat bone marrow by Percoll and identified by osteogenic differentiation in vitro. The cell-penetrating peptides labeled with fluorescein-5-isothiocyanate and gadolinium were synthesized by a solid-phase peptide synthesis method and the relaxivity of cell-penetrating peptide-gadolinium paramagnetic conjugate on 400 MHz nuclear magnetic resonance was 5.7311 ± 0.0122 mmol-1 s-1, higher than that of diethylenetriamine pentaacetic acid gadolinium (p < 0.05). Fluorescein imaging confirmed that this new peptide could internalize into the cytoplasm and nucleus. Gadolinium was efficiently internalized into mesenchymal stem cells by the peptide in a time- or concentration-dependent fashion, resulting in intercellular T1 relaxation enhancement, which was obviously detected by 1.5 T magnetic resonance imaging. Cytotoxicity assay and flow cytometric analysis showed the intercellular contrast medium incorporation did not affect cell viability and membrane potential gradient. The research in vitro suggests that the newly constructed peptides could be a vector for tracking mesenchymal stem cells

  13. Cell-penetrating peptides; chemical modification, mechanism of uptake and formulation development

    OpenAIRE

    Ezzat, Kariem

    2012-01-01

    Gene therapy holds the promise of revolutionizing the way we treat diseases. By using recombinant DNA and oligonucleotides (ONs), gene functions can be restored, altered or silenced according to the therapeutic need. However, gene therapy approaches require the delivery of large and charged nucleic acid-based molecules to their intracellular targets across the plasma membrane, which is inherently impermeable to such molecules. In this thesis, two chemically modified cell-penetrating peptides ...

  14. Peptide Internalization Enabled by Folding: Triple Helical Cell-Penetrating Peptides

    OpenAIRE

    Shinde, Aparna; Feher, Katie M.; Hu, Chloe; Slowinska, Katarzyna

    2014-01-01

    Cell-Penetrating Peptides (CPPs) are known as efficient transporters of molecular cargo across cellular membranes. Their properties make them ideal candidates for in vivo applications. However, challenges in development of effective CPPs still exist: CPPs are often fast degraded by proteases and large concentration of CPPs required for cargo transporting can cause cytotoxicity. It was previously shown that restricting peptide flexibility can improve peptide stability against enzymatic degrada...

  15. Cell-Penetrating Ability of Peptide Hormones: Key Role of Glycosaminoglycans Clustering

    Directory of Open Access Journals (Sweden)

    Armelle Tchoumi Neree

    2015-11-01

    Full Text Available Over the last two decades, the potential usage of cell-penetrating peptides (CPPs for the intracellular delivery of various molecules has prompted the identification of novel peptidic identities. However, cytotoxic effects and unpredicted immunological responses have often limited the use of various CPP sequences in the clinic. To overcome these issues, the usage of endogenous peptides appears as an appropriate alternative approach. The hormone pituitary adenylate-cyclase-activating polypeptide (PACAP38 has been recently identified as a novel and very efficient CPP. This 38-residue polycationic peptide is a member of the secretin/glucagon/growth hormone-releasing hormone (GHRH superfamily, with which PACAP38 shares high structural and conformational homologies. In this study, we evaluated the cell-penetrating ability of cationic peptide hormones in the context of the expression of cell surface glycosaminoglycans (GAGs. Our results indicated that among all peptides evaluated, PACAP38 was unique for its potent efficiency of cellular uptake. Interestingly, the abilities of the peptides to reach the intracellular space did not correlate with their binding affinities to sulfated GAGs, but rather to their capacity to clustered heparin in vitro. This study demonstrates that the uptake efficiency of a given cationic CPP does not necessarily correlate with its affinity to sulfated GAGs and that its ability to cluster GAGs should be considered for the identification of novel peptidic sequences with potent cellular penetrating properties.

  16. Parametric analysis of neutron streaming through major penetrations in the 0.914 m TFTR test cell floor

    International Nuclear Information System (INIS)

    Neutron streaming through penetrations in the 0.914 m TFTR test cell floor has two distinct features: (1) the oblique angle of incidence; and (2) the high order of anisotropy in the angular distribution for incident neutrons with energies > 10 keV. The effects of these features on the neutron streaming into the TFTR basement were studied parametrically for isolated penetrations. Variations with respect to the source energies, angular distributions, and sizes of the penetrations were made. The results form a data base from which the spatial distribution of the neutron flux in the basement due to multiple penetrations may be evaluated

  17. Penetration of fosfomycin into IPEC-J2 cells in the presence or absence of deoxynivalenol.

    Directory of Open Access Journals (Sweden)

    Guadalupe Martínez

    Full Text Available Fosfomycin (FOS is an antibiotic used in pig farms for treatment and prevention of infections caused by resistant bacteria during the post-weaning period. Antibiotics and non-toxic concentrations of mycotoxins, such as deoxynivalenol (DON are frequently found in the diet of animals. These compounds can establish interactions in the intestinal tract, which could affect and/or modify the penetration of FOS to enterocytes. The aim of this study was to determine the penetration of FOS into IPECJ-2 cells, a cell line derived from the small intestine of piglets, in the presence and absence of DON. The results from this study showed that there was statistically significant difference in the intracellular concentration of FOS between cells incubated with 580 µg/ml FOS and cells incubated with 580 µg/ml FOS and 1 µg/ml DON. The Cmax of the intracellular antibiotic in the culture plates incubated with FOS in absence of DON was 45.81 µg/ml with a tmax of 4 h. When IPEC-2 cells were incubated with FOS and DON the Cmax was 20.06 µg/ml and the tmax was 30 min. It is concluded that the non-toxic concentration of DON on IPEC-J2 cells after short-term exposure, interferes with the pharmacokinetics of the antibiotic.

  18. Potent inhibition of late stages of hepadnavirus replication by a modified cell penetrating peptide

    DEFF Research Database (Denmark)

    Abdul, Fabien; Ndeboko, Bénédicte; Buronfosse, Thierry;

    2012-01-01

    of Duck Hepatitis B Virus (DHBV), a reference model for human HBV. Amongst twelve CatLip peptides we identified Deca-(Arg)8 having a particularly potent antiviral activity, leading to a drastic inhibition of viral particle secretion without detectable toxicity. Inhibition of virion secretion......Cationic cell-penetrating peptides (CPPs) and their lipid domain-conjugates (CatLip) are agents for the delivery of (uncharged) biologically active molecules into the cell. Using infection and transfection assays we surprisingly discovered that CatLip peptides were able to inhibit replication...

  19. Cell penetrating peptide-based polyplexes shelled with polysaccharide to improve stability and gene transfection

    Science.gov (United States)

    Li, Wenyu; Liu, Yajie; Du, Jianwei; Ren, Kefeng; Wang, Youxiang

    2015-04-01

    Cell-penetrating peptides (CPP) have been widely developed as a strategy to enhance cell penetrating ability and transfection. In this work, octa-arginine modified dextran gene vector with pH-sensitivity was developed via host-guest interactions. α-Cyclodextrin was modified with octa-arginine (CDR), which had excellent cell penetrating ability. Dextran was selected as a backbone and modified with azobenzene as guest units by acid-labile imine bonds (Az-I-Dex). The supramolecular polymer CDR/Az-I-Dex with high a C/A molar ratio (molar ratio of CD on CDR to Az on Az-I-Dex) was unfavorable for DNA condensation. The dextran shell of CDR/Az-I-Dex/DNA polyplexes improved the stability under physiological conditions. However, once treated with acetate buffer (pH 5.4) for 3 h, large aggregates formed rapidly due to the cleavage of the dextran shell. As expected, the vector had cell viability of 80% even when the CDR concentration increased to 100 μg mL-1. Moreover, due to the effective cellular uptake efficiency, CDR/Az-I-Dex/DNA polyplexes had 6-300 times higher transfection efficiency than CDR/DNA polyplexes. It was even higher than high molecular weight PLL-based polyplexes of HEK293 T cells. Importantly, chloroquine as an endosomal escape agent could not improve the transfection of CDR/Az-I-Dex/DNA polyplexes, which indicated that the CDR/Az-I-Dex supramolecular polymer had its own ability for endosomal escape. These results suggested that the CPP-based polyplexes shelled with polysaccharide can be promising non-viral gene delivery carriers.Cell-penetrating peptides (CPP) have been widely developed as a strategy to enhance cell penetrating ability and transfection. In this work, octa-arginine modified dextran gene vector with pH-sensitivity was developed via host-guest interactions. α-Cyclodextrin was modified with octa-arginine (CDR), which had excellent cell penetrating ability. Dextran was selected as a backbone and modified with azobenzene as guest units by acid

  20. Dimerization of 30Kc19 protein in the presence of amphiphilic moiety and importance of Cys-57 during cell penetration.

    Science.gov (United States)

    Park, Hee Ho; Sohn, Youngsoo; Yeo, Ji Woo; Park, Ju Hyun; Lee, Hong Jai; Ryu, Jina; Rhee, Won Jong; Park, Tai Hyun

    2014-12-01

    Recently, the recombinant 30Kc19 protein, originating from silkworm hemolymph of Bombyx mori has attracted attention due to its cell-penetrating property and potential application as a protein delivery system. However, this observation of penetration across cell membrane has raised questions concerning the interaction of the protein-lipid bilayer. Here, we report a dimerization propensity of the 30Kc19 protein in the presence of amphiphilic moieties; sodium dodecyl sulfate (SDS) or phospholipid. Native PAGE showed that the 30Kc19 monomer formed a dimer when SDS or phospholipid was present. In the glutathione-S-transferase (GST) pull-down assay, supplementation of the 30Kc19 protein to mammalian cell culture medium showed dimerization and penetration; due to phospholipids at the cell membrane, the main components of the lipid bilayer. Mutagenesis was performed, and penetration was observed by 30Kc19 C76A and not 30Kc19 C57A, which meant that the presence of cysteine at position 57 (Cys-57) is involved in dimerization of the 30Kc19 at the cell membrane during penetration. We anticipate application of the native 30Kc19 protein with high cell-penetrating efficiency for delivery of cargos to various cell types. The intracellular cargo delivery using the 30Kc19 protein is a non-virus derived (e.g. TAT) delivery method, which can open up new approaches for the delivery of therapeutics in bioindustries, such as pharma- and cosmeceuticals.

  1. Single wall carbon nanotubes enter cells by endocytosis and not membrane penetration

    Directory of Open Access Journals (Sweden)

    Lösche Mathias

    2011-09-01

    Full Text Available Abstract Background Carbon nanotubes are increasingly being tested for use in cellular applications. Determining the mode of entry is essential to control and regulate specific interactions with cells, to understand toxicological effects of nanotubes, and to develop nanotube-based cellular technologies. We investigated cellular uptake of Pluronic copolymer-stabilized, purified ~145 nm long single wall carbon nanotubes (SWCNTs through a series of complementary cellular, cell-mimetic, and in vitro model membrane experiments. Results SWCNTs localized within fluorescently labeled endosomes, and confocal Raman spectroscopy showed a dramatic reduction in SWCNT uptake into cells at 4°C compared with 37°C. These data suggest energy-dependent endocytosis, as shown previously. We also examined the possibility for non-specific physical penetration of SWCNTs through the plasma membrane. Electrochemical impedance spectroscopy and Langmuir monolayer film balance measurements showed that Pluronic-stabilized SWCNTs associated with membranes but did not possess sufficient insertion energy to penetrate through the membrane. SWCNTs associated with vesicles made from plasma membranes but did not rupture the vesicles. Conclusions These measurements, combined, demonstrate that Pluronic-stabilized SWCNTs only enter cells via energy-dependent endocytosis, and association of SWCNTs to membrane likely increases uptake.

  2. Distinct behaviour of the homeodomain derived cell penetrating peptide penetratin in interaction with different phospholipids.

    Directory of Open Access Journals (Sweden)

    Ofelia Maniti

    Full Text Available BACKGROUND: Penetratin is a protein transduction domain derived from the homeoprotein Antennapedia. Thereby it is currently used as a cell penetrating peptide to introduce diverse molecules into eukaryotic cells, and it could also be involved in the cellular export of transcription factors. Moreover, it has been shown that it is able to act as an antimicrobial agent. The mechanisms involved in all these processes are quite controversial. METHODOLOGY/PRINCIPAL FINDINGS: In this article, we report spectroscopic, calorimetric and biochemical data on the penetratin interaction with three different phospholipids: phosphatidylcholine (PC and phosphatidylethanolamine (PE to mimic respectively the outer and the inner leaflets of the eukaryotic plasma membrane and phosphatidylglycerol (PG to mimic the bacterial membrane. We demonstrate that with PC, penetratin is able to form vesicle aggregates with no major change in membrane fluidity and presents no well defined secondary structure organization. With PE, penetratin aggregates vesicles, increases membrane rigidity and acquires an α-helical structure. With PG membranes, penetratin does not aggregate vesicles but decreases membrane fluidity and acquires a structure with both α-helical and β-sheet contributions. CONCLUSIONS/SIGNIFICANCE: These data from membrane models suggest that the different penetratin actions in eukaryotic cells (membrane translocation during export and import and on prokaryotes may result from different peptide and lipid structural arrangements. The data suggest that, for eukaryotic cell penetration, penetratin does not acquire classical secondary structure but requires a different conformation compared to that in solution.

  3. Cationic cell-penetrating peptides as vehicles for siRNA delivery.

    Science.gov (United States)

    Beloor, Jagadish; Zeller, Skye; Choi, Chang Seon; Lee, Sang-Kyung; Kumar, Priti

    2015-01-01

    RNA interference mediated gene silencing has tremendous applicability in fields ranging from basic biological research to clinical therapy. However, delivery of siRNA across the cell membrane into the cytoplasm, where the RNA silencing machinery is located, is a significant hurdle in most primary cells. Cell-penetrating peptides (CPPs), peptides that possess an intrinsic ability to translocate across cell membranes, have been explored as a means to achieve cellular delivery of siRNA. Approaches using CPPs by themselves or through incorporation into other siRNA delivery platforms have been investigated with the intent of improving cytoplasmic delivery. Here, we review the utilization of CPPs for siRNA delivery with a focus on strategies developed to enhance cellular uptake, endosomal escape and cytoplasmic localization of CPP/siRNA complexes.

  4. Relationships between Cargo, Cell Penetrating Peptides and Cell Type for Uptake of Non-Covalent Complexes into Live Cells

    Directory of Open Access Journals (Sweden)

    Andrea-Anneliese Keller

    2013-02-01

    Full Text Available Modulating signaling pathways for research and therapy requires either suppression or expression of selected genes or internalization of proteins such as enzymes, antibodies, nucleotide binding proteins or substrates including nucleoside phosphates and enzyme inhibitors. Peptides, proteins and nucleotides are transported by fusing or conjugating them to cell penetrating peptides or by formation of non-covalent complexes. The latter is often preferred because of easy handling, uptake efficiency and auto-release of cargo into the live cell. In our studies complexes are formed with labeled or readily detectable cargoes for qualitative and quantitative estimation of their internalization. Properties and behavior of adhesion and suspension vertebrate cells as well as the protozoa Leishmania tarentolae are investigated with respect to proteolytic activity, uptake efficiency, intracellular localization and cytotoxicity. Our results show that peptide stability to membrane-bound, secreted or intracellular proteases varies between different CPPs and that the suitability of individual CPPs for a particular cargo in complex formation by non-covalent interactions requires detailed studies. Cells vary in their sensitivity to increasing concentrations of CPPs. Thus, most cells can be efficiently transduced with peptides, proteins and nucleotides with intracellular concentrations in the low micromole range. For each cargo, cell type and CPP the optimal conditions must be determined separately.

  5. Poly(NIPAm-AMPS) nanoparticles for targeted delivery of anti-inflammatory cell penetrating peptides

    Science.gov (United States)

    Bartlett, Rush Lloyd, II

    Inflammatory diseases such as osteoarthritis and rheumatoid arthritis cause $127.8 billion in US healthcare expenditures each year and are the cause of disability for 27% of disabled persons in the United States. Current treatment options rarely halt disease progression and often result in significant unwanted and debilitating side effects. Our laboratory has previously developed a family of cell penetrating peptides (CPPs) which inhibit the activity of mitogen activated protein kinase activate protein kinase 2 (MK2). MK2 mediates the inflammatory response by activating Tristetraprline (TTP). Once activated, TTP rapidly stabilizes AU rich regions of pro-inflammatory cytokine mRNA which allows translation of pro-inflammatory cytokines to occur. Blocking MK2 with our labs CPPs yields a decrease in inflammatory activity but CPPs by are highly non specific and prone to rapid enzymatic degradation in vivo.. In order to increase the potency of MK2 inhibiting CPPs we have developed a novel nanoparticle drug carrier composed of poly(N-isopropylacrylamide-co-2-acrylamido-2-methyl-1-propanesulfonic acid). This drug carrier has been shown to have preliminary efficacy in vitro and ex vivo for suppressing pro-inflammatory cytokine production when releasing CPPs. This thesis will present progress made on three aims: Specific Aim 1) Create and validate a NIPAm based drug delivery system that mimics the binding and release previously observed between cell penetrating peptides and glycosaminoglycans. Specific Aim 2) Engineer degradability into poly(NIPAm-AMPS) nanoparticles to enable more drug to be released and qualify that system in vitro. Specific Aim 3) Validate poly(NIPAm-AMPS) nanoparticles for targeted drug delivery in an ex vivo inflammatory model. Overall we have developed a novel anionic nanoparticle system that is biocompatible and efficient at loading and releasing cell penetrating peptides to inflamed tissue. Once loaded with a CPP the nanoparticle drug complex is

  6. Cell-Penetrating Peptides: A Comparative Study on Lipid Affinity and Cargo Delivery Properties

    Directory of Open Access Journals (Sweden)

    Paolo Ruzza

    2010-03-01

    Full Text Available A growing number of natural and/or synthetic peptides with cell membrane penetrating capability have been identified and described in the past years. These molecules have been considered promising tools for delivering bioactive compounds into various cell types. Although the mechanism of uptake is still unclear, it is reasonable to assume that the relative contribute of each proposed mechanism could differ for the same peptide, depending on experimental protocol and cargo molecule composition. In this work we try to connect the capability to interact with model lipid membrane and structural and chemical characteristics of CPPs in order to obtain a biophysical classification that predicts the behavior of CPP-cargo molecules in cell systems. Indeed, the binding with cell membrane is one of the primary step in the interaction of CPPs with cells, and consequently the studies on model membrane could become important for understanding peptide-membrane interaction on a molecular level, explaining how CPPs may translocate a membrane without destroying it and how this interactions come into play in shuttling CPPs via different routes with different efficiency. We analyzed by CD and fluorescence spectroscopies the binding properties of six different CPPs (kFGF, Nle54-Antp and Tat derived peptides, and oligoarginine peptides containing 6, 8 or 10 residues in absence or presence of the same cargo peptide (the 392-401pTyr396 fragment of HS1 protein. The phospholipid binding properties were correlated to the conformational and chemical characteristics of peptides, as well as to the cell penetrating properties of the CPP-cargo conjugates. Results show that even if certain physico-chemical properties (conformation, positive charge govern CPP capability to interact with the model membrane, these cannot fully explain cell-permeability properties.

  7. Therapeutic Potential of Cell Penetrating Peptides (CPPs) and Cationic Polymers for Chronic Hepatitis B

    DEFF Research Database (Denmark)

    Ndeboko, Bénédicte; Lemamy, Guy Joseph; Nielsen, Peter E;

    2015-01-01

    hepatitis B virus (DHBV), a reference model for human HBV infection. The in vivo administration of PNA-CPP conjugates to neonatal ducklings showed that they reached the liver and inhibited DHBV replication. Interestingly, our results indicated also that a modified CPP (CatLip) alone, in the absence of its......Chronic hepatitis B virus (HBV) infection remains a major health problem worldwide. Because current anti-HBV treatments are only virostatic, there is an urgent need for development of alternative antiviral approaches. In this context, cell-penetrating peptides (CPPs) and cationic polymers...

  8. SAP(E) - A cell-penetrating polyproline helix at lipid interfaces.

    Science.gov (United States)

    Franz, Johannes; Lelle, Marco; Peneva, Kalina; Bonn, Mischa; Weidner, Tobias

    2016-09-01

    Cell-penetrating peptides (CPPs) are short membrane-permeating amino acid sequences that can be used to deliver cargoes, e.g. drugs, into cells. The mechanism for CPP internalization is still subject of ongoing research. An interesting family of CPPs is the sweet arrow peptides - SAP(E) - which are known to adopt a polyproline II helical secondary structure. SAP(E) peptides stand out among CPPs because they carry a net negative charge while most CPPs are positively charged, the latter being conducive to electrostatic interaction with generally negatively charged membranes. For SAP(E)s, an internalization mechanism has been proposed, based on polypeptide aggregation on the cell surface, followed by an endocytic uptake. However, this process has not yet been observed directly - since peptide-membrane interactions are inherently difficult to monitor on a molecular scale. Here, we use sum frequency generation (SFG) vibrational spectroscopy to investigate molecular interactions of SAP(E) with differently charged model membranes, in both mono- and bi-layer configurations. The data suggest that the initial binding mechanism is accompanied by structural changes of the peptide. Also, the peptide-model membrane interaction depends on the charge of the lipid headgroup with phosphocholine being a favorable binding site. Moreover, while direct penetration has also been observed for some CPPs, the spectroscopy reveals that for SAP(E), its interaction with model membranes remains limited to the headgroup region, and insertion into the hydrophobic core of the lipid layer does not occur.

  9. SAP(E) - A cell-penetrating polyproline helix at lipid interfaces.

    Science.gov (United States)

    Franz, Johannes; Lelle, Marco; Peneva, Kalina; Bonn, Mischa; Weidner, Tobias

    2016-09-01

    Cell-penetrating peptides (CPPs) are short membrane-permeating amino acid sequences that can be used to deliver cargoes, e.g. drugs, into cells. The mechanism for CPP internalization is still subject of ongoing research. An interesting family of CPPs is the sweet arrow peptides - SAP(E) - which are known to adopt a polyproline II helical secondary structure. SAP(E) peptides stand out among CPPs because they carry a net negative charge while most CPPs are positively charged, the latter being conducive to electrostatic interaction with generally negatively charged membranes. For SAP(E)s, an internalization mechanism has been proposed, based on polypeptide aggregation on the cell surface, followed by an endocytic uptake. However, this process has not yet been observed directly - since peptide-membrane interactions are inherently difficult to monitor on a molecular scale. Here, we use sum frequency generation (SFG) vibrational spectroscopy to investigate molecular interactions of SAP(E) with differently charged model membranes, in both mono- and bi-layer configurations. The data suggest that the initial binding mechanism is accompanied by structural changes of the peptide. Also, the peptide-model membrane interaction depends on the charge of the lipid headgroup with phosphocholine being a favorable binding site. Moreover, while direct penetration has also been observed for some CPPs, the spectroscopy reveals that for SAP(E), its interaction with model membranes remains limited to the headgroup region, and insertion into the hydrophobic core of the lipid layer does not occur. PMID:27237727

  10. Cell penetrating peptide delivery of splice directing oligonucleotides as a treatment for Duchenne muscular dystrophy.

    Science.gov (United States)

    Betts, Corinne A; Wood, Matthew J A

    2013-01-01

    Duchenne muscular dystrophy is a severe, X-linked muscle wasting disorder caused by the absence of an integral structural protein called dystrophin. This is caused by mutations or deletions in the dystrophin gene which disrupt the reading frame, thereby halting the production of a functional protein. A number of potential therapies have been investigated for the treatment of this disease including utrophin upregulation, 'stop-codon read through' aminoglycosides and adeno-associated virus gene replacement as well as stem cell therapy. However, the most promising treatment to date is the use of antisense oligonucleotides which cause exon skipping by binding to a specific mRNA sequence, skipping the desired exon, thereby restoring the reading frame and producing a truncated yet functional protein. The results from recent 2'OMePS and morpholino clinical trials have renewed hope for Duchenne patients; however in vivo studies in a mouse model, mdx, have revealed low systemic distribution and poor delivery of oligonucleotides to affected tissues such as the brain and heart. However a variety of cell penetrating peptides directly conjugated to antisense oligonucleotides have been shown to enhance delivery in Duchenne model systems with improved systemic distribution and greater efficacy compared to 'naked' antisense oligonucleotides. These cell penetrating peptides, combined with an optimised dose and dosing regimen, as well as thorough toxicity profile have the potential to be developed into a promising treatment which may be progressed to clinical trial. PMID:23140454

  11. Selective mono-radioiodination and characterization of a cell-penetrating peptide. L-Tyr-maurocalcine

    Energy Technology Data Exchange (ETDEWEB)

    Ahmadi, Mitra; Bacot, Sandrine; Perret, Pascale; Riou, Laurent; Ghezzi, Catherine [Universite Joseph Fourier, Grenoble (France); INSERM U1039, Grenoble (France). Radiopharmaceutiques Biocliniques; Poillot, Cathy; Cestele, Sandrine [INSERM U836, Grenoble (France). Grenoble Inst. of Neuroscience; Universite Joseph Fourier, Grenoble (France); Desruet, Marie-Dominique [INSERM U1039, Grenoble (France). Radiopharmaceutiques Biocliniques; Couvet, Morgane; Bourgoin, Sandrine; Seve, Michel [CRI-INSERM U823, Grenoble (France). Inst. of Albert Bonniot; Universite Joseph Fourier, Grenoble (France); Waard, Michel de [INSERM U836, Grenoble (France). Grenoble Inst. of Neuroscience; Universite Joseph Fourier, Grenoble (France); Smartox Biotechnologies, Grenoble (France)

    2014-07-01

    Mono-and poly-iodinated peptides form frequently during radioiodination procedures. However, the formation of a single species in its mono-iodinated form is essential for quantitative studies such as determination of tissue concentration or image quantification. Therefore, the aim of the present study was to define the optimal experimental conditions in order to exclusively obtain the mono-iodinated form of L-maurocalcine (L-MCa). L-MCa is an animal venom toxin which was shown to act as a cell-penetrating peptide. In order to apply the current direct radioiodination technique using oxidative agents including chloramine T, Iodo-Gen {sup registered} or lactoperoxidase, an analogue of this peptide containing a tyrosine residue (Tyr-L-MCa) was synthesized and was shown to fold/oxidize properly. The enzymatic approach using lactoperoxidase/H{sub 2}O{sub 2} was found to be the best method for radioiodination of Tyr-L-MCa. MALDI-TOF mass spectrometry analyses were then used for identification of the chromatographic eluting components of the reaction mixtures. We observed that the production of different radioiodinated species depended upon the reaction conditions. Our results successfully described the experimental conditions of peptide radioiodination allowing the exclusive production of the mono-iodinated form with high radiochemical purity and without the need for a purification step. Mono-radioiodination of L-Tyr-MCa will be crucial for future quantitative studies, investigating the mechanism of cell penetration and in vivo biodistribution.

  12. Selective mono-radioiodination and characterization of a cell-penetrating peptide. L-Tyr-maurocalcine

    International Nuclear Information System (INIS)

    Mono-and poly-iodinated peptides form frequently during radioiodination procedures. However, the formation of a single species in its mono-iodinated form is essential for quantitative studies such as determination of tissue concentration or image quantification. Therefore, the aim of the present study was to define the optimal experimental conditions in order to exclusively obtain the mono-iodinated form of L-maurocalcine (L-MCa). L-MCa is an animal venom toxin which was shown to act as a cell-penetrating peptide. In order to apply the current direct radioiodination technique using oxidative agents including chloramine T, Iodo-Gen registered or lactoperoxidase, an analogue of this peptide containing a tyrosine residue (Tyr-L-MCa) was synthesized and was shown to fold/oxidize properly. The enzymatic approach using lactoperoxidase/H2O2 was found to be the best method for radioiodination of Tyr-L-MCa. MALDI-TOF mass spectrometry analyses were then used for identification of the chromatographic eluting components of the reaction mixtures. We observed that the production of different radioiodinated species depended upon the reaction conditions. Our results successfully described the experimental conditions of peptide radioiodination allowing the exclusive production of the mono-iodinated form with high radiochemical purity and without the need for a purification step. Mono-radioiodination of L-Tyr-MCa will be crucial for future quantitative studies, investigating the mechanism of cell penetration and in vivo biodistribution.

  13. p53 Aggregates penetrate cells and induce the co-aggregation of intracellular p53.

    Directory of Open Access Journals (Sweden)

    Karolyn J Forget

    Full Text Available Prion diseases are unique pathologies in which the infectious particles are prions, a protein aggregate. The prion protein has many particular features, such as spontaneous aggregation, conformation transmission to other native PrP proteins and transmission from an individual to another. Protein aggregation is now frequently associated to many human diseases, for example Alzheimer's disease, Parkinson's disease or type 2 diabetes. A few proteins associated to these conformational diseases are part of a new category of proteins, called prionoids: proteins that share some, but not all, of the characteristics associated with prions. The p53 protein, a transcription factor that plays a major role in cancer, has recently been suggested to be a possible prionoid. The protein has been shown to accumulate in multiple cancer cell types, and its aggregation has also been reproduced in vitro by many independent groups. These observations suggest a role for p53 aggregates in cancer development. This study aims to test the «prion-like» features of p53. Our results show in vitro aggregation of the full length and N-terminally truncated protein (p53C, and penetration of these aggregates into cells. According to our findings, the aggregates enter cells using macropinocytosis, a non-specific pathway of entry. Lastly, we also show that once internalized by the cell, p53C aggregates can co-aggregate with endogenous p53 protein. Together, these findings suggest prion-like characteristics for p53 protein, based on the fact that p53 can spontaneously aggregate, these aggregates can penetrate cells and co-aggregate with cellular p53.

  14. Cell-Penetrating Peptide as a Means of Directing the Differentiation of Induced-Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Taku Kaitsuka

    2015-11-01

    Full Text Available Protein transduction using cell-penetrating peptides (CPPs is useful for the delivery of large protein molecules, including some transcription factors. This method is safer than gene transfection methods with a viral vector because there is no risk of genomic integration of the exogenous DNA. Recently, this method was reported as a means for the induction of induced pluripotent stem (iPS cells, directing the differentiation into specific cell types and supporting gene editing/correction. Furthermore, we developed a direct differentiation method to obtain a pancreatic lineage from mouse and human pluripotent stem cells via the protein transduction of three transcription factors, Pdx1, NeuroD, and MafA. Here, we discuss the possibility of using CPPs as a means of directing the differentiation of iPS cells and other stem cell technologies.

  15. Cell-Penetrating Peptide as a Means of Directing the Differentiation of Induced-Pluripotent Stem Cells.

    Science.gov (United States)

    Kaitsuka, Taku; Tomizawa, Kazuhito

    2015-11-06

    Protein transduction using cell-penetrating peptides (CPPs) is useful for the delivery of large protein molecules, including some transcription factors. This method is safer than gene transfection methods with a viral vector because there is no risk of genomic integration of the exogenous DNA. Recently, this method was reported as a means for the induction of induced pluripotent stem (iPS) cells, directing the differentiation into specific cell types and supporting gene editing/correction. Furthermore, we developed a direct differentiation method to obtain a pancreatic lineage from mouse and human pluripotent stem cells via the protein transduction of three transcription factors, Pdx1, NeuroD, and MafA. Here, we discuss the possibility of using CPPs as a means of directing the differentiation of iPS cells and other stem cell technologies.

  16. Reduced activity of TAFI (thrombin-activatable fibrinolysis inhibitor) in acute promyelocytic leukaemia

    NARCIS (Netherlands)

    Meijers, JCM; Oudijk, EJD; Mosnier, LO; Nieuwenhuis, HK; Fijnheer, R; Bouma, Bonno N.; Bos, R

    2000-01-01

    Acute promyelocytic leukaemia (APL) is a disease that is distinguished from other leukaemias by the high potential for early haemorrhagic death. Several processes are involved, such as disseminated intravascular coagulation and hyperfibrinolysis. Recently, TAFI (thrombin-activatable fibrinolysis inh

  17. Cellular uptake of antisense oligonucleotides after complexing or conjugation with cell-penetrating model peptides.

    Science.gov (United States)

    Oehlke, J; Birth, P; Klauschenz, E; Wiesner, B; Beyermann, M; Oksche, A; Bienert, M

    2002-08-01

    The uptake by mammalian cells of phosphorothioate oligonucleotides was compared with that of their respective complexes or conjugates with cationic, cell-penetrating model peptides of varying helix-forming propensity and amphipathicity. An HPLC-based protocol for the synthesis and purification of disulfide bridged conjugates in the 10-100 nmol range was developed. Confocal laser scanning microscopy (CLSM) in combination with gel-capillary electrophoresis and laser induced fluorescence detection (GCE-LIF) revealed cytoplasmic and nuclear accumulationin all cases. The uptake differences between naked oligonucleotides and their respective peptide complexes or conjugates were generally confined to one order of magnitude. No significant influence of the structural properties of the peptide components upon cellular uptake was found. Our results question the common belief that the increased biological activity of oligonucleotides after derivatization with membrane permeable peptides may be primarily due to improved membrane translocation.

  18. Metabolic cleavage of cell-penetrating peptides in contact with epithelial models

    DEFF Research Database (Denmark)

    Tréhin, Rachel; Nielsen, Hanne Mørck; Jahnke, Heinz-Georg;

    2004-01-01

    We assessed the metabolic degradation kinetics and cleavage patterns of some selected CPP (cell-penetrating peptides) after incubation with confluent epithelial models. Synthesis of N-terminal CF [5(6)-carboxyfluorescein]-labelled CPP, namely hCT (human calcitonin)-derived sequences, Tat(47-57) and...... penetratin(43-58), was through Fmoc (fluoren-9-ylmethoxycarbonyl) chemistry. Metabolic degradation kinetics of the tested CPP in contact with three cell-cultured epithelial models, MDCK (Madin-Darby canine kidney), Calu-3 and TR146, was evaluated by reversed-phase HPLC. Identification of the resulting...... models and the CPP. The Calu-3 model exhibited the highest proteolytic activity. The patterns of metabolic cleavage of hCT(9-32) were similar in all three models. Initial cleavage of this peptide occurred at the N-terminal domain, possibly by endopeptidase activity yielding both the N- and the C...

  19. Modeling the endosomal escape of cell-penetrating peptides using a transmembrane pH gradient.

    Science.gov (United States)

    Madani, Fatemeh; Abdo, Rania; Lindberg, Staffan; Hirose, Hisaaki; Futaki, Shiroh; Langel, Ulo; Gräslund, Astrid

    2013-04-01

    Cell-penetrating peptides (CPPs) can internalize into cells with covalently or non-covalently bound biologically active cargo molecules, which by themselves are not able to pass the cell membrane. Direct penetration and endocytosis are two main pathways suggested for the cellular uptake of CPPs. Cargo molecules which have entered the cell via an endocytotic pathway must be released from the endosome before degradation by enzymatic processes and endosomal acidification. Endosomal entrapment seems to be a major limitation in delivery of these molecules into the cytoplasm. Bacteriorhodopsin (BR) asymmetrically introduced into large unilamellar vesicles (LUVs) was used to induce a pH gradient across the lipid bilayer. By measuring pH outside the LUVs, we observed light-induced proton pumping mediated by BR from the outside to the inside of the LUVs, creating an acidic pH inside the LUVs, similar to the late endosomes in vivo. Here we studied the background mechanism(s) of endosomal escape. 20% negatively charged LUVs were used as model endosomes with incorporated BR into the membrane and fluorescein-labeled CPPs entrapped inside the LUVs, together with a fluorescence quencher. The translocation of different CPPs in the presence of a pH gradient across the membrane was studied. The results show that the light-induced pH gradient induced by BR facilitates vesicle membrane translocation, particularly for the intermediately hydrophobic CPPs, and much less for hydrophilic CPPs. The presence of chloroquine inside the LUVs or addition of pyrenebutyrate outside the LUVs destabilizes the vesicle membrane, resulting in significant changes of the pH gradient across the membrane.

  20. Regulation of tissue inflammation by thrombin-activatable carboxypeptidase B (or TAFI)

    OpenAIRE

    Leung, Lawrence L. K.; Myles, Timothy; Nishimura, Toshihiko; Song, Jason J.; Robinson, William H.

    2008-01-01

    Thrombin-activatable procarboxypeptidase B (proCPB or thrombin-activatable fibrinolysis inhibitor or TAFI) is a plasma procarboxypeptidase that is activated by the thrombin-thrombomodulin complex on the vascular endothelial surface. The activated CPB removes the newly exposed carboxyl terminal lysines in the partially digested fibrin clot, diminishes tissue plasminogen activator and plasminogen binding, and protects the clot from premature lysis. We have recently shown that CPB is catalytical...

  1. Cell membrane penetration and mitochondrial targeting by platinum-decorated ceria nanoparticles.

    Science.gov (United States)

    Torrano, Adriano A; Herrmann, Rudolf; Strobel, Claudia; Rennhak, Markus; Engelke, Hanna; Reller, Armin; Hilger, Ingrid; Wixforth, Achim; Bräuchle, Christoph

    2016-07-01

    In this work we investigate the interaction between endothelial cells and nanoparticles emitted by catalytic converters. Although catalyst-derived particles are recognized as growing burden added to environmental pollution, very little is known about their health impact. We use platinum-decorated ceria nanoparticles as model compounds for the actual emitted particles and focus on their fast uptake and association with mitochondria, the cell's powerhouse. Using live-cell imaging and electron microscopy we clearly show that 46 nm platinum-decorated ceria nanoparticles can rapidly penetrate cell membranes and reach the cytosol. Moreover, if suitably targeted, these particles are able to selectively attach to mitochondria. These results are complemented by cytotoxicity assays, thus providing insights into the biological effects of these particles on cells. Interestingly, no permanent membrane disruption or any other significant adverse effects on cells were observed. The unusual uptake behavior observed for 46 nm nanoparticles was not observed for equivalent but larger 143 nm and 285 nm platinum-decorated particles. Our results demonstrate a remarkable particle size effect in which particles smaller than ∼50-100 nm escape the usual endocytic pathway and translocate directly into the cytosol, while particles larger than ∼150 nm are internalized by conventional endocytosis. Since the small particles are able to bypass endocytosis they could be explored as drug and gene delivery vehicles. Platinum-decorated nanoparticles are therefore highly interesting in the fields of nanotoxicology and nanomedicine. PMID:27341699

  2. Cell-Penetrating Peptides Selectively Cross the Blood-Brain Barrier In Vivo.

    Directory of Open Access Journals (Sweden)

    Sofie Stalmans

    Full Text Available Cell-penetrating peptides (CPPs are a group of peptides, which have the ability to cross cell membrane bilayers. CPPs themselves can exert biological activity and can be formed endogenously. Fragmentary studies demonstrate their ability to enhance transport of different cargoes across the blood-brain barrier (BBB. However, comparative, quantitative data on the BBB permeability of different CPPs are currently lacking. Therefore, the in vivo BBB transport characteristics of five chemically diverse CPPs, i.e. pVEC, SynB3, Tat 47-57, transportan 10 (TP10 and TP10-2, were determined. The results of the multiple time regression (MTR analysis revealed that CPPs show divergent BBB influx properties: Tat 47-57, SynB3, and especially pVEC showed very high unidirectional influx rates of 4.73 μl/(g × min, 5.63 μl/(g × min and 6.02 μl/(g × min, respectively, while the transportan analogs showed a negligible to low brain influx. Using capillary depletion, it was found that 80% of the influxed peptides effectively reached the brain parenchyma. Except for pVEC, all peptides showed a significant efflux out of the brain. Co-injection of pVEC with radioiodinated bovine serum albumin (BSA did not enhance the brain influx of radiodionated BSA, indicating that pVEC does not itself significantly alter the BBB properties. A saturable mechanism could not be demonstrated by co-injecting an excess dose of non-radiolabeled CPP. No significant regional differences in brain influx were observed, with the exception for pVEC, for which the regional variations were only marginal. The observed BBB influx transport properties cannot be correlated with their cell-penetrating ability, and therefore, good CPP properties do not imply efficient brain influx.

  3. Cell-Penetrating Peptides Selectively Cross the Blood-Brain Barrier In Vivo.

    Science.gov (United States)

    Stalmans, Sofie; Bracke, Nathalie; Wynendaele, Evelien; Gevaert, Bert; Peremans, Kathelijne; Burvenich, Christian; Polis, Ingeborgh; De Spiegeleer, Bart

    2015-01-01

    Cell-penetrating peptides (CPPs) are a group of peptides, which have the ability to cross cell membrane bilayers. CPPs themselves can exert biological activity and can be formed endogenously. Fragmentary studies demonstrate their ability to enhance transport of different cargoes across the blood-brain barrier (BBB). However, comparative, quantitative data on the BBB permeability of different CPPs are currently lacking. Therefore, the in vivo BBB transport characteristics of five chemically diverse CPPs, i.e. pVEC, SynB3, Tat 47-57, transportan 10 (TP10) and TP10-2, were determined. The results of the multiple time regression (MTR) analysis revealed that CPPs show divergent BBB influx properties: Tat 47-57, SynB3, and especially pVEC showed very high unidirectional influx rates of 4.73 μl/(g × min), 5.63 μl/(g × min) and 6.02 μl/(g × min), respectively, while the transportan analogs showed a negligible to low brain influx. Using capillary depletion, it was found that 80% of the influxed peptides effectively reached the brain parenchyma. Except for pVEC, all peptides showed a significant efflux out of the brain. Co-injection of pVEC with radioiodinated bovine serum albumin (BSA) did not enhance the brain influx of radiodionated BSA, indicating that pVEC does not itself significantly alter the BBB properties. A saturable mechanism could not be demonstrated by co-injecting an excess dose of non-radiolabeled CPP. No significant regional differences in brain influx were observed, with the exception for pVEC, for which the regional variations were only marginal. The observed BBB influx transport properties cannot be correlated with their cell-penetrating ability, and therefore, good CPP properties do not imply efficient brain influx.

  4. Porous silicon-cell penetrating peptide hybrid nanocarrier for intracellular delivery of oligonucleotides.

    Science.gov (United States)

    Rytkönen, Jussi; Arukuusk, Piret; Xu, Wujun; Kurrikoff, Kaido; Langel, Ulo; Lehto, Vesa-Pekka; Närvänen, Ale

    2014-02-01

    The largest obstacle to the use of oligonucleotides as therapeutic agents is the delivery of these large and negatively charged biomolecules through cell membranes into intracellular space. Mesoporous silicon (PSi) is widely recognized as a potential material for drug delivery purposes due to its several beneficial features like large surface area and pore volume, high loading capacity, biocompatibility, and biodegradability. In the present study, PSi nanoparticles stabilized by thermal oxidation or thermal carbonization and subsequently modified by grafting aminosilanes on the surface are utilized as an oligonucleotide carrier. Splice correcting oligonucleotides (SCOs), a model oligonucleotide drug, were loaded into the positively charged PSi nanoparticles with a loading degree as high as 14.3% (w/w). Rapid loading was achieved by electrostatic interactions, with the loading efficiencies reaching 100% within 5 min. The nanoparticles were shown to deliver and release SCOs, in its biologically active form, inside cells when formulated together with cell penetrating peptides (CPP). The biological effect was monitored with splice correction assay and confocal microscopy utilizing HeLa pLuc 705 cells. Furthermore, the use of PSi carrier platform in oligonucleotide delivery did not reduce the cell viability. Additionally, the SCO-CPP complexes formed in the pores of the carrier were stabilized against proteolytic digestion. The advantageous properties of protecting and releasing the cargo and the possibility to further functionalize the carrier surface make the hybrid nanoparticles a potential system for oligonucleotide delivery.

  5. Comparative mechanisms of protein transduction mediated by cell-penetrating peptides in prokaryotes.

    Science.gov (United States)

    Liu, Betty Revon; Huang, Yue-Wern; Aronstam, Robert S; Lee, Han-Jung

    2015-04-01

    Bacterial and archaeal cell envelopes are complex multilayered barriers that serve to protect these microorganisms from their extremely harsh and often hostile environments. Import of exogenous proteins and nanoparticles into cells is important for biotechnological applications in prokaryotes. In this report, we demonstrate that cell-penetrating peptides (CPPs), both bacteria-expressed nona-arginine peptide (R9) and synthetic R9 (SR9), are able to deliver noncovalently associated proteins or quantum dots into four representative species of prokaryotes: cyanobacteria (Synechocystis sp. PCC 6803), bacteria (Escherichia coli DH5α and Arthrobacter ilicis D-50), and archaea (Thermus aquaticus). Although energy-dependent endocytosis is generally accepted as a hallmark that distinguishes eukaryotes from prokaryotes, cellular uptake of uncomplexed green fluorescent protein (GFP) by cyanobacteria was mediated by classical endocytosis. Mechanistic studies revealed that macropinocytosis plays a critical and major role in CPP-mediated protein transduction in all four prokaryotes. Membrane damage was not observed when cyanobacterial cells were treated with R9/GFP complexes, nor was cytotoxicity detected when bacteria or archaea were treated with SR9/QD complexes in the presence of macropinocytic inhibitors. These results indicate that the uptake of protein is not due to a compromise of membrane integrity in cyanobacteria, and that CPP can be an effective and safe carrier for membrane trafficking in prokaryotic cells. Our investigation provides important new insights into the transport of exogenous proteins and nanoparticles across the complex membrane systems of prokaryotes.

  6. Cell-penetrating TAT peptide in drug delivery systems: Proteolytic stability requirements

    Science.gov (United States)

    Koren, Erez; Apte, Anjali; Sawant, Rupa R.; Grunwald, Jacob; Torchilin, Vladimir P.

    2012-01-01

    The stability and activity of the HIV cell-penetrating TAT peptide (TATp) on the surface of TATp-modified micelles and liposomes in relation to its proteolytic cleavage was investigated. TATp moieties were attached to the surface of these nanocarriers using TATp modified with a conjugate of phosphatidyl ethanolamine with a ‘short’ PEG (PEG-PE). Following pre-incubation with trypsin, elastase, or collagenase, the proteolytic stability of TATp on the surface of these modified carriers was studied by HPLC with fluorescence detection using fluorenylmethyl chloroformate (FMOC) labeling. All tested enzymes produced a dose-dependent cleavage of TATp as shown by the presence of TATp Arg-Arg fragments. Inhibition of TATp cleavage occurred when these TATp-micelles were modified by the addition of longer PEG-PE blocks, indicating an effective shielding of TATp from proteolysis by these blocks. TATp-modified carriers were also tested for their ability to accumulate in EL-4, HeLa, and B16-F10 cells. Trypsin treatment of TATp-modified liposomes and micelles resulted in decreased uptake and cell interaction, as measured by fluorescence microscopy and fluorescence-activated cell sorting techniques. Furthermore, a decrease in the cytotoxicity of TATp-modified liposomes loaded with doxorubicin (Doxil) was observed following trypsin treatment. In conclusion, steric shielding of TATp is essential to ensure its in vivo therapeutic function. PMID:21438724

  7. Graphene microsheets enter cells through spontaneous membrane penetration at edge asperities and corner sites.

    Science.gov (United States)

    Li, Yinfeng; Yuan, Hongyan; von dem Bussche, Annette; Creighton, Megan; Hurt, Robert H; Kane, Agnes B; Gao, Huajian

    2013-07-23

    Understanding and controlling the interaction of graphene-based materials with cell membranes is key to the development of graphene-enabled biomedical technologies and to the management of graphene health and safety issues. Very little is known about the fundamental behavior of cell membranes exposed to ultrathin 2D synthetic materials. Here we investigate the interactions of graphene and few-layer graphene (FLG) microsheets with three cell types and with model lipid bilayers by combining coarse-grained molecular dynamics (MD), all-atom MD, analytical modeling, confocal fluorescence imaging, and electron microscopic imaging. The imaging experiments show edge-first uptake and complete internalization for a range of FLG samples of 0.5- to 10-μm lateral dimension. In contrast, the simulations show large energy barriers relative to kBT for membrane penetration by model graphene or FLG microsheets of similar size. More detailed simulations resolve this paradox by showing that entry is initiated at corners or asperities that are abundant along the irregular edges of fabricated graphene materials. Local piercing by these sharp protrusions initiates membrane propagation along the extended graphene edge and thus avoids the high energy barrier calculated in simple idealized MD simulations. We propose that this mechanism allows cellular uptake of even large multilayer sheets of micrometer-scale lateral dimension, which is consistent with our multimodal bioimaging results for primary human keratinocytes, human lung epithelial cells, and murine macrophages.

  8. Cell membrane penetration and mitochondrial targeting by platinum-decorated ceria nanoparticles

    Science.gov (United States)

    Torrano, Adriano A.; Herrmann, Rudolf; Strobel, Claudia; Rennhak, Markus; Engelke, Hanna; Reller, Armin; Hilger, Ingrid; Wixforth, Achim; Bräuchle, Christoph

    2016-07-01

    In this work we investigate the interaction between endothelial cells and nanoparticles emitted by catalytic converters. Although catalyst-derived particles are recognized as growing burden added to environmental pollution, very little is known about their health impact. We use platinum-decorated ceria nanoparticles as model compounds for the actual emitted particles and focus on their fast uptake and association with mitochondria, the cell's powerhouse. Using live-cell imaging and electron microscopy we clearly show that 46 nm platinum-decorated ceria nanoparticles can rapidly penetrate cell membranes and reach the cytosol. Moreover, if suitably targeted, these particles are able to selectively attach to mitochondria. These results are complemented by cytotoxicity assays, thus providing insights into the biological effects of these particles on cells. Interestingly, no permanent membrane disruption or any other significant adverse effects on cells were observed. The unusual uptake behavior observed for 46 nm nanoparticles was not observed for equivalent but larger 143 nm and 285 nm platinum-decorated particles. Our results demonstrate a remarkable particle size effect in which particles smaller than ~50-100 nm escape the usual endocytic pathway and translocate directly into the cytosol, while particles larger than ~150 nm are internalized by conventional endocytosis. Since the small particles are able to bypass endocytosis they could be explored as drug and gene delivery vehicles. Platinum-decorated nanoparticles are therefore highly interesting in the fields of nanotoxicology and nanomedicine.In this work we investigate the interaction between endothelial cells and nanoparticles emitted by catalytic converters. Although catalyst-derived particles are recognized as growing burden added to environmental pollution, very little is known about their health impact. We use platinum-decorated ceria nanoparticles as model compounds for the actual emitted particles and

  9. Endothelial cell counts after Descemet’s stripping automated endothelial keratoplasty versus penetrating keratoplasty in Asian eyes

    Directory of Open Access Journals (Sweden)

    Ang M

    2012-04-01

    Full Text Available Marcus Ang1,2, Jodhbir S Mehta1–4, Arundhati Anshu1,2, Hon Kiat Wong5, Hla M Htoon2, Donald Tan1–31Singapore National Eye Centre, 2Singapore Eye Research Institute, 3Department of Ophthalmology, National University Health Systems, 4Department of Clinical Sciences, Duke-NUS Graduate Medical School, 5Department of Ophthalmology, Tan Tock Seng Hospital, SingaporeBackground: The purpose of this study was to compare endothelial cell counts after Descemet’s stripping automated endothelial keratoplasty (DSAEK and penetrating keratoplasty in Asian eyes.Methods: This was a retrospective study of patients from our prospective Singapore Corneal Transplant Study cohort who received corneal transplantation in 2006–2008. We compared eyes that underwent DSAEK or penetrating keratoplasty for Fuchs’ endothelial dystrophy or pseudophakic and aphakic bullous keratopathy. Clinical data, and donor and recipient characteristics were recorded. Of 241 patients who met our inclusion criteria, 68 underwent DSAEK and 173 underwent penetrating keratoplasty. The main outcome measure was endothelial cell loss at 1 year. Secondary outcome measures were graft survival and visual outcomes at 1-year follow-up.Results: There were no significant differences in baseline characteristics of patients between the treatment groups. Percent endothelial cell loss at 1-year follow-up was greater in penetrating keratoplasty eyes (40.9% ± 2.9% compared with DSAEK eyes (22.4% ± 2.3%; P < 0.001. DSAEK-treated eyes had significantly superior uncorrected visual acuity (mean difference = 0.42 ± 0.0059; P < 0.001 and best spectacle-corrected visual acuity (mean difference = 0.14 ± 0.032; P < 0.001 as compared with penetrating keratoplasty-treated eyes. Penetrating keratoplasty-treated eyes had worse astigmatism as compared with DSAEK-treated eyes (-3.0 ± 2.1 versus -1.7 ± 0.8; P < 0.001. Graft survival at 1 year was comparable in both groups, ie, 66/68 (97.0% DSAEK-treated eyes

  10. Generation of GFP Native Protein for Detection of Its Intracellular Uptake by Cell-Penetrating Peptides.

    Science.gov (United States)

    Kadkhodayan, S; Sadat, S M; Irani, S; Fotouhi, F; Bolhassani, A

    2016-01-01

    Different types of lipid- and polymer-based vectors have been developed to deliver proteins into cells, but these methods showed relatively poor efficiency. Recently, a group of short, highly basic peptides known as cell-penetrating peptides (CPPs) were used to carry polypeptides and proteins into cells. In this study, expression and purification of GFP protein was performed using the prokaryotic pET expression system. We used two amphipathic CPPs (Pep-1 and CADY-2) as a novel delivery system to transfer the GFP protein into cells. The morphological features of the CPP/GFP complexes were studied by scanning electron microscopy (SEM), Zetasizer, and SDS-PAGE. The efficiency of GFP transfection using Pep-1 and CADY-2 peptides and TurboFect reagent was compared with FITC-antibody protein control delivered by these transfection vehicles in the HEK-293T cell line. SEM data confirmed formation of discrete nanoparticles with a diameter of below 300 nm. Moreover, formation of the complexes was detected using SDS-PAGE as two individual bands, indicating non-covalent interaction. The size and homogeneity of Pep-1/GFP and CADY-2/GFP complexes were dependent on the ratio of peptide/cargo formulations, and responsible for their biological efficiency. The cells transfected by Pep-1/GFP and CADY-2/GFP complexes at a molar ratio of 20 : 1 demonstrated spreading green regions using fluorescent microscopy. Flow cytometry results showed that the transfection efficiency of Pep-based nanoparticles was similar to CADY-based nanoparticles and comparable with TurboFect-protein complexes. These data open an efficient way for future therapeutic purposes. PMID:27516189

  11. Conjugation to the cell-penetrating peptide TAT potentiates the photodynamic effect of carboxytetramethylrhodamine.

    Directory of Open Access Journals (Sweden)

    Divyamani Srinivasan

    Full Text Available BACKGROUND: Cell-penetrating peptides (CPPs can transport macromolecular cargos into live cells. However, the cellular delivery efficiency of these reagents is often suboptimal because CPP-cargo conjugates typically remain trapped inside endosomes. Interestingly, irradiation of fluorescently labeled CPPs with light increases the release of the peptide and its cargos into the cytosol. However, the mechanism of this phenomenon is not clear. Here we investigate the molecular basis of the photo-induced endosomolytic activity of the prototypical CPPs TAT labeled to the fluorophore 5(6-carboxytetramethylrhodamine (TMR. METHODOLOGY/PRINCIPAL FINDINGS: We report that TMR-TAT acts as a photosensitizer that can destroy membranes. TMR-TAT escapes from endosomes after exposure to moderate light doses. However, this is also accompanied by loss of plasma membrane integrity, membrane blebbing, and cell-death. In addition, the peptide causes the destruction of cells when applied extracellularly and also triggers the photohemolysis of red blood cells. These photolytic and photocytotoxic effects were inhibited by hydrophobic singlet oxygen quenchers but not by hydrophilic quenchers. CONCLUSIONS/SIGNIFICANCE: Together, these results suggest that TAT can convert an innocuous fluorophore such as TMR into a potent photolytic agent. This effect involves the targeting of the fluorophore to cellular membranes and the production of singlet oxygen within the hydrophobic environment of the membranes. Our findings may be relevant for the design of reagents with photo-induced endosomolytic activity. The photocytotoxicity exhibited by TMR-TAT also suggests that CPP-chromophore conjugates could aid the development of novel Photodynamic Therapy agents.

  12. CpG expedites regression of local and systemic tumors when combined with activatable nanodelivery.

    Science.gov (United States)

    Kheirolomoom, Azadeh; Ingham, Elizabeth S; Mahakian, Lisa M; Tam, Sarah M; Silvestrini, Matthew T; Tumbale, Spencer K; Foiret, Josquin; Hubbard, Neil E; Borowsky, Alexander D; Murphy, William J; Ferrara, Katherine W

    2015-12-28

    Ultrasonic activation of nanoparticles provides the opportunity to deliver a large fraction of the injected dose to insonified tumors and produce a complete local response. Here, we evaluate whether the local and systemic response to chemotherapy can be enhanced by combining such a therapy with locally-administered CpG as an immune adjuvant. In order to create stable, activatable particles, a complex between copper and doxorubicin (CuDox) was created within temperature-sensitive liposomes. Whereas insonation of the CuDox liposomes alone has been shown to produce a complete response in murine breast cancer after 8 treatments of 6 mg/kg delivered over 4 weeks, combining this treatment with CpG resolved local cancers within 3 treatments delivered over 7 days. Further, contralateral tumors regressed as a result of the combined treatment, and survival was extended in systemic disease. In both the treated and contralateral tumor site, the combined treatment increased leukocytes and CD4+ and CD8+ T-effector cells and reduced myeloid-derived suppressor cells (MDSCs). Taken together, the results suggest that this combinatorial treatment significantly enhances the systemic efficacy of locally-activated nanotherapy. PMID:26471394

  13. Design and screening of a glial cell-specific, cell penetrating peptide for therapeutic applications in multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    Corey Heffernan

    Full Text Available Multiple Sclerosis (MS is an autoimmune, neurodegenerative disease of the central nervous system (CNS characterized by demyelination through glial cell loss. Current and proposed therapeutic strategies to arrest demyelination and/or promote further remyelination include: (i modulation of the host immune system; and/or (ii transplantation of myelinating/stem or progenitor cells to the circulation or sites of injury. However, significant drawbacks are inherent with both approaches. Cell penetrating peptides (CPP are short amino acid sequences with an intrinsic ability to translocate across plasma membranes, and theoretically represent an attractive vector for delivery of therapeutic peptides or nanoparticles to glia to promote cell survival or remyelination. The CPPs described to date are commonly non-selective in the cell types they transduce, limiting their therapeutic application in vivo. Here, we describe a theoretical framework for design of a novel CPP sequence that selectively transduces human glial cells (excluding non-glial cell types, and conduct preliminary screens of purified, recombinant CPPs with immature and matured human oligodendrocytes and astrocytes, and two non-glial cell types. A candidate peptide, termed TD2.2, consistently transduced glial cells, was significantly more effective at transducing immature oligodendrocytes than matured progeny, and was virtually incapable of transducing two non-glial cell types: (i human neural cells and (ii human dermal fibroblasts. Time-lapse confocal microscopy confirms trafficking of TD2.2 (fused to EGFP to mature oligodendrocytes 3-6 hours after protein application in vitro. We propose selectivity of TD2.2 for glial cells represents a new therapeutic strategy for the treatment of glial-related disease, such as MS.

  14. Nanocarriers Conjugated with Cell Penetrating Peptides: New Trojan Horses by Modern Ulysses.

    Science.gov (United States)

    Zappavigna, Silvia; Misso, Gabriella; Falanga, Annarita; Perillo, Emiliana; Novellino, Ettore; Galdiero, Massimiliano; Grieco, Paolo; Caraglia, Michele; Galdiero, Stefania

    2016-01-01

    Nanomedicine has opened the way to the design of more efficient diagnostics and therapeutics. Moreover, recent literature has illustrated the use of short cationic and/or amphipathic peptides, known as cell-penetrating peptides (CPPs), for mediating advanced drug delivery. CPPs exploit their ability to enter cells and enhance the uptake of many cargoes ranging from small molecules to proteins. The distinctive properties of nanocarriers (NC) based systems provide unforeseen benefits over pure drugs for biomedical applications and constitute a challenging research field particularly focused on imaging and delivery; nonetheless, several problems have to be overcome to make them a viable option in clinic. The use of CPPs improves significantly their delivery to specific intracellular targets and thus readily contributes to their use both for effective tumor therapy and gene therapy. A key issue is related to their mechanism of uptake, because although classical CPPs enhance NCs' uptake, the entry mechanism involves the endocytic pathway, which means that the delivered material is sequestered within vesicles and only a small amount will escape from this environment and reach the desired target. In this review, we will summarize recent advances in the use of CPP for enhanced delivery of nanocarriers, nucleic acids, and drugs, we will discuss their uptake mechanisms and we will describe novel approaches to improve endosomal escape of internalized nanosystems. PMID:27087493

  15. Cell-penetrating recombinant peptides for potential use in agricultural pest control applications.

    Science.gov (United States)

    Hughes, Stephen R; Dowd, Patrick F; Johnson, Eric T

    2012-09-28

    Several important areas of interest intersect in a class of peptides characterized by their highly cationic and partly hydrophobic structure. These molecules have been called cell-penetrating peptides (CPPs) because they possess the ability to translocate across cell membranes. This ability makes these peptides attractive candidates for delivery of therapeutic compounds, especially to the interior of cells. Compounds with characteristics similar to CPPs and that, in addition, have antimicrobial properties are being investigated as antibiotics with a reduced risk of causing resistance. These CPP-like membrane-acting antimicrobial peptides (MAMPs) are α-helical amphipathic peptides that interact with and perturb cell membranes to produce their antimicrobial effects. One source of MAMPs is spider venom. Because these compounds are toxic to insects, they also show promise for development as biological agents for control of insecticide-resistant agricultural pests. Spider venom is a potential source of novel insect-specific peptide toxins. One example is the small amphipathic α-helical peptide lycotoxin-1 (Lyt-1 or LCTX) from the wolf spider (Lycosa carolinensis). One side of the α-helix has mostly hydrophilic and the other mainly hydrophobic amino acid residues. The positive charge of the hydrophilic side interacts with negatively charged prokaryotic membranes and the hydrophobic side associates with the membrane lipid bilayer to permeabilize it. Because the surface of the exoskeleton, or cuticle, of an insect is highly hydrophobic, to repel water and dirt, it would be expected that amphipathic compounds could permeabilize it. Mutagenized lycotoxin 1 peptides were produced and expressed in yeast cultures that were fed to fall armyworm (Spodoptera frugiperda) larvae to identify the most lethal mutants. Transgenic expression of spider venom toxins such as lycotoxin-1 in plants could provide durable insect resistance.

  16. Cell-Penetrating Recombinant Peptides for Potential Use in Agricultural Pest Control Applications

    Directory of Open Access Journals (Sweden)

    Eric T. Johnson

    2012-09-01

    Full Text Available Several important areas of interest intersect in a class of peptides characterized by their highly cationic and partly hydrophobic structure. These molecules have been called cell-penetrating peptides (CPPs because they possess the ability to translocate across cell membranes. This ability makes these peptides attractive candidates for delivery of therapeutic compounds, especially to the interior of cells. Compounds with characteristics similar to CPPs and that, in addition, have antimicrobial properties are being investigated as antibiotics with a reduced risk of causing resistance. These CPP-like membrane-acting antimicrobial peptides (MAMPs are α-helical amphipathic peptides that interact with and perturb cell membranes to produce their antimicrobial effects. One source of MAMPs is spider venom. Because these compounds are toxic to insects, they also show promise for development as biological agents for control of insecticide-resistant agricultural pests. Spider venom is a potential source of novel insect-specific peptide toxins. One example is the small amphipathic α-helical peptide lycotoxin-1 (Lyt-1 or LCTX from the wolf spider (Lycosa carolinensis. One side of the α-helix has mostly hydrophilic and the other mainly hydrophobic amino acid residues. The positive charge of the hydrophilic side interacts with negatively charged prokaryotic membranes and the hydrophobic side associates with the membrane lipid bilayer to permeabilize it. Because the surface of the exoskeleton, or cuticle, of an insect is highly hydrophobic, to repel water and dirt, it would be expected that amphipathic compounds could permeabilize it. Mutagenized lycotoxin 1 peptides were produced and expressed in yeast cultures that were fed to fall armyworm (Spodoptera frugiperda larvae to identify the most lethal mutants. Transgenic expression of spider venom toxins such as lycotoxin-1 in plants could provide durable insect resistance.

  17. The targeted transduction of MMP-overexpressing tumor cells by ACPP-HPMA copolymer-coated adenovirus conjugates.

    Directory of Open Access Journals (Sweden)

    Shuhua Li

    Full Text Available We have designed and tested a new way to selectively deliver HPMA polymer-coated adenovirus type 5 (Ad5 particles into matrix metalloproteinase (MMP-overexpressing tumor cells. An activatable cell penetrating peptide (ACPP was designed and attached to the reactive 4-nitrophenoxy groups of HPMA polymers by the C-terminal amino acid (asparagine, N. ACPPs are activatable cell penetrating peptides (CPPs with a linker between polycationic and polyanionic domains, and MMP-mediated cleavage releases the CPP portion and its attached cargo to enable cell entry. Our data indicate that the transport of these HPMA polymer conjugates by a single ACPP molecule to the cytoplasm occurs via a nonendocytotic and concentration-independent process. The uptake was observed to finish within 20 minutes by inverted fluorescence microscopy. In contrast, HPMA polymer-coated Ad5 without ACPPs was internalized solely by endocytosis. The optimal formulation was not affected by the presence of Ad5 neutralizing antibodies during transduction, and ACPP/polymer-coated Ad5 also retained high targeting capability to several MMP-overexpressing tumor cell types. For the first time, ACPP-mediated cytoplasmic delivery of polymer-bound Ad5 to MMP-overexpressing tumor cells was demonstrated. These findings are significant, as they demonstrate the use of a polymer-based system for the targeted delivery into MMP-overexpressing solid tumors and highlight how to overcome major cellular obstacles to achieve intracellular macromolecular delivery.

  18. Cell-penetrating compounds preferentially bind glycosaminoglycans over plasma membrane lipids in a charge density- and stereochemistry-dependent manner.

    Science.gov (United States)

    Prevette, Lisa E; Benish, Nicolas C; Schoenecker, Amber R; Braden, Kristin J

    2015-12-01

    Cell-penetrating compounds (CPCs) are often conjugated to drugs and genes to facilitate cellular uptake. We hypothesize that the electrostatic interaction between the positively charged amines of the cell-penetrating compounds and the negatively charged glycosaminoglycans (GAGs) extending from cell surfaces is the initiating step in the internalization process. The interactions of generation 5 PAMAM dendrimer, Tat peptide and 25 kDa linear PEI with four different GAGs have been studied using isothermal titration calorimetry to elucidate structure-function relationships that could lead to improved drug and gene delivery methods to a wide variety of cell types. Detailed thermodynamic analysis has determined that CPC-GAG binding constants range from 8.7×10(3) to 2.4×10(6)M(-1) and that affinity is dependent upon GAG charge density and stereochemistry and CPC molecular weight. The effect of GAG composition on affinity is likely due to hydrogen bonding between CPC amines and amides and GAG hydroxyl and amine groups. These results were compared to the association of CPCs with lipid vesicles of varying composition as model plasma membranes to finally clarify the relative importance of each cell surface component in initial cell recognition. CPC-lipid affinity increases with anionic lipid content, but GAG affinity is higher for all cell-penetrating compounds, confirming the role these heterogeneous polysaccharides play in cellular association and clustering.

  19. Virulence Potential of Activatable Shiga Toxin 2d–Producing Escherichia coli Isolates from Fresh Produce

    Science.gov (United States)

    Melton-Celsa, Angela R.; O'Brien, Alison D.; Feng, Peter C. H.

    2016-01-01

    Shiga toxin (Stx)–producing Escherichia coli (STEC) strains are food- and waterborne pathogens that are often transmitted via beef products or fresh produce. STEC strains cause both sporadic infections and outbreaks, which may result in hemorrhagic colitis and hemolytic uremic syndrome. STEC strains may elaborate Stx1, Stx2, and/or subtypes of those toxins. Epidemiological evidence indicates that STEC that produce subtypes Stx2a, Stx2c, and/or Stx2d are more often associated with serious illness. The Stx2d subtype becomes more toxic to Vero cells after incubation with intestinal mucus or elastase, a process named “activation.” Stx2d is not generally found in the E. coli serotypes most commonly connected to STEC outbreaks. However, STEC strains that are stx2d positive can be isolated from foods, an occurrence that gives rise to the question of whether those food isolates are potential human pathogens. In this study, we examined 14 STEC strains from fresh produce that were stx2d positive and found that they all produced the mucus-activatable Stx2d and that a subset of the strains tested were virulent in streptomycin-treated mice. PMID:26555533

  20. Virulence Potential of Activatable Shiga Toxin 2d-Producing Escherichia coli Isolates from Fresh Produce.

    Science.gov (United States)

    Melton-Celsa, Angela R; O'Brien, Alison D; Feng, Peter C H

    2015-11-01

    Shiga toxin (Stx)-producing Escherichia coli (STEC) strains are food- and waterborne pathogens that are often transmitted via beef products or fresh produce. STEC strains cause both sporadic infections and outbreaks, which may result in hemorrhagic colitis and hemolytic uremic syndrome. STEC strains may elaborate Stx1, Stx2, and/or subtypes of those toxins. Epidemiological evidence indicates that STEC that produce subtypes Stx2a, Stx2c, and/or Stx2d are more often associated with serious illness. The Stx2d subtype becomes more toxic to Vero cells after incubation with intestinal mucus or elastase, a process named "activation." Stx2d is not generally found in the E. coli serotypes most commonly connected to STEC outbreaks. However, STEC strains that are stx2d positive can be isolated from foods, an occurrence that gives rise to the question of whether those food isolates are potential human pathogens. In this study, we examined 14 STEC strains from fresh produce that were stx2d positive and found that they all produced the mucus-activatable Stx2d and that a subset of the strains tested were virulent in streptomycin-treated mice.

  1. Enhanced penetration into 3D cell culture using two and three layered gold nanoparticles

    Directory of Open Access Journals (Sweden)

    Engl

    2013-10-01

    Full Text Available Christopher G England,1 Thomas Priest,2 Guandong Zhang,2 Xinghua Sun,2 Dhruvinkumar N Patel,2 Lacey R McNally,3,4 Victor van Berkel,4,5 André M Gobin,2 Hermann B Frieboes1,2,41Department of Pharmacology and Toxicology, 2Department of Bioengineering, 3Department of Medicine, 4James Graham Brown Cancer Center, 5Department of Surgery, University of Louisville, KY, USAAbstract: Nano-scale particles sized 10–400 nm administered systemically preferentially extravasate from tumor vasculature due to the enhanced permeability and retention effect. Therapeutic success remains elusive, however, because of inhomogeneous particle distribution within tumor tissue. Insufficient tumor vascularization limits particle transport and also results in avascular hypoxic regions with non-proliferating cells, which can regenerate tissue after nanoparticle-delivered cytotoxicity or thermal ablation. Nanoparticle surface modifications provide for increasing tumor targeting and uptake while decreasing immunogenicity and toxicity. Herein, we created novel two layer gold-nanoshell particles coated with alkanethiol and phosphatidylcholine, and three layer nanoshells additionally coated with high-density-lipoprotein. We hypothesize that these particles have enhanced penetration into 3-dimensional cell cultures modeling avascular tissue when compared to standard poly(ethylene glycol (PEG-coated nanoshells. Particle uptake and distribution in liver, lung, and pancreatic tumor cell cultures were evaluated using silver-enhancement staining and hyperspectral imaging with dark field microscopy. Two layer nanoshells exhibited significantly higher uptake compared to PEGylated nanoshells. This multilayer formulation may help overcome transport barriers presented by tumor vasculature, and could be further investigated in vivo as a platform for targeted cancer therapies.Keywords: cancer nanotherapy, tumor hypoxia, nanovector transport

  2. Studies on penetration of antibiotic in bacterial cells in space conditions (7-IML-1)

    Science.gov (United States)

    Tixador, R.

    1992-01-01

    The Cytos 2 experiment was performed aboard Salyut 7 in order to test the antibiotic sensitivity of bacteria cultivated in vitro in space. An increase of the Minimal Inhibitory Concentration (MIC) in the inflight cultures (i.e., an increase of the antibiotic resistance) was observed. Complementary studies of the ultrastructure showed a thickening of the cell envelope. In order to confirm the results of the Cytos 2 experiment, we performed the ANTIBIO experiment during the D1 mission to try to differentiate, by means of the 1 g centrifuge in the Biorack, between the biological effects of cosmic rays and those caused by microgravity conditions. The originality of this experiment was in the fact that it was designed to test the antibiotic sensitivity of bacteria cultivated in vitro during the orbital phase of the flight. The results show an increase in resistance to Colistin in in-flight bacteria. The MIC is practically double in the in-flight cultures. A cell count of living bacteria in the cultures containing the different Colistin concentrations showed a significant difference between the cultures developed during space flight and the ground based cultures. The comparison between the 1 g and 0 g in-flight cultures show similar behavior for the two sets. Nevertheless, a small difference between the two sets of ground based control cultures was noted. The cultures developed on the ground centrifuge (1.4 g) present a slight decrease in comparison with the cultures developed in the static rack (1 g). In order to approach the mechanisms of the increase of antibiotic resistance on bacteria cultivated in vitro in space, we have proposed the study on penetration of antibiotics in bacterial cells in space conditions. This experiment was selected for the International Microgravity Laboratory 1 (IML-1) mission.

  3. Cellular uptake but low permeation of human calcitonin-derived cell penetrating peptides and Tat(47-57) through well-differentiated epithelial models

    DEFF Research Database (Denmark)

    Tréhin, Rachel; Krauss, Ulrike; Beck-Sickinger, Annette G;

    2004-01-01

    To investigate whether cell penetrating peptides (CPP) derived from human calcitonin (hCT) possess, in addition to cellular uptake, the capacity to deliver their cargo through epithelial barriers.......To investigate whether cell penetrating peptides (CPP) derived from human calcitonin (hCT) possess, in addition to cellular uptake, the capacity to deliver their cargo through epithelial barriers....

  4. CPPsite 2.0: a repository of experimentally validated cell-penetrating peptides.

    Science.gov (United States)

    Agrawal, Piyush; Bhalla, Sherry; Usmani, Salman Sadullah; Singh, Sandeep; Chaudhary, Kumardeep; Raghava, Gajendra P S; Gautam, Ankur

    2016-01-01

    CPPsite 2.0 (http://crdd.osdd.net/raghava/cppsite/) is an updated version of manually curated database (CPPsite) of cell-penetrating peptides (CPPs). The current version holds around 1850 peptide entries, which is nearly two times than the entries in the previous version. The updated data were curated from research papers and patents published in last three years. It was observed that most of the CPPs discovered/ tested, in last three years, have diverse chemical modifications (e.g. non-natural residues, linkers, lipid moieties, etc.). We have compiled this information on chemical modifications systematically in the updated version of the database. In order to understand the structure-function relationship of these peptides, we predicted tertiary structure of CPPs, possessing both modified and natural residues, using state-of-the-art techniques. CPPsite 2.0 also maintains information about model systems (in vitro/in vivo) used for CPP evaluation and different type of cargoes (e.g. nucleic acid, protein, nanoparticles, etc.) delivered by these peptides. In order to assist a wide range of users, we developed a user-friendly responsive website, with various tools, suitable for smartphone, tablet and desktop users. In conclusion, CPPsite 2.0 provides significant improvements over the previous version in terms of data content. PMID:26586798

  5. Biofunctionalized nanoparticles with pH-responsive and cell penetrating blocks for gene delivery

    Science.gov (United States)

    Gaspar, V. M.; Marques, J. G.; Sousa, F.; Louro, R. O.; Queiroz, J. A.; Correia, I. J.

    2013-07-01

    Bridging the gap between nanoparticulate delivery systems and translational gene therapy is a long sought after requirement in nanomedicine-based applications. However, recent developments regarding nanoparticle functionalization have brought forward the ability to synthesize materials with biofunctional moieties that mimic the evolved features of viral particles. Herein we report the versatile conjugation of both cell penetrating arginine and pH-responsive histidine moieties into the chitosan polymeric backbone, to improve the physicochemical characteristics of the native material. Amino acid coupling was confirmed by 2D TOCSY NMR and Fourier transform infrared spectroscopy. The synthesized chitosan-histidine-arginine (CH-H-R) polymer complexed plasmid DNA biopharmaceuticals, and spontaneously assembled into stable 105 nm nanoparticles with spherical morphology and positive surface charge. The functionalized delivery systems were efficiently internalized into the intracellular compartment, and exhibited remarkably higher transfection efficiency than unmodified chitosan without causing any cytotoxic effect. Additional findings regarding intracellular trafficking events reveal their preferential escape from degradative lysosomal pathways and nuclear localization. Overall, this assembly of nanocarriers with bioinspired moieties provides the foundations for the design of efficient and customizable materials for cancer gene therapy.

  6. Effects of Tryptophan Content and Backbone Spacing on the Uptake Efficiency of Cell-Penetrating Peptides

    KAUST Repository

    Rydberg, Hanna A.

    2012-07-10

    Cell-penetrating peptides (CPPs) are able to traverse cellular membranes and deliver macromolecular cargo. Uptake occurs through both endocytotic and nonendocytotic pathways, but the molecular requirements for efficient internalization are not fully understood. Here we investigate how the presence of tryptophans and their position within an oligoarginine influence uptake mechanism and efficiency. Flow cytometry and confocal fluorescence imaging are used to estimate uptake efficiency, intracellular distribution and toxicity in Chinese hamster ovarian cells. Further, membrane leakage and lipid membrane affinity are investigated. The peptides contain eight arginine residues and one to four tryptophans, the tryptophans positioned either at the N-terminus, in the middle, or evenly distributed along the amino acid sequence. Our data show that the intracellular distribution varies among peptides with different tryptophan content and backbone spacing. Uptake efficiency is higher for the peptides with four tryptophans in the middle, or evenly distributed along the peptide sequence, than for the peptide with four tryptophans at the N-terminus. All peptides display low cytotoxicity except for the one with four tryptophans at the N-terminus, which was moderately toxic. This finding is consistent with their inability to induce efficient leakage of dye from lipid vesicles. All peptides have comparable affinities for lipid vesicles, showing that lipid binding is not a decisive parameter for uptake. Our results indicate that tryptophan content and backbone spacing can affect both the CPP uptake efficiency and the CPP uptake mechanism. The low cytotoxicity of these peptides and the possibilities of tuning their uptake mechanism are interesting from a therapeutic point of view. © 2012 American Chemical Society.

  7. Development of a Cell-penetrating Peptide that Exhibits Responsive Changes in its Secondary Structure in the Cellular Environment.

    Science.gov (United States)

    Yamashita, Hiroko; Kato, Takuma; Oba, Makoto; Misawa, Takashi; Hattori, Takayuki; Ohoka, Nobumichi; Tanaka, Masakazu; Naito, Mikihiko; Kurihara, Masaaki; Demizu, Yosuke

    2016-01-01

    Cell-penetrating peptides (CPP) are received a lot of attention as an intracellular delivery tool for hydrophilic molecules such as drugs, proteins, and DNAs. We designed and synthesized nona-arginine analogues 1-5 [FAM-β-Ala-(l-Arg-l-Arg-l-Pro)3-(Gly)3-NH2 (1), FAM-β-Ala-(l-Arg-l-Arg-l-Pro(NH2))3-(Gly)3-NH2 (2), FAM-β-Ala-(l-Arg-l-Arg-l-Pro(Gu))3-(Gly)3-NH2 (3), FAM-β-Ala-(l-Arg)2-(l-Pro(Gu))2-(l-Arg)4-l-Pro(Gu)-(Gly)3-NH2 (4), and FAM-β-Ala-(l-Arg)6-(l-Pro(Gu))3-(Gly)3-NH2 (5)] containing l-proline (l-Pro) or cationic proline derivatives (l-Pro(NH2) and l-Pro(Gu)), and investigated their cell-penetrating abilities. Interestingly, only peptide 3 having the side-chain guanidinyl l-Pro(Gu) exhibited a secondary structural change in cellular environment. Specifically, peptide 3 formed a random structure in hydrophilic conditions, whereas it formed a helical structure under amphipathic conditions. Furthermore, during cellular permeability tests, peptide 3 demonstrated greater cell-penetrating activity than other peptides and effectively transported plasmid DNA into HeLa cells. Thus, l-Pro(Gu)-containing peptide 3 may be a useful candidate as a gene delivery carrier. PMID:27609319

  8. Development of a Cell-penetrating Peptide that Exhibits Responsive Changes in its Secondary Structure in the Cellular Environment

    Science.gov (United States)

    Yamashita, Hiroko; Kato, Takuma; Oba, Makoto; Misawa, Takashi; Hattori, Takayuki; Ohoka, Nobumichi; Tanaka, Masakazu; Naito, Mikihiko; Kurihara, Masaaki; Demizu, Yosuke

    2016-01-01

    Cell-penetrating peptides (CPP) are received a lot of attention as an intracellular delivery tool for hydrophilic molecules such as drugs, proteins, and DNAs. We designed and synthesized nona-arginine analogues 1–5 [FAM-β-Ala-(l-Arg-l-Arg-l-Pro)3-(Gly)3-NH2 (1), FAM-β-Ala-(l-Arg-l-Arg-l-ProNH2)3-(Gly)3-NH2 (2), FAM-β-Ala-(l-Arg-l-Arg-l-ProGu)3-(Gly)3-NH2 (3), FAM-β-Ala-(l-Arg)2-(l-ProGu)2-(l-Arg)4-l-ProGu-(Gly)3-NH2 (4), and FAM-β-Ala-(l-Arg)6-(l-ProGu)3-(Gly)3-NH2 (5)] containing l-proline (l-Pro) or cationic proline derivatives (l-ProNH2 and l-ProGu), and investigated their cell-penetrating abilities. Interestingly, only peptide 3 having the side-chain guanidinyl l-ProGu exhibited a secondary structural change in cellular environment. Specifically, peptide 3 formed a random structure in hydrophilic conditions, whereas it formed a helical structure under amphipathic conditions. Furthermore, during cellular permeability tests, peptide 3 demonstrated greater cell-penetrating activity than other peptides and effectively transported plasmid DNA into HeLa cells. Thus, l-ProGu-containing peptide 3 may be a useful candidate as a gene delivery carrier. PMID:27609319

  9. Conjugation of a cell-penetrating peptide to parathyroid hormone affects its structure, potency, and transepithelial permeation

    DEFF Research Database (Denmark)

    Kristensen, Mie; de Groot, Anne Marit; Berthelsen, Jens;

    2015-01-01

    hormone, i.e. PTH(1-34), and to evaluate the effect with regards to secondary structure, potency in Saos-2 cells, immunogenicity, safety as well as the transepithelial permeation across monolayers by using the Caco-2 cell culture model. Further, co-administration of CPP and PTH(1-34) as an alternative......Delivery of therapeutic peptides and proteins by the use of cell-penetrating peptides (CPPs) as carriers has been suggested as a feasible strategy. The aim of the present study was to investigate the effect of conjugating a series of well-known CPPs to the biologically active part of parathyroid...

  10. Effect of the infectious laryngotracheitis virus (ILTV) glycoprotein G on virus attachment, penetration, growth curve and direct cell-to-cell spread

    Institute of Scientific and Technical Information of China (English)

    SUN Zhaogang; ZHANG Manfu

    2005-01-01

    The secreted alphaherpesvirus glycoprotein G (gG) works differently from other proteins. Analysis of the role of ILTV gG in virus attachment, penetration, direct cell-to-cell spread (CTCS) and the growth curve showed that gG or its antibody had no effect on ILTV attachment and penetration and that the gG antibody reduced the virus plaque size and the one-step growth curve on chicken embryo liver (CEL) cells, but gG did not affect the virus plaque size or the one-step growth curve on CEL cells. Laser scanning confocal microscopy (LSCM) detection showed that ILTV gG is located in the perinuclear region and the membrane of the CEL cells. These results suggested that ILTV gG might contribute to direct cell-to-cell transmission.

  11. Dataset on FAP-induced emergence of spontaneous metastases and on the preparation of activatable FAP-targeting immunoliposomes to detect the metastases.

    Science.gov (United States)

    Tansi, Felista L; Rüger, Ronny; Böhm, Claudia; Kontermann, Roland E; Teichgraeber, Ulf K; Fahr, Alfred; Hilger, Ingrid

    2016-12-01

    The underlying data demonstrates that fibroblast activation protein (FAP) paves the way for fibrosarcoma cells, which require the proteolysis of the extracellular matrix (ECM) and basement membranes to intravasate from implanted subcutaneous primary tumors into blood vessels, be transported to distant organs where they extravasate from the blood vessels, reattach and proliferate to metastases. The data additionally shows that FAP, when overexpressed on fibrosarcoma cells induces their invasion and formation of spontaneous metastases in multiple organs, particularly after subcutaneous co-implantation of the FAP-expressing and wildtype fibrosarcoma. The raw and processed data presented herein is related to a research article entitled "Potential of activatable FAP-targeting immunoliposomes in intraoperative imaging of spontaneous metastases" (F.L. Tansi, R. Rüger, C. Böhm, R.E. Kontermann, U.K. Teichgraeber, A. Fahr, I. Hilger, 2016) [1]. Furthermore, evidence for the detection of FAP-expressing tumor cells and cells of the tumor stroma by activatable FAP-targeting liposomes is presented in this dataset.

  12. Follicular penetration and targeting.

    Science.gov (United States)

    Lademann, Jürgen; Otberg, Nina; Jacobi, Ute; Hoffman, Robert M; Blume-Peytavi, Ulrike

    2005-12-01

    In the past, intercellular penetration was assumed to be the most important penetration pathway of topically applied substances. First hints that follicular penetration needs to be taken into consideration were confirmed by recent investigations, presented during the workshop "Follicular Penetration and Targeting" at the 4th Intercontinental Meeting of Hair Research Societies", in Berlin 2004. Hair follicles represent an efficient reservoir for the penetration of topically applied substances with subsequent targeting of distinct cell populations, e.g., nestin-expressing follicular bulge cells. The volume of this reservoir can be determined by differential stripping technology. The follicular penetration processes are significantly influenced by the state of the follicular infundibulum; recent experimental investigations could demonstrate that it is essential to distinguish between open and closed hair follicles. Topically applied substances can only penetrate into open hair follicle. Knowledge of follicular penetration is of high clinical relevance for functional targeting of distinct follicular regions. Human hair follicles show a hair-cycle-dependent variation of the dense neuronal and vascular network. Moreover, during hair follicle cycling with initiation of anagen, newly formed vessels occur. Thus, the potential of nestin-expressing hair follicle stem cells to form neurons and blood vessels was investigated.

  13. Effects of Lipid Composition on the Entry of Cell-Penetrating Peptide Oligoarginine into Single Vesicles.

    Science.gov (United States)

    Sharmin, Sabrina; Islam, Md Zahidul; Karal, Mohammad Abu Sayem; Alam Shibly, Sayed Ul; Dohra, Hideo; Yamazaki, Masahito

    2016-08-01

    The cell-penetrating peptide R9, an oligoarginine comprising nine arginines, has been used to transport biological cargos into cells. However, the mechanisms underlying its translocation across membranes remain unclear. In this report, we investigated the entry of carboxyfluorescein (CF)-labeled R9 (CF-R9) into single giant unilamellar vesicles (GUVs) of various lipid compositions and the CF-R9-induced leakage of a fluorescent probe, Alexa Fluor 647 hydrazide (AF647), using a method developed recently by us. First, we investigated the interaction of CF-R9 with dioleoylphosphatidylglycerol (DOPG)/dioleoylphosphatidylcholine (DOPC) GUVs containing AF647 and small DOPG/DOPC vesicles. The fluorescence intensity of the GUV membrane due to CF-R9 (i.e., the rim intensity) increased with time to a steady-state value, and then the fluorescence intensity of the membranes of the small vesicles in the GUV lumen increased without leakage of AF647. This result indicates that CF-R9 entered the GUV lumen from the outside by translocating across the lipid membrane without forming pores through which AF647 could leak. The fraction of entry of CF-R9 at 6 min in the absence of pore formation, Pentry (6 min), increased with an increase in CF-R9 concentration, but the CF-R9 concentration in the lumen was low. We obtained similar results for dilauroyl-PG (DLPG)/ditridecanoyl-PC (DTPC) (2/8) GUVs. The values of Pentry (6 min) of CF-R9 for DLPG/DTPC (2/8) GUVs were larger than those obtained with DOPG/DOPC (2/8) GUVs at the same CF-R9 concentrations. In contrast, a high concentration of CF-R9 induced pores in DLPG/DTPC (4/6) GUVs through which CF-R9 entered the GUV lumen, so the CF-R9 concentration in the lumen was higher. However, CF-R9 could not enter DOPG/DOPC/cholesterol (2/6/4) GUVs. Analysis of the rim intensity showed that CF-R9 was located only in the outer monolayer of the DOPG/DOPC/cholesterol (2/6/4) GUVs. On the basis of analyses of these results, we discuss the elementary

  14. riDOM, a cell-penetrating peptide. Interaction with DNA and heparan sulfate.

    Science.gov (United States)

    Québatte, Gabriela; Kitas, Eric; Seelig, Joachim

    2013-09-19

    DNA condensation in the presence of polycationic molecules is a well-known phenomenon exploited in gene delivery. riDOM (retro-inverso dioleoylmelittin) is a cell-penetrating peptide with excellent transporter properties for DNA. It is a chimeric molecule where ri-melittin is fused to dioleoylphosphoethanolamine. The physical-chemical properties of riDOM in solution and in the presence of DNA and heparan sulfate were investigated with spectroscopic and thermodynamic methods. Dynamic light scattering shows that riDOM in solution aggregates to well-defined nanoparticles with a diameter of ∼13 nm and a ζ-potential of 22 mV, composed of about 220-270 molecules. Binding of riDOM to DNA was studied with dynamic light scattering, ζ-potential measurements, and isothermal titration calorimetry and was compared with authentic melittin-DNA interaction. riDOM binds tightly to DNA with a microscopic binding constant of 5 × 10(7) M(-1) and a stoichiometry of 12 riDOM per 10 DNA base pairs. In the complex the DNA double strand is completely shielded by the more hydrophobic riDOM molecules. Authentic melittin binds to DNA with a much lower binding constant of 5 × 10(6) M(-1) and lower stoichiometry of 5 melittin per 10 DNA base pairs. The binding enthalpies for riDOM and melittin are small and the binding reactions are entropy-driven. Sulfated glycosaminoglycans such as heparan sulfate are also linear molecules with a negative charge. riDOM binding to heparan sulfate on cell surfaces can therefore interfere with DNA-riDOM binding. riDOM-heparan sulfate complex formation was characterized by isothermal titration calorimetry and spectroscopic methods. The binding constant of riDOM for heparan sulfate is K ≈ 2 × 10(6) M(-1). Authentic melittin has a similar binding constant but riDOM shows a 3-fold higher packing density on heparan sulfate than the distinctly smaller melittin.

  15. The role of thrombin-activatable fibrinolysis inhibitor in diabetic wound healing

    NARCIS (Netherlands)

    C.J.N. Verkleij; J.J.T.H. Roelofs; S.R. Havik; J.C.M. Meijers; P.F. Marx

    2010-01-01

    Introduction: One of the major complications in patients with diabetes mellitus is impaired wound healing. The fibrinolytic system is involved in parts of the wound healing process and deficiency of thrombin-activatable fibrinolysis inhibitor (TAFI) results in delayed wound closure. Moreover, levels

  16. Quantitative & qualitative analysis of endothelial cells of donor cornea before & after penetrating keratoplasty in different pathological conditions

    Directory of Open Access Journals (Sweden)

    Aruna K.R. Gupta

    2016-01-01

    Full Text Available Background & objectives: Endothelial cells of the donor cornea are known to be affected quantitatively and qualitatively in different pathological conditions after penetrating keratoplasty (PK and this has direct effect on the clarity of vision obtained after PK. This study was undertaken to analyze the qualitative and quantitative changes in donor endothelial cells before and after PK in different pathological conditions. Methods: A prospective investigational analysis of 100 consecutive donor corneas used for penetrating keratoplasty between June 2006 and June 2008, was conducted. The patients were evaluated on the first day, at the end of first week, first month, third and six months and one year. Results: A decrease was observed in endothelial cell count in all pathological conditions. After one year of follow up the loss was 33.1 per cent in corneal opacity, 45.9 per cent in acute infective keratitis (AIK, 58.5 per cent in regrafts, 28.5 per cent in pseudophakic bullous keratopathy (PBK, 37 per cent in descemetocele, 27 per cent in keratoconus and 35.5 per cent in aphakic bullous keratopathy (ABK cases. Interpretation & conclusions: The endothelial cell loss was highest in regraft cases which was significant (P<0.05, while the least endothelial cell loss was seen in keratoconus cases. The cell loss was associated with increase in coefficient of variation (CV, i.e. polymegathism and pleomorphism. Inspite of this polymegathism and pleomorphism, the clarity of the graft was maintained.

  17. Enhancing tumor-specific intracellular delivering efficiency of cell-penetrating peptide by fusion with a peptide targeting to EGFR.

    Science.gov (United States)

    Nguyen, Long The; Yang, Xu-Zhong; Du, Xuan; Wang, Jia-Wei; Zhang, Rui; Zhao, Jian; Wang, Fu-Jun; Dong, Yang; Li, Peng-Fei

    2015-05-01

    Cell-penetrating peptides (CPPs) are well known as intracellular delivery vectors. However, unsatisfactory delivery efficiency and poor specificity are challenging barriers to CPP applications at the clinical trial stage. Here, we showed that S3, an EGFR-binding domain derived from vaccinia virus growth factor, when fused to a CPP such as HBD or TAT can substantially enhance its internalization efficiency and tumor selectivity. The uptake of S3-HBD (S3H) recombinant molecule by tumor cells was nearly 80 folds increased compared to HBD alone. By contrast, the uptake of S3H by non-neoplastic cells still remained at a low level. The specific recognition between S3 and its receptor, EGFR, as well as between HBD and heparan sulfate proteoglycans on the cell surface was essential for these improvements, suggesting a syngeneic effect between the two functional domains in conjugation. This syngeneic effect is likely similar to that of the heparin-binding epidermal growth factor, which is highly abundant particularly in metastatic tumors. The process that S3H entered cells was dependent on time, dosage, and energy, via macropinocytosis pathway. With excellent cell-penetrating efficacy and a novel tumor-targeting ability, S3H appears as a promising candidate vector for targeted anti-cancer drug delivery. PMID:25655386

  18. Enhanced anticancer activity of nanopreparation containing an MMP2-sensitive PEG-drug conjugate and cell-penetrating moiety.

    Science.gov (United States)

    Zhu, Lin; Wang, Tao; Perche, Federico; Taigind, Anton; Torchilin, Vladimir P

    2013-10-15

    In response to the challenges of cancer chemotherapeutics, including poor physicochemical properties, low tumor targeting, insufficient tumor cell internalization/bioavailability, and side effects, we developed a unique tumor-targeted micellar drug-delivery platform. Using paclitaxel as a model therapeutic, a nanopreparation composed of a matrix metalloproteinase 2 (MMP2)-sensitive self-assembly PEG 2000-paclitaxel conjugate (as a prodrug and MMP 2-sensitive moiety), transactivating transcriptional activator peptide-PEG1000-phosphoethanolamine (PE) (a cell-penetrating enhancer), and PEG1000-PE (a nanocarrier building block) was prepared. Several major drug delivery strategies, including self-assembly, PEGylation, the enhanced permeability and retention effect, stimulus sensitivity, a cell-penetrating moiety, and the concept of prodrug, were used in design of this nanoparticle in a collaborative manner. The nanopreparation allowed superior cell internalization, cytotoxicity, tumor targeting, and antitumor efficacy in vitro and in vivo over its nonsensitive counterpart, free paclitaxel and conventional micelles. This uniquely engineered nanoparticle has potential for effective intracellular delivery of drug into cancer cells. PMID:24062440

  19. Recombinant expression and purification of a MAP30-cell penetrating peptide fusion protein with higher anti-tumor bioactivity.

    Science.gov (United States)

    Lv, Qiang; Yang, Xu-Zhong; Fu, Long-Yun; Lu, Yv-Ting; Lu, Yan-Hua; Zhao, Jian; Wang, Fu-Jun

    2015-07-01

    MAP30 (Momordica Antiviral Protein 30 Kd), a single-stranded type-I ribosome inactivating protein, possesses versatile biological activities including anti-tumor abilities. However, the low efficiency penetrating into tumor cells hampers the tumoricidal effect of MAP30. This paper describes MAP30 fused with a human-derived cell penetrating peptide HBD which overcome the low uptake efficiency by tumor cells and exhibits higher anti-tumor bioactivity. MAP30 gene was cloned from the genomic DNA of Momordica charantia and the recombinant plasmid pET28b-MAP30-HBD was established and transferred into Escherichia coli BL21 (DE3). The recombinant MAP30-HBD protein (rMAP30-HBD) was expressed in a soluble form after being induced by 0.5mM IPTG for 14h at 15°C. The recombinant protein was purified to greater than 95% purity with Ni-NTA affinity chromatography. The rMAP30-HBD protein not only has topological inactivation and protein translation inhibition activity but also showed significant improvements in cytotoxic activity compared to that of the rMAP30 protein without HBD in the tested tumor cell lines, and induced higher apoptosis rates in HeLa cells analyzed by Annexin V-FITC with FACS. This paper demonstrated a new method for improving MAP30 protein anti-tumor activity and might have potential applications in cancer therapy area.

  20. Recombinant expression and purification of a MAP30-cell penetrating peptide fusion protein with higher anti-tumor bioactivity.

    Science.gov (United States)

    Lv, Qiang; Yang, Xu-Zhong; Fu, Long-Yun; Lu, Yv-Ting; Lu, Yan-Hua; Zhao, Jian; Wang, Fu-Jun

    2015-07-01

    MAP30 (Momordica Antiviral Protein 30 Kd), a single-stranded type-I ribosome inactivating protein, possesses versatile biological activities including anti-tumor abilities. However, the low efficiency penetrating into tumor cells hampers the tumoricidal effect of MAP30. This paper describes MAP30 fused with a human-derived cell penetrating peptide HBD which overcome the low uptake efficiency by tumor cells and exhibits higher anti-tumor bioactivity. MAP30 gene was cloned from the genomic DNA of Momordica charantia and the recombinant plasmid pET28b-MAP30-HBD was established and transferred into Escherichia coli BL21 (DE3). The recombinant MAP30-HBD protein (rMAP30-HBD) was expressed in a soluble form after being induced by 0.5mM IPTG for 14h at 15°C. The recombinant protein was purified to greater than 95% purity with Ni-NTA affinity chromatography. The rMAP30-HBD protein not only has topological inactivation and protein translation inhibition activity but also showed significant improvements in cytotoxic activity compared to that of the rMAP30 protein without HBD in the tested tumor cell lines, and induced higher apoptosis rates in HeLa cells analyzed by Annexin V-FITC with FACS. This paper demonstrated a new method for improving MAP30 protein anti-tumor activity and might have potential applications in cancer therapy area. PMID:25797209

  1. Cell penetrable humanized-VH/V(H)H that inhibit RNA dependent RNA polymerase (NS5B) of HCV.

    Science.gov (United States)

    Thueng-in, Kanyarat; Thanongsaksrikul, Jeeraphong; Srimanote, Potjanee; Bangphoomi, Kunan; Poungpair, Ornnuthchar; Maneewatch, Santi; Choowongkomon, Kiattawee; Chaicumpa, Wanpen

    2012-01-01

    NS5B is pivotal RNA dependent RNA polymerase (RdRp) of HCV and NS5B function interfering halts the virus infective cycle. This work aimed to produce cell penetrable humanized single domain antibodies (SdAb; VH/V(H)H) that interfere with the RdRp activity. Recombinant NS5BΔ55 of genotype 3a HCV with de novo RNA synthetic activity was produced and used in phage biopanning for selecting phage clones that displayed NS5BΔ55 bound VH/V(H)H from a humanized-camel VH/V(H)H display library. VH/V(H)H from E. coli transfected with four selected phage clones inhibited RdRp activity when tested by ELISA inhibition using 3'di-cytidylate 25 nucleotide directed in vitro RNA synthesis. Deduced amino acid sequences of two clones showed V(H)H hallmark and were designated V(H)H6 and V(H)H24; other clones were conventional VH, designated VH9 and VH13. All VH/V(H)H were linked molecularly to a cell penetrating peptide, penetratin. The cell penetrable VH9, VH13, V(H)H6 and V(H)H24 added to culture of Huh7 cells transfected with JHF-1 RNA of genotype 2a HCV reduced the amounts of RNA intracellularly and in culture medium implying that they inhibited the virus replication. VH/V(H)H mimotopes matched with residues scattered on the polymerase fingers, palm and thumb which were likely juxtaposed to form conformational epitopes. Molecular docking revealed that the antibodies covered the RdRp catalytic groove. The transbodies await further studies for in vivo role in inhibiting HCV replication.

  2.   Cell Penetrating Peptoids (CPPos: Synthesis of a Small Combinatorial Library by Using IRORI MiniKans

    Directory of Open Access Journals (Sweden)

    Dominik K. Kölmel

    2012-11-01

    Full Text Available Cell penetrating peptoids (CPPos are potent mimics of the corresponding cell penetrating peptides (CPPs. The synthesis of diverse oligomeric libraries that display a variety of backbone scaffolds and side-chain appendages are a very promising source of novel CPPos, which can be used to either target different cellular organelles or even different tissues and organs. In this study we established the submonomer-based solid phase synthesis of a “proof of principle” peptoid library in IRORI MiniKans to expand the amount for phenotypic high throughput screens of CPPos. The library consisting of tetrameric peptoids [oligo(N-alkylglycines] was established on Rink amide resin in a split and mix approach with hydrophilic and hydrophobic peptoid side chains. All CPPos of the presented library were labeled with rhodamine B to allow for the monitoring of cellular uptake by fluorescent confocal microscopy. Eventually, all the purified peptoids were subjected to live cell imaging to screen for CPPos with organelle specificity. While highly charged CPPos enter the cells by endocytosis with subsequent endosomal release, critical levels of lipophilicity allow other CPPos to specifically localize to mitochondria once a certain lipophilicity threshold is reached.

  3. Cell-penetrating peptide-doxorubicin conjugate loaded NGR-modified nanobubbles for ultrasound triggered drug delivery.

    Science.gov (United States)

    Lin, Wen; Xie, Xiangyang; Deng, Jianping; Liu, Hui; Chen, Ying; Fu, Xudong; Liu, Hong; Yang, Yang

    2016-01-01

    A new drug-targeting system for CD13(+) tumors has been developed, based on ultrasound-sensitive nanobubbles (NBs) and cell-permeable peptides (CPPs). Here, the CPP-doxorubicin conjugate (CPP-DOX) was entrapped in the asparagine-glycine-arginine (NGR) peptide modified NB (CPP-DOX/NGR-NB) and the penetration of CPP-DOX was temporally masked; local ultrasound stimulation could trigger the CPP-DOX release from NB and activate its penetration. The CPP-DOX/NGR-NBs had particle sizes of about 200 nm and drug entrapment efficiency larger than 90%. In vitro release results showed that over 85% of the encapsulated DOX or CPP-DOX would release from NBs in the presence of ultrasound, while less than 1.5% of that (30 min) without ultrasound. Cell experiments showed the higher cellular CPP-DOX uptake of CPP-DOX/NGR-NB among the various NB formulations in Human fibrosarcoma cells (HT-1080, CD13(+)). The CPP-DOX/NGR-NB with ultrasound treatment exhibited an increased cytotoxic activity than the one without ultrasound. In nude mice xenograft of HT-1080 cells, CPP-DOX/NGR-NB with ultrasound showed a higher tumor inhibition effect (3.1% of T/C%, day 24), longer median survival time (50 days) and excellent body safety compared with the normal DOX injection group. These results indicate that the constructed vesicle would be a promising drug delivery system for specific cancer treatment.

  4. Intracellular delivery of cell-penetrating peptide-transcriptional factor fusion protein and its role in selective osteogenesis

    Directory of Open Access Journals (Sweden)

    Suh JS

    2014-03-01

    Full Text Available Jin Sook Suh,1,* Jue Yeon Lee,2,* Yoon Jung Choi,1 Hyung Keun You,3 Seong-Doo Hong,4 Chong Pyoung Chung,2 Yoon Jeong Park1,2 1Dental Regenerative Biotechnology, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, 2Central Research Institute, Nano Intelligent Biomedical Engineering Corporation (NIBEC, Seoul, 3Department of Periodontology, College of Dentistry, Wonkwang University, Iksan, 4Department of Oral Pathology, School of Dentistry, Seoul National University, Seoul, Republic of Korea *These authors contributed equally to this work Abstract: Protein-transduction technology has been attempted to deliver macromolecular materials, including protein, nucleic acids, and polymeric drugs, for either diagnosis or therapeutic purposes. Herein, fusion protein composed of an arginine-rich cell-penetrating peptide, termed low-molecular-weight protamine (LMWP, and a transcriptional coactivator with a PDZ-binding motif (TAZ protein was prepared and applied in combination with biomaterials to increase bone-forming capacity. TAZ has been recently identified as a specific osteogenic stimulating transcriptional coactivator in human mesenchymal stem cell (hMSC differentiation, while simultaneously blocking adipogenic differentiation. However, TAZ by itself cannot penetrate the cells, and thus needs a transfection tool for translocalization. The LMWP-TAZ fusion proteins were efficiently translocalized into the cytosol of hMSCs. The hMSCs treated with cell-penetrating LMWP-TAZ exhibited increased expression of osteoblastic genes and protein, producing significantly higher quantities of mineralized matrix compared to free TAZ. In contrast, adipogenic differentiation of the hMSCs was blocked by treatment of LMWP-TAZ fusion protein, as reflected by reduced marker-protein expression, adipocyte fatty acid-binding protein 2, and peroxisome proliferator-activated receptor-γ messenger ribonucleic acid levels. LMWP-TAZ was applied in

  5. Cell-Penetrating, Guanidinium-Rich Oligophosphoesters: Effective and Versatile Molecular Transporters for Drug and Probe Delivery.

    Science.gov (United States)

    McKinlay, Colin J; Waymouth, Robert M; Wender, Paul A

    2016-03-16

    The design, synthesis, and biological evaluation of a new family of highly effective cell-penetrating molecular transporters, guanidinium-rich oligophosphoesters, are described. These unique transporters are synthesized in two steps, irrespective of oligomer length, by the organocatalytic ring-opening polymerization (OROP) of 5-membered cyclic phospholane monomers followed by oligomer deprotection. Varying the initiating alcohol results in a wide variety of cargo attachment strategies for releasable or nonreleasable transporter applications. Initiation of oligomerization with a fluorescent probe produces, upon deprotection, a transporter-probe conjugate that is shown to readily enter multiple cell lines in a dose-dependent manner. These new transporters are superior in cell uptake to previously studied guanidinium-rich oligocarbonates and oligoarginines, showing over 2-fold higher uptake than the former and 6-fold higher uptake than the latter. Initiation with a protected thiol gives, upon deprotection, thiol-terminated transporters which can be thiol-click conjugated to a variety of probes, drugs and other cargos as exemplified by the conjugation and delivery of the model probe fluorescein-maleimide and the medicinal agent paclitaxel (PTX) into cells. Of particular significance given that drug resistance is a major cause of chemotherapy failure, the PTX-transporter conjugate, designed to evade Pgp export and release free PTX after cell entry, shows efficacy against PTX-resistant ovarian cancer cells. Collectively this study introduces a new and highly effective class of guanidinium-rich cell-penetrating transporters and methodology for their single-step conjugation to drugs and probes, and demonstrates that the resulting drug/probe-conjugates readily enter cells, outperforming previously reported guanidinium-rich oligocarbonates and peptide transporters. PMID:26900771

  6. 细胞穿透肽的研究进展%Recent advances in research on cell penetrating peptides

    Institute of Scientific and Technical Information of China (English)

    李凤英; 何京; 钟海军

    2013-01-01

    细胞穿透肽(cell penetrating peptides,CPP)是一类能够通过生物膜进入细胞的短肽,它具有一些其他运载系统无法比拟的优点:低浓度条件下,可以穿过细胞膜进入细胞并且不会对膜造成明显破坏和损伤;能介导各种物质包括小分子、核酸、蛋白多肽以及纳米粒子等入胞;高效、低毒.本文就CPP的分类、与载物的连接方式、穿膜机制、应用和常用研究方法等方面进行系统的综述,并对CPP的临床应用前景进行展望.%Cell penetrating peptides (CPP) are a class of peptides of less than 35 amino acids with a remarkable capacity for membrane transportation.Compared with other delivery systems,the CPP-based system has several advantages.These peptides are able to penetrate into cells at low micromolar concentrations in vitro and in vivo without causing significant membrane damage.Furthermore,they constitute very promising tools for cellular import of various substances including small molecules,nucleic acids,proteins and nanoparticles.In this review,the categories,chemical linkage with cargoes,cellular uptake mechanism,applications and research methods of CPP are systematically summarized.The research future of CPP,especially the future of its clinical application,is also introduced.

  7. Combined effect of a peptide–morpholino oligonucleotide conjugate and a cell-penetrating peptide as an antibiotic

    OpenAIRE

    Wesolowski, Donna; Alonso, Dulce; Altman, Sidney

    2013-01-01

    A cell-penetrating peptide (CPP)–morpholino oligonucleotide (MO) conjugate (PMO) that has an antibiotic effect in culture had some contaminating CPPs in earlier preparations. The mixed conjugate had gene-specific and gene-nonspecific effects. An improved purification procedure separates the PMO from the free CPP and MO. The gene-specific effects are a result of the PMO, and the nonspecific effects are a result of the unlinked, unreacted CPP. The PMO and the CPP can be mixed together, as has b...

  8. Transduction of proteins into leishmania tarentolae by formation of non-covalent complexes with cell-penetrating peptides.

    Science.gov (United States)

    Keller, Andrea-Anneliese; Breitling, Reinhard; Hemmerich, Peter; Kappe, Katarina; Braun, Maria; Wittig, Berith; Schaefer, Buerk; Lorkowski, Stefan; Reissmann, Siegmund

    2014-02-01

    Cell-penetrating peptides (CPPs) are used to transport peptides, proteins, different types of ribonucleic acids (or mimics of these molecules), and DNA into live cells, both plant and mammalian. Leishmania belongs to the class of protozoa having, in comparison to mammalian cells, a different lipid composition of the membrane, proteoglycans on the surface, and signal pathways. We investigated the uptake of two different and easily detectable proteins into the non-pathogenic strain Leishmania tarentolae. From the large number of CPPs available, six and a histone were chosen specifically for their ability to form non-covalent complexes. For Leishmania we used the enzyme β-galactosidase and fluorescent labeled bovine serum albumin as cargoes. The results are compared to similar internalization studies using mammalian cells [Mussbach et al., ]. Leishmania cells can degrade CPPs by a secreted and membrane-bound chymotrypsin-like protease. Both cargo proteins were internalized with sufficient efficiency and achieved intramolecular concentrations similar to mammalian cells. The transport efficiencies of the CPPs differed from each other, and showed a different rank order for both cargoes. The intracellular distribution of fluorescent-labeled bovine serum albumin showed highest concentrations in the nucleus and kinetoplast. Leishmania are susceptible to high concentrations of some CPPs, although comparably dissimilar to mammalian cells. MPG-peptides are more cytotoxic in Leishmania than in mammalian cells, acting as antimicrobial peptides. Our results contribute to a better understanding of molecular interactions in Leishmania cells and possibly to new treatments of leishmaniasis.

  9. A novel cell-penetrating peptide derived from WT1 enhances p53 activity, induces cell senescence and displays antimelanoma activity in xeno- and syngeneic systems☆

    Science.gov (United States)

    Massaoka, Mariana H.; Matsuo, Alisson L.; Figueiredo, Carlos R.; Girola, Natalia; Faria, Camyla F.; Azevedo, Ricardo A.; Travassos, Luiz R.

    2014-01-01

    The Wilms tumor protein 1 (WT1) transcription factor has been associated in malignant melanoma with cell survival and metastasis, thus emerging as a candidate for targeted therapy. A lysine–arginine rich peptide, WT1-pTj, derived from the ZF domain of WT1 was evaluated as an antitumor agent against A2058 human melanoma cells and B16F10-Nex2 syngeneic murine melanoma. Peptide WT1-pTj quickly penetrated human melanoma cells and induced senescence, recognized by increased SA-β-galactosidase activity, enhanced transcriptional activity of p53, and induction of the cell cycle inhibitors p21 and p27. Moreover, the peptide bound to p53 and competed with WT1 protein for binding to p53. WT1-pTj treatment led to sustained cell growth suppression, abrogation of clonogenicity and G2/M cell cycle arrest. Notably, in vivo studies showed that WT1-pTj inhibited both the metastases and subcutaneous growth of murine melanoma in syngeneic mice, and prolonged the survival of nude mice challenged with human melanoma cells. The 27-amino acid cell-penetrating WT1-derived peptide, depends on C3 and H16 for effective antimelanoma activity, inhibits proliferation of WT1-expressing human tumor cell lines, and may have an effective role in the treatment of WT1-expressing malignancies. PMID:24490140

  10. A novel cell-penetrating peptide derived from WT1 enhances p53 activity, induces cell senescence and displays antimelanoma activity in xeno- and syngeneic systems

    Directory of Open Access Journals (Sweden)

    Mariana H. Massaoka

    2014-01-01

    Full Text Available The Wilms tumor protein 1 (WT1 transcription factor has been associated in malignant melanoma with cell survival and metastasis, thus emerging as a candidate for targeted therapy. A lysine–arginine rich peptide, WT1-pTj, derived from the ZF domain of WT1 was evaluated as an antitumor agent against A2058 human melanoma cells and B16F10-Nex2 syngeneic murine melanoma. Peptide WT1-pTj quickly penetrated human melanoma cells and induced senescence, recognized by increased SA-β-galactosidase activity, enhanced transcriptional activity of p53, and induction of the cell cycle inhibitors p21 and p27. Moreover, the peptide bound to p53 and competed with WT1 protein for binding to p53. WT1-pTj treatment led to sustained cell growth suppression, abrogation of clonogenicity and G2/M cell cycle arrest. Notably, in vivo studies showed that WT1-pTj inhibited both the metastases and subcutaneous growth of murine melanoma in syngeneic mice, and prolonged the survival of nude mice challenged with human melanoma cells. The 27-amino acid cell-penetrating WT1-derived peptide, depends on C3 and H16 for effective antimelanoma activity, inhibits proliferation of WT1-expressing human tumor cell lines, and may have an effective role in the treatment of WT1-expressing malignancies.

  11. Cell- and nuclear-penetrating anti-dsDNA autoantibodies have multiple arginines in CDR3 of VH and increase cellular level of pERK and Bcl-2 in mesangial cells.

    Science.gov (United States)

    Im, Sae-Ran; Im, Sun-Woo; Chung, Hee-Yong; Pravinsagar, Pavithra; Jang, Young-Ju

    2015-10-01

    Investigation of characteristics of cell- and nuclear-penetrating anti-double stranded (ds)DNA autoantibodies (autoAbs) is important to understand pathogenesis of lupus nephritis, but has not been clearly explored. The present study reports that three anti-dsDNA monoclonal autoAbs, which contain more than two arginine residues in their CDR3s of variable heavy domain (VH), penetrated into living murine mesangial cells and the cell nuclei. However, an anti-dsDNA monoclonal Ab (mAb) having only one arginine in the CDR3-VH did not penetrate cells. To assess the contribution of antigen-binding sites, especially the VH, in cell- and nuclear-penetration, we evaluated the characteristics of recombinant single chain Fv(scFv), VH, and variable light domain (VL) of a penetrating mAb. The scFv and VH domain, containing arginine in CDR3-VH maintained the ability to penetrate cells and the cell nuclei, whereas the VL domain, having no arginine in CDR3, did not penetrate cells. The penetratingm Abs, scFv, and VH activated ERK and increased cellular protein levels of Bcl-2, whereas the non-penetrating Ab and VL did not. The cell survival was decreased by the penetrating mAbs, scFv and VH, not by the non-penetrating mAb and VL. The data indicate that an antigen-binding site is required for cell-penetration and that positively-charged arginine residues in CDR3-VH contribute to the cell- and nuclear-penetrating ability of a subset of anti-dsDNA autoAbs. Furthermore,the nuclear-penetrating anti-dsDNA autoAbs could possibly function as a pathogenic factor for lupus nephritis by up-regulating ERK activation and Bcl-2 production in mesangial cells. The cell- and nuclear-penetrating VH domain may be exploited as a vehicle for the intra cellular delivery of various useful molecules.

  12. Conjugation of doxorubicin to cell penetrating peptides sensitizes human breast MDA-MB 231 cancer cells to endogenous TRAIL-induced apoptosis.

    OpenAIRE

    Aroui, Sonia; Brahim, Souhir; Hamelin, Jocelyne; De Waard, Michel; Bréard, Jacqueline; Kenani, Abderraouf

    2009-01-01

    International audience Previous work from our laboratory has shown that coupling doxorubicin (Dox) to cell penetrating peptides (Dox-CPPs) is a good strategy to overcome Dox resistance in MDA-MB 231 breast cancer cells. We also reported that, in contrast to unconjugated Dox-induced cell death, the increase in apoptotic response does not involve the mitochondrial apoptotic pathway. In this study, we demonstrate that both Dox and Dox-CPPs can increase the density of the TRAIL receptors DR4 a...

  13. Cell-penetration by Co(III)cyclen-based peptide-cleaving catalysts selective for pathogenic proteins of amyloidoses.

    Science.gov (United States)

    Chei, Woo Suk; Lee, Joo-Won; Kim, Jae Bum; Suh, Junghun

    2010-07-15

    Derivatives of the Co(III) complex of 1,4,7,10-tetraazacyclododecane (cyclen) with various organic pendants have been reported as target-selective peptide-cleaving catalysts, which can be exploited as catalytic drugs. In order to provide a firm basis for the catalytic drugs based on Co(III)cyclen, the ability of the Co(III)cyclen-containing peptide-cleaving catalysts to penetrate animal cells such as mouse fibroblast NIH-3T 3 or human embryonic kidney (HEK) 293 cells is demonstrated in the present study. Since the catalysts destroy pathogenic proteins for amyloidoses, results of the present study are expected to initiate extensive efforts to obtain therapeutically safe catalytic drugs for amyloidoses such as Alzheimer's disease, type 2 diabetes mellitus, Parkinson's disease, Huntington's disease, mad cow disease, and so on. PMID:20542701

  14. Stearylated antimicrobial peptide [D]-K6L9 with cell penetrating property for efficient gene transfer.

    Science.gov (United States)

    Zhang, Wei; Song, Jingjing; Liang, Ranran; Zheng, Xin; Chen, Jianbo; Li, Guolin; Zhang, Bangzhi; Wang, Kairong; Yan, Xiang; Wang, Rui

    2013-08-01

    Stearyl-cell penetrating peptides (CPPs) have been proved to be efficient nonviral gene vectors. Due to the similarities between antimicrobial peptides and CPPs, we constructed a novel type of gene vectors by introducing stearyl moiety to the N-terminus of antimicrobial peptide [D]-K6L9. In this study, stearyl-[D]-K6L9 delivered plasmids into cells by clathrin- and caveolin-mediated endocytosis. Gratifyingly, stearyl-[D]-K6L9 exhibited high transfection efficiency and almost reached the level of Lipofectamine 2000. Taken together, the combination of the stearyl moiety with [D]-K6L9 provides a novel framework for the development of excellent nonviral gene vectors. PMID:23727033

  15. Applications and Challenges for Use of Cell-Penetrating Peptides as Delivery Vectors for Peptide and Protein Cargos

    DEFF Research Database (Denmark)

    Kristensen, Mie; Birch, Ditlev; Mørck Nielsen, Hanne

    2016-01-01

    -penetrating peptides (CPPs) constitute a promising tool and have shown applications for peptide and protein delivery into cells as well as across various epithelia and the blood-brain barrier (BBB). CPP-mediated delivery of peptides and proteins may be pursued via covalent conjugation of the CPP to the cargo peptide...... or protein or via physical complexation obtained by simple bulk-mixing of the CPP with its cargo. Both approaches have their pros and cons, and which is the better choice likely relates to the physicochemical properties of the CPP and its cargo as well as the route of administration, the specific...... barrier and the target cell. Besides the physical barrier, a metabolic barrier must be taken into consideration when applying peptide-based delivery vectors, such as the CPPs, and stability-enhancing strategies are commonly employed to prolong the CPP half-life. The mechanisms by which CPPs translocate...

  16. Peptide based DNA nanocarriers incorporating a cell-penetrating peptide derived from neurturin protein and poly-L-lysine dendrons.

    Science.gov (United States)

    Rosli, Nurlina; Christie, Michelle P; Moyle, Peter M; Toth, Istvan

    2015-05-15

    Multicomponent gene delivery systems incorporating cell-penetrating peptides (CPP) from the human neurturin protein (NRTN-30, NRTN(132-161); NRTN-17, NRTN(145-161)) and a poly-l-lysine (PLL) dendron, were synthesized and characterized for plasmid DNA (pDNA) delivery. Acetylated NRTN peptides (Ac-CPP) and peptides conjugated to a PLL dendron (DEN-CPP) efficiently condensed and stabilized pDNA. Complexes between pDNA and DEN-CPP formed smaller and more stable nanoparticles. Flow cytometry experiments showed that pDNA-DEN-CPPs were taken up more efficiently into HeLa cells. There was also no significant difference between NRTN-30 and NRTN-17 for pDNA uptake, indicating that the truncated peptide alone is sufficient as a CPP for pDNA delivery.

  17. Cell-penetrating peptide derived from human eosinophil cationic protein inhibits mite allergen Der p 2 induced inflammasome activation.

    Directory of Open Access Journals (Sweden)

    Sheng-Jie Yu

    Full Text Available Newly discovered cell penetration peptides derived from human eosinophil cationic proteins (CPPecp have the characteristic of cell internalization, but the effect of CPPecp on immunomodulation has not been clarified. House dust mite (HDM major allergen, Der p 2, can induce proinflammatory cytokine production which contributes to airway inflammation and allergic asthma. However, the mechanism of Der p 2 on NLRP3 inflammasome activation remains unclear. The aim of this study was to investigate the immunomodulatory effect of CPPecp on inhibition of Der p 2 induced inflammasome activation. We showed that proinflammatory cytokines IL-1β, IL-6 and IL-8 were significantly upregulated in peripheral blood mononuclear cells (PBMCs derived from HDM allergic patients after Der p 2 stimulation. Expression of NLRP3, ASC, Caspase-1, IL-1β and Caspase-1 activity was upregulated in THP-1 cells after Der p 2 stimulation. Proinflammatory cytokine production, NLRP3 inflammasome activation and caspase-1 activity were downregulated in THP-1 cells and CD14+ cells co-cultured with Der p 2 and CPPecp. The immunomodulatory effect of CPPecp was through upregulation of IFN-α production but not induction of autophagy. These results suggested Der p 2 plays an important role in NLRP3 inflammasome activation and CPPecp has the potential to be a novel anti-inflammatory agent for allergic inflammation treatment in the future.

  18. Effect of chirality and length on the penetrability of single-walled carbon nanotubes into lipid bilayer cell membranes.

    Science.gov (United States)

    Skandani, A Alipour; Zeineldin, R; Al-Haik, M

    2012-05-22

    The ability of carbon nanotubes to enter the cell membrane acting as drug-delivery vehicles has yielded a plethora of experimental investigations, mostly with inconclusive results because of the wide spectra of carbon nanotube structures. Because of the virtual impossibility of synthesizing CNTs with distinct chirality, we report a parametric study on the use of molecular dynamics to provide better insight into the effect of the carbon nanotube chirality and the aspect ratio on the interaction with a lipid bilayer membrane. The simulation results indicated that a single-walled carbon nanotube utilizes different time-evolving mechanisms to facilitate their internalization within the membrane. These mechanisms comprise both penetration and endocytosis. It was observed that carbon nanotubes with higher aspect ratios penetrate the membrane faster whereas shorter nanotubes undergo significant rotation during the final stages of endocytosis. Furthermore, nanotubes with lower chiral indices developed significant adhesion with the membrane. This adhesion is hypothesized to consume some of the carbon nanotube energy, thus resulting in longer times for the nanotube to translocate through the membrane.

  19. Enhancement of intracellular concentration and biological activity of PNA after conjugation with a cell-penetrating synthetic model peptide.

    Science.gov (United States)

    Oehlke, Johannes; Wallukat, Gerd; Wolf, Yvonne; Ehrlich, Angelika; Wiesner, Burkhard; Berger, Hartmut; Bienert, Michael

    2004-07-01

    In order to evaluate the ability of the cell-penetrating alpha-helical amphipathic model peptide KLALKLALKALKAALKLA-NH(2) (MAP) to deliver peptide nucleic acids (PNAs) into mammalian cells, MAP was covalently linked to the 12-mer PNA 5'-GGAGCAGGAAAG-3' directed against the mRNA of the nociceptin/orphanin FQ receptor. The cellular uptake of both the naked PNA and its MAP-conjugate was studied by means of capillary electrophoresis combined with laser-induced fluorescence detection, confocal laser scanning microscopy and fluorescence-activated cell sorting. Incubation with the fluorescein-labelled PNA-peptide conjugate led to three- and eightfold higher intracellular concentrations in neonatal rat cardiomyocytes and CHO cells, respectively, than found after exposure of the cells to the naked PNA. Correspondingly, pretreatment of spontaneously-beating neonatal rat cardiomyocytes with the PNA-peptide conjugate and the naked PNA slowed down the positive chronotropic effect elicited by the neuropeptide nociceptin by 10- and twofold, respectively. The main reasons for the higher bioavailability of the PNA-peptide conjugate were found to be a more rapid cellular uptake in combination with a lowered re-export and resistance against influences of serum.

  20. Distal phenylalanine modification for enhancing cellular delivery of fluorophores, proteins and quantum dots by cell penetrating peptides.

    Science.gov (United States)

    Sayers, E J; Cleal, K; Eissa, N G; Watson, P; Jones, A T

    2014-12-10

    For cell penetrating peptides (CPPs) to fulfil their promise as effective delivery vectors we need a better understanding of their mechanisms of cell binding and uptake. This is especially the case when they are linked to different types of cargo. Here we describe new studies based on our previous findings suggesting that, for peptide-CPP chimeras, distal hydrophobic residues upstream of the CPP sequence can have profound effects on the way they interact with cells. We studied peptides bearing an N-terminal Glycine or Phenylalanine linked via a neutral and flexible bridging group, SGSGSGSG, to three well-studied CPPs: octaarginine, penetratin and TP10. Using a combination of flow cytometry, live-cell imaging and image analysis we examined the effects of this single amino acid change on binding and uptake of Alexa488-fluorophore, bovine serum albumin and quantum dot cargoes. The influence of the glycine-phenylalanine switch for fluorophore delivery was most dramatic in TP10, increasing cellular uptake by 4.4 and 9.9 fold in non-adherent and adherent cells, respectively. Only penetratin showed effective uptake of bovine serum albumin with the phenylalanine variant showing an increase of 1.6 fold over the glycine variant. The uptake of quantum dots was most efficiently demonstrated by octaarginine, with the glycine variant increasing uptake 4.8 fold and the phenylalanine variant increasing uptake 9.5 fold over quantum dots alone. Overall the data demonstrate that hydrophobicity distal to the CPP could be utilised to enhance their capacity to bind to the cell membrane and deliver a range of macromolecules to the insides of cells.

  1. Ultra-small lipid nanoparticles promote the penetration of coenzyme Q10 in skin cells and counteract oxidative stress.

    Science.gov (United States)

    Lohan, Silke B; Bauersachs, Sonja; Ahlberg, Sebastian; Baisaeng, Nuttakorn; Keck, Cornelia M; Müller, Rainer H; Witte, Ellen; Wolk, Kerstin; Hackbarth, Steffen; Röder, Beate; Lademann, Jürgen; Meinke, Martina C

    2015-01-01

    UV irradiation leads to the formation of reactive oxygen species (ROS). An imbalance between the antioxidant system and ROS can lead to cell damage, premature skin aging or skin cancer. To counteract these processes, antioxidants such as coenzyme Q10 (CoQ10) are contained in many cosmetics. To improve and optimize cell/tissue penetration properties of the lipophilic CoQ10, ultra-small lipid nanoparticles (usNLC) were developed. The antioxidant effectiveness of CoQ10-loaded usNLC compared to conventional nanocarriers was investigated in the human keratinocyte cell line HaCaT. Using confocal laser scanning microscopy investigations of the carriers additionally loaded with nile red showed a clear uptake into cells and their distribution within the cytoplasm. By use of the XTT cell viability test, CoQ10 concentrations of 10-50 μg/ml were shown to be non-toxic, and the antioxidant potential of 10 μg/ml CoQ10 loaded usNLC in the HaCaT cells was analyzed via electron paramagnetic resonance spectroscopy after cellular exposure to UVA (1J/cm(2)) and UVB (18 mJ/cm(2)) irradiation. In comparison with the CoQ10-loaded conventional carriers, usNLC-CoQ10 demonstrated the strongest reduction of the radical formation; reaching up to 23% compared to control cells without nanocarrier treatment. Therefore, usNLC-CoQ10 are very suitable to increase the antioxidant potential of skin.

  2. Cell Penetrable Human scFv Specific to Middle Domain of Matrix Protein-1 Protects Mice from Lethal Influenza

    Directory of Open Access Journals (Sweden)

    Fonthip Dong-din-on

    2015-01-01

    Full Text Available A new anti-influenza remedy that can tolerate the virus antigenic variation is needed. Influenza virus matrix protein-1 (M1 is highly conserved and pivotal for the virus replication cycle: virus uncoating, assembly and budding. An agent that blocks the M1 functions should be an effective anti-influenza agent. In this study, human scFv that bound to recombinant M1 middle domain (MD and native M1 of A/H5N1 was produced. Phage mimotope search and computerized molecular docking revealed that the scFv bound to the MD conformational epitope formed by juxtaposed helices 7 and 9 of the M1. The scFv was linked molecularly to a cell penetrable peptide, penetratin (PEN. The PEN-scFv (transbody, when used to treat the cells pre-infected with the heterologous clade/subclade A/H5N1 reduced the viral mRNA intracellularly and in the cell culture fluids. The transbody mitigated symptom severity and lung histopathology of the H5N1 infected mice and caused reduction of virus antigen in the tissues as well as extricated the animals from the lethal challenge in a dose dependent manner. The transbody specific to the M1 MD, either alone or in combination with the cognate human scFvs specific to other influenza virus proteins, should be an effective, safe and mutation tolerable anti-influenza agent.

  3. Spotlight on Human LL-37, an Immunomodulatory Peptide with Promising Cell-Penetrating Properties

    Directory of Open Access Journals (Sweden)

    Marc Ferdinand Lensink

    2010-11-01

    Full Text Available Cationic antimicrobial peptides are major components of innate immunity and help control the initial steps of the infectious process. They are expressed not only by immunocytes, but also by epithelial cells. They share an amphipathic secondary structure with a polar cationic site, which explains their tropism for prokaryote membranes and their hydrophobic site contributing to the destructuration of these membranes. LL-37 is the only cationic antimicrobial peptide derived from human cathelicidin. LL-37 can also cross the plasma membrane of eukaryotic cells, probably through special domains of this membrane called lipid rafts. This transfer could be beneficial in the context of vaccination: the activation of intracellular toll-like receptors by a complex formed between CpG oligonucleotides and LL-37 could conceivably play a major role in the building of a cellular immunity involving NK cells.

  4. Two functional motifs define the interaction, internalization and toxicity of the cell-penetrating antifungal peptide PAF26 on fungal cells.

    Directory of Open Access Journals (Sweden)

    Alberto Muñoz

    Full Text Available The synthetic, cell penetrating hexapeptide PAF26 (RKKWFW is antifungal at low micromolar concentrations and has been proposed as a model for cationic, cell-penetrating antifungal peptides. Its short amino acid sequence facilitates the analysis of its structure-activity relationships using the fungal models Neurospora crassa and Saccharomyces cerevisiae, and human and plant pathogens Aspergillus fumigatus and Penicillium digitatum, respectively. Previously, PAF26 at low fungicidal concentrations was shown to be endocytically internalized, accumulated in vacuoles and then actively transported into the cytoplasm where it exerts its antifungal activity. In the present study, two PAF26 derivatives, PAF95 (AAAWFW and PAF96 (RKKAAA, were designed to characterize the roles of the N-terminal cationic and the C-terminal hydrophobic motifs in PAF26's mode-of-action. PAF95 and PAF96 exhibited substantially reduced antifungal activity against all the fungi analyzed. PAF96 localized to fungal cell envelopes and was not internalized by the fungi. In contrast, PAF95 was taken up into vacuoles of N. crassa, wherein it accumulated and was trapped without toxic effects. Also, the PAF26 resistant Δarg1 strain of S. cerevisiae exhibited increased PAF26 accumulation in vacuoles. Live-cell imaging of GFP-labelled nuclei in A. fumigatus showed that transport of PAF26 from the vacuole to the cytoplasm was followed by nuclear breakdown and dissolution. This work demonstrates that the amphipathic PAF26 possesses two distinct motifs that allow three stages in its antifungal action to be defined: (i its interaction with the cell envelope; (ii its internalization and transport to vacuoles mediated by the aromatic hydrophobic domain; and (iii its transport from vacuoles to the cytoplasm. Significantly, cationic residues in PAF26 are important not only for the electrostatic attraction and interaction with the fungal cell but also for transport from the vacuole to the

  5. Cell penetrating silica nanoparticles doped with two-photon absorbing fluorophores

    NARCIS (Netherlands)

    Bertazza, Loris; Celotti, Lucia; Fabbrini, Graziano; Loi, Maria Antonietta; Maggini, Michele; Mancin, Fabrizio; Marcuz, Silvia; Menna, Enzo; Muccini, Michele; Tonellato, Umberto

    2006-01-01

    Organosilica nanoparticles, doped with two-photon absorbing distyrylbenzene derivatives, were prepared and studied as cell staining agents. Two dyes were used, bearing either two peripheral dimethylamino groups or one dimethylamino and one cyano group. Due to the internal charge transfer character o

  6. Research Progress of Cell -penetrating Peptides Applied in Vaccine%穿膜肽在疫苗中应用的研究进展

    Institute of Scientific and Technical Information of China (English)

    范兴琼; 吴双林; 刘成倩; 易建中

    2012-01-01

    Since the discovery of the function of protein transduction, cell- penetrating peptides (CPP) provides a novel approach for gene therapy and vaccine development. According to recent studies on the CPP species, structure, membrane -penetrating mechanisms and relative applications, cell -penetrating peptides applied in the DNA vaccine and tumor vaccine were reviewed and prospected.%穿膜肽自发现以来,因其独特的蛋白转导功能,为基因治疗和新型疫苗的开发等开辟了新的途径。通过综述穿膜肽的种类、结构特点、作用机制及其在疫苗中的应用,为进一步研发DNA疫苗和肿瘤疫苗奠定基础。

  7. Penetration testing

    OpenAIRE

    Zemaníková, Martina

    2010-01-01

    This work focuses on the practical demonstration of design penetration testing. Testing is carried out based on the order of a particular entity/subject which is at their request anonymous. The work is divided into two parts, theoretical and practical. We will be made in first part familiarize with the process and techniques of testing, as is now used in ethical hacking, then we can find in the end appropriate recommendations 6 how to prevent and fight against it in order to protect the...

  8. [Cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy].

    Science.gov (United States)

    Tan, Jiao; Wang, Ya-Ping; Wang, Hui-Xin; Liang, Jian-Ming; Zhang, Meng; Sun, Xun; Huang, Yong-Zhuo

    2014-12-01

    To develop a cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy, we prepared the AVPI-LMWP/pTRAIL self-assembled complexes containing a therapeutic combination of peptide drug AVPI and DNA drug TRAIL. The chimeric apoptotic peptide AVPI-LMWP was synthesized using the standard solid-phase synthesis. The cationic AVPI-LMWP could condense pTRAIL by electrostatic interaction. The physical-chemical properties of the AVPI-LMWP/pTRAIL complexes were characterized. The cellular uptake efficiency and the inhibitory activity of the AVPI-LMWP/pTRAIL complexes on tumor cell were also performed. The results showed that the AVPI-LMWP/pTRAIL complexes were successfully prepared by co-incubation. With the increase of mass ratio (AVPI-LMWP/DNA), the particle size was decreased and the zeta potential had few change. Agarose gel electrophoresis showed that AVPI-LMWP could fully bind and condense pTRAIL at a mass ratio above 15:1. Cellular uptake efficiency was improved along with the increased ratio of W(AVPI-LMWP)/WpTRAIL. The in vitro cytotoxicity experiments demonstrated that the AVPI-LMWP/pTRAIL (W:W = 20:1) complexes was significantly more effective than the pTRAIL, AVPI-LMWP alone or LMWP/pTRAIL complexes on inhibition of HeLa cell growth. Our studies indicated that the AVPI-LMWP/pTRAIL co-delivery system could deliver plasmid into HeLa cell and induce tumor cell apoptosis efficiently, which showed its potential in cancer therapy using combination of apoptoic peptide and gene drugs.

  9. The use of nanoscale fluorescence microscopic to decipher cell wall modifications during fungal penetration

    OpenAIRE

    Dorothea eEllinger; Christian eVoigt

    2014-01-01

    Plant diseases are one of the most studied subjects in the field of plant science due to their impact on crop yield and food security. Our increased understanding of plant–pathogen interactions was mainly driven by the development of new techniques that facilitated analyses on a subcellular and molecular level. The development of labeling technologies, which allowed the visualization and localization of cellular structures and proteins in live cell imaging, promoted the use of fluorescence an...

  10. Titanium Dioxide Nanoparticle Penetration into the Skin and Effects on HaCaT Cells.

    Science.gov (United States)

    Crosera, Matteo; Prodi, Andrea; Mauro, Marcella; Pelin, Marco; Florio, Chiara; Bellomo, Francesca; Adami, Gianpiero; Apostoli, Pietro; De Palma, Giuseppe; Bovenzi, Massimo; Campanini, Marco; Filon, Francesca Larese

    2015-08-07

    Titanium dioxide nanoparticles (TiO2NPs) suspensions (concentration 1.0 g/L) in synthetic sweat solution were applied on Franz cells for 24 h using intact and needle-abraded human skin. Titanium content into skin and receiving phases was determined. Cytotoxicity (MTT, AlamarBlue(®) and propidium iodide, PI, uptake assays) was evaluated on HaCat keratinocytes after 24 h, 48 h, and seven days of exposure. After 24 h of exposure, no titanium was detectable in receiving solutions for both intact and damaged skin. Titanium was found in the epidermal layer after 24 h of exposure (0.47 ± 0.33 μg/cm(2)) while in the dermal layer, the concentration was below the limit of detection. Damaged skin, in its whole, has shown a similar concentration (0.53 ± 0.26 μg/cm(2)). Cytotoxicity studies on HaCaT cells demonstrated that TiO2NPs induced cytotoxic effects only at very high concentrations, reducing cell viability after seven days of exposure with EC50s of 8.8 × 10(-4) M (MTT assay), 3.8 × 10(-5) M (AlamarBlue(®) assay), and 7.6 × 10(-4) M (PI uptake, index of a necrotic cell death). Our study demonstrated that TiO2NPs cannot permeate intact and damaged skin and can be found only in the stratum corneum and epidermis. Moreover, the low cytotoxic effect observed on human HaCaT keratinocytes suggests that these nano-compounds have a potential toxic effect at the skin level only after long-term exposure.

  11. Enhanced cellular delivery of cell-penetrating peptide-peptide nucleic acid conjugates by photochemical internalization

    DEFF Research Database (Denmark)

    Shiraishi, Takehiko; Nielsen, Peter E

    2011-01-01

    )) or tetraphenylporphyrin tetrasulfonic acid (TPPS). Cellular uptake of the PNA conjugates were evaluated by using a sensitive cellular method with HeLa pLuc705 cells based on the splicing correction of luciferase gene by targeting antisense oligonucleotides to a cryptic splice site of the mutated luciferase gene....... The cellular efficacy of CPP conjugates were evaluated by measuring luciferase activity as a result of splicing correction and was also confirmed by RT-PCR analysis of luciferase pre-mRNA....

  12. Titanium Dioxide Nanoparticle Penetration into the Skin and Effects on HaCaT Cells

    Directory of Open Access Journals (Sweden)

    Matteo Crosera

    2015-08-01

    Full Text Available Titanium dioxide nanoparticles (TiO2NPs suspensions (concentration 1.0 g/L in synthetic sweat solution were applied on Franz cells for 24 h using intact and needle-abraded human skin. Titanium content into skin and receiving phases was determined. Cytotoxicity (MTT, AlamarBlue® and propidium iodide, PI, uptake assays was evaluated on HaCat keratinocytes after 24 h, 48 h, and seven days of exposure. After 24 h of exposure, no titanium was detectable in receiving solutions for both intact and damaged skin. Titanium was found in the epidermal layer after 24 h of exposure (0.47 ± 0.33 μg/cm2 while in the dermal layer, the concentration was below the limit of detection. Damaged skin, in its whole, has shown a similar concentration (0.53 ± 0.26 μg/cm2. Cytotoxicity studies on HaCaT cells demonstrated that TiO2NPs induced cytotoxic effects only at very high concentrations, reducing cell viability after seven days of exposure with EC50s of 8.8 × 10−4 M (MTT assay, 3.8 × 10−5 M (AlamarBlue® assay, and 7.6 × 10−4 M (PI uptake, index of a necrotic cell death. Our study demonstrated that TiO2NPs cannot permeate intact and damaged skin and can be found only in the stratum corneum and epidermis. Moreover, the low cytotoxic effect observed on human HaCaT keratinocytes suggests that these nano-compounds have a potential toxic effect at the skin level only after long-term exposure.

  13. Antitumor activity of tripterine via cell-penetrating peptide-coated nanostructured lipid carriers in a prostate cancer model

    Directory of Open Access Journals (Sweden)

    Yuan L

    2013-11-01

    Full Text Available Ling Yuan,1 Congyan Liu,2 Yan Chen,2 Zhenhai Zhang,2 Lei Zhou,1 Ding Qu2 1Department of Pharmaceutics, School of Pharmacy, Jiangsu University, Zhenjiang, Jiangsu, 2Key Laboratory of New Drug Delivery System of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Nanjing, Jiangsu, People's Republic of China Background: The purpose of this study was to evaluate the antitumor effect of cell-penetrating peptide-coated tripterine-loaded nanostructured lipid carriers (CT-NLC on prostate tumor cells in vitro and in vivo. Methods: CT-NLC were developed to improve the hydrophilicity of tripterine. The antiproliferative effects of CT-NLC, tripterine-loaded nanostructured lipid carriers (T-NLC, and free tripterine in a human prostatic carcinoma cell line (PC-3 and a mouse prostate carcinoma cell line (RM-1 were evaluated using an MTT assay. The advantage of CT-NLC over T-NLC and free tripterine with regard to antitumor activity in vivo was evaluated in a prostate tumor-bearing mouse model. The induced tumor necrosis factor-alpha and interleukin-6 cytokine content was investigated by enzyme-linked immunosorbent assay to determine the effect of CT-NLC, T-NLC, and free tripterine on immune responses. Histologic and TUNEL assays were carried out to investigate the mechanisms of tumor necrosis and apoptosis. Results: CT-NLC, T-NLC, and free tripterine showed high antiproliferative activity in a dose-dependent manner, with an IC50 of 0.60, 0.81, and 1.02 µg/mL in the PC-3 cell line and 0.41, 0.54, and 0.89 µg/mL in the RM-1 cell line after 36 hours. In vivo, the tumor inhibition rates for cyclophosphamide, high-dose (4 mg/kg and low-dose (2 mg/kg tripterine, high-dose (4 mg/kg and low-dose (2 mg/kg T-NLC, high-dose (4 mg/kg and low-dose (2 mg/kg CT-NLC were 76.51%, 37.07%, 29.53%, 63.56%, 48.25%, 72.68%, and 54.50%, respectively, showing a dose-dependent pattern. The induced tumor necrosis factor-alpha and interleukin-6 cytokine content

  14. Study on the Penetrability of PEP-1-P27mt for Cell Membranes in Vitro

    Institute of Scientific and Technical Information of China (English)

    YAN Shirong; ZHU Minglei; QIU Fangcheng; WANG Lilin; QU Shen

    2007-01-01

    Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, prokaryotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS transformed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentrations in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological protocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.

  15. Comparison of Franz cells and microdialysis for assessing salicylic acid penetration through human skin.

    Science.gov (United States)

    Leveque, N; Makki, S; Hadgraft, J; Humbert, Ph

    2004-01-28

    The purpose of the present study was to compare Franz cells (FC) and microdialysis (MD) for monitoring the skin absorption of salicylic acid (SA). The influence of pH on SA flux was also assessed by these two techniques. Excised abdominal human skin was used in the experiments. SA was dissolved in phosphate buffer solutions of pH 2, 5 and 7 (2 mg/ml). SA concentrations in the receptor FC solutions and in MD samples were assessed by high performance liquid chromatography (HPLC). The results demonstrate that the flux of SA decreased with increased pH. The profiles permeation determined by Franz cells and microdialysis were similar. However, whatever the pH, the SA flux was higher with microdialysis than with Franz cells. The results showed that SA percutaneous permeation conformed to the pH partition hypothesis. The flux of SA was different when it was determined by the two techniques. The collect of SA, by these two techniques is different. The results of the two techniques are compared and discussed.

  16. Antibody tumor penetration

    Science.gov (United States)

    Thurber, Greg M.; Schmidt, Michael M.; Wittrup, K. Dane

    2009-01-01

    Antibodies have proven to be effective agents in cancer imaging and therapy. One of the major challenges still facing the field is the heterogeneous distribution of these agents in tumors when administered systemically. Large regions of untargeted cells can therefore escape therapy and potentially select for more resistant cells. We present here a summary of theoretical and experimental approaches to analyze and improve antibody penetration in tumor tissue. PMID:18541331

  17. State of the Art in the Studies on Crotamine, a Cell Penetrating Peptide from South American Rattlesnake

    Directory of Open Access Journals (Sweden)

    Irina Kerkis

    2014-01-01

    Full Text Available Animal venoms comprise a naturally selected cocktail of bioactive peptides/proteins and other molecules, each of which playing a defined role thanks to the highly specific interactions with diverse molecular targets found in the prey. Research focused on isolation, structural, and functional characterizations of novel natural biologics (bioactive peptides/proteins from natural sources has a long way to go through from the basic science to clinical applications. Herein, we overview the structural and functional characteristics of the myoneurotoxin crotamine, firstly isolated from the South American rattlesnake venom. Crotamine is the first venom peptide classified as a natural cell penetrating and antimicrobial peptide (CPP and AMP with a more pronounced antifungal activity. In contrast to other known natural CPPs and AMPs, crotamine demonstrates a wide spectrum of biological activities with potential biotechnological and therapeutic values. More recent studies have demonstrated the selective in vitro anticancer activity of crotamine. In vivo, using a murine melanoma model, it was shown that crotamine delays tumor implantation, inhibits tumor cells proliferation, and also increases the survival of mice engrafted with subcutaneous melanoma. The structural and functional properties and also the possible biotechnological applications of minimized molecules derived from crotamine are also discussed.

  18. Improvement on light penetrability and microalgae biomass production by periodically pre-harvesting Chlorella vulgaris cells with culture medium recycling.

    Science.gov (United States)

    Huang, Yun; Sun, Yahui; Liao, Qiang; Fu, Qian; Xia, Ao; Zhu, Xun

    2016-09-01

    To improve light penetrability and biomass production in batch cultivation, a cultivation mode that periodically pre-harvesting partial microalgae cells from suspension with culture medium recycling was proposed. By daily pre-harvesting 30% microalgae cells from the suspension, the average light intensity in the photobioreactor (PBR) was enhanced by 27.05-122.06%, resulting in a 46.48% increase in total biomass production than that cultivated in batch cultivation without pre-harvesting under an incident light intensity of 160μmolm(-2)s(-1). Compared with the semi-continuous cultivation with 30% microalgae suspension daily replaced with equivalent volume of fresh medium, nutrients and water input was reduced by 60% in the proposed cultivation mode but with slightly decrease (12.82%) in biomass production. No additional nutrient was replenished when culture medium recycling. Furthermore, higher pre-harvesting ratios (40%, 60%) and lower pre-harvesting frequencies (every 2, 2.5days) were not advantageous for the pre-harvesting cultivation mode. PMID:27289058

  19. Synergistic Enhancement of Antitumor Efficacy by PEGylated Multi-walled Carbon Nanotubes Modified with Cell-Penetrating Peptide TAT

    Science.gov (United States)

    Hu, Shanshan; Wang, Tong; Pei, Xibo; Cai, He; Chen, Junyu; Zhang, Xin; Wan, Qianbing; Wang, Jian

    2016-10-01

    In the present study, a cell-penetrating peptide, the transactivating transcriptional factor (TAT) domain from HIV, was linked to PEGylated multi-walled carbon nanotubes (MWCNTs) to develop a highly effective antitumor drug delivery system. FITC was conjugated on MWCNTs-polyethylene glycol (PEG) and MWCNTs-PEG-TAT to provide fluorescence signal for tracing the cellular uptake of the nanocarrier. After loaded with an anticancer agent, doxorubicin (DOX) via π - π stacking interaction, the physicochemical characteristics, release profile and biological evaluation of the obtained nano-sized drug carrier were investigated. The DOX loaded MWCNTs-PEG and MWCNTs-PEG-TAT drug carriers both displayed appropriate particle size, excellent stability, high drug loading, and pH-dependent drug release profile. Nevertheless, compared with DOX-MWCNTs-PEG, DOX-MWCNTs-PEG-TAT showed improved cell internalization, intracellular distribution and potentiated anticancer efficacy due to the TAT-mediated membrane translocation, endosomal escape and nuclear targeting. Furthermore, the therapeutic efficacy of DOX was not compromised after being conjugated with MWCNTs-PEG-TAT and the proposed nanocarrier was also confirmed to have a good biocompatibility. In conclusion, our results suggested that the unique combination of TAT and MWCNTs as a multifunctional drug delivery system might be a powerful tool for improved anticancer drug development.

  20. Electroporation-based delivery of cell-penetrating peptide conjugates of peptide nucleic acids for antisense inhibition of intracellular bacteria.

    Science.gov (United States)

    Ma, Sai; Schroeder, Betsy; Sun, Chen; Loufakis, Despina Nelie; Cao, Zhenning; Sriranganathan, Nammalwar; Lu, Chang

    2014-10-01

    Cell penetrating peptides (CPPs) have been used for a myriad of cellular delivery applications and were recently explored for delivery of antisense agents such as peptide nucleic acids (PNAs) for bacterial inhibition. Although these molecular systems (i.e. CPP-PNAs) have shown ability to inhibit growth of bacterial cultures in vitro, they show limited effectiveness in killing encapsulated intracellular bacteria in mammalian cells such as macrophages, presumably due to difficulty involved in the endosomal escape of the reagents. In this report, we show that electroporation delivery dramatically increases the bioavailability of CPP-PNAs to kill Salmonella enterica serovar Typhimurium LT2 inside macrophages. Electroporation delivers the molecules without involving endocytosis and greatly increases the antisense effect. The decrease in the average number of Salmonella per macrophage under a 1200 V cm(-1) and 5 ms pulse was a factor of 9 higher than that without electroporation (in an experiment with a multiplicity of infection of 2 : 1). Our results suggest that electroporation is an effective approach for a wide range of applications involving CPP-based delivery. The microfluidic format will allow convenient functional screening and testing of PNA-based reagents for antisense applications.

  1. Biochemical characterization of bovine plasma thrombin-activatable fibrinolysis inhibitor (TAFI)

    DEFF Research Database (Denmark)

    Valnickova, Zuzana; Thaysen-Andersen, Morten; Højrup, Peter;

    2009-01-01

    BACKGROUND: TAFI is a plasma protein assumed to be an important link between coagulation and fibrinolysis. The three-dimensional crystal structures of authentic mature bovine TAFI (TAFIa) in complex with tick carboxypeptidase inhibitor, authentic full lenght bovine plasma thrombin-activatable fib......BACKGROUND: TAFI is a plasma protein assumed to be an important link between coagulation and fibrinolysis. The three-dimensional crystal structures of authentic mature bovine TAFI (TAFIa) in complex with tick carboxypeptidase inhibitor, authentic full lenght bovine plasma thrombin...

  2. Recurrent invasive squamous cell carcinoma of the ocular surface requiring penetrating therapeutic sclerokeratoplasty

    Directory of Open Access Journals (Sweden)

    Mark J. Mannis

    2012-12-01

    Full Text Available Purpose: We review a case of invasive squamous cell carcinoma invading the cornea to discuss optimal management. Methods:  Observational case report with histopathologic analysis. Results: Histopathology demonstrates corneal invasion by the tumor that appears to have been completely excised with a large therapeutic keratoplasty and adjuvant cryotherapy. Conclusions: Successful management of ocular surface squamous neoplasia (OSSN requires removal of identifiably abnormal tissue without disruption of normal protective architecture, careful histopathologic analysis, and the employment of adjuvant therapy at the time of or subsequent to surgical excision.

  3. Titanium Dioxide Nanoparticle Penetration into the Skin and Effects on HaCaT Cells

    OpenAIRE

    Matteo Crosera; Andrea Prodi; Marcella Mauro; Marco Pelin; Chiara Florio; Francesca Bellomo; Gianpiero Adami; Pietro Apostoli; Giuseppe Palma; Massimo Bovenzi; Marco Campanini; Francesca Larese Filon

    2015-01-01

    Titanium dioxide nanoparticles (TiO2NPs) suspensions (concentration 1.0 g/L) in synthetic sweat solution were applied on Franz cells for 24 h using intact and needle-abraded human skin. Titanium content into skin and receiving phases was determined. Cytotoxicity (MTT, AlamarBlue® and propidium iodide, PI, uptake assays) was evaluated on HaCat keratinocytes after 24 h, 48 h, and seven days of exposure. After 24 h of exposure, no titanium was detectable in receiving solutions for both intact an...

  4. Functionalization with C-terminal cysteine enhances transfection efficiency of cell-penetrating peptides through dimer formation

    International Nuclear Information System (INIS)

    Highlights: ► Reversible CPP dimerisation is a simple yet efficient strategy to improve delivery. ► Dimer formation enhances peptiplex stability, resulting in increased transfection. ► By dimerisation, the CPP EB1 even gain endosomal escape properties while lowering cytotoxicity. -- Abstract: Cell-penetrating peptides have the ability to stimulate uptake of macromolecular cargo in mammalian cells in a non-toxic manner and therefore hold promise as efficient and well tolerated gene delivery vectors. Non-covalent peptide-DNA complexes (“peptiplexes”) enter cells via endocytosis, but poor peptiplex stability and endosomal entrapment are considered as main barriers to peptide-mediated delivery. We explore a simple, yet highly efficient, strategy to improve the function of peptide-based vectors, by adding one terminal cysteine residue. This allows the peptide to dimerize by disulfide bond formation, increasing its affinity for nucleic acids by the “chelate effect” and, when the bond is reduced intracellularly, letting the complex dissociate to deliver the nucleic acid. By introducing a single C-terminal cysteine in the classical CPP penetratin and the penetratin analogs PenArg and EB1, we show that this minor modification greatly enhances the transfection capacity for plasmid DNA in HEK293T cells. We conclude that this effect is mainly due to enhanced thermodynamic stability of the peptiplexes as endosome-disruptive chloroquine is still required for transfection and the effect is more pronounced for peptides with lower inherent DNA condensation capacity. Interestingly, for EB1, addition of one cysteine makes the peptide able to mediate transfection in absence of chloroquine, indicating that dimerisation can also improve endosomal escape properties. Further, the cytotoxicity of EB1 peptiplexes is considerably reduced, possibly due to lower concentration of free peptide dimer resulting from its stronger binding to DNA.

  5. The spacer arm length in cell-penetrating peptides influences chitosan/siRNA nanoparticle delivery for pulmonary inflammation treatment

    Science.gov (United States)

    Jeong, Eun Ju; Choi, Moonhwan; Lee, Jangwook; Rhim, Taiyoun; Lee, Kuen Yong

    2015-11-01

    Although chitosan and its derivatives have been frequently utilized as delivery vehicles for small interfering RNA (siRNA), it is challenging to improve the gene silencing efficiency of chitosan-based nanoparticles. In this study, we hypothesized that controlling the spacer arm length between a cell-penetrating peptide (CPP) and a nanoparticle could be critical to enhancing the cellular uptake as well as the gene silencing efficiency of conventional chitosan/siRNA nanoparticles. A peptide consisting of nine arginine units (R9) was used as a CPP, and the spacer arm length was controlled by varying the number of glycine units between the peptide (R9Gn) and the nanoparticle (n = 0, 4, and 10). Various physicochemical characteristics of R9Gn-chitosan/siRNA nanoparticles were investigated in vitro. Increasing the spacing arm length did not significantly affect the complex formation between R9Gn-chitosan and siRNA. However, R9G10-chitosan was much more effective in delivering genes both in vitro and in vivo compared with non-modified chitosan (without the peptide) and R9-chitosan (without the spacer arm). Chitosan derivatives modified with oligoarginine containing a spacer arm can be considered as potential delivery vehicles for various genes.Although chitosan and its derivatives have been frequently utilized as delivery vehicles for small interfering RNA (siRNA), it is challenging to improve the gene silencing efficiency of chitosan-based nanoparticles. In this study, we hypothesized that controlling the spacer arm length between a cell-penetrating peptide (CPP) and a nanoparticle could be critical to enhancing the cellular uptake as well as the gene silencing efficiency of conventional chitosan/siRNA nanoparticles. A peptide consisting of nine arginine units (R9) was used as a CPP, and the spacer arm length was controlled by varying the number of glycine units between the peptide (R9Gn) and the nanoparticle (n = 0, 4, and 10). Various physicochemical characteristics of

  6. Intercellular imaging by a polyarginine derived cell penetrating peptide labeled magnetic resonance contrast agent,diethylenetriamine pentaacetic acid gadolinium

    Institute of Scientific and Technical Information of China (English)

    GUO You-min; LIU Min; YANG Jun-le; GUO Xiao-juan; WANG Si-cen; DUAN Xiao-yi; WANG Peng

    2007-01-01

    Background The cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro. Methods Fluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T1WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide reduction assay (MTT). Results The molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW=2163.34, Gd-DTPA-CPP MW=2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T1 weighted imaging (T1WI) signal, and that the T1WI signal intensity was increasing in a time-dependent manner (r=0.972, P=0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P=0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F=0.006, P=1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group. Conclusions The newly constructed CPP based on polyarginine can translocate cells by carrying FITC

  7. Development of an Optimized Activatable MMP-14 targeted SPECT Imaging Probe

    OpenAIRE

    Watkins, Gregory A.; Jones, Ella Fung; Shell, M. Scott; VanBrocklin, Henry F.; Pan, Mei-Hsiu; Hanrahan, Stephen M.; Feng, Jin Jin; He, Jiang; Sounni, Nor Eddine; Dill, Ken A.; Contag, Christopher H.; Coussens, Lisa M; Franc, Benjamin L.

    2008-01-01

    Matrix Metalloproteinase-14 (MT1-MMP or MMP-14) is a membrane-associated protease implicated in a variety of tissue remodeling processes and a molecular hallmark of select metastatic cancers. The ability to detect MMP-14 in vivo would be useful in studying its role in pathologic processes and may potentially serve as a guide for the development of targeted molecular therapies. Four MMP-14 specific probes containing a positively charged cell penetrating peptide (CPP) d-arginine octamer (r8) li...

  8. The use of nanoscale fluorescence microscopic to decipher cell wall modifications during fungal penetration

    Directory of Open Access Journals (Sweden)

    Dorothea eEllinger

    2014-06-01

    Full Text Available Plant diseases are one of the most studied subjects in the field of plant science due to their impact on crop yield and food security. Our increased understanding of plant–pathogen interactions was mainly driven by the development of new techniques that facilitated analyses on a subcellular and molecular level. The development of labeling technologies, which allowed the visualization and localization of cellular structures and proteins in live cell imaging, promoted the use of fluorescence and laser-scanning microscopy in the field of plant–pathogen interactions. Recent advances in new microscopic technologies opened their application in plant science and in the investigation of plant diseases. In this regard, in planta Förster/Fluorescence resonance energy transfer has demonstrated to facilitate the measurement of protein-protein interactions within the living tissue, supporting the analysis of regulatory pathways involved in plant immunity and putative host-pathogen interactions on a nanoscale level. Localization microscopy, an emerging, non-invasive microscopic technology, will allow investigations with a nanoscale resolution leading to new possibilities in the understanding of molecular processes.

  9. A novel cell penetrating peptide carrier for the delivery of nematocidal proteins drug

    Science.gov (United States)

    Kim, Jea Hyun

    Nematodes have recently become a primary source of harmful diseases to the environment that inflict harsh damages to pine trees and marine species. However, nematodes cannot be killed by normal pesticides or chemicals due to their thick outer protective layer mainly composed of collagen and cuticles. Thus, a novel approach to trigger intracellular delivery of chemicals through the layers of nematodes is required. In this study, the selection of the novel CPP was carefully progressed through protein database and serial digested fragmentation, internalization of each amino sequence was analyzed through flow cytometry and confocal microscope. As one of the most effective CPP material, JH 1.6 was compared with other major CPPs and its cellular toxicity was investigated. Furthermore, JH 1.6 was attached to various RNA, DNA, and proteins and internalization efficiency was evaluated for mammalian cells. To examine its effects on nematodes in vivo, JH 1.6 was conjugated with nematocidal protein - botulinum neurotoxin (BnT) and treated in C.elegans as a model animal. The results showed that JH 1.6 had high relative internalization rate and low cellular toxicity compared to other major CPP such as TAT and GV1001 peptides.

  10. Effective Penetration of Cell-permeable Peptide Mimic of Tyrosine Residue 654 Domain of β-catenin into Human Renal Tubular Epithelial Cells

    Institute of Scientific and Technical Information of China (English)

    ZENG Rui; XU Gang; HAN Min; LIU Wei; LIU Xiaocheng

    2007-01-01

    Phosphorylation of β-catenin tyrosine residue 654 plays an important role in the epithelial to myofibroblast transition (EMT). Introducing mimic peptide of tyrosine residue 654 domain of β-catenin into cells may influence phosphorylation of β-catenin tyrosine residue 654. To deliver this mimic peptide into renal epithelial cells, we used penetratin as a vector, which is a novel cell perme-able peptide, to deliver hydrophilic molecules into cells. A tyrosine 654 residue domain mimic pep-tide of β-catenin (PM) with fused penetratin was constructed, purified and then detected for the pene-tration of the mimic peptide into human renal tubular epithelial cells (HK-2). The results showed that purified fusion mimic peptide could efficiently and rapidly translocate into human renal tubular epithelial cells. It is concluded that a cell-permeable peptides mimic of tyrosine residue 654 domain of β-catenin was successfully obtained, which may provide a useful reagent for interfering the human renal tubular epithelial-mesenchymal transition.

  11. Targeting the EGFR/PCNA signaling suppresses tumor growth of triple-negative breast cancer cells with cell-penetrating PCNA peptides.

    Directory of Open Access Journals (Sweden)

    Yung-Luen Yu

    Full Text Available Tyrosine 211 (Y211 phosphorylation of proliferation cell nuclear antigen (PCNA coincides with pronounced cancer cell proliferation and correlates with poor survival of breast cancer patients. In epidermal growth factor receptor (EGFR tyrosine kinase inhibitor (TKI-resistant cells, both nuclear EGFR (nEGFR expression and PCNA Y211 phosphorylation are increased. Moreover, the resistance to EGFR TKI is a major clinical problem in treating EGFR-overexpressing triple-negative breast cancer (TNBC. Thus, effective treatment to combat resistance is urgently needed. Here, we show that treatment of cell-penetrating PCNA peptide (CPPP inhibits growth and induces apoptosis of human TNBC cells. The Y211F CPPP specifically targets EGFR and competes directly for PCNA tyrosine Y211 phosphorylation and prevents nEGFR from binding PCNA in vivo; it also suppresses tumor growth by sensitizing EGFR TKI resistant cells, which have enhanced nEGFR function and abrogated classical EGFR membrane signaling. Furthermore, we identify an active motif of CPPP, RFLNFF (RF6 CPPP, which is necessary and sufficient to inhibit TKI-resistant TNBC cell growth of orthotopic implanted tumor in mice. Finally, the activity of its synthetic retro-inverted derivative, D-RF6 CPPP, on an equimolar basis, is more potent than RF6 CPPP. Our study reveals a drug candidate with translational potential for the future development of safe and effective therapeutic for EGFR TKI resistance in TNBC.

  12. Curb Challenges of the “Trojan Horse” Approach: Smart Strategies in Achieving Effective yet Safe Cell-penetrating Peptide-based Drug Delivery

    OpenAIRE

    Huang, Yongzhuo; Jiang, Yifan; Wang, Huiyuan; Wang, Jianxin; Shin, Meong Cheol; Byun, Youngro; He, Huining; Liang, Yanqin; Yang, Victor C.

    2013-01-01

    Cell-penetrating peptide (CPP)-mediated intracellular drug delivery system, often specifically termed as “the Trojan horse approach”, has become the “holy grail” in achieving effective delivery of macromolecular compounds such as proteins, DNA, siRNAs, and drug carriers. It is characterized by the unique cell- (or receptor-), temperature-, and payload-independent mechanisms, therefore offering potent means to improve poor cellular uptake of a variety of macromolecular drugs. Nevertheless, thi...

  13. Development of a market penetration forecasting model for Hydrogen Fuel Cell Vehicles considering infrastructure and cost reduction effects

    International Nuclear Information System (INIS)

    In order to cope with climate change, the development and deployment of Hydrogen Fuel Cell Vehicles (HFCVs) is becoming more important. In this study, we developed a forecasting model for HFCVs based on the generalized Bass diffusion model and a simulation model using system dynamics. Through the developed model, we could forecast that the saturation of HFCVs in Korea can be moved up 12 years compared with the US. A sensitivity analysis on external variables such as price reduction rates of HFCVs and number of hydrogen refueling stations is also conducted. The results of this study can give insights on the effects of external variables on the market penetration of HFCVs, and the developed model can also be applied to other studies in analyzing the diffusion effects of HFCVs. - Highlights: → A forecasting model for HFCVs was developed using the generalized Bass diffusion model. → A simulation model using system dynamics was also developed. → The empirical study shows that the infrastructure is an important factor to the initial purchase. → The results of this study can promote research related to the diffusion of innovation.

  14. Hemocompatible poly(NIPAm-MBA-AMPS) colloidal nanoparticles as carriers of anti-inflammatory cell penetrating peptides.

    Science.gov (United States)

    Bartlett, Rush L; Medow, Matthew R; Panitch, Alyssa; Seal, Brandon

    2012-04-01

    Anionic copolymer systems containing sulfated monomers have great potential for delivery of cationic therapeutics, but N-isopropylacrylamide (NIPAm) 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) copolymer nanoparticles have seen limited characterization to date with regard to physical properties relevant to loading and release of therapeutics. Characterization of polymeric nanoparticles incorporating AMPS showed an increased size and decreased thermodynamic swelling ratios of AMPS containing particles as compared to NIPAm nanoparticles lacking AMPS. Particles with increasing AMPS addition showed an increased propensity for uniformity, intraparticle colloidal stability, and drug loading capacity. Peptide encapsulated in particles was shielded from peptide degradation in serum. Particles were shown not impede blood coagulation or to cause hemolysis. This study has demonstrated that AMPS incorporation into traditional NIPAm nanoparticles presents a tunable parameter for changing particle LCST, size, swelling ratio, ζ potential, and cationic peptide loading potential. This one-pot synthesis results in a thermosensitive anionic nanoparticle system that is a potentially useful platform to deliver cationic cell penetrating peptides.

  15. Binding characteristics of thrombin-activatable fibrinolysis inhibitor to streptococcal surface collagen-like proteins A and B

    NARCIS (Netherlands)

    Seron, Mercedes Valls; Plug, Tom; Marquart, J. Arnoud; Marx, Pauline F.; Herwald, Heiko; de Groot, Philip G.; Meijers, Joost C. M.

    2011-01-01

    Streptococcus pyogenes is the causative agent in a wide range of diseases in humans. Thrombin-activatable fibrinolysis inhibitor (TAFI) binds to collagen-like proteins ScIA and ScIB at the surface of S. pyogenes. Activation of TAFI at this surface redirects inflammation from a transient to chronic s

  16. Binding characteristics of thrombin-activatable fibrinolysis inhibitor to streptococcal surface collagen-like proteins A and B

    NARCIS (Netherlands)

    M.V. Serón; T. Plug; J.A. Marquart; P.F. Marx; H. Herwald; P.G. de Groot; J.C.M. Meijers

    2011-01-01

    Streptococcus pyogenes is the causative agent in a wide range of diseases in humans. Thrombin-activatable fibrinolysis inhibitor (TAFI) binds to collagen-like proteins SclA and SclB at the surface of S. pyogenes. Activation of TAFI at this surface redirects inflammation from a transient to chronic s

  17. Modeling the endosomal escape of cell-penetrating peptides: transmembrane pH gradient driven translocation across phospholipid bilayers.

    Science.gov (United States)

    Magzoub, Mazin; Pramanik, Aladdin; Gräslund, Astrid

    2005-11-15

    Cell-penetrating peptides (CPPs) are able to mediate the efficient cellular uptake of a wide range of cargoes. Internalization of a number of CPPs requires uptake by endocytosis, initiated by binding to anionic cell surface heparan sulfate (HS), followed by escape from endosomes. To elucidate the endosomal escape mechanism, we have modeled the process for two CPPs: penetratin (pAntp) and the N-terminal signal peptide of the unprocessed bovine prion protein (bPrPp). Large unilamellar phospholipid vesicles (LUVs) were produced encapsulating either peptide, and an ionophore, nigericin, was used to create a transmembrane pH gradient (DeltapH(mem), inside acidic) similar to the one arising in endosomes in vivo. In the absence of DeltapH(mem), no pAntp escape from the LUVs is observed, while a fraction of bPrPp escapes. In the presence of DeltapH(mem), a significant amount of pAntp escapes and an even higher degree of bPrPp escape takes place. These results, together with the differences in kinetics of escape, indicate different escape mechanisms for the two peptides. A minimum threshold peptide concentration exists for the escape of both peptides. Coupling of the peptides to a cargo reduces the fraction escaping, while complexation with HS significantly hinders the escape. Fluorescence correlation spectroscopy results show that during the escape process the LUVs are intact. Taken together, these results suggest a model for endosomal escape of CPPs: DeltapH(mem)-mediated mechanism, following dissociation from HS of the peptides, above a minimum threshold peptide concentration, in a process that does not involve lysis of the vesicles.

  18. Tumor penetrating peptides

    Directory of Open Access Journals (Sweden)

    Tambet eTeesalu

    2013-08-01

    Full Text Available Tumor-homing peptides can be used to deliver drugs into tumors. Phage library screening in live mice has recently identified homing peptides that specifically recognize the endothelium of tumor vessels, extravasate, and penetrate deep into the extravascular tumor tissue. The prototypic peptide of this class, iRGD (CRGDKGPDC, contains the integrin-binding RGD motif. RGD mediates tumor homing through binding to αv integrins, which are selectively expressed on various cells in tumors, including tumor endothelial cells. The tumor-penetrating properties of iRGD are mediated by a second sequence motif, R/KXXR/K. This C-end Rule (or CendR motif is active only when the second basic residue is exposed at the C-terminus of the peptide. Proteolytic processing of iRGD in tumors activates the cryptic CendR motif, which then binds to neuropilin-1 activating an endocytic bulk transport pathway through tumor tissue. Phage screening has also yielded tumor-penetrating peptides that function like iRGD in activating the CendR pathway, but bind to a different primary receptor. Moreover, novel tumor-homing peptides can be constructed from tumor-homing motifs, CendR elements and protease cleavage sites. Pathologies other than tumors can be targeted with tissue-penetrating peptides, and the primary receptor can also be a vascular zip code of a normal tissue. The CendR technology provides a solution to a major problem in tumor therapy, poor penetration of drugs into tumors. The tumor-penetrating peptides are capable of taking a payload deep into tumor tissue in mice, and they also penetrate into human tumors ex vivo. Targeting with these peptides specifically increases the accumulation in tumors of a variety of drugs and contrast agents, such as doxorubicin, antibodies and nanoparticle-based compounds. Remarkably the drug to be targeted does not have to be coupled to the peptide; the bulk transport system activated by the peptide sweeps along any compound that is

  19. Redoxable heteronanocrystals functioning magnetic relaxation switch for activatable T1 and T2 dual-mode magnetic resonance imaging.

    Science.gov (United States)

    Kim, Myeong-Hoon; Son, Hye-Young; Kim, Ga-Yun; Park, Kwangyeol; Huh, Yong-Min; Haam, Seungjoo

    2016-09-01

    T1/T2 dual-mode magnetic resonance (MR) contrast agents (DMCAs) have gained much attention because of their ability to improve accuracy by providing two pieces of complementary information with one instrument. However, most of these agents are "always ON" systems that emit MR contrast regardless of their interaction with target cells or biomarkers, which may result in poor target-to-background ratios. Herein, we introduce a rationally designed magnetic relaxation switch (MGRS) for an activatable T1/T2 dual MR imaging system. Redox-responsive heteronanocrystals, consisting of a superparamagnetic Fe3O4 core and a paramagnetic Mn3O4 shell, are synthesized through seed-mediated growth and subsequently surface-modified with polysorbate 80. The Mn3O4 shell acts as both a protector of Fe3O4 in aqueous environments to attenuate T2 relaxation and as a redoxable switch that can be activated in intracellular reducing environments by glutathione. This simultaneously generates large amounts of magnetically decoupled Mn(2+) ions and allows Fe3O4 to interact with the water protons. This smart nanoplatform shows an appropriate hydrodynamic size for the EPR effect (10-100 nm) and demonstrates biocompatibility. Efficient transitions of OFF/ON dual contrast effects are observed by in vitro imaging and MR relaxivity measurements. The ability to use these materials as DMCAs is demonstrated via effective passive tumor targeting for T1- and T2-weighted MR imaging in tumor-bearing mice. PMID:27281684

  20. PREPARATION OF CHEMICAL AND PHYSICAL CONJUGATES OF SELF-ASSEMBLING NANOPARTICLES WITH CELL-PENETRATING PEPTIDE AND DOXORUBICIN

    Directory of Open Access Journals (Sweden)

    Zhadyra Sagykyzy Shagyrova

    2015-09-01

    Full Text Available Abstract: Nano-sized carriers can help to reduce toxicity and improve clinical efficacy of drugs. Virus-like particles (VLPs are biocompatible and biodegradable self-assembling nanoparticles, which show great promise as carriers for substances for targeted delivery and controlled release. Either chemical conjugation of physical incorporation without formation of covalent bonds is possible to load substances of interest into VLPs.Objectives: To produce VLPs from recombinant viral capsid protein (HBcAg and test feasibility of methods of formation of chemical and physical conjugates of VLPs with substances of pharmacological interest.Methods: Virus-like particles composed from recombinant hepatitis B core antigen (HBcAg were produced by recombinant expression in E.coli and purified by successive centrifugation through sucrose gradients. Peptide transportan 10 was synthesized and used for carbodiimide (EDC-mediated conjugation to VLPs. Doxorubicin (DOX was loaded into the nucleic acid-containing VLPs to form physical conjugate.Results: VLPs with chemically attached moieties of cell-penetrating peptide transportan 10 were produced. The conjugate was examined in SDS-PAGE to confirm presence of conjugation products. Conjugation efficiency (molar ration peptide/protein in the conjugate reaches 0.5:1 (i.e. 50% of protein chains have one attached peptide moiety. The nucleic acid-containing VLPs can be loaded with the DOX forming stable non-covalent physical conjugate.Conclusion: Recombinantly expressed VLPs allow easy attaching of small molecules making them a convenient platform to develop drug carriers.

  1. Region-Dependent Role of Cell-Penetrating Peptides in Insulin Absorption Across the Rat Small Intestinal Membrane.

    Science.gov (United States)

    Khafagy, El-Sayed; Iwamae, Ruisha; Kamei, Noriyasu; Takeda-Morishita, Mariko

    2015-11-01

    We have reported that the cell-penetrating peptide (CPP) penetratin acts as a potential absorption enhancer in oral insulin delivery systems and that this action occurs through noncovalent intermolecular interactions. However, the region-dependent role of CPPs in intestinal insulin absorption has not been clarified. To identify the intestinal region where CPPs have the most effect in increasing insulin absorption, the region-dependent action of penetratin was investigated using in situ closed intestinal loops in rats. The order of the insulin area under the insulin concentration-time curve (AUC) increase effect by L-penetratin was ileum > jejunum > duodenum > colon. By contrast, the AUC order after coadministration of insulin with D-penetratin was colon > duodenum ≥ jejunum and ileum. We also compared the effects of the L- and D-forms of penetratin, R8, and PenetraMax on ileal insulin absorption. Along with the CPPs used in this study, L- and D-PenetraMax produced the largest insulin AUCs. An absorption study using ilea pretreated with CPPs showed that PenetraMax had no irreversible effect on the intestinal epithelial membrane. The degradation of insulin in the presence of CPPs was assessed in rat intestinal enzymatic fluid. The half-life (t 1/2) of insulin increased from 14.5 to 23.7 and 184.7 min in the presence of L- and D-PenetraMax, respectively. These enzymatic degradation-resistant effects might contribute partly to the increased ileal absorption of insulin induced by D-PenetraMax. In conclusion, this study demonstrated that the ability of the L- and D-forms of penetratin to increase intestinal insulin absorption was maximal in the ileum and the colon, respectively, and that D-PenetraMax is a powerful but transient enhancer of oral insulin absorption.

  2. Hierarchy of Specific Lipid-Peptide Interactions Produces the Activity of Cell-penetrating and Cell-permeating Peptides

    Science.gov (United States)

    Davis, Matthew; Parente, Daniel; Gordon, Vernita; Mishra, Abhijit; Schmidt, Nathan; Yang, Lihua; Coridan, Robert; Som, Abhigyan; Tew, Gregory; Wong, Gerard

    2008-03-01

    Protein transduction domains can cross cell membranes with high efficiency, even when carrying a variety of cargos, and thus has strong biotechnological potential. The molecular mechanism of entry, however, is not well understood. We use small-angle x-ray scattering (SAXS) and confocal microscopy to systematically study the interaction of the TAT and ANTP PTD with model membranes of variable composition. Their membrane transduction activity requires the presence of both PE and PS lipids in the membrane. Antimicrobial peptides (AMP's) are cationic amphiphiles that comprise a key component of innate immunity. Synthetic analogs of AMP's, such as the family of phenylene ethynylene antimicrobial oligomers (AMO's), recently demonstrated broad-spectrum antimicrobial activity, but the underlying molecular mechanism is unknown. PE lipid greatly enhances permeating activity of AMO in these membranes, showing the importance of specific lipid composition for the activity of cell-permeating peptides. Since bacterial cell membranes are richer in PE lipids than are eukaryotic cell membranes, this may indicate a mechanism for antimicrobial specificity.

  3. Identification of a Short Cell-Penetrating Peptide from Bovine Lactoferricin for Intracellular Delivery of DNA in Human A549 Cells.

    Science.gov (United States)

    Liu, Betty R; Huang, Yue-Wern; Aronstam, Robert S; Lee, Han-Jung

    2016-01-01

    Cell-penetrating peptides (CPPs) have been shown to deliver cargos, including protein, DNA, RNA, and nanomaterials, in fully active forms into live cells. Most of the CPP sequences in use today are based on non-native proteins that may be immunogenic. Here we demonstrate that the L5a CPP (RRWQW) from bovine lactoferricin (LFcin), stably and noncovalently complexed with plasmid DNA and prepared at an optimal nitrogen/phosphate ratio of 12, is able to efficiently enter into human lung cancer A549 cells. The L5a CPP delivered a plasmid containing the enhanced green fluorescent protein (EGFP) coding sequence that was subsequently expressed in cells, as revealed by real-time PCR and fluorescent microscopy at the mRNA and protein levels, respectively. Treatment with calcium chloride increased the level of gene expression, without affecting CPP-mediated transfection efficiency. Zeta-potential analysis revealed that positively electrostatic interactions of CPP/DNA complexes correlated with CPP-mediated transport. The L5a and L5a/DNA complexes were not cytotoxic. This biomimetic LFcin L5a represents one of the shortest effective CPPs and could be a promising lead peptide with less immunogenic for DNA delivery in gene therapy.

  4. The feasibility of a targeted ultrasound contrast agent carrying genes and cell-penetrating peptides to hypoxic HUVEC

    International Nuclear Information System (INIS)

    Objective: To prepare an anti-P-selectin targeted ultrasound contrast agent carrying genes and cell-penetrating peptides (CPP) and to investigate its feasibility of delivery to hypoxic human umbilical vein endothelial cells (HUVEC). Methods: Anti-P-selectin targeted ultrasound contrast agent carrying a green fluorescent protein gene (pEGFP-N1) and CPP was prepared by mechanical vibration and carbodiimide techniques. The appearance, distribution, concentration and diameter of the ultrasound contrast agent were measured. The gene and CPP distribution on the agent was investigated using confocal laser scanning microscopy (CLSM). The efficiency of the ultrasound contrast agent to carry the gene and CPP was investigated by fluorospectrophotometry. HUVEC were cultured in vitro and hypoxic HUVEC were prepared using hydrogen peroxide (H2O2). Hypoxic HUVEC were randomly assigned targeted ultrasound contrast agents and non-targeted ultrasound contrast agents for transfection. The transfection effect of green fluorescent protein in the two groups was observed using fluorescence microscopy and flow cytometry. T-test and linear correlation analysis were used for statistical analysis. Results: The average diameter of anti-P-selectin targeted ultrasound contrast agents carrying gene and CPP was (2.15 ±0.36) μm and the concentration was (1.58 ± 0.23) × 107/ml.The results of CLSM showed that gene and CPP were distributed on the shell of the agent. The gene encapsulation efficiency was 28% (y=0.932x-0.09, r=0.993, P<0.05), and the CPP encapsulation efficiency was 25% (y=5.875x-0.81, r=0.987, P<0.05). EGFP expression was observed using fluorescence microscopy in targeted ultrasound contrast agents and non-targeted ultrasound contrast agents. The average transfection efficiencies of targeted ultrasound contrast agents and non-targeted ultrasound contrast agents were (18.74 ± 0.47) % and (15.34 ± 0.22) % after 24 h (t=10.923, P<0.001). Conclusions: The in vitro studies showed

  5. Differential neuroprotective potential of CRMP2 peptide aptamers conjugated to cationic, hydrophobic, and amphipathic cell penetrating peptides

    Directory of Open Access Journals (Sweden)

    Aubin eMoutal

    2015-01-01

    Full Text Available The microtubule-associated axonal specification collapsin response mediator protein 2 (CRMP2 is a novel target for neuroprotection. A CRMP2 peptide (TAT-CBD3 conjugated to the HIV transactivator of transcription (TAT protein’s cationic cell penetrating peptide motif (CPP protected neurons in the face of toxic levels of Ca2+ influx leaked in via N-methyl-D-aspartate receptor (NMDAR hyperactivation. Here we tested whether replacing the hydrophilic TAT motif with alternative cationic (nona-arginine (R9, hydrophobic (membrane transport sequence (MTS of k-fibroblast growth factor or amphipathic (model amphipathic peptide (MAP CPPs could be superior to the neuroprotection bestowed by TAT-CBD3. In giant plasma membrane vesicles (GPMVs derived from cortical neurons, the peptides translocated across plasma membranes with similar efficiencies. Cortical neurons, acutely treated with peptides prior to a toxic glutamate challenge, demonstrated enhanced efflux of R9-CBD3 compared to others. R9-CBD3 inhibited N-methyl-D-aspartate (NMDA-evoked Ca2+ influx to a similar extent as TAT-CBD3 while MTS-CBD3 was ineffective which correlated with the ability of R9- and TAT-CBD3, but not MTS-CBD3, to block NMDAR interaction with CRMP2. Unrestricted Ca2+ influx through NMDARs leading to delayed calcium dysregulation and neuronal cell death was blocked by all peptides but MAP-CBD3. When applied acutely for 10 minutes, R9-CBD3 was more effective than TAT-CBD3 at neuroprotection while MTS- and MAP-CBD3 were ineffective. In contrast, long-term (> 24 hours treatment with MTS-CBD3 conferred neuroprotection where TAT-CBD3 failed. Neither peptide altered surface trafficking of NMDARs. Neuroprotection conferred by MTS-CBD3 peptide is likely due to its increased uptake coupled with decreased efflux when compared to TAT-CBD3. Overall, our results demonstrate that altering CPPs can bestow differential neuroprotective potential onto the CBD3 cargo.

  6. Thrombin-activatable fibrinolysis inhibitor activity in healthy and diseased dogs

    DEFF Research Database (Denmark)

    Jessen, Lisbeth Rem; Wiinberg, Bo; Kjelgaard-Hansen, Mads;

    2010-01-01

    Background: In people, increased thrombin-activatable fibrinolysis inhibitor (TAFI) antigen has been associated with increased risk of thrombosis, and decreased TAFI may contribute to bleeding diathesis. TAFI activity in dogs has been described in experimental models, but not in dogs...... with spontaneous disease. Objective: The aim of this study was to compare TAFI activity in healthy dogs with TAFI activity in dogs with spontaneous disease. Methods: Plasma samples from 20 clinically healthy Beagles and from 35 dogs with various diseases were analyzed using a commercial chromogenic assay...... that measured TAFI activity relative to activity in standardized pooled human plasma. Results: Median TAFI activity for the 20 Beagles was 46.1% (range 32.2-70.8%) compared with 62.6% (29.1-250%) for the 35 diseased dogs, and 14/35 (40%) had TAFI activities >the upper limit for controls. The highest individual...

  7. DNA-histone complexes as ligands amplify cell penetration and nuclear targeting of anti-DNA antibodies via energy-independent mechanisms.

    Science.gov (United States)

    Zannikou, Markella; Bellou, Sofia; Eliades, Petros; Hatzioannou, Aikaterini; Mantzaris, Michael D; Carayanniotis, George; Avrameas, Stratis; Lymberi, Peggy

    2016-01-01

    We have generated three monoclonal cell-penetrating antibodies (CPAbs) from a non-immunized lupus-prone (NZB × NZW)F1 mouse that exhibited high anti-DNA serum titres. These CPAbs are polyreactive because they bind to DNA and other cellular components, and localize mainly in the nucleus of HeLa cells, albeit with a distinct nuclear labelling profile. Herein, we have examined whether DNA-histone complexes (DHC) binding to CPAbs, before cell entry, could modify the cell penetration of CPAbs or their nuclear staining properties. By applying confocal microscopy and image analysis, we found that extracellular binding of purified CPAbs to DHC significantly enhanced their subsequent cell-entry, both in terms of percentages of positively labelled cells and fluorescence intensity (internalized CPAb amount), whereas there was a variable effect on their nuclear staining profile. Internalization of CPAbs, either alone or bound to DHC, remained unaltered after the addition of endocytosis-specific inhibitors at 37° or assay performance at 4°, suggesting the involvement of energy-independent mechanisms in the internalization process. These findings assign to CPAbs a more complex pathogenetic role in systemic lupus erythematosus where both CPAbs and nuclear components are abundant.

  8. Structure analysis and conformational transitions of the cell penetrating peptide transportan 10 in the membrane-bound state.

    Directory of Open Access Journals (Sweden)

    Susanne Fanghänel

    Full Text Available Structure analysis of the cell-penetrating peptide transportan 10 (TP10 revealed an exemplary range of different conformations in the membrane-bound state. The bipartite peptide (derived N-terminally from galanin and C-terminally from mastoparan was found to exhibit prominent characteristics of (i amphiphilic α-helices, (ii intrinsically disordered peptides, as well as (iii β-pleated amyloid fibrils, and these conformational states become interconverted as a function of concentration. We used a complementary approach of solid-state (19F-NMR and circular dichroism in oriented membrane samples to characterize the structural and dynamical behaviour of TP10 in its monomeric and aggregated forms. Nine different positions in the peptide were selectively substituted with either the L- or D-enantiomer of 3-(trifluoromethyl-bicyclopent-[1.1.1]-1-ylglycine (CF3-Bpg as a reporter group for (19F-NMR. Using the L-epimeric analogs, a comprehensive three-dimensional structure analysis was carried out in lipid bilayers at low peptide concentration, where TP10 is monomeric. While the N-terminal region is flexible and intrinsically unstructured within the plane of the lipid bilayer, the C-terminal α-helix is embedded in the membrane with an oblique tilt angle of ∼ 55° and in accordance with its amphiphilic profile. Incorporation of the sterically obstructive D-CF3-Bpg reporter group into the helical region leads to a local unfolding of the membrane-bound peptide. At high concentration, these helix-destabilizing C-terminal substitutions promote aggregation into immobile β-sheets, which resemble amyloid fibrils. On the other hand, the obstructive D-CF3-Bpg substitutions can be accommodated in the flexible N-terminus of TP10 where they do not promote aggregation at high concentration. The cross-talk between the two regions of TP10 thus exerts a delicate balance on its conformational switch, as the presence of the α-helix counteracts the tendency of the

  9. Penetration through the Skin Barrier

    DEFF Research Database (Denmark)

    Nielsen, Jesper Bo; Benfeldt, Eva; Holmgaard, Rikke

    2016-01-01

    . During recent decades, the latter has received increased attention as a route for intentionally delivering drugs to patients. This has stimulated research in methods for sampling, measuring and predicting percutaneous penetration. Previous chapters have described how different endogenous, genetic...... and exogenous factors may affect barrier characteristics. The present chapter introduces the theory for barrier penetration (Fick's law), and describes and discusses different methods for measuring the kinetics of percutaneous penetration of chemicals, including in vitro methods (static and flow......-through diffusion cells) as well as in vivo methods (microdialysis and microperfusion). Then follows a discussion with examples of how different characteristics of the skin (age, site and integrity) and of the penetrants (size, solubility, ionization, logPow and vehicles) affect the kinetics of percutaneous...

  10. Folic Acid-Targeted and Cell Penetrating Peptide-Mediated Theranostic Nanoplatform for High-Efficiency Tri-Modal Imaging-Guided Synergistic Anticancer Phototherapy.

    Science.gov (United States)

    Li, Na; Li, Tingting; Liu, Chen; Ye, Shiyi; Liang, Jiangong; Han, Heyou

    2016-05-01

    A novel nanomaterial with precisely-defined size and shape, biocompatible composition, and excellent stability, which can integrate multi modal targeted imaging and therapy into a single system for visualized therapeutics, has recently attracted significant research interest. Here, we developed a multifunctional nanoplatform based on silica-coated 4-mercaptobenzoic acid-modified gold nanorods (Au NRs) decorated with gold nanoclusters rich in the photosensitizer Ce6 (Au-Ce6 NCs). The nanoparticles also comprised folic acid and cell penetrating peptide molecules anchored on the surface, obtaining the Au@SiO2@Au-cell penetrating peptide nanocomposite. The Au-Ce6 NCs enhanced the photophysical stability, provided numerous bonding sites and offered a large surface-area and interior space to achieve a high drug loading efficiency (up to 55%). The anchored folic acid and cell penetrating peptide synergistically enhanced the efficiency of uptake of nanocomposites by HeLa cells (up to 70.7%) and improved therapeutic efficacy. The nanocomposite also has good water-solubility, excellent biocompatibility, and long-term stability against illumination and exposure to pH 3-12, thus facilitating their bioapplications in cancer theranostics. Here, the nanocomposite was established for high-resolution and noninvasive tri-modal surface-enhanced Raman spectrum/dark-field/fluorescence imaging-guided high-efficiency synergistic photodynamic/photothermal therapy of cancer. Our studies demonstrate that the multifunctional nanocomposite has the potential as a novel and sensitive contrast agent for complementary and synergistic theranostics in the clinic. PMID:27305812

  11. Novel cell-penetrating peptide-loaded nanobubbles synergized with ultrasound irradiation enhance EGFR siRNA delivery for triple negative Breast cancer therapy.

    Science.gov (United States)

    Jing, Hui; Cheng, Wen; Li, Shouqiang; Wu, Bolin; Leng, Xiaoping; Xu, Shouping; Tian, Jiawei

    2016-10-01

    The lack of safe and effective gene delivery strategies remains a bottleneck for cancer gene therapy. Here, we describe the synthesis, characterization, and application of cell-penetrating peptide (CPP)-loaded nanobubbles (NBs), which are characterized by their safety, strong penetrating power and high gene loading capability for gene delivery. An epidermal growth factor receptor (EGFR)-targeted small interfering RNA (siEGFR) was transfected into triple negative breast cancer (TNBC) cells via prepared CPP-NBs synergized with ultrasound-targeted microbubble destruction (UTMD) technology. Fluorescence microscopy showed that siEGFR and CPP were loaded on the shells of the NBs. The transfection efficiency and cell proliferation levels were evaluated by FACS and MTT assays, respectively. In addition, in vivo experiments showed that the expression of EGFR mRNA and protein could be efficiently downregulated and that the growth of a xenograft tumor derived from TNBC cells could be inhibited. Our results indicate that CPP-NBs carrying siEGFR could potentially be used as a promising non-viral gene vector that can be synergized with UTMD technology for efficient TNBC therapy. PMID:27388967

  12. Identification of a cell-penetrating peptide domain from human beta-defensin 3 and characterization of its anti-inflammatory activity

    Directory of Open Access Journals (Sweden)

    Lee JY

    2015-08-01

    Full Text Available Jue Yeon Lee,1,* Jin Sook Suh,2,* Jung Min Kim,1 Jeong Hwa Kim,1 Hyun Jung Park,1 Yoon Jeong Park,1,2 Chong Pyoung Chung1 1Central Research Institute, Nano Intelligent Biomedical Engineering Corporation (NIBEC, Chungcheongbuk-do, Republic of Korea; 2Dental Regenerative Biotechnology, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Republic of Korea *These authors contributed equally to this work Abstract: Human beta-defensins (hBDs are crucial factors of intrinsic immunity that function in the immunologic response to a variety of invading enveloped viruses, bacteria, and fungi. hBDs can cause membrane depolarization and cell lysis due to their highly cationic nature. These molecules participate in antimicrobial defenses and the control of adaptive and innate immunity in every mammalian species and are produced by various cell types. The C-terminal 15-mer peptide within hBD3, designated as hBD3-3, was selected for study due to its cell- and skin-penetrating activity, which can induce anti-inflammatory activity in lipopolysaccharide-treated RAW 264.7 macrophages. hBD3-3 penetrated both the outer membrane of the cells and mouse skin within a short treatment period. Two other peptide fragments showed poorer penetration activity compared to hBD3-3. hBD3-3 inhibited the lipopolysaccharide-induced production of inducible nitric oxide synthase, nitric oxide, and secretory cytokines, such as interleukin-6 and tumor necrosis factor in a concentration-dependent manner. Moreover, hBD3-3 reduced the interstitial infiltration of polymorphonuclear leukocytes in a lung inflammation model. Further investigation also revealed that hBD3-3 downregulated nuclear factor kappa B-dependent inflammation by directly suppressing the degradation of phosphorylated-IκBα and by downregulating active nuclear factor kappa B p65. Our findings indicate that hBD3-3 may be conjugated with drugs of interest to ensure their proper translocation to

  13. Penetration of Rifampin through Staphylococcus epidermidis Biofilms

    OpenAIRE

    Zheng, Zhilan; Philip S. Stewart

    2002-01-01

    Rifampin penetrated biofilms formed by Staphylococcus epidermidis but failed to effectively kill the bacteria. Penetration was demonstrated by a simple diffusion cell bioassay and by transmission electron microscopic observation of antibiotic-affected cells at the distal edge of the biofilm.

  14. Penetration through the Skin Barrier.

    Science.gov (United States)

    Nielsen, Jesper Bo; Benfeldt, Eva; Holmgaard, Rikke

    2016-01-01

    The skin is a strong and flexible organ with barrier properties essential for maintaining homeostasis and thereby human life. Characterizing this barrier is the ability to prevent some chemicals from crossing the barrier while allowing others, including medicinal products, to pass at varying rates. During recent decades, the latter has received increased attention as a route for intentionally delivering drugs to patients. This has stimulated research in methods for sampling, measuring and predicting percutaneous penetration. Previous chapters have described how different endogenous, genetic and exogenous factors may affect barrier characteristics. The present chapter introduces the theory for barrier penetration (Fick's law), and describes and discusses different methods for measuring the kinetics of percutaneous penetration of chemicals, including in vitro methods (static and flow-through diffusion cells) as well as in vivo methods (microdialysis and microperfusion). Then follows a discussion with examples of how different characteristics of the skin (age, site and integrity) and of the penetrants (size, solubility, ionization, logPow and vehicles) affect the kinetics of percutaneous penetration. Finally, a short discussion of the advantages and challenges of each method is provided, which will hopefully allow the reader to improve decision making and treatment planning, as well as the evaluation of experimental studies of percutaneous penetration of chemicals. PMID:26844902

  15. Combined HLA matched limbal stem cells allograft with amniotic membrane transplantation as a prophylactic surgical procedure to prevent corneal graft rejection after penetrating keratoplasty: case report

    Directory of Open Access Journals (Sweden)

    Paolo Capozzi

    2014-09-01

    Full Text Available Purpose. To determine if the use of combined HLA matched limbal stem cells allograft with amniotic membrane transplantation (AMT is a safe and effective prophylactic surgical procedure to prevent corneal graft after penetrating keratoplasty (PK. Methods. We report the case of a 17 years old patient with a history of congenital glaucoma, trabeculectomy and multiple corneal graft rejections, presenting total limbal cell deficiency. To reduce the possibility of graft rejection in the left eye after a new PK, a two step procedure was performed. At first the patient underwent a combined HLA matched limbal stem cells allograft (LAT and AMT and then, 10 months later, a new PK. Results. During 12 months of follow-up, the corneal graft remained stable and smooth, with no sign of graft rejection. Conclusions. In our patient, the prophylactic use of LAT from HLA-matched donors and AMT before PK, may result in a better prognosis of corneal graft survival.

  16. Fusion of cell-penetrating peptides to thermally responsive biopolymer improves tumor accumulation of p21 peptide in a mouse model of pancreatic cancer

    Directory of Open Access Journals (Sweden)

    Walker LR

    2014-10-01

    Full Text Available Leslie R Walker,1 Jung Su Ryu,1 Eddie Perkins,2 Lacey R McNally,3 Drazen Raucher1 1Department of Biochemistry, 2Department of Neurosurgery, University of Mississippi Medical Center, Jackson, MS, USA; 3Division of Hematology and Oncology, University of Louisville, Louisville, KY, USAAbstract: Current therapies for the treatment of pancreatic cancer are limited. The limitations of this type of treatment are abundant. The majority of chemotherapeutic agents used in clinics are highly toxic to both tumor cells and normal tissues due to the lack of specificity. Resistance can develop due to overexposure of these agents. To address these issues, these agents must be made more exclusive toward the tumor site. We have developed a macromolecular carrier based on the sequence of the biopolymer elastin-like polypeptide (ELP that is able to aggregate upon reaching the externally heated tumor environment. This carrier is specific to the tumor as it only aggregates at the heated tumor site. ELP is soluble below its transition temperature but will aggregate when the temperature is raised above its transition temperature. ELP was modified by p21, a cell cycle inhibitory peptide, and the addition of Bac, a cell-penetrating peptide with nuclear localization capabilities. In this study, p21-ELP-Bac and its control, ELP-p21, were used in cell proliferation studies using the pancreatic cancer cell lines Panc-1, MiaPaca-2, and S2013. ELP-p21 had little effect on proliferation, while the half maximal inhibitory concentration of p21-ELP-Bac was ~30 µM. As translocation across the plasma membrane is a limiting step for delivery of macromolecules, these polypeptides were utilized in a pancreatic xenograft model to study the plasma clearance, biodistribution, tumor accumulation, and tumor reduction capabilities of the polypeptide with and without a cell-penetrating peptide.Keywords: elastin-like polypeptide, peptide, targeted drug delivery, macromolecule

  17. Biochemical characterization of bovine plasma thrombin-activatable fibrinolysis inhibitor (TAFI

    Directory of Open Access Journals (Sweden)

    Kristensen Torsten

    2009-05-01

    Full Text Available Abstract Background TAFI is a plasma protein assumed to be an important link between coagulation and fibrinolysis. The three-dimensional crystal structures of authentic mature bovine TAFI (TAFIa in complex with tick carboxypeptidase inhibitor, authentic full lenght bovine plasma thrombin-activatable fibrinolysis inhibitor (TAFI, and recombinant human TAFI have recently been solved. In light of these recent advances, we have characterized authentic bovine TAFI biochemically and compared it to human TAFI. Results The four N-linked glycosylation sequons within the activation peptide were all occupied in bovine TAFI, similar to human TAFI, while the sequon located within the enzyme moiety of the bovine protein was non-glycosylated. The enzymatic stability and the kinetic constants of TAFIa differed somewhat between the two proteins, as did the isoelectric point of TAFI, but not TAFIa. Equivalent to human TAFI, bovine TAFI was a substrate for transglutaminases and could be proteolytically cleaved by trypsin or thrombin/solulin complex, although small differences in the fragmentation patterns were observed. Furthermore, bovine TAFI exhibited intrinsic activity and TAFIa attenuated tPA-mediated fibrinolysis similar to the human protein. Conclusion The findings presented here suggest that the properties of these two orthologous proteins are similar and that conclusions reached using the bovine TAFI may be extrapolated to the human protein.

  18. Fusion of EBV with the surface of receptor-negative human hepatoma cell line Li7A permits virus penetration and infection.

    Science.gov (United States)

    Lisi, A; Pozzi, D; Carloni, G; Da Villa, G; Iacovacci, S; Valli, M B; Grimaldi, S

    1995-01-01

    Our preliminary data suggest that Epstein-Barr virus (EBV) is able to bind to and fuse with the surface membranes of hepatoma cell line Li7A. In order to obtain further evidence, we utilized the relief of rhodamine fluorescence to monitor whether fusion would also take place when Li7A cells were exposed to experimental conditions such as neutral or low pH. It is well known that for some viruses, protonation in the endosomal compartment is needed to trigger the fusion. We show, furthermore, that the rate and extent of fusion are not affected by pretreatment of the cells with agents known to elevate the lysosomal and ensodomal pH, such as chloroquine or NH4Cl (lysosomotropic agent). By indirect immunofluorescence assay, in addition, we confirmed the binding of the EBV to the Li7A cell surface membrane. We attempted finally to correlate the above processes with successful infection of Li7A cells by EBV detected using the polymerase chain reaction technique. In spite of the apparent lack of viral receptor CD21, these nonlymphoid cells appeared susceptible to EBV penetration and infection through fusion with the plasma membrane at the surface of the cells. PMID:8539493

  19. An efficient PEGylated liposomal nanocarrier containing cell-penetrating peptide and pH-sensitive hydrazone bond for enhancing tumor-targeted drug delivery

    Directory of Open Access Journals (Sweden)

    Ding Y

    2015-10-01

    Full Text Available Yuan Ding,1,* Dan Sun,1,* Gui-Ling Wang,1 Hong-Ge Yang,1 Hai-Feng Xu,1 Jian-Hua Chen,2 Ying Xie,1,3 Zhi-Qiang Wang4 1Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, School of Pharmaceutical Sciences, Peking University, Beijing, 2School of Medicine, Jianghan University, Wuhan, 3State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, People’s Republic of China; 4Department of Chemistry and Biochemistry, Kent State University Geauga, Burton, OH, USA *These authors contributed equally to this work Abstract: Cell-penetrating peptides (CPPs as small molecular transporters with abilities of cell penetrating, internalization, and endosomal escape have potential prospect in drug delivery systems. However, a bottleneck hampering their application is the poor specificity for cells. By utilizing the function of hydration shell of polyethylene glycol (PEG and acid sensitivity of hydrazone bond, we constructed a kind of CPP-modified pH-sensitive PEGylated liposomes (CPPL to improve the selectivity of these peptides for tumor targeting. In CPPL, CPP was directly attached to liposome surfaces via coupling with stearate (STR to avoid the hindrance of PEG as a linker on the penetrating efficiency of CPP. A PEG derivative by conjugating PEG with STR via acid-degradable hydrazone bond (PEG2000-Hz-STR, PHS was synthesized. High-performance liquid chromatography and flow cytometry demonstrated that PHS was stable at normal neutral conditions and PEG could be completely cleaved from liposome surface to expose CPP under acidic environments in tumor. An optimal CPP density on liposomes was screened to guaranty a maximum targeting efficiency on tumor cells as well as not being captured by normal cells that consequently lead to a long circulation in blood. In vitro and in vivo studies indicated, in 4 mol% CPP of lipid modified system, that CPP exerted higher efficiency on internalizing the liposomes into

  20. Cell-Penetrating Poly(disulfide) Assisted Intracellular Delivery of Mesoporous Silica Nanoparticles for Inhibition of miR-21 Function and Detection of Subsequent Therapeutic Effects.

    Science.gov (United States)

    Yu, Changmin; Qian, Linghui; Ge, Jingyan; Fu, Jiaqi; Yuan, Peiyan; Yao, Samantha C L; Yao, Shao Q

    2016-08-01

    The design of drug delivery systems capable of minimal endolysosomal trapping, controlled drug release, and real-time monitoring of drug effect is highly desirable for personalized medicine. Herein, by using mesoporous silica nanoparticles (MSNs) coated with cell-penetrating poly(disulfide)s and a fluorogenic apoptosis-detecting peptide (DEVD-AAN), we have developed a platform that could be uptaken rapidly by mammalian cells via endocytosis-independent pathways. Subsequent loading of these MSNs with small molecule inhibitors and antisense oligonucleotides resulted in intracellular release of these drugs, leading to combination inhibition of endogenous miR-21 activities which was immediately detectable by the MSN surface-coated peptide using two-photon fluorescence microscopy. PMID:27325284

  1. Cell-penetrating superoxide dismutase attenuates oxidative stress-induced senescence by regulating the p53-p21Cip1 pathway and restores osteoblastic differentiation in human dental pulp stem cells

    Directory of Open Access Journals (Sweden)

    Park YJ

    2012-09-01

    Full Text Available Yoon Jung Choi,1,* Jue Yeon Lee,2,* Chong Pyoung Chung,2 Yoon Jeong Park,1,21Craniomaxillofacial Reconstructive Sciences, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Republic of Korea; 2Research Institute, Nano Intelligent Biomedical Engineering, Seoul, Republic of Korea*These authors contributed equally to this workBackground: Human dental pulp stem cells (DPSCs have potential applications in tissue regeneration because of their convenient cell harvesting procedures and multipotent capacity. However, the tissue regenerative potential of DPSCs is known to be negatively regulated by aging in long-term culture and under oxidative stress. With an aim of reducing cellular senescence and oxidative stress in DPSCs, an intracellular delivery system for superoxide dismutase 1 (SOD1 was developed. We conjugated SOD1 with a cell-penetrating peptide known as low-molecular weight protamine (LMWP, and investigated the effect of LMWP-SOD1 conjugates on hydrogen peroxide-induced cellular senescence and osteoblastic differentiation.Results: LMWP-SOD1 significantly attenuated enlarged and flattened cell morphology and increased senescence-associated β-galactosidase activity. Under the same conditions, LMWP-SOD1 abolished activation of the cell cycle regulator proteins, p53 and p21Cip1, induced by hydrogen peroxide. In addition, LMWP-SOD1 reversed the inhibition of osteoblastic differentiation and downregulation of osteogenic gene markers induced by hydrogen peroxide. However, LMWP-SOD1 could not reverse the decrease in odontogenesis caused by hydrogen peroxide.Conclusion: Overall, cell-penetrating LMWP-SOD1 conjugates are effective for attenuation of cellular senescence and reversal of osteoblastic differentiation of DPSCs caused by oxidative stress inhibition. This result suggests potential application in the field of antiaging and tissue engineering to overcome the limitations of senescent stem cells.Keywords: superoxide

  2. In vivo near-infrared fluorescence imaging of FAP-expressing tumors with activatable FAP-targeted, single-chain Fv-immunoliposomes.

    Science.gov (United States)

    Rüger, Ronny; Tansi, Felista L; Rabenhold, Markus; Steiniger, Frank; Kontermann, Roland E; Fahr, Alfred; Hilger, Ingrid

    2014-07-28

    Molecular and cellular changes that precede the invasive growth of solid tumors include the release of proteolytic enzymes and peptides in the tumor stroma, the recruitment of phagocytic and lymphoid infiltrates and alteration of the extracellular matrix. The reactive tumor stroma consists of a large number of myofibroblasts, characterized by high expression of fibroblast activation protein alpha (FAP). FAP, a type-II transmembrane sialoglycoprotein is an attractive target in diagnosis and therapy of several pathologic disorders especially cancer. In the underlying work, a fluorescence-activatable liposome (fluorescence-quenched during circulation and fluorescence activation upon cellular uptake), bearing specific single-chain Fv fragments directed against FAP (scFv'FAP) was developed, and its potential for use in fluorescence diagnostic imaging of FAP-expressing tumor cells was evaluated by whole body fluorescence imaging. The liposomes termed anti-FAP-IL were prepared via post-insertion of ligand-phospholipid-conjugates into preformed DY-676-COOH-containing liposomes. The anti-FAP-IL revealed a homogeneous size distribution and showed specific interaction and binding with FAP-expressing cells in vitro. The high level of fluorescence quenching of the near-infrared fluorescent dye sequestered in the aqueous interior of the liposomes enables fluorescence imaging exclusively upon uptake and degradation by cells, which results in fluorescence activation. Only FAP-expressing cells were able to take up and activate fluorescence of anti-FAP-IL in vitro. Furthermore, anti-FAP-IL accumulated selectively in FAP-expressing xenograft models in vivo, as demonstrated by blocking experiments using free scFv'FAP. The local tumor fluorescence intensities were in agreement with the intrinsic degree of FAP-expression in different xenograft models. Thus, anti-FAP-IL can serve as a suitable in vivo diagnostic tool for pathological disorders accompanied by high FAP-expression.

  3. Intestinal Cell Tight Junctions Limit Invasion of Candida albicans through Active Penetration and Endocytosis in the Early Stages of the Interaction of the Fungus with the Intestinal Barrier.

    Directory of Open Access Journals (Sweden)

    Marianne Goyer

    Full Text Available C. albicans is a commensal yeast of the mucous membranes in healthy humans that can also cause disseminated candidiasis, mainly originating from the digestive tract, in vulnerable patients. It is necessary to understand the cellular and molecular mechanisms of the interaction of C. albicans with enterocytes to better understand the basis of commensalism and pathogenicity of the yeast and to improve the management of disseminated candidiasis. In this study, we investigated the kinetics of tight junction (TJ formation in parallel with the invasion of C. albicans into the Caco-2 intestinal cell line. Using invasiveness assays on Caco-2 cells displaying pharmacologically altered TJ (i.e. differentiated epithelial cells treated with EGTA or patulin, we were able to demonstrate that TJ protect enterocytes against invasion of C. albicans. Moreover, treatment with a pharmacological inhibitor of endocytosis decreased invasion of the fungus into Caco-2 cells displaying altered TJ, suggesting that facilitating access of the yeast to the basolateral side of intestinal cells promotes endocytosis of C. albicans in its hyphal form. These data were supported by SEM observations of differentiated Caco-2 cells displaying altered TJ, which highlighted membrane protrusions engulfing C. albicans hyphae. We furthermore demonstrated that Als3, a hypha-specific C. albicans invasin, facilitates internalization of the fungus by active penetration and induced endocytosis by differentiated Caco-2 cells displaying altered TJ. However, our observations failed to demonstrate binding of Als3 to E-cadherin as the trigger mechanism of endocytosis of C. albicans into differentiated Caco-2 cells displaying altered TJ.

  4. Penetrating abdominal trauma.

    Science.gov (United States)

    Henneman, P L

    1989-08-01

    The management of patients with penetrating abdominal trauma is outlined in Figure 1. Patients with hemodynamic instability, evisceration, significant gastrointestinal bleeding, peritoneal signs, gunshot wounds with peritoneal violation, and type 2 and 3 shotgun wounds should undergo emergency laparotomy. The initial ED management of these patients includes airway management, monitoring of cardiac rhythm and vital signs, history, physical examination, and placement of intravenous lines. Blood should be obtained for initial hematocrit, type and cross-matching, electrolytes, and an alcohol level or drug screen as needed. Initial resuscitation should utilize crystalloid fluid replacement. If more than 2 liters of crystalloid are needed to stabilize an adult (less in a child), blood should be given. Group O Rh-negative packed red blood cells should be immediately available for a patient in impending arrest or massive hemorrhage. Type-specific blood should be available within 15 minutes. A patient with penetrating thoracic and high abdominal trauma should receive a portable chest x-ray, and a hemo- or pneumothorax should be treated with tube thoracostomy. An unstable patient with clinical signs consistent with a pneumothorax, however, should receive a tube thoracostomy prior to obtaining roentgenographic confirmation. If time permits, a nasogastric tube and Foley catheter should be placed, and the urine evaluated for blood (these procedures can be performed in the operating room). If kidney involvement is suspected because of hematuria or penetrating trauma in the area of a kidney or ureter in a patient requiring surgery, a single-shot IVP should be performed either in the ED or the operating room. An ECG is important in patients with possible cardiac involvement and in patients over the age of 40 going to the operating room. Tetanus status should be updated, and appropriate antibiotics covering bowel flora should be given. Operative management should rarely be delayed

  5. Design and synthesis of phospholipase C and A2-activatable near-infrared fluorescent smart probes.

    Science.gov (United States)

    Popov, Anatoliy V; Mawn, Theresa M; Kim, Soungkyoo; Zheng, Gang; Delikatny, E James

    2010-10-20

    The primary focus of this work was to develop activatable probes suitable for in vivo detection of phospholipase activity. Phospholipases (PLs) are ubiquitous enzymes that perform a number of critical regulatory functions. They catalyze phospholipid breakdown and are categorized as A(1), A(2) (PLA(2)), C (PLC), and D (PLD) based on their site of action. Here, we report the design, synthesis, and characterization of self-quenching reporter probes that release fluorescent moieties upon cleavage with PLA(2) or PLC. A series of phospholipids were synthesized bearing the NIR fluorophore pyropheophorbide a (Pyro) at the sn-2 position. Fluorescence quenching was achieved by attachment of either a positively charged black hole quencher-3 (BHQ-3) to the phospholipid headgroup or another neutral Pyro moiety at the sn-1 position. The specificity to different phospholipases was modulated by insertion of spacers (C(6), C(12)) between Pyro and the lipid backbone. The specificity of the quenched fluorescent phospholipids was assayed on a plate reader against a number of phospholipases and compared with two commercial probes bearing the visible fluorophore BODIPY. While PyroC(6)-PyroC(6)-PtdCho revealed significant background fluorescence, and a 10% fluorescence increase under the action of PLA(2), Pyro-PtdEtn-BHQ demonstrated high selective sensitivity to PLC, particularly to the PC-PLC isoform, and its sensitivity to PLA(2) was negligible due to steric hindrance at the sn-2 position. In contrast, the C(12)-spacered PyroC(12)-PtdEtn-BHQ demonstrated a remarkable selectivity for PLA(2) and the best relative PLA(2)/PLC sensitivity, significantly outperforming previously known probes. These results open an avenue for future in vivo experiments and for new probes to detect PL activity. PMID:20882956

  6. Quantification of Cell-Penetrating Peptide Associated with Polymeric Nanoparticles Using Isobaric-Tagging and MALDI-TOF MS/MS

    Science.gov (United States)

    Chiu, Jasper Z. S.; Tucker, Ian G.; McDowell, Arlene

    2016-11-01

    High sensitivity quantification of the putative cell-penetrating peptide di-arginine-histidine (RRH) associated with poly (ethyl-cyanoacrylate) (PECA) nanoparticles was achieved without analyte separation, using a novel application of isobaric-tagging and high matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometry. Isobaric-tagging reaction equilibrium was reached after 5 min, with 90% or greater RRH peptide successfully isobaric-tagged after 60 min. The accuracy was greater than 90%, which indicates good reliability of using isobaric-tagged RRH as an internal standard for RRH quantification. The sample intra- and inter-spot coefficients of variations were less than 11%, which indicate good repeatability. The majority of RRH peptides in the nanoparticle formulation were physically associated with the nanoparticles (46.6%), whereas only a small fraction remained unassociated (13.7%). The unrecovered RRH peptide (~40%) was assumed to be covalently associated with PECA nanoparticles.

  7. Activatable albumin-photosensitizer nanoassemblies for triple-modal imaging and thermal-modulated photodynamic therapy of cancer.

    Science.gov (United States)

    Hu, Dehong; Sheng, Zonghai; Gao, Guanhui; Siu, Fungming; Liu, Chengbo; Wan, Qian; Gong, Ping; Zheng, Hairong; Ma, Yifan; Cai, Lintao

    2016-07-01

    Photodynamic therapy (PDT) is a noninvasive and effective approach for cancer treatment. The main bottlenecks of clinical PDT are poor selectivity of photosensitizer and inadequate oxygen supply resulting in serious side effects and low therapeutic efficiency. Herein, a thermal-modulated reactive oxygen species (ROS) strategy using activatable human serum albumin-chlorin e6 nanoassemblies (HSA-Ce6 NAs) for promoting PDT against cancer is developed. Through intermolecular disulfide bond crosslinking and hydrophobic interaction, Ce6 photosensitizer is effectively loaded into the HSA NAs, and the obtained HSA-Ce6 NAs exhibit excellent reduction response, as well as enhanced tumor accumulation and retention. By the precision control of the overall body temperature instead of local tumor temperature increasing from 37 °C to 43 °C, the photosensitization reaction rate of HSA-Ce6 NAs increases 20%, and the oxygen saturation of tumor tissue raise 52%, significantly enhancing the generation of ROS for promoting PDT. Meanwhile, the intrinsic fluorescence and photoacoustic properties, and the chelating characteristic of porphyrin ring can endow the HSA-Ce6 NAs with fluorescence, photoacoustic and magnetic resonance triple-modal imaging functions. Upon irradiation of low-energy near-infrared laser, the tumors are completely suppressed without tumor recurrence and therapy-induced side effects. The robust thermal-modulated ROS strategy combined with albumin-based activatable nanophotosensitizer is highly potential for multi-modal imaging-guided PDT and clinical translation. PMID:27061266

  8. Functional study of p38 mitogen-activated protein kinase based on cell-penetrating peptide delivery system

    Institute of Scientific and Technical Information of China (English)

    Liping Yang; Yongming Yao; Zhiyong Sheng; Xiaomei Zhu; Yong Jiang

    2009-01-01

    Objective p38 Mitogen-activated protein kinase (MAPK) is a crossing center of various pathways. In this study, protein transduction system based on human immunodeficiency virus (HIV)-1 transactivator of transcription (TAT), which is an efficient delivery peptide of the foreign proteins into cells, was employed to study p38 MAPK functions in eukaryotic cells. Methods p38 And its dominant negative form, p38AF, were constructed into pET-His-TAT vector correctly to verify that the recombinant plasmids were well-founded through restriction enzyme digestion and DNA sequencing. The two proteins, His-TAT-p38 and His-TAT-p38AF, were expressed and purified in Escherichia coli by SDS-PAGE. Then they were incubated with ECV304 cells respectively and readily transduced into cells in a time-dependent and dose-dependent manner. The cells were stimulated by sorbitol. Activating transcription factor (ATF) 2 phosphorylation level was checked using Western blot to assess the activity of endogenous p38. Results Compared with controls, it was found that His-TAT-p38 increased the level ofATF2 phosphorylation in sorbitol-stimulated ECV304 cells, while His-TAT-p38AF inhibited it, indicating p38 MAPK protein delivery system based on TAT was constructed successfully. TAT-p38 and its dominant negative form possessed high biological activity after transduction into ECV304 cells by TAT protein delivery system. The results showed that p38AF fused with TAT could inhibit the transduction of endogenous p38 signal pathway in part, and other pathway might regulate p38 phosphorylation. Conclusions Our study provides a novel pathway to inhibit p38 signal pathway and establish a new method to study p38 function.

  9. The effect of genetic variants in the thrombin activatable fibrinolysis inhibitor (TAFI) gene on TAFI-antigen levels, clot lysis time and the risk of venous thrombosis

    NARCIS (Netherlands)

    C.H. Martini (C.); A. Brandts (A.); E.L.E. de Bruijne; A. van Hylckama Vlieg (Astrid); F.W.G. Leebeek (Frank); T. Lisman (Ton); F.R. Rosendaal (Frits)

    2006-01-01

    textabstractThrombin activatable fibrinolysis inhibitor (TAFI) is an important inhibitor of fibrinolysis. High TAFI antigen levels are associated with an increased risk of deep venous thrombosis (DVT). Because TAFI levels are partly determined genetically, we assessed the association between three T

  10. Effect of long-term hormone replacement therapy on tissue factor pathway inhibitor and thrombin activatable fibrinolysis inhibitor in healthy postmenopausal women: a randomized controlled study

    DEFF Research Database (Denmark)

    Bladbjerg, E-M; Madsen, J S; Kristensen, S R;

    2003-01-01

    arterial tissue factor expression and higher thrombin formation, and changes in tissue factor pathway coagulation inhibitor (TFPI) and thrombin activatable fibrinolysis inhibitor (TAFI) may be deleterious. Healthy postmenopausal women (n = 719) were randomized to hormone therapy [n = 357; opposed (n = 290...

  11. Parallel Synthesis of Cell-Penetrating Peptide Conjugates of PMO Toward Exon Skipping Enhancement in Duchenne Muscular Dystrophy

    NARCIS (Netherlands)

    O'Donovan, Liz; Okamoto, Itaru; Arzumanov, Andrey A.; Williams, Donna L.; Deuss, Peter; Gait, Michael J.

    2015-01-01

    We describe two new methods of parallel chemical synthesis of libraries of peptide conjugates of phosphorodiamidate morpholino oligonucleotide (PMO) cargoes on a scale suitable for cell screening prior to in vivo analysis for therapeutic development. The methods represent an extension of the SELecti

  12. Hair follicle targeting, penetration enhancement and Langerhans cell activation make cyanoacrylate skin surface stripping a promising delivery technique for transcutaneous immunization with large molecules and particle-based vaccines.

    Science.gov (United States)

    Vogt, Annika; Hadam, Sabrina; Deckert, Iliane; Schmidt, Julia; Stroux, Andrea; Afraz, Zahra; Rancan, Fiorenza; Lademann, Jürgen; Combadiere, Behazine; Blume-Peytavi, Ulrike

    2015-01-01

    Transcutaneous immunization (TCI) requires targeting of a maximum number of skin antigen-presenting cells as non-invasive as possible on small skin areas. In two clinical trials, we introduced cyanoacrylate skin surface stripping (CSSS) as a safe method for TCI. Here, using ex vivo human skin, we demonstrate that one CSSS procedure removed only 30% of stratum corneum, but significantly increased the penetration of 200 nm polystyrene particles deep into vellus and intermediate hair follicles from where they could not been retrieved by conventional tape stripping. Two subsequent CSSS had no striking additional effect. CSSS increased particle penetration in superficial stratum corneum and induced Langerhans cell activation. Formulation in amphiphilic ointment or massage did not substantially influences the interfollicular penetration profiles. Hair follicle (HF) targeting by CSSS could become a highly effective tool for TCI when combined with carrier-based delivery and is gaining new attention as our understanding on the HF immune system increases.

  13. Hair follicle targeting, penetration enhancement and Langerhans cell activation make cyanoacrylate skin surface stripping a promising delivery technique for transcutaneous immunization with large molecules and particle-based vaccines.

    Science.gov (United States)

    Vogt, Annika; Hadam, Sabrina; Deckert, Iliane; Schmidt, Julia; Stroux, Andrea; Afraz, Zahra; Rancan, Fiorenza; Lademann, Jürgen; Combadiere, Behazine; Blume-Peytavi, Ulrike

    2015-01-01

    Transcutaneous immunization (TCI) requires targeting of a maximum number of skin antigen-presenting cells as non-invasive as possible on small skin areas. In two clinical trials, we introduced cyanoacrylate skin surface stripping (CSSS) as a safe method for TCI. Here, using ex vivo human skin, we demonstrate that one CSSS procedure removed only 30% of stratum corneum, but significantly increased the penetration of 200 nm polystyrene particles deep into vellus and intermediate hair follicles from where they could not been retrieved by conventional tape stripping. Two subsequent CSSS had no striking additional effect. CSSS increased particle penetration in superficial stratum corneum and induced Langerhans cell activation. Formulation in amphiphilic ointment or massage did not substantially influences the interfollicular penetration profiles. Hair follicle (HF) targeting by CSSS could become a highly effective tool for TCI when combined with carrier-based delivery and is gaining new attention as our understanding on the HF immune system increases. PMID:25382068

  14. A single-cell technique for the measurement of membrane potential, membrane conductance, and the efflux of rapidly penetrating solutes in Amphiuma erythrocytes.

    Science.gov (United States)

    Stoner, L C; Kregenow, F M

    1980-10-01

    We describe a single-cell technique for measuring membrane potential, membrane resistance, and the efflux of rapidly penetrating solutes such as Cl and H2O. Erythrocytes from Amphiuma means were aspirated into a Sylgard (Dow Corning Corp.)-coated capillary. The aspirated cell separated a solution within the capillary from a solution in the bath. Each of these two solutions was contiguous with approximately 5% of the total membrane surface. Microelectrodes placed concentrically within the capillary permit the measurement of intracellular voltage, specific membrane resistance, and the electrical seal between the two solutions. The intracellular voltage averaged -17.7 mV (pH 7.6) and changed as either intra- or extracellular chloride was varied. The average specific membrane resistance measured by passing current across the exposed membrane surface was 110 ohm-cm2. 36Cl and tritiated H2O fluxes (0.84 +/- 0.05 x 10(-6) M . cm-2 . min-1 and 6.4 +/- 1.5 x 10(-3) M . cm-2 . min-1, respectively) were determined by noting the rate at which isotope leaves the cell and crosses the membrane exposed to the bath. Our measured values for the flux of 36Cl and tritiated H2O approximate reported values for free-floating cells. 36Cl efflux, in addition, is inhibited by 4-acetamido-4'-isothiocyano-stilbene 2,2'-disulfonic acid (SITS) and furosemide, known inhibitors of the anion exchange mechanism responsible for the rapid anion fluxes of red blood cells. One can also demonstrate directly that > 89% of 36Cl efflux is "electrically silent" by analyzing the flux in the presence of an imposed transcellular voltage.

  15. A Multilayered Cell Culture Model for Transport Study in Solid Tumors: Evaluation of Tissue Penetration of Polyethyleneimine Based Cationic Micelles

    OpenAIRE

    Miura, Seiji; Suzuki, Hidenori; Bae, You Han

    2014-01-01

    Limited drug distribution is partially responsible for the efficacy gap between preclinical and clinical studies of nano-sized drug carriers for cancer therapy. In this study, we examined the transport behavior of cationic micelles formed from a triblock copolymer of poly(D,L-lactide-co-glycolide)-block-branched polyethyleneimine-block-poly(D,L-lactide-co-glycolide) using a unique in vitro tumor model composed of a multilayered cell culture (MCC) and an Ussing chamber system. The Cy3-labeled ...

  16. Parallel Synthesis of Cell-Penetrating Peptide Conjugates of PMO Toward Exon Skipping Enhancement in Duchenne Muscular Dystrophy

    OpenAIRE

    O'Donovan, Liz; Okamoto, Itaru; Arzumanov, Andrey A; Williams, Donna L.; Deuss, Peter; Gait, Michael J.

    2015-01-01

    We describe two new methods of parallel chemical synthesis of libraries of peptide conjugates of phosphorodiamidate morpholino oligonucleotide (PMO) cargoes on a scale suitable for cell screening prior to in vivo analysis for therapeutic development. The methods represent an extension of the SELection of PEPtide CONjugates (SELPEPCON) approach previously developed for parallel peptide-peptide nucleic acid (PNA) synthesis. However, these new methods allow for the utilization of commercial PMO ...

  17. Sucrose esters increase drug penetration, but do not inhibit p-glycoprotein in caco-2 intestinal epithelial cells.

    Science.gov (United States)

    Kiss, Lóránd; Hellinger, Éva; Pilbat, Ana-Maria; Kittel, Ágnes; Török, Zsolt; Füredi, András; Szakács, Gergely; Veszelka, Szilvia; Sipos, Péter; Ózsvári, Béla; Puskás, László G; Vastag, Monika; Szabó-Révész, Piroska; Deli, Mária A

    2014-10-01

    Sucrose fatty acid esters are increasingly used as excipients in pharmaceutical products, but few data are available on their toxicity profile, mode of action, and efficacy on intestinal epithelial models. Three water-soluble sucrose esters, palmitate (P-1695), myristate (M-1695), laurate (D-1216), and two reference absorption enhancers, Tween 80 and Cremophor RH40, were tested on Caco-2 cells. Caco-2 monolayers formed a good barrier as reflected by high transepithelial resistance and positive immunostaining for junctional proteins claudin-1, ZO-1, and β-catenin. Sucrose esters in nontoxic concentrations significantly reduced resistance and impedance, and increased permeability for atenolol, fluorescein, vinblastine, and rhodamine 123 in Caco-2 monolayers. No visible opening of the tight junctions was induced by sucrose esters assessed by immunohistochemistry and electron microscopy, but some alterations were seen in the structure of filamentous actin microfilaments. Sucrose esters fluidized the plasma membrane and enhanced the accumulation of efflux transporter ligands rhodamine 123 and calcein AM in epithelial cells, but did not inhibit the P-glycoprotein (P-gp)-mediated calcein AM accumulation in MES-SA/Dx5 cell line. These data indicate that in addition to their dissolution-increasing properties sucrose esters can enhance drug permeability through both the transcellular and paracellular routes without inhibiting P-gp.

  18. A mouse model for inducible overexpression of Prdm14 results in rapid-onset and highly penetrant T-cell acute lymphoblastic leukemia (T-ALL

    Directory of Open Access Journals (Sweden)

    Brandi L. Carofino

    2013-11-01

    PRDM14 functions in embryonic stem cell (ESC maintenance to promote the expression of pluripotency-associated genes while suppressing differentiation genes. Expression of PRDM14 is tightly regulated and typically limited to ESCs and primordial germ cells; however, aberrant expression is associated with tumor initiation in a wide variety of human cancers, including breast cancer and leukemia. Here, we describe the generation of a Cre-recombinase-inducible mouse model for the spatial and temporal control of Prdm14 misexpression [ROSA26 floxed-stop Prdm14 (R26PR]. When R26PR is mated to either of two Cre lines, Mx1-cre or MMTV-cre, mice develop early-onset T-cell acute lymphoblastic leukemia (T-ALL with median overall survival of 41 and 64 days for R26PR;Mx1-cre and R26PR;MMTV-cre, respectively. T-ALL is characterized by the accumulation of immature single-positive CD8 cells and their widespread infiltration. Leukemia is preceded by a dramatic expansion of cells resembling hematopoietic stem cells and lymphoid-committed progenitors prior to disease onset, accompanied by a blockage in B-cell differentiation at the early pro-B stage. Rapid-onset PRDM14-induced T-ALL requires factors that are present in stem and progenitor cells: R26PR;dLck-cre animals, which express Prdm14 starting at the double-positive stage of thymocyte development, do not develop disease. PRDM14-induced leukemic cells contain high levels of activated NOTCH1 and downstream NOTCH1 targets, including MYC and HES1, and are sensitive to pharmacological inhibition of NOTCH1 with the γ-secretase inhibitor DAPT. Greater than 50% of human T-ALLs harbor activating mutations in NOTCH1; thus, our model carries clinically relevant molecular aberrations. The penetrance, short latency and involvement of the NOTCH1 pathway will make this hematopoietic R26PR mouse model ideal for future studies on disease initiation, relapse and novel therapeutic drug combinations. Furthermore, breeding R26PR to additional Cre

  19. Structural Elucidation of the Cell-Penetrating Penetratin Peptide in Model Membranes at the Atomic Level: Probing Hydrophobic Interactions in the Blood-Brain Barrier.

    Science.gov (United States)

    Bera, Swapna; Kar, Rajiv K; Mondal, Susanta; Pahan, Kalipada; Bhunia, Anirban

    2016-09-01

    Cell-penetrating peptides (CPPs) have shown promise in nonpermeable therapeutic drug delivery, because of their ability to transport a variety of cargo molecules across the cell membranes and their noncytotoxicity. Drosophila antennapedia homeodomain-derived CPP penetratin (RQIKIWFQNRRMKWKK), being rich in positively charged residues, has been increasingly used as a potential drug carrier for various purposes. Penetratin can breach the tight endothelial network known as the blood-brain barrier (BBB), permitting treatment of several neurodegenerative maladies, including Alzheimer's disease, Parkinson's disease, and Huntington's disease. However, a detailed structural understanding of penetratin and its mechanism of action is lacking. This study defines structural features of the penetratin-derived peptide, DK17 (DRQIKIWFQNRRMKWKK), in several model membranes and describes a membrane-induced conformational transition of the DK17 peptide in these environments. A series of biophysical experiments, including high-resolution nuclear magnetic resonance spectroscopy, provides the three-dimensional structure of DK17 in different membranes mimicking the BBB or total brain lipid extract. Molecular dynamics simulations support the experimental results showing preferential binding of DK17 to particular lipids at atomic resolution. The peptide conserves the structure of the subdomain spanning residues Ile6-Arg11, despite considerable conformational variation in different membrane models. In vivo data suggest that the wild type, not a mutated sequence, enters the central nervous system. Together, these data highlight important structural and functional attributes of DK17 that could be utilized in drug delivery for neurodegenerative disorders. PMID:27532224

  20. Peptide nucleic acid (PNA) cell penetrating peptide (CPP) conjugates as carriers for cellular delivery of antisense oligomers

    DEFF Research Database (Denmark)

    Shiraishi, Takehiko; Nielsen, Peter E

    2012-01-01

    splicing correction of the mutated luciferase gene in the HeLa pLuc705 cell line, reporting cellular (nuclear) uptake of the antisense PNA via luciferase activity measurement. Carrier CPP-PNA constructs were studied in terms of construct modification (with octaarginine and/or decanoic acid) and carrier PNA...... that the carrier might facilitate endosomal escape. Furthermore, 50% downregulation of luciferase expression at 60 nM siRNA was obtained using this carrier CPP-PNA delivery strategy (with CQ co-treatment) for a single stranded antisense RNA targeting normal luciferase mRNA. These results indicated that CPP...

  1. Parallel synthesis of cell-penetrating peptide conjugates of PMO toward exon skipping enhancement in Duchenne muscular dystrophy.

    Science.gov (United States)

    O'Donovan, Liz; Okamoto, Itaru; Arzumanov, Andrey A; Williams, Donna L; Deuss, Peter; Gait, Michael J

    2015-02-01

    We describe two new methods of parallel chemical synthesis of libraries of peptide conjugates of phosphorodiamidate morpholino oligonucleotide (PMO) cargoes on a scale suitable for cell screening prior to in vivo analysis for therapeutic development. The methods represent an extension of the SELection of PEPtide CONjugates (SELPEPCON) approach previously developed for parallel peptide-peptide nucleic acid (PNA) synthesis. However, these new methods allow for the utilization of commercial PMO as cargo with both C- and N-termini unfunctionalized. The synthetic methods involve conjugation in solution phase, followed by rapid purification via biotin-streptavidin immobilization and subsequent reductive release into solution, avoiding the need for painstaking high-performance liquid chromatography purifications. The synthesis methods were applied for screening of PMO conjugates of a 16-member library of variants of a 10-residue ApoE peptide, which was suggested for blood-brain barrier crossing. In this work the conjugate library was tested in an exon skipping assay using skeletal mouse mdx cells, a model of Duchene's muscular dystrophy where higher activity peptide-PMO conjugates were identified compared with the starting peptide-PMO. The results demonstrate the power of the parallel synthesis methods for increasing the speed of optimization of peptide sequences in conjugates of PMO for therapeutic screening. PMID:25412073

  2. Metasploit penetration testing cookbook

    CERN Document Server

    Agarwal, Monika

    2013-01-01

    This book follows a Cookbook style with recipes explaining the steps for penetration testing with WLAN, VOIP, and even cloud computing. There is plenty of code and commands used to make your learning curve easy and quick.This book targets both professional penetration testers as well as new users of Metasploit, who wish to gain expertise over the framework and learn an additional skill of penetration testing, not limited to a particular OS. The book requires basic knowledge of scanning, exploitation, and the Ruby language.

  3. Penetration testing with Perl

    CERN Document Server

    Berdeaux, Douglas

    2014-01-01

    If you are an expert Perl programmer interested in penetration testing or information security, this guide is designed for you. However, it will also be helpful for you even if you have little or no Linux shell experience.

  4. Prolongation of life in rats with malignant glioma by intranasal siRNA/drug codelivery to the brain with cell-penetrating peptide-modified micelles.

    Science.gov (United States)

    Kanazawa, Takanori; Morisaki, Kazuki; Suzuki, Shohei; Takashima, Yuuki

    2014-05-01

    New therapeutic strategies are required to develop candidate drugs and ensure efficient delivery of these drugs to the brain and the central nervous system (CNS). Small interfering RNA (siRNA)-based therapies have been investigated as potential novel approaches for the treatment of brain disorders. Previously, we showed that Tat, a cell-penetrating peptide derived from HIV-Tat, and the modified block copolymers (MPEG-PCL-Tat) can form stable complexes with siRNA or can be loaded with an anticancer drug and efficiently deliver the drugs to the brain tissue via intranasal delivery. In this study, to develop a novel, efficient, and safe therapeutic strategy for managing brain disorders, we used MPEG-PCL-Tat micelles with a nose-to-brain delivery system to investigate its therapeutic effects on a rat model of malignant glioma using siRNA with a Raf-1 (siRaf-1)/camptothecin (CPT) codelivery system. MPEG-PCL-Tat and CPT-loaded MPEG-PCL-Tat can form a stable complex with siRNA with a particle size from 60 to 200 nm and a positive charge at N/P ratios up to 5. Additionally, MPEG-PCL-Tat/siRaf-1 and CPT-loaded MPEG-PCL-Tat/siRaf-1 have fostered cell death in rat glioma cells after the high cellular uptake of siRaf-1/drug by the MPEG-PCL-Tat carrier. Furthermore, compared to the unloaded MPEG-PCL-Tat/siRaf-1 complex, a CPT-loaded MPEG-PCL-Tat/siRaf-1 complex achieved the high therapeutic effect because of the additive effects of CPT and siRaf-1. These results indicate that drug/siRNA codelivery using MPEG-PCL-Tat nanomicelles with nose-to-brain delivery is an excellent therapeutic approach for brain and CNS diseases.

  5. Penetration of short fluorescence-labeled peptides into the nucleus in HeLa cells and in vitro specific interaction of the peptides with deoxyribooligonucleotides and DNA.

    Science.gov (United States)

    Fedoreyeva, L I; Kireev, I I; Khavinson, V Kh; Vanyushin, B F

    2011-11-01

    Marked fluorescence in cytoplasm, nucleus, and nucleolus was observed in HeLa cells after incubation with each of several fluorescein isothiocyanate-labeled peptides (epithalon, Ala-Glu-Asp-Gly; pinealon, Glu-Asp-Arg; testagen, Lys-Glu-Asp-Gly). This means that short biologically active peptides are able to penetrate into an animal cell and its nucleus and, in principle they may interact with various components of cytoplasm and nucleus including DNA and RNA. It was established that various initial (intact) peptides differently affect the fluorescence of the 5,6-carboxyfluorescein-labeled deoxyribooligonucleotides and DNA-ethidium bromide complexes. The Stern-Volmer constants characterizing the degree of fluorescence quenching of various single- and double-stranded fluorescence-labeled deoxyribooligonucleotides with short peptides used were different depending on the peptide primary structures. This indicates the specific interaction between short biologically active peptides and nucleic acid structures. On binding to them, the peptides discriminate between different nucleotide sequences and recognize even their cytosine methylation status. Judging from corresponding constants of the fluorescence quenching, the epithalon, pinealon, and bronchogen (Ala-Glu-Asp-Leu) bind preferentially with deoxyribooligonucleotides containing CNG sequence (CNG sites are targets for cytosine DNA methylation in eukaryotes). Epithalon, testagen, and pinealon seem to preferentially bind with CAG- but bronchogen with CTG-containing sequences. The site-specific interactions of peptides with DNA can control epigenetically the cell genetic functions, and they seem to play an important role in regulation of gene activity even at the earliest stages of life origin and in evolution.

  6. Polypyrrole-based nanotheranostics for activatable fluorescence imaging and chemo/photothermal dual therapy of triple-negative breast cancer

    Science.gov (United States)

    Park, Dongjin; Ahn, Kyung-Ohk; Jeong, Kyung-Chae; Choi, Yongdoo

    2016-05-01

    Here, we fabricated polypyrrole nanoparticles (PPys) (termed HA10-PPy, HA20-PPy, and HA40-PPy) doped with different average molecular weight hyaluronic acids (HAs) (10, 20, and 40 kDa, respectively), and evaluated the effect of molecular weight of doped HA on photothermal induction, fluorescence quenching, and drug loading efficiencies. Doxorubicin-loaded HA-doped PPys (DOX@HA-PPys) could be used for imaging and therapy of triple-negative breast cancer (TNBC). Fluorescence turn-on, stimuli-responsive drug release, and photo-induced heating of DOX@HA-PPys enabled not only activatable fluorescence imaging but also subsequent chemo/photothermal dual therapy for TNBC. In particular, we illustrated the potential usefulness of the photothermal effect of the nanoparticles for overcoming chemoresistance in TNBC.

  7. Thermodynamics of cell-penetrating HIV1 TAT peptide insertion into PC/PS/CHOL model bilayers through transmembrane pores: the roles of cholesterol and anionic lipids.

    Science.gov (United States)

    Hu, Yuan; Patel, Sandeep

    2016-08-10

    Efficient delivery of pharmaceutically active molecules across cellular membranes using cell penetrating peptides (CPPs), such as the cationic human immunodeficiency virus-1 trans-acting activator of transcription peptide (HIV-1 TAT), continues to attract scientific attention in drug design and disease treatment. Experimental results show that the TAT peptide is not only capable of directly penetrating the biological membrane in a passive manner, but also forming physical, membrane-spanning pores that may facilitate transport. Experiments further show that anionic lipids accelerate peptide permeation within a range of mole percentage composition. In this work, we explored the structures and translocation thermodynamics of the cationic TAT peptide across a series of DPPC/DPPS model membranes with the presence of 0-30 mol% cholesterol. We computed the potentials of the mean force by using umbrella sampling molecular dynamics simulations coupled to the Martini coarse-grained force field. We systematically investigated the roles of cholesterol and anionic lipids (membrane surface charge) in TAT peptide translocation. In qualitative agreement with experimental findings, the barrier heights were significantly reduced in the presence of anionic lipids. A toroidal hydrophilic pore was strongly suggested by membrane structure analysis. Cholesterol stabilizes the liquid-ordered (Lo) phase of membranes and increases the elastic stiffness of bilayers. Consequently, it hinders transmembrane pore formation and thus modulates solute permeability, since the liquid-ordered phase suppresses reorientation of the lipid molecules on simulation time scales. Though cholesterol contributes marginally to the total free energy associated with peptide permeation, the coordination of cholesterol to the peptide weakens more favorable peptide-lipid interactions. The addition of the anionic lipid DPPS to the neutral DPPC bilayer leads to the emergence and further enhancement of an interfacially

  8. Deployable Wireless Camera Penetrators

    Science.gov (United States)

    Badescu, Mircea; Jones, Jack; Sherrit, Stewart; Wu, Jiunn Jeng

    2008-01-01

    A lightweight, low-power camera dart has been designed and tested for context imaging of sampling sites and ground surveys from an aerobot or an orbiting spacecraft in a microgravity environment. The camera penetrators also can be used to image any line-of-sight surface, such as cliff walls, that is difficult to access. Tethered cameras to inspect the surfaces of planetary bodies use both power and signal transmission lines to operate. A tether adds the possibility of inadvertently anchoring the aerobot, and requires some form of station-keeping capability of the aerobot if extended examination time is required. The new camera penetrators are deployed without a tether, weigh less than 30 grams, and are disposable. They are designed to drop from any altitude with the boost in transmitting power currently demonstrated at approximately 100-m line-of-sight. The penetrators also can be deployed to monitor lander or rover operations from a distance, and can be used for surface surveys or for context information gathering from a touch-and-go sampling site. Thanks to wireless operation, the complexity of the sampling or survey mechanisms may be reduced. The penetrators may be battery powered for short-duration missions, or have solar panels for longer or intermittent duration missions. The imaging device is embedded in the penetrator, which is dropped or projected at the surface of a study site at 90 to the surface. Mirrors can be used in the design to image the ground or the horizon. Some of the camera features were tested using commercial "nanny" or "spy" camera components with the charge-coupled device (CCD) looking at a direction parallel to the ground. Figure 1 shows components of one camera that weighs less than 8 g and occupies a volume of 11 cm3. This camera could transmit a standard television signal, including sound, up to 100 m. Figure 2 shows the CAD models of a version of the penetrator. A low-volume array of such penetrator cameras could be deployed from an

  9. Study on Penetration Characteristics of Tungsten Cylindrical Penetrator

    Energy Technology Data Exchange (ETDEWEB)

    Jo, Jong Hyun; Lee, Young Shin; Kim, Jae Hoon [Chungnam Nat' l Univ., Daejeon (Korea, Republic of); Bae, Yong Woon [Agency for Defense Development, Daejeon (Korea, Republic of)

    2013-09-15

    The design of missile require extremely small warheads that must be highly efficient and lethal. The penetration characteristics of each penetrator and the total number of penetrators on the warhead are obvious key factors that influence warhead lethality. The design of the penetrator shape and size are directly related to the space and weight of the warhead. The design of the penetrator L/D was directly related to the space and weight of the warhead. L and D are the length and the diameter of the projectile, respectively. The AUTODYN-3a code was used to study the effect of penetrator penetration. The objective of numerical analysis was to determine the penetration characteristics of penetrator produced by hypervelocity impacts under different initial conditions such as initial velocity, obliquity angle and L/D of penetrator. The residual velocity and residual mass were decreased with increasing initial impact velocity under L/D{<=}4.

  10. Ground penetrating radar

    CERN Document Server

    Daniels, David J

    2004-01-01

    Ground-penetrating radar has come to public attention in recent criminal investigations, but has actually been a developing and maturing remote sensing field for some time. In the light of recent expansion of the technique to a wide range of applications, the need for an up-to-date reference has become pressing. This fully revised and expanded edition of the best-selling Surface-Penetrating Radar (IEE, 1996) presents, for the non-specialist user or engineer, all the key elements of this technique, which span several disciplines including electromagnetics, geophysics and signal processing. The

  11. Session: Hard Rock Penetration

    Energy Technology Data Exchange (ETDEWEB)

    Tennyson, George P. Jr.; Dunn, James C.; Drumheller, Douglas S.; Glowka, David A.; Lysne, Peter

    1992-01-01

    This session at the Geothermal Energy Program Review X: Geothermal Energy and the Utility Market consisted of five presentations: ''Hard Rock Penetration - Summary'' by George P. Tennyson, Jr.; ''Overview - Hard Rock Penetration'' by James C. Dunn; ''An Overview of Acoustic Telemetry'' by Douglas S. Drumheller; ''Lost Circulation Technology Development Status'' by David A. Glowka; ''Downhole Memory-Logging Tools'' by Peter Lysne.

  12. Photoacoustic Imaging: Semiconducting Oligomer Nanoparticles as an Activatable Photoacoustic Probe with Amplified Brightness for In Vivo Imaging of pH (Adv. Mater. 19/2016).

    Science.gov (United States)

    Miao, Qingqing; Lyu, Yan; Ding, Dan; Pu, Kanyi

    2016-05-01

    Despite the great potential of photoacoustic imaging in the life sciences, the development of smart activatable photoacoustic probes remains elusive. On page 3662, K. Pu and co-workers report a facile nanoengineering approach based on semiconducting oligomer nano-particles to develop ratiometric photoacoustic probes with amplified brightness and enhanced sensing capability for accurate photoacoustic mapping of pH in the tumors of living mice.

  13. The cell-penetrating peptide domain from human heparin-binding epidermal growth factor-like growth factor (HB-EGF) has anti-inflammatory activity in vitro and in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jue-Yeon; Seo, Yoo-Na; Park, Hyun-Jung [Research Center, Nano Intelligent Biomedical Engineering Corporation (NIBEC), Seoul (Korea, Republic of); Park, Yoon-Jeong, E-mail: parkyj@snu.ac.kr [Research Center, Nano Intelligent Biomedical Engineering Corporation (NIBEC), Seoul (Korea, Republic of); Department of Dental Regenerative Biotechnology, Dental Research Institute and School of Dentistry, Seoul National University, Seoul (Korea, Republic of); Chung, Chong-Pyoung, E-mail: ccpperio@snu.ac.kr [Research Center, Nano Intelligent Biomedical Engineering Corporation (NIBEC), Seoul (Korea, Republic of); Department of Periodontology, College of Dentistry, Seoul National University, Seoul (Korea, Republic of)

    2012-03-23

    Highlights: Black-Right-Pointing-Pointer HBP sequence identified from HB-EGF has cell penetration activity. Black-Right-Pointing-Pointer HBP inhibits the NF-{kappa}B dependent inflammatory responses. Black-Right-Pointing-Pointer HBP directly blocks phosphorylation and degradation of I{kappa}B{alpha}. Black-Right-Pointing-Pointer HBP inhibits nuclear translocation of NF-{kappa}B p65 subunit. -- Abstract: A heparin-binding peptide (HBP) sequence from human heparin-binding epidermal growth factor-like growth factor (HB-EGF) was identified and was shown to exhibit cell penetration activity. This cell penetration induced an anti-inflammatory reaction in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. HBP penetrated the cell membrane during the 10 min treatment and reduced the LPS-induced production of nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cytokines (TNF-{alpha} and IL-6) in a concentration-dependent manner. Additionally, HBP inhibited the LPS-induced upregulation of cytokines, including TNF-{alpha} and IL-6, and decreased the interstitial infiltration of polymorphonuclear leukocytes in a lung inflammation model. HBP inhibited NF-{kappa}B-dependent inflammatory responses by directly blocking the phosphorylation and degradation of I{kappa}B{alpha} and by subsequently inhibiting the nuclear translocation of the p65 subunit of NF-{kappa}B. Taken together, this novel HBP may be potentially useful candidate for anti-inflammatory treatments and can be combined with other drugs of interest to transport attached molecules into cells.

  14. Percutaneous Penetration - Methodological Considerations

    DEFF Research Database (Denmark)

    Holmgaard, Rikke; Benfeldt, Eva; Nielsen, Jesper B

    2014-01-01

    Review discusses some of the existing and well-known experimental in vitro and in vivo methods for studies of percutaneous penetration together with some more recent and promising methods. After this, some considerations and recommendations about advantages and limitations of the different methods...

  15. Penetration of erythromycin through Staphylococcus epidermidis biofilm

    Institute of Scientific and Technical Information of China (English)

    LIN Mao-hu; HE Lei; GAO Jie; LIU Yun-xi; SUO Ji-jiang; XING Yu-bin; JIA Ning

    2013-01-01

    Background The catheter related infection caused by Staphylococcus epiderrnidis biofilm is increasing and difficult to treat by antimicrobial chemotherapy.The properties of biofilms that give rise to antibiotic resistance are only partially understood.This study aimed to elucidate the penetration of erythromycin through Staphylococcus epidermidis biofilm.Methods The penetration ratio of erythromycin through Staphylococcus epidermidis biofilms of 1457,1457-msrA,and wild isolate S68 was detected by biofilm penetration model at different time points according to the standard regression curve.The RNNDNA ratio and the cell density within the biofilms were observed by confocal laser microscope and transmission electromicroscope,respectively.Results The penetration ratios of erythromycin through the biofilms of 1457,1457-msrA,and S68 after cultivation for 36 hours were 0.93,0.55 and 0.4,respectively.The erythromycin penetration ratio through 1457 biofilm (0.58 after 8 hours)was higher than that through the other two (0.499 and 0.31 after 24 hours).Lower growth rate of the cells in biofilm was shown,with reduction of RNA/DNA proportion observed by confocal laser microscope through acridine orange stain.Compared with the control group observed by transmission electrmicroscope,the cell density of biofilm air face was lower than that of agar face,with more cell debris.Conclusions Erythromycin could penetrate to the Staphylococcus epidermidis biofilm,but could not kill the cells thoroughly.The lower growth rate of the cells within biofilm could help decreasing the erythromycin susceptibility.

  16. Prevention of Escherichia coli K1 Penetration of the Blood-Brain Barrier by Counteracting the Host Cell Receptor and Signaling Molecule Involved in E. coli Invasion of Human Brain Microvascular Endothelial Cells▿

    OpenAIRE

    Zhu, Longkun; Pearce, Donna; Kim, Kwang Sik

    2010-01-01

    Escherichia coli meningitis is an important cause of mortality and morbidity, and a key contributing factor is our incomplete understanding of the pathogenesis of E. coli meningitis. We have shown that E. coli penetration into the brain requires E. coli invasion of human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. E. coli invasion of HBMEC involves its interaction with HBMEC receptors, such as E. coli cytotoxic necrotizing factor 1 (CNF1) interacti...

  17. Cellular penetration and nuclear importation properties of {sup 111}In-labeled and {sup 123}I-labeled HIV-1 tat peptide immunoconjugates in BT-474 human breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Cornelissen, Bart [Division of Nuclear Medicine, University Health Network, Toronto, ON, M5S 3M2 (Canada); Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada); Hu, Meiduo [Division of Nuclear Medicine, University Health Network, Toronto, ON, M5S 3M2 (Canada); Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada); McLarty, Kristin [Division of Nuclear Medicine, University Health Network, Toronto, ON, M5S 3M2 (Canada); Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada); Costantini, Dan [Division of Nuclear Medicine, University Health Network, Toronto, ON, M5S 3M2 (Canada); Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada); Reilly, Raymond M. [Division of Nuclear Medicine, University Health Network, Toronto, ON, M5S 3M2 (Canada) and Department of Medical Imaging, University of Toronto, Toronto, ON, M5S 3M2 (Canada) and Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, M5S 3M2 (Canada) and Toronto General Research Institute, University Health Network, Toronto, ON, M5S 3M2 (Canada)]. E-mail: raymond.reilly@utoronto.ca

    2007-01-15

    Introduction: Our objective was to compare the cell penetration and nuclear importation properties of {sup 111}In-labeled and {sup 123}I-labeled immunoconjugates (ICs) composed of 16-mer peptides (GRKKRRQRRRPPQGYG) derived from HIV-1 transactivator of transcription (tat) protein and anti-mouse IgG (mIgG) in BT-474 breast cancer (BC) cells. Methods: [{sup 111}In]tat ICs were constructed by site-specific conjugation of tat peptides to NaIO{sub 4} {sup -}-oxidized carbohydrates in the Fc domain of diethylenetriaminepentaacetic-acid-modified anti-mIgG antibodies. Immunoreactivity against mIgG was assessed in a competition assay. The kinetics of the accumulation of [{sup 111}In]anti-mIgG-tat IC and [{sup 123}I]anti-mIgG-tat ICs in BT-474 cells and the elimination of radioactivity from cells, cytoplasm or nuclei were determined. The effects of excess tat peptides or NH{sub 4}Cl (an inhibitor of endosomal acidification) on cellular uptake and nuclear importation of [{sup 111}In]anti-mIgG-tat were measured. Results: [{sup 111}In]anti-mIgG-tat was >97% radiochemically pure and exhibited preserved immunoreactivity with mIgG epitopes. [{sup 123}I]Anti-mIgG-tat penetrated BT-474 cells more rapidly than [{sup 111}In]anti-mIgG-tat ICs and achieved a 1.5-fold to a 2-fold higher uptake in cells and nuclei. Cell penetration and nuclear uptake of [{sup 111}In]anti-mIgG-tat were inhibited by excess tat peptides and NH{sub 4}Cl. Elimination of radioactivity from BT-474 cells and nuclei was more rapid and complete for {sup 123}I-labeled than for {sup 111}In-labeled anti-mIgG-tat ICs. Conclusion: Tat peptides derived from HIV-1 tat protein promoted the penetration and nuclear uptake of radioactivity following the incubation of {sup 111}In-labeled and {sup 123}I-labeled anti-mIgG antibodies with BT-474 human BC cells. {sup 111}In-labeled tat ICs are feasible for inserting radionuclides into cancer cells with potential for targeting intracellular and, particularly, nuclear epitopes for

  18. Ground-penetrating rada

    Science.gov (United States)

    Thuma, W. R.

    The theory and applications of digital Ground-Penetrating Radar were discussed at a 5-day seminar held at the China University of Geosciences in Wuhan, People's Republic of China, in April. Cohosted by the Department of Applied Geophysics and Canada-China Geoscience, more than 60 senior geophysicists, engineers, technical specialists, university professors and researchers attended.Focus of the meeting was the expanded uses of the new deep-penetrating fully digital PulseEKKO, which is gaining wide acceptance around the world. Attendees showed intense interest in this new and unique technology. Applications covered were groundwater and mineral exploration; engineering, construction and toxic waste site surveying; tunnel and underground mine probing for potential geological hazards, blind ore zones, karst cavities and solution pathways; and locating buried objects such as petroleum storage tanks, unexploded bombs and archeological remains.

  19. Python penetration testing essentials

    CERN Document Server

    Mohit

    2015-01-01

    If you are a Python programmer or a security researcher who has basic knowledge of Python programming and want to learn about penetration testing with the help of Python, this book is ideal for you. Even if you are new to the field of ethical hacking, this book can help you find the vulnerabilities in your system so that you are ready to tackle any kind of attack or intrusion.

  20. Market penetration of biodiesel

    Directory of Open Access Journals (Sweden)

    Kenneth R. Szulczyk, Bruce A. McCarl

    2010-01-01

    Full Text Available This research examines in detail the technology and economics of substituting biodiesel for diesel #2. This endeavor examines three areas. First, the benefits of biodiesel are examined, and the technical problems of large-scale implementation. Second, the biodiesel production possibilities are examined for soybean oil, corn oil, tallow, and yellow grease, which are the largest sources of feedstocks for the United States. Examining in detail the production possibilities allows to identity the extent of technological change, production costs, byproducts, and greenhouse gas (GHG emissions. Finally, a U.S. agricultural model, FASOMGHG was used to predict market penetration of biodiesel, given technological progress, variety of technologies and feedstocks, market interactions, energy prices, and carbon dioxide equivalent prices. FASOMGHG has several interesting results. First, diesel fuel prices have an expansionary impact on the biodiesel industry. The higher the diesel fuel prices, the more biodiesel is produced. However, given the most favorable circumstances, the maximum biodiesel market penetration is 9% in 2030 with a wholesale diesel price of $4 per gallon. Second, the two dominant sources of biodiesel are from corn and soybeans. Sources like tallow and yellow grease are more limited, because they are byproducts of other industries. Third, GHG prices have an expansionary impact on the biodiesel prices, because biodiesel is quite GHG efficient. Finally, U.S. government subsidies on biofuels have an expansionary impact on biodiesel production, and increase market penetration at least an additional 3%.

  1. Conjugation of cell-penetrating peptides with poly(lactic-co-glycolic acid-polyethylene glycol nanoparticles improves ocular drug delivery

    Directory of Open Access Journals (Sweden)

    Vasconcelos A

    2015-01-01

    Full Text Available Aimee Vasconcelos,1 Estefania Vega,2 Yolanda Pérez,3 María J Gómara,1 María Luisa García,2 Isabel Haro1 1Unit of Synthesis and Biomedical Applications of Peptides, Department of Biomedical Chemistry, Institute for Advanced Chemistry of Catalonia, Consejo Superior de Investigaciones Científicas (IQAC-CSIC, 2Department of Physical Chemistry, Institute of Nanoscience and Nanotechnology, Faculty of Pharmacy, University of Barcelona, 3Nuclear Magnetic Resonance Unit, IQAC-CSIC, Barcelona, Spain Abstract: In this work, a peptide for ocular delivery (POD and human immunodeficiency virus transactivator were conjugated with biodegradable poly(lactic-co-glycolic acid (PGLA–polyethylene glycol (PEG-nanoparticles (NPs in an attempt to improve ocular drug bioavailability. The NPs were prepared by the solvent displacement method following two different pathways. One involved preparation of PLGA NPs followed by PEG and peptide conjugation (PLGA-NPs-PEG-peptide; the other involved self-assembly of PLGA-PEG and the PLGA-PEG-peptide copolymer followed by NP formulation. The conjugation of the PEG and the peptide was confirmed by a colorimetric test and proton nuclear magnetic resonance spectroscopy. Flurbiprofen was used as an example of an anti-inflammatory drug. The physicochemical properties of the resulting NPs (morphology, in vitro release, cell viability, and ocular tolerance were studied. In vivo anti-inflammatory efficacy was assessed in rabbit eyes after topical instillation of sodium arachidonate. Of the formulations developed, the PLGA-PEG-POD NPs were the smaller particles and exhibited greater entrapment efficiency and more sustained release. The positive charge on the surface of these NPs, due to the conjugation with the positively charged peptide, facilitated penetration into the corneal epithelium, resulting in more effective prevention of ocular inflammation. The in vitro toxicity of the NPs developed was very low; no ocular irritation

  2. Large-scale preparation of active caspase-3 in E. coli by designing its thrombin-activatable precursors

    Directory of Open Access Journals (Sweden)

    Park Sung

    2008-12-01

    Full Text Available Abstract Background Caspase-3, a principal apoptotic effector that cleaves the majority of cellular substrates, is an important medicinal target for the treatment of cancers and neurodegenerative diseases. Large amounts of the protein are required for drug discovery research. However, previous efforts to express the full-length caspase-3 gene in E. coli have been unsuccessful. Results Overproducers of thrombin-activatable full-length caspase-3 precursors were prepared by engineering the auto-activation sites of caspase-3 precursor into a sequence susceptible to thrombin hydrolysis. The engineered precursors were highly expressed as soluble proteins in E. coli and easily purified by affinity chromatography, to levels of 10–15 mg from 1 L of E. coli culture, and readily activated by thrombin digestion. Kinetic evaluation disclosed that thrombin digestion enhanced catalytic activity (kcat/KM of the precursor proteins by two orders of magnitude. Conclusion A novel method for a large-scale preparation of active caspase-3 was developed by a strategic engineering to lack auto-activation during expression with amino acid sequences susceptible to thrombin, facilitating high-level expression in E. coli. The precursor protein was easily purified and activated through specific cleavage at the engineered sites by thrombin, generating active caspase-3 in high yields.

  3. Chiral, J-Aggregate-Forming Dyes for Alternative Signal Modulation Mechanisms in Self-Immolative Enzyme-Activatable Optical Probes.

    Science.gov (United States)

    Sloniec-Myszk, Jagoda; Resch-Genger, Ute; Hennig, Andreas

    2016-02-11

    Enzyme-activatable optical probes are important for future advances in cancer imaging, but may easily suffer from low signal-to-background ratios unless not optimized. To address this shortcoming, numerous mechanisms to modulate the fluorescence signal have been explored. We report herein newly synthesized probes based on self-immolative linkers containing chiral J-aggregate-forming dyes. Signal modulation by formation of chiral J-aggregates is yet unexplored in optical enzyme probe design. The comprehensive characterization of the probes by absorption, CD, fluorescence, and time-resolved fluorescence spectroscopy revealed dye-dye interactions not observed for the free dyes in solution as well as dye-protein interactions with the enzyme. This suggested that J-aggregate formation is challenging to achieve with current probe design and that interactions of the dyes with the enzyme may interfere with achieving high signal-to-background ratios. The detailed understanding of the interactions provided herein provides valuable guidelines for the future design of similar probes.

  4. Microbial Penetration through Nutrient-Saturated Berea Sandstone.

    Science.gov (United States)

    Jenneman, G E; McInerney, M J; Knapp, R M

    1985-08-01

    Penetration times and penetration rates for a motile Bacillus strain growing in nutrient-saturated Berea sandstone cores were determined. The rate of penetration was essentially independent of permeabilities above 100 mdarcys and rapidly declined for permeabilities below 100 mdarcys. It was found that these penetration rates could be grouped into two statistically distinct classes consisting of rates for permeabilities above 100 mdarcys and rates for those below 100 mdarcys. Instantaneous penetration rates were found to be zero order with respect to core length for cores with permeabilities above 100 mdarcys and first order with respect to core length for cores with permeabilities below 100 mdarcys. The maximum observed penetration rate was 0.47 cm . h, and the slowest was 0.06 cm . h; however, these rates may be underestimates of the true penetration rate, since the observed rates included the time required for growth in the flask as well as the core. The relationship of penetration time to the square of the length of the core suggested that cells penetrated high-permeability cores as a band and low-permeability cores in a diffuse fashion. The motile Enterobacter aerogenes strain penetrated Berea sandstone cores three to eight times faster than did the nonmotile Klebsiella pneumoniae strain when cores of comparable length and permeability were used. A penetration mechanism based entirely on motility predicted penetration times that were in agreement with the observed penetration times for motile strains. The fact that nonmotile strains penetrated the cores suggested that filamentous or unrestricted growth, or both, may also be important.

  5. Market penetration of ethanol

    International Nuclear Information System (INIS)

    This research examines in detail the technology and economics of substituting ethanol for gasoline. This endeavor examines three issues. First, the benefits of ethanol/gasoline blends are examined, and then the technical problems of large-scale implementation of ethanol. Second, ethanol production possibilities are examined in detail from a variety of feedstocks and technologies. The feedstocks are the starch/sugar crops and crop residues, while the technologies are corn wet mill, dry grind, and lignocellulosic fermentation. Examining in detail the production possibilities allows the researchers to identity the extent of technological change, production costs, byproducts, and GHG emissions. Finally, a U.S. agricultural model, FASOMGHG, is updated which predicts the market penetration of ethanol given technological progress, variety of technologies and feedstocks, market interactions, energy prices, and GHG prices. FASOMGHG has several interesting results. First, gasoline prices have a small expansionary impact on the U.S. ethanol industry. Both agricultural producers' income and cost both increase with higher energy prices. If wholesale gasoline is $4 per gallon, the predicted ethanol market penetration attains 53% of U.S. gasoline consumption in 2030. Second, the corn wet mill remains an important industry for ethanol production, because this industry also produces corn oil, which could be converted to biodiesel. Third, GHG prices expand the ethanol industry. However, the GHG price expands the corn wet mill, but has an ambiguous impact on lignocellulosic ethanol. Feedstocks for lignocellulosic fermentation can also be burned with coal to generate electricity. Both industries are quite GHG efficient. Finally, U.S. government subsidies on biofuels have an expansionary impact on ethanol production, but may only increase market penetration by an additional 1% in 2030, which is approximately 6 billion gallons. (author)

  6. An Overview of Penetration Testing

    Directory of Open Access Journals (Sweden)

    Aileen G. Bacudio

    2011-12-01

    Full Text Available Penetration testing is a series of activities undertaken to identify and exploit security vulnerabilities. It helps confirm the effectiveness or ineffectiveness of the security measures that have been implemented. This paper provides an overview of penetration testing. It discusses the benefits, the strategies and the methodology of conducting penetration testing. The methodology of penetration testing includes three phases: test preparation, test and test analysis. The test phase involves the following steps: information gathering, vulnerability analysis, and vulnerability exploit. This paper further illustrates how to apply this methodology to conduct penetration testing on two example web applications.

  7. Enhanced chlorhexidine skin penetration with eucalyptus oil

    Directory of Open Access Journals (Sweden)

    Worthington Tony

    2010-09-01

    Full Text Available Abstract Background Chlorhexidine digluconate (CHG is a widely used skin antiseptic, however it poorly penetrates the skin, limiting its efficacy against microorganisms residing beneath the surface layers of skin. The aim of the current study was to improve the delivery of chlorhexidine digluconate (CHG when used as a skin antiseptic. Method Chlorhexidine was applied to the surface of donor skin and its penetration and retention under different conditions was evaluated. Skin penetration studies were performed on full-thickness donor human skin using a Franz diffusion cell system. Skin was exposed to 2% (w/v CHG in various concentrations of eucalyptus oil (EO and 70% (v/v isopropyl alcohol (IPA. The concentration of CHG (μg/mg of skin was determined to a skin depth of 1500 μm by high performance liquid chromatography (HPLC. Results The 2% (w/v CHG penetration into the lower layers of skin was significantly enhanced in the presence of EO. Ten percent (v/v EO in combination with 2% (w/v CHG in 70% (v/v IPA significantly increased the amount of CHG which penetrated into the skin within 2 min. Conclusion The delivery of CHG into the epidermis and dermis can be enhanced by combination with EO, which in turn may improve biocide contact with additional microorganisms present in the skin, thereby enhancing antisepsis.

  8. The Effect of Bacteria Penetration on Chalk Permeability

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Shapiro, Alexander; Nielsen, Sidsel Marie;

    into reservoirs, however, a complete understanding of the penetration behavior of bacteria is lacking, especially in chalk formations where the pore throat sizes are almost comparable with the sizes of bacteria vegetative cells. This study investigates the penetration of bacteria into chalk. Two bacteria types...

  9. Different patterns of nuclear and mitochondrial penetration by the G3 PAMAM dendrimer and its biotin–pyridoxal bioconjugate BC-PAMAM in normal and cancer cells in vitro

    Science.gov (United States)

    Uram, Łukasz; Szuster, Magdalena; Filipowicz, Aleksandra; Gargasz, Krzysztof; Wołowiec, Stanisław; Wałajtys-Rode, Elżbieta

    2015-01-01

    The intracellular localization and colocalization of a fluorescently labeled G3 amine-terminated cationic polyamidoamine (PAMAM) dendrimer and its biotin–pyridoxal (BC-PAMAM) bioconjugate were investigated in a concentration-dependent manner in normal human fibroblast (BJ) and squamous epithelial carcinoma (SCC-15) cell lines. After 24 hours treatment, both cell lines revealed different patterns of intracellular dendrimer accumulation depending on their cytotoxic effects. Cancer cells exhibited much higher (20-fold) tolerance for native PAMAM treatment than fibroblasts, whereas BC-PAMAM was significantly toxic only for fibroblasts at 50 µM concentration. Fibroblasts accumulated the native and bioconjugated dendrimers in a concentration-dependent manner at nontoxic range of concentration, with significantly lower bioconjugate loading. After reaching the cytotoxicity level, fluorescein isothiocyanate-PAMAM accumulation remains at high, comparable level. In cancer cells, native PAMAM loading at higher, but not cytotoxic concentrations, was kept at constant level with a sharp increase at toxic concentration. Mander’s coefficient calculated for fibroblasts and cancer cells confirmed more efficient native PAMAM penetration as compared to BC-PAMAM. Significant differences in nuclear dendrimer penetration were observed for both cell lines. In cancer cells, PAMAM signals amounted to ~25%–35% of the total nuclei area at all investigated concentrations, with lower level (15%–25%) observed for BC-PAMAM. In fibroblasts, the dendrimer nuclear signal amounted to 15% at nontoxic and up to 70% at toxic concentrations, whereas BC-PAMAM remained at a lower concentration-dependent level (0.3%–20%). Mitochondrial localization of PAMAM and BC-PAMAM revealed similar patterns in both cell lines, depending on the extracellular dendrimer concentration, and presented significantly lower signals from BC-PAMAM, which correlated well with the cytotoxicity. PMID:26379435

  10. Penetration Dynamics of Earth Penetration Warhead into Composite Target Media

    Directory of Open Access Journals (Sweden)

    P. K. Roy

    1987-07-01

    Full Text Available Attempts have been made to develop a suitable computer code that can find solutions to the axi-symmetric penetration of an Earth Penetrating Warhead yielding complete space-time histories of the resistive force offered by the target medium. The consequent warhead deceleration and velocity reduction, the resulting axial compressive stress developed in warhead casing as the penetration process progresses into the composite target media consisting of hard concrete of specified thickness followed by earth soil have been discussed.

  11. Au@Ag/Au nanoparticles assembled with activatable aptamer probes as smart ``nano-doctors'' for image-guided cancer thermotherapy

    Science.gov (United States)

    Shi, Hui; Ye, Xiaosheng; He, Xiaoxiao; Wang, Kemin; Cui, Wensi; He, Dinggeng; Li, Duo; Jia, Xuekun

    2014-07-01

    Although nanomaterial-based theranostics have increased positive expectations from cancer treatment, it remains challenging to develop in vivo ``nano-doctors'' that provide high-contrast image-guided site-specific therapy. Here we designed an activatable theranostic nanoprobe (ATNP) via self-assembly of activatable aptamer probes (AAPs) on Au@Ag/Au nanoparticles (NPs). As both quenchers and heaters, novel Au@Ag/Au NPs were prepared, showing excellent fluorescence quenching and more effective near-infrared photothermal therapy than Au nanorods. The AAP comprised a thiolated aptamer and a fluorophore-labeled complementary DNA; thus, the ATNP with quenched fluorescence in the free state could realize signal activation through target binding-induced conformational change of the AAP, and then achieve on-demand treatment under image-guided irradiation. By using S6 aptamer as the model, in vitro and in vivo studies of A549 lung cancer verified that the ATNP greatly improved imaging contrast and specific destruction, suggesting a robust and versatile theranostic strategy for personalized medicine in future.Although nanomaterial-based theranostics have increased positive expectations from cancer treatment, it remains challenging to develop in vivo ``nano-doctors'' that provide high-contrast image-guided site-specific therapy. Here we designed an activatable theranostic nanoprobe (ATNP) via self-assembly of activatable aptamer probes (AAPs) on Au@Ag/Au nanoparticles (NPs). As both quenchers and heaters, novel Au@Ag/Au NPs were prepared, showing excellent fluorescence quenching and more effective near-infrared photothermal therapy than Au nanorods. The AAP comprised a thiolated aptamer and a fluorophore-labeled complementary DNA; thus, the ATNP with quenched fluorescence in the free state could realize signal activation through target binding-induced conformational change of the AAP, and then achieve on-demand treatment under image-guided irradiation. By using S6 aptamer as

  12. Membrane damage as first and DNA as the secondary target for anti-candidal activity of antimicrobial peptide P7 derived from cell-penetrating peptide ppTG20 against Candida albicans.

    Science.gov (United States)

    Li, Lirong; Song, Fengxia; Sun, Jin; Tian, Xu; Xia, Shufang; Le, Guowei

    2016-06-01

    P7, a peptide analogue derived from cell-penetrating peptide ppTG20, possesses antibacterial and antitumor activities without significant hemolytic activity. In this study, we investigated the antifungal effect of P7 and its anti-Candida acting mode in Candida albicans. P7 displayed antifungal activity against the reference C. albicans (MIC = 4 μM), Aspergilla niger (MIC = 32 μM), Aspergillus flavus (MIC = 8 μM), and Trichopyton rubrum (MIC = 16 μM). The effect of P7 on the C. albicans cell membrane was examined by investigating the calcein leakage from fungal membrane models made of egg yolk l-phosphatidylcholine/ergosterol (10 : 1, w/w) liposomes. P7 showed potent leakage effects against fungal liposomes similar to Melittin-treated cells. C. albicans protoplast regeneration assay demonstrated that P7 interacted with the C. albicans plasma membrane. Flow cytometry of the plasma membrane potential and integrity of C. albicans showed that P7 caused 60.9 ± 1.8% depolarization of the membrane potential of intact C. albicans cells and caused 58.1 ± 3.2% C. albicans cell membrane damage. Confocal laser scanning microscopy demonstrated that part of FITC-P7 accumulated in the cytoplasm. DNA retardation analysis was also performed, which showed that P7 interacted with C. albicans genomic DNA after penetrating the cell membrane, completely inhibiting the migration of genomic DNA above the weight ratio (peptide : DNA) of 6. Our results indicated that the plasma membrane was the primary target, and DNA was the secondary intracellular target of the mode of action of P7 against C. albicans. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. PMID:27197902

  13. Penetration Tester's Open Source Toolkit

    CERN Document Server

    Faircloth, Jeremy

    2011-01-01

    Great commercial penetration testing tools can be very expensive and sometimes hard to use or of questionable accuracy. This book helps solve both of these problems. The open source, no-cost penetration testing tools presented do a great job and can be modified by the user for each situation. Many tools, even ones that cost thousands of dollars, do not come with any type of instruction on how and in which situations the penetration tester can best use them. Penetration Tester's Open Source Toolkit, Third Edition, expands upon existing instructions so that a professional can get the most accura

  14. Liquid Crystalline Nanodispersions Functionalized with Cell-Penetrating Peptides for Topical Delivery of Short-Interfering RNAs: A Proposal for Silencing a Pro-Inflammatory Cytokine in Cutaneous Diseases.

    Science.gov (United States)

    Petrilli, R; Eloy, J O; Praça, F S G; Del Ciampo, J O; Fantini, M A C; Fonseca, M J V; Bentley, M V L B

    2016-05-01

    Short-interfering RNAs (siRNAs) are a potential strategy for the treatment of cutaneous diseases. In this context, liquid crystalline nanoparticles functionalized with specific proteins and peptide-transduction domains (PTDs), which act as penetration enhancers, are a promising carrier for siRNA delivery through the skin. Herein, hexagonal phase liquid crystal nanoparticles based on monoolein (MO) and/or oleic acid (OA) containing (or lacking) the cationic polymer polyethylenimine (PEI) and the cationic lipid oleylamine (OAM) were functionalized with the membrane transduction peptides transcriptional activator (TAT) or penetratin (PNT). These nanoparticles were complexed with siRNA and characterized by particle size, polydispersity, zeta potential, complexation efficiency and siRNA release. The formulations containing cationic agents presented positive zeta potentials, sizes on the nanometer scale, and complexed siRNAs at concentrations of 10 μM; these agents were successfully released in a heparin competition assay. Cell culture studies demonstrated that nanoparticles composed of MO:OA:PEI functionalized with TAT were the most efficient at transfecting L929 cells, and the uptake efficiency was enhanced by TAT peptide functionalization. Thereafter, the selected formulations were evaluated for in vivo skin irritation, penetration and in vivo efficacy using a chemically induced inflammatory animal model. These nanoparticles did not irritate the skin and provided higher siRNA penetration and delivery into the skin than control formulations. Additionally, efficacy studies in the animal model showed that the association of TAT with the nanodispersion provided higher suppression of tumor necrosis factor (TNF)-α. Thus, the development of liquid crystalline nanodispersions containing TAT may lead to improved topical siRNA delivery for the treatment of inflammatory skin diseases. PMID:27305826

  15. An Earth Penetrating Modeling Assessment

    Energy Technology Data Exchange (ETDEWEB)

    Stokes, E; Yarrington, P; Glenn, L

    2005-06-21

    Documentation of a study to assess the capability of computer codes to predict lateral loads on earth penetrating projectiles under conditions of non-normal impact. Calculations simulated a set of small scale penetration tests into concrete targets with oblique faces at angles of 15 and 30 degrees to the line-of-flight. Predictive codes used by the various calculational teams cover a wide range of modeling approaches from approximate techniques, such as cavity expansion, to numerical methods, such as finite element codes. The modeling assessment was performed under the auspices of the Phenomenology Integrated Product Team (PIPT) for the Robust Nuclear Earth Penetrator Program (RNEP). Funding for the penetration experiments and modeling was provided by multiple earth penetrator programs.

  16. Penetration of Photovoltaics in Greece

    Directory of Open Access Journals (Sweden)

    Eugenia Giannini

    2015-06-01

    Full Text Available Recently, an interesting experiment was completed in Greece concerning photovoltaic penetration into the electricity production sector. Based on the relevant laws and in accordance to the related European directives, an explosive penetration process was completed in less than three years, resulting in a 7% share of photovoltaics in electricity production instead of the previous negligible share. The legislation was based on licensing simplification and generous feed-in-tariffs. This approach transformed photovoltaic technology from a prohibitively expensive to a competitive one. This work aims to summarize the relevant legislation and illustrate its effect on the resulting penetration. A sigmoid-shape penetration was observed which was explained by a pulse-type driving force. The return on investment indicator was proposed as an appropriate driving force, which incorporates feed-in-tariffs and turnkey-cost. Furthermore, the resulting surcharge on the electricity price due to photovoltaic penetration was also analyzed.

  17. Imaging Primary Mouse Sarcomas After Radiation Therapy Using Cathepsin-Activatable Fluorescent Imaging Agents

    Energy Technology Data Exchange (ETDEWEB)

    Cuneo, Kyle C. [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States); Mito, Jeffrey K.; Javid, Melodi P. [Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States); Ferrer, Jorge M. [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Kim, Yongbaek [Department of Clinical Pathology, College of Veterinary Medicine, Seoul National University, Seoul (Korea, Republic of); Lee, W. David [The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Bawendi, Moungi G. [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Brigman, Brian E. [Department of Orthopedic Surgery, Duke University School of Medicine, Durham, North Carolina (United States); Kirsch, David G., E-mail: david.kirsch@duke.edu [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States); Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States)

    2013-05-01

    Purpose: Cathepsin-activated fluorescent probes can detect tumors in mice and in canine patients. We previously showed that these probes can detect microscopic residual sarcoma in the tumor bed of mice during gross total resection. Many patients with soft tissue sarcoma (STS) and other tumors undergo radiation therapy (RT) before surgery. This study assesses the effect of RT on the ability of cathepsin-activated probes to differentiate between normal and cancerous tissue. Methods and Materials: A genetically engineered mouse model of STS was used to generate primary hind limb sarcomas that were treated with hypofractionated RT. Mice were injected intravenously with cathepsin-activated fluorescent probes, and various tissues, including the tumor, were imaged using a hand-held imaging device. Resected tumor and normal muscle samples were harvested to assess cathepsin expression by Western blot. Uptake of activated probe was analyzed by flow cytometry and confocal microscopy. Parallel in vitro studies using mouse sarcoma cells were performed. Results: RT of primary STS in mice and mouse sarcoma cell lines caused no change in probe activation or cathepsin protease expression. Increasing radiation dose resulted in an upward trend in probe activation. Flow cytometry and immunofluorescence showed that a substantial proportion of probe-labeled cells were CD11b-positive tumor-associated immune cells. Conclusions: In this primary murine model of STS, RT did not affect the ability of cathepsin-activated probes to differentiate between tumor and normal muscle. Cathepsin-activated probes labeled tumor cells and tumor-associated macrophages. Our results suggest that it would be feasible to include patients who have received preoperative RT in clinical studies evaluating cathepsin-activated imaging probes.

  18. Enhancing Targeted Tumor Treatment by Near IR Light-Activatable Photodynamic–Photothermal Synergistic Therapy

    OpenAIRE

    Fan, Zhen; Dai, Xuemei; Lu, Yuefeng; Yu, Eugene; Brahmbatt, Nupur; Carter, NaTasha; Tchouwou, Christine; Singh, Anant Kumar; Jones, Yolanda; Yu, Hongtao; Ray, Paresh Chandra

    2014-01-01

    For several decades, cancer has been one of the most life-threatening diseases. For enhancing anticancer efficiency with minimum side effects, combination therapy is envisioned. The current manuscript reports for the first time the development of a methylene blue (MB) bound nanoplatform, which is capable of delivering targeted diagnostic and combined synergistic photothermal and photodynamic treatment of cancer. Experimental data found that, once the nanoparticle binds with the target cell su...

  19. An albumin nanocomplex-based endosomal pH-activatable on/off probe system.

    Science.gov (United States)

    Lee, Changkyu; Lee, Seunghyun; Thao, Le Quang; Hwang, Ha Shin; Kim, Jong Oh; Lee, Eun Seong; Oh, Kyung Taek; Shin, Beom Soo; Choi, Han-Gon; Youn, Yu Seok

    2016-08-01

    Albumin has gained considerable interest as a material for fabricating nanoparticulate systems due to its biomedical advantages, such as biocompatibility and chemical functionality. Here, we report a new pH-sensitive albumin nanocomplex prototype with a zinc-imidazole coordination bond. Albumin was conjugated with 1-(3-aminopropyl)imidazole and mPEG10kDa-NHS, and the resulting albumin conjugate (PBI) was then modified with either Cy5.5 or BHQ-3. The newly formed albumin nanocomplex (C/BQ-PBI Zn NCs: ∼116nm) system was facilely self-assembled around pH 7.4 in the presence of Zn(2+), but it quickly disassembled in an acidic environment (∼pH 5.0). Based on this pH-sensitivity, C/BQ-PBI Zn NCs emitted strong near-infrared fluorescence and released Zn(2+), turning "off" at pH ∼7.4 (e.g., plasma) and "on" at pH ∼5.0 (e.g., endo/lysosomes in tumor cells) on account of fluorescence resonance energy transfer. C/BQ-PBI Zn NCs displayed significant cytotoxicity due to an increase in cellular Zn(2+) in response to endosomal pH (∼5.0) in breast cancer MCF-7 cells and lung adenocarcinoma A549 cells. Particularly, confocal laser scanning microscopic images showed a strong fluorescence signal caused by the disassembly of C/BQ-PBI Zn NCs in the endosomal region of MCF-7 cells. Based on these results, we believe that this albumin nanocomplex is an attractive biocompatible tumor targeting probe carrier for the theranostic purpose. PMID:27108210

  20. Comparison of mechanisms and cellular uptake of cell-penetrating peptide on different cell lines%不同细胞系对细胞穿透肽的摄取和机制比较

    Institute of Scientific and Technical Information of China (English)

    马冬旭; 齐宪荣

    2010-01-01

    细胞穿透肽(cell-penetrating peptide,CPP)作为一种潜在的药物输送高效转运载体一直得到研究者的广泛关注.本文中采用4种肿瘤细胞系(MCF-7、MDA-MB-231、C6和B16F10)分别摄取异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记的CPP,观察到CPP入胞,并具有时间和浓度的依赖性,同时发现了C6细胞对CPP的胞吐作用,其胞吐动力学符合零级方程;在低温(4℃)和内吞抑制剂存在条件下探讨了CPP入胞的机制.低温条件对CPP的入胞未产生抑制作用;肝素钠作为细胞表面硫酸糖蛋白受体抑制剂对CPP的入胞有较强抑制作用,肝素组对CPP的摄取只达到对照组的3%~10%;而氯丙嗪、氯喹和N-乙酰基-N-异丙基阿米洛利[5-(N-ethyL-N-isopropyl)-amiloride,EIPA]对CPP的入胞影响不大.本研究表明,CPP穿透细胞没有选择性,即缺乏细胞特异性,但CPP的摄取量与细胞种类有关.硫酸蛋白聚糖的吸附介导在CPP穿透细胞中发挥了重要作用.

  1. Cooperative Transmembrane Penetration of Nanoparticles

    Science.gov (United States)

    Zhang, Haizhen; Ji, Qiuju; Huang, Changjin; Zhang, Sulin; Yuan, Bing; Yang, Kai; Ma, Yu-qiang

    2015-01-01

    Physical penetration of lipid bilayer membranes presents an alternative pathway for cellular delivery of nanoparticles (NPs) besides endocytosis. NPs delivered through this pathway could reach the cytoplasm, thereby opening the possibility of organelle-specific targeting. Herein we perform dissipative particle dynamics simulations to elucidate the transmembrane penetration mechanisms of multiple NPs. Our simulations demonstrate that NPs’ translocation proceeds in a cooperative manner, where the interplay of the quantity and surface chemistry of the NPs regulates the translocation efficiency. For NPs with hydrophilic surfaces, the increase of particle quantity facilitates penetration, while for NPs with partly or totally hydrophobic surfaces, the opposite highly possibly holds. Moreover, a set of interesting cooperative ways, such as aggregation, aggregation-dispersion, and aggregation-dispersion-reaggregation of the NPs, are observed during the penetration process. We find that the penetration behaviors of multiple NPs are mostly dominated by the changes of the NP-membrane force components in the membrane plane direction, in addition to that in the penetration direction, suggesting a different interaction mechanism between the multiple NPs and the membrane compared with the one-NP case. These results provide a fundamental understanding in the underlying mechanisms of cooperative penetration of NPs, and shed light on the NP-based drug and gene delivery. PMID:26013284

  2. Deep ocean model penetrator experiments

    International Nuclear Information System (INIS)

    Preliminary trials of experimental model penetrators in the deep ocean have been conducted as an international collaborative exercise by participating members (national bodies and the CEC) of the Engineering Studies Task Group of the Nuclear Energy Agency's Seabed Working Group. This report describes and gives the results of these experiments, which were conducted at two deep ocean study areas in the Atlantic: Great Meteor East and the Nares Abyssal Plain. Velocity profiles of penetrators of differing dimensions and weights have been determined as they free-fell through the water column and impacted the sediment. These velocity profiles are used to determine the final embedment depth of the penetrators and the resistance to penetration offered by the sediment. The results are compared with predictions of embedment depth derived from elementary models of a penetrator impacting with a sediment. It is tentatively concluded that once the resistance to penetration offered by a sediment at a particular site has been determined, this quantity can be used to sucessfully predict the embedment that penetrators of differing sizes and weights would achieve at the same site

  3. Highly activatable and environment-insensitive optical highlighters for selective spatiotemporal imaging of target proteins.

    Science.gov (United States)

    Kobayashi, Tomonori; Komatsu, Toru; Kamiya, Mako; Campos, Cláudia; González-Gaitán, Marcos; Terai, Takuya; Hanaoka, Kenjiro; Nagano, Tetsuo; Urano, Yasuteru

    2012-07-11

    Optical highlighters are photoactivatable fluorescent molecules that exhibit pronounced changes in their spectral properties in response to irradiation with light of a specific wavelength and intensity. Here, we present a novel design strategy for a new class of caged BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) fluorophores, based on the use of photoremovable protecting groups (PRPGs) with high reduction potentials that serve as both a photosensitive unit and a fluorescence quencher via photoinduced electron transfer (PeT). 2,6-Dinitrobenzyl (DNB)-caged BODIPY was efficiently photoactivated, with activation ratios exceeding 600-fold in aqueous solutions. We then combined this photoactivatable fluorophore with a SNAP (mutant of O(6)-alkylguanine DNA alkyltransferase) ligand to obtain a small-molecule-based optical highlighter for visualization of protein dynamics, using the well-established SNAP tag technology. As proof of concept, we demonstrate spatiotemporal imaging of the fusion protein of epidermal growth factor receptor (EGFR) with SNAP tag in living cells. We also demonstrate highlighting of cells of interest in live zebrafish embryos, using the fusion protein of histone 2A with SNAP tag.

  4. Percutaneous penetration through slightly damaged skin

    DEFF Research Database (Denmark)

    Nielsen, Jesper B

    2005-01-01

    Guidelines for experimental studies of percutaneous penetration prescribe optimal barrier integrity of the skin. The barrier integrity of the skin exposed in occupational or household situations is, however, not always ideal, and skin problems are among the most dominant reasons for absence from...... work. We have therefore evaluated an experimental model for percutaneous penetration through slightly damaged skin. The influence of a slight damage to the skin was evaluated using five pesticides covering a wide range of solubilities. We used an experimental model with static diffusion cells mounted...... with human skin. A slight damage to the barrier integrity was induced by pre-treatment of the skin with sodium lauryl sulphate (SLS) before pesticide exposure. The experimental model with 3 h pre-treatment with SLS (0.1% or 0.3%) assured a significant but controlled damage to the barrier integrity, a damage...

  5. Cement penetration after patella venting.

    Science.gov (United States)

    Jones, Christopher W; Lam, Li-On; Butler, Adam; Wood, David J; Walsh, William R

    2009-01-01

    There is a high rate of patellofemoral complications following total knee arthroplasty. Optimization of the cement-bone interface by venting and suction of the tibial plateau has been shown to improve cement penetration. Our study was designed to investigate if venting the patella prior to cementing improved cement penetration. Ten paired cadaver patellae were allocated prior to resurfacing to be vented or non-vented. Bone mineral density (BMD) was measured by DEXA scanning. In vented specimens, a 1.6 mm Kirschner wire was used to breach the anterior cortex at the center. Specimens were resurfaced with standard Profix instrumentation and Versabond bone cement (Smith and Nephew PLC, UK). Cement penetration was assessed from Faxitron and sectioned images by a digital image software package (ImageJ V1.38, NIH, USA). Wilcoxon rank sum test was used to assess the difference in cement penetration between groups. The relationship between BMD and cement penetration was analyzed by Pearson correlation coefficient. There was a strong negative correlation between peak BMD and cement penetration when analyzed independent of experimental grouping (r(2)=-0.812, p=0.004). Wilcoxon rank sum testing demonstrated no significant difference (rank sum statistic W=27, p=0.579) in cement penetration between vented (10.53%+/-4.66; mean+/-std dev) and non-vented patellae (11.51%+/-6.23; mean+/-std dev). Venting the patella using a Kirschner wire does not have a significant effect on the amount of cement penetration achieved in vitro using Profix instrumentation and Versabond cement. PMID:19010682

  6. Novel approach for characterizing the specific shunt resistance caused by the penetration of the front contact through the p-n junction in solar cell

    Institute of Scientific and Technical Information of China (English)

    Zhang Lucheng; Shen Hui

    2009-01-01

    on process was completely compatible with the industrial silicon fabrication sequence, which was of great convenience. The measurement results give informations on the solar cell structure, material ingredients, and process parameters.

  7. Expression and purification of Tat-GFP fusion protein and its cell membrane penetrating activity%Tat-GFP融合蛋白的表达纯化及其穿膜活性

    Institute of Scientific and Technical Information of China (English)

    关新刚; 苏维恒; 于欣; 佟海滨; 孙新

    2014-01-01

    Objective To obtain the Tat-GFP fusion proteins with penetrating activity and labeled with green fluorescence protein (GFP), and to explore the cell membrane penetrating activity of Tat-GFP in MCF-7 cells. Methods The plasmid pET-24a-Tat-GFP was transformed into Escherichia coli BL21 cells. Different concentrations (0.5 and 1.0 mmol · L-1 ) of isopropyl-β-D-thiogalactopyranoside (IPTG ) and cell culture temperatures (22℃ and 37℃)were used to optimize the protein expression.The Tat-GFP proteins in supernatant were purified using Ni-IDA resins. Western blotting analysis was used to identify the Tat-GFP protein, and confocal laser scanning microscope (CLSM ) was used to examine the cell penetration of Tat-GFP protein. Results There was no significant difference in the Tat-GFP protein production induced by 0.5 and 1.0 mmol·L-1 IPTG;however,the low temperature (22℃)-induced BL21 cells expressed more Tat-GFP proteins than that at 37℃ induction.The Western blotting analysis results showed that GFP antibody could specifically recognize the proteins in PVDF membranes in dose-dependent manner;the CLSM results indicated the distribution of green fluorescence in cytoplasm and nucleus of MCF-7 cells.Conclusion The Tat-GFP protein highly expresses in the supenatant of Escherichia coli i BL2 1 cells at low temperature;the obtained Tat-GFP protein with green fluorescence preserves the cell penetrating activity.%目的:获得具备穿膜活性与绿色荧光蛋白(GFP)标记的 Tat-GFP 融合蛋白,探讨 Tat-GFP 在MCF-7细胞中的跨膜转运特性。方法:应用 pET-24a-Tat-GFP质粒转化大肠杆菌 BL21感受态细胞,检测不同异丙基硫代半乳糖苷(IPTG)浓度(0.5和1.0 mmol·L-1)和不同温度(22℃和37℃)诱导融合蛋白的表达情况;利用 Ni-IDA树脂亲和纯化Tat-GFP蛋白,利用 GFP特异性抗体采用 Western blotting法分析洗脱液中的蛋白;激光共聚焦荧光显微镜下检测Tat-GFP融合蛋白

  8. Case Report: Penetrating Cardiac Injury

    Directory of Open Access Journals (Sweden)

    Adem Grbolar

    2013-10-01

    Full Text Available Summary: Penetrating cardiac injurys caused by gunshots and penetrating tools have high mortality rates. The way of injury, how the cardiac area is effected and the presence of cardiac tamponadecauses mortality in different rates. However the better treatment quality of hospitals, increasingoperative techniques, and internel care unit quality has not been change during the years. Searching the literature, we want to present a 42 years old male patient whowas injured by knife and had a 1 cm skin wound on chest with cardiac tamponade. After sternotomy a 7 cm laseration was observed in heart. Cardioraphy was performed.

  9. Development of an antisense RNA delivery system using conjugates of the MS2 bacteriophage capsids and HIV-1 TAT cell-penetrating peptide.

    Science.gov (United States)

    Wei, Baojun; Wei, Yuxiang; Zhang, Kuo; Wang, Jing; Xu, Ruihuan; Zhan, Sien; Lin, Guigao; Wang, Wei; Liu, Min; Wang, Lunan; Zhang, Rui; Li, Jinming

    2009-05-01

    RNA-based therapeutic strategies are used widely due to their highly specific mode of action. However, the major obstacle in any RNA-based therapy is cellular delivery and stability in the cells. The self-assembly of the MS2 bacteriophage capsids has been used to develop virus-like particles (VLPs) for drug delivery. In this study, we utilized the heterobifunctional crosslinker, sulfosuccinimidyl-4-(p-maleimidophenyl)-butyrate (sulfo-SMPB), to conjugate the human immunodeficiency virus-1 (HIV-1) Tat peptide and MS2 VLPs; the antisense RNA against the 5'-untranslated region (UTR) and the internal ribosome entry site (IRES) of the hepatitis C virus (HCV) was packaged into these particles by using a two-plasmid coexpression system. The MS2 VLPs conjugated with the Tat peptide were then transferred into Huh-7 cells containing an HCV reporter system. The packaged antisense RNA showed an inhibitory effect on the translation of HCV. This paper describes our initial results with this system using the Tat peptide. PMID:18823738

  10. Simulation of laser penetration efficiency

    Science.gov (United States)

    Semak, V. V.; Miller, T. F.

    2013-09-01

    The results of numerical simulation of laser beam interaction with a hypothetical metallic material with properties similar to a steel alloy are reported. The numerical simulation was performed using a physical model that includes detailed consideration of surface evaporation, evaporative cooling of the surface and evaporation recoil induced melt ejection. The laser beam ‘penetration’ is considered in terms of melting through the sample or drilling through the sample due to both evaporation and recoil ejection of material. As a demonstration of the predictive capabilities of the model, the average velocity of penetration through a material with steel-like properties is numerically predicted for various laser interaction parameters such as, laser beam radius, laser pulse duration (including CW regime), laser pulse energy and pulse repetition. In particular, the average penetration velocities through a sample due to melting are compared for pulsed and CW lasers of the same power. For the sake of another demonstration of penetration simulation, the temporal dynamics of the position of melt front relative to the sample surface irradiated by a laser beam was computed for different laser pulse repetition rates and constant average laser power. An illustration of the penetration efficiency (W parameter) defined as the amount of energy per unit volume delivered into a target in order to achieve either melting of drilling through a target wall is shown in a wide range of laser pulse parameters covering regimes corresponding to domination of melting through and drilling through.

  11. The Penetration of Shaped Charges

    Institute of Scientific and Technical Information of China (English)

    Wu Mingde

    1996-01-01

    @@ In order to get the best perforating effect, many facts have to be considered. Perforating job is often design by computer. A perforating engineer has to decide the perforating technology and procedure, the gun and charge system, the shot density and phasing, the penetration and hole size, etc.

  12. FAA Fluorescent Penetrant Laboratory Inspections

    Energy Technology Data Exchange (ETDEWEB)

    WINDES,CONNOR L.; MOORE,DAVID G.

    2000-08-02

    The Federal Aviation Administration Airworthiness Assurance NDI Validation Center currently assesses the capability of various non-destructive inspection (NDI) methods used for analyzing aircraft components. The focus of one such exercise is to evaluate the sensitivity of fluorescent liquid penetrant inspection. A baseline procedure using the water-washable fluorescent penetrant method defines a foundation for comparing the brightness of low cycle fatigue cracks in titanium test panels. The analysis of deviations in the baseline procedure will determine an acceptable range of operation for the steps in the inspection process. The data also gives insight into the depth of each crack and which step(s) of the inspection process most affect penetrant sensitivities. A set of six low cycle fatigue cracks produced in 6.35-mm thick Ti-6Al-4V specimens was used to conduct the experiments to produce sensitivity data. The results will document the consistency of the crack readings and compare previous experiments to find the best parameters for water-washable penetrant.

  13. Stylet penetration activities by aphids

    NARCIS (Netherlands)

    Tjallingii, W.F.

    1985-01-01

    The composition of stylet penetration behaviour in aphids and its possible role in food-plant selection is the subject of these studies. Evidence is presented that the labium is devoid of external chemoreceptors (Chapter 1). In addition to other morphological data (Wensler & Filshie, 1969; Wensler,

  14. Projectile penetration into ballistic gelatin.

    Science.gov (United States)

    Swain, M V; Kieser, D C; Shah, S; Kieser, J A

    2014-01-01

    Ballistic gelatin is frequently used as a model for soft biological tissues that experience projectile impact. In this paper we investigate the response of a number of gelatin materials to the penetration of spherical steel projectiles (7 to 11mm diameter) with a range of lower impacting velocities (number of predictive relationships available in the literature, it is found that over the range of projectiles and compositions used, the results fit a simple relationship that takes into account the projectile diameter, the threshold velocity for penetration into the gelatin and a value of the shear modulus of the gelatin estimated from the threshold velocity for penetration. The normalised depth is found to fit the elastic Froude number when this is modified to allow for a threshold impact velocity. The normalised penetration data are found to best fit this modified elastic Froude number with a slope of 1/2 instead of 1/3 as suggested by Akers and Belmonte (2006). Possible explanations for this difference are discussed. PMID:24184862

  15. Simulation of laser penetration efficiency

    International Nuclear Information System (INIS)

    The results of numerical simulation of laser beam interaction with a hypothetical metallic material with properties similar to a steel alloy are reported. The numerical simulation was performed using a physical model that includes detailed consideration of surface evaporation, evaporative cooling of the surface and evaporation recoil induced melt ejection. The laser beam ‘penetration’ is considered in terms of melting through the sample or drilling through the sample due to both evaporation and recoil ejection of material. As a demonstration of the predictive capabilities of the model, the average velocity of penetration through a material with steel-like properties is numerically predicted for various laser interaction parameters such as, laser beam radius, laser pulse duration (including CW regime), laser pulse energy and pulse repetition. In particular, the average penetration velocities through a sample due to melting are compared for pulsed and CW lasers of the same power. For the sake of another demonstration of penetration simulation, the temporal dynamics of the position of melt front relative to the sample surface irradiated by a laser beam was computed for different laser pulse repetition rates and constant average laser power. An illustration of the penetration efficiency (W parameter) defined as the amount of energy per unit volume delivered into a target in order to achieve either melting of drilling through a target wall is shown in a wide range of laser pulse parameters covering regimes corresponding to domination of melting through and drilling through. (paper)

  16. Industrial Penetration and Internet Intensity

    NARCIS (Netherlands)

    C-L. Chang (Chia-Lin); M.J. McAleer (Michael); Y-C. Wu (Yu-Chieh)

    2016-01-01

    textabstractThis paper investigates the effect of industrial penetration and internet intensity for Taiwan manufacturing firms, and analyses whether the relationships are substitutes or complements. The sample observations are based on 153,081 manufacturing plants, and covers 26 two-digit industry c

  17. LeciPlex, invasomes, and liposomes: A skin penetration study.

    Science.gov (United States)

    Shah, Sanket M; Ashtikar, Mukul; Jain, Ankitkumar S; Makhija, Dinesh T; Nikam, Yuvraj; Gude, Rajiv P; Steiniger, Frank; Jagtap, Aarti A; Nagarsenker, Mangal S; Fahr, Alfred

    2015-07-25

    The present study compares three vesicular systems, cationic LeciPlex, invasomes, and conventional liposomes for their ability to deliver drugs deep into the skin. Skin penetration ability of the three vesicular systems was studied for two drugs namely idebenone (antioxidant/anticancer) and azelaic acid (antiacne). All systems showed sizes in nanometer range with small polydispersity indices. Vesicular systems were characterized by CryoTEM studies to understand the differences in morphology of the vesicular systems. Ex vivo human skin penetration studies suggested a pattern in penetration of drugs in different layers of the skin: LeciPlex showed higher penetration for idebenone whereas invasomes showed higher penetration of azelaic acid. Ex vivo study using a fluorescent dye (DiI) was performed to understand the differences in the penetration behavior of the three vesicular systems on excised human skin. In vitro cytotoxicity studies on B16F10 melanoma cell lines revealed, when loaded with idebenone, LeciPlex formulations had the superior activity followed by invasomes and liposomes. In vitro antimicrobial study of azelaic acid loaded systems on Propionibacterium acne revealed high antimicrobial activity for DDAB leciplex followed by almost equal activity for invasomes and CTAB LeciPlex followed by liposomes. Whereas antiacne efficacy study in rats for azelaic acid loaded systems, invasomes exhibited the best antiacne efficacy followed by liposomes and LeciPlex.

  18. Akon - A Penetrator for Europa

    Science.gov (United States)

    Jones, Geraint

    2016-04-01

    Jupiter's moon Europa is one of the most intriguing objects in our Solar System. This 2000km-wide body has a geologically young solid water ice crust that is believed to cover a global ocean of liquid water. The presence of this ocean, together with a source of heating through tidal forces, make Europa a conceivable location for extraterrestrial life. The science case for exploring all aspects of this icy world is compelling. NASA has selected the Europa Mission (formerly Europa Clipper) to study Europa in detail in the 2020s through multiple flybys, and ESA's JUICE mission will perform two flybys of the body in the 2030s. The US agency has extended to the European Space Agency an invitation to provide a contribution to their mission. European scientists interested in Europa science and exploration are currently organizing themselves, in the framework of a coordinated Europa M5 Inititative to study concurrently the main options for this ESA contribution, from a simple addition of individual instruments to the NASA spacecraft, to a lander to investigate Europa's surface in situ. A high speed lander - a penetrator - is by far the most promising technology to achieve this latter option within the anticipated mass constraints, and studies of such a hard lander, many funded by ESA, are now at an advanced level. An international team to formally propose an Europa penetrator to ESA in response to the anticipated ESA M5 call is growing. The working title of this proposal is Akon (Άκων), named after the highly accurate javelin gifted to Europa by Zeus in ancient Greek mythology. We present plans for the Akon penetrator, which would impact Europa's surface at several hundred metres per second, and travel up to several metres into the moon's subsurface. To achieve this, the penetrator would be delivered to the surface by a dedicated descent module, to be destroyed on impact following release of the penetrator above the surface. It is planned that the instruments to be

  19. Assessing high wind energy penetration

    DEFF Research Database (Denmark)

    Tande, J.O.

    1995-01-01

    expanding the grid connected wind power capacity in Praia, the capital of Cape Verde. The currently installed 1 MW of wind power is estimated to supply close to 10% of the electric energy consumption in 1996. Increasing the wind energy penetration to a higher level is considered viable as the project......In order to convincingly promote installing wind power capacity as a substantial part of the energy supply system, a set of careful analyses must be undertaken. This paper applies a case study concentrated on assessing the cost/benefit of high wind energy penetration. The case study considers...... settings are close to ideal, including a very capable national utility company, Electra, a conventional power supply system based on imported heavy fuel and gas oil, and favourable wind conditions with an estimated annual average of 9.3 m/s at the hub height of the wind turbines. With the applied case...

  20. Correlation Between Cone Penetration Rate And Measured Cone Penetration Parameters In Silty Soils

    DEFF Research Database (Denmark)

    Poulsen, Rikke; Nielsen, Benjaminn Nordahl; Ibsen, Lars Bo

    2013-01-01

    This paper shows, how a change in cone penetration rate affects the cone penetration measurements, hence the cone resistance, pore pressure, and sleeve friction in silty soil. The standard rate of penetration is 20 mm/s, and it is generally accepted that undrained penetration occurs in clay while...... penetration tests with varying penetration rates conducted at a test site where the subsoil primary consists of sandy silt. It is shown how a reduced penetration rate influences the cone penetration measurements e.g. the cone resistance, pore pressure, and sleeve friction....

  1. Evaluation of drug penetration with cationic micelles and their penetration mechanism using an in vitro tumor model.

    Science.gov (United States)

    Suzuki, Hidenori; Bae, You Han

    2016-08-01

    Elevated interstitial fluid pressure (IFP) and abnormal extracellular matrix (ECM) are major factors causing significant barriers to penetration of nanomedicines in solid tumors. To better understand the barriers, various in vitro tumor models including multicellular spheroids and multilayered cell cultures (MCCs) have been developing. Recently, we have established a unique in vitro tumor model composed of a MCC and an Ussing chamber system which is modified to add a hydraulic pressure gradient through the MCC. In this study, we evaluated the drug penetration ability of cationic micelles using the unique in vitro tumor model. The doxorubicin (DOX)-loaded cationic micelles, which are formed from a triblock copolymer of poly(d,l-lactide-co-glycolide)-block-branched polyethyleneimine-block-poly(d,l-lactide-co-glycolide), deeply penetrated and released the DOX throughout the MCC against convectional flow caused by a hydraulic pressure gradient. The studies using endocytosis markers and inhibitors showed that the micelles utilized mainly macropinocytosis as an internalization pathway. Furthermore, the penetration was apparently inhibited by treatment with exocytosis inhibitor Exo1. These results suggest that the active penetration of the cationic micelles is induced by iterative transcytosis via macropinocytosis and exocytosis. Our findings could be beneficial information to improve intratumoral penetration in new nanomedicines for solid tumors. PMID:27182814

  2. PENETRATION ENHANCEMENT OF MEDICINAL AGENTS

    OpenAIRE

    Sharma Ganesh N.; Sanadya Jyotsana; Kaushik Avinash; Dwivedi Abha

    2012-01-01

    Many current therapeutic agents like antibiotics, ionizable and peptide drugs are impermeable or do not possess the requisite physicochemical properties for efficient transport through outer tissue barrier to attain therapeutic blood level. For this reason the delivery of such drugs through barriers is currently one of the major interests in pharmaceutical research. Penetration enhancers or promoters are agents that have no therapeutic properties of their own but can transport the sorption of...

  3. 经穿膜肽与PEG修饰的核糖体失活蛋白Gelonin抗肿瘤作用的研究%Study on cell-penetrating peptide modified and PEGylated ribosome inactive protein Gelonin

    Institute of Scientific and Technical Information of China (English)

    张娅洁; 王慧媛; 陈应之; 汤懿斯; 杨志民; 黄永焯

    2015-01-01

    Objective:To improve anti-tumor effect of Gelonin, the plant-sourced RIP is modified by chemically conjugating a cell-penetrating peptide and polyethylene glycol (PEG). Methods:Purified protein was obtained after being performed on FPLC (fast protein liquid chromatography) Superdex75 column. Cytotoxicity was detected by MTT assay. The cellular uptake by HT1080 cells was studied by using inverted fluorescence microscopy and flow cytometry. In-vivo imaging technology was utilized for investigation of the in-vivo drug distribution in the HT1080 tumor-bearing mice. Results:The modified product was purified by using gel filtration chromatergraphy. Moreover, compared with native Gelonin, the cytotoxicity of modified protein was increased, especially in HT1080, presumably due to the enhanced cellular uptake. The in-vivo imaging results suggested that drug accumulation in tumor was improved by PEGylation. Conclusion:Modified Gelonin can improve cell penetration and cytotoxicity in tumor cells. PEGylation can increase tumor accumulation of the protein drug, and thereby enhance its anti-tumor effect.%目的:通过对核糖体失活蛋白Gelonin进行化学修饰,利用穿膜肽和聚乙二醇(PEG)偶联来提高其到达肿瘤部位和进入肿瘤细胞的能力,使Gelonin更高效地发挥抑瘤作用. 方法:利用FPLC Superdex75分子筛预装柱纯化系统对所修饰的Gelonin进行纯化后,在不同细胞系测试细胞毒性;通过倒置荧光显微镜、流式细胞分析技术等对药物进入纤维肉瘤细胞HT1080的能力进行评价;采用小动物活体成像技术考察药物体系在荷瘤动物体内的分布情况. 结果:采用分子筛色谱纯化可以得到纯度相对较高的修饰产物,其毒性较无修饰的Gelonin强,且在HT1080细胞系作用最明显;细胞摄取结果显示,与未修饰的Gelonin相比,该药物体系具有更高的细胞摄取效率;动物成像结果表明,PEG5000修饰可以改变Gelonin在动物体内的分布情况,

  4. Cardiac Penetrating Injuries and Pseudoaneurysm

    Institute of Scientific and Technical Information of China (English)

    CHEN Shifeng

    2002-01-01

    Objective To discuss the early diagnosis and treatment of cardiac penetrating injuries and pseudoaneurysm. Methods 18 cases of cardiac penetrating injuries, in which 2 cases were complicated with pseudoaneurysm, were diagnosed by emergency operation and color Doppler echocardiography between May 1973 and Dec. 2001 in our hospital. The basis for emergency operation is the injured path locating in cardiac dangerous zone, severe shock or pericardial tamponade. ResultsAmong 18 cases of this study, 17 cases underwent emergency operation. During the operation, 11 cases were found injured in right ventricle, 2 cases were found injured in right atrium, 1 case was found injured in pulmonary artery,4 cases were found injured in left ventricle, 2 cases were found complicated with pseudoaneurysm. 17cases underwent cardiac repair including 1 case of rupture of aneurysm. 1 case underwent elective aneurysm resection. In whole group, 15 cases survived(83.33% ), 3 cases died( 16.67%). The cause of death is mainly hemorrhagic shock. Conclusion Highly suspicious cardiac penetrating injuries or hemopericaridium should undergo direct operative exploration. Pseudoaneurysm should be resected early,which can prevent severe complications.

  5. Combined logging/penetration methods

    International Nuclear Information System (INIS)

    A method is described for studying important soil and rock properties by simultaneous penetration and logging measurements. In penetration measurements, a BORROS Co. probe with a coned tip, 35.6 mm in diameter, a point angle of 60 deg, and with a friction roller 150 cm2 in area was dipped in the soil at a constant speed. Tip resistance and skin friction were measured. At the same time, gamma logging was carried out to determine natural radioactivity, gamma-gamma logging to determine bulk density, neutron-neutron logging to determine bulk density and porosity, and inclinometry. An original probe fitted with a NaI(Tl) scintillation detector with a crystal 25 mm in diameter and 100 mm in length was developed for gamma logging. For gamma-gamma logging, a 137Cs emitter probe was used with the emitter/detector distance of 45 cm. A NaI(Tl) crystal 18 mm in diameter and 50 mm in length was used for radiation detection. For neutron-neutron logging, a NIV 2 probe was purchased in the USSR and modified for simultaneous penetration measurements. A Soviet-made UMI 30 inclinometer with a modified case was used for inclinometry. (Ha)

  6. Network Penetration Testing and Research

    Science.gov (United States)

    Murphy, Brandon F.

    2013-01-01

    This paper will focus the on research and testing done on penetrating a network for security purposes. This research will provide the IT security office new methods of attacks across and against a company's network as well as introduce them to new platforms and software that can be used to better assist with protecting against such attacks. Throughout this paper testing and research has been done on two different Linux based operating systems, for attacking and compromising a Windows based host computer. Backtrack 5 and BlackBuntu (Linux based penetration testing operating systems) are two different "attacker'' computers that will attempt to plant viruses and or NASA USRP - Internship Final Report exploits on a host Windows 7 operating system, as well as try to retrieve information from the host. On each Linux OS (Backtrack 5 and BlackBuntu) there is penetration testing software which provides the necessary tools to create exploits that can compromise a windows system as well as other operating systems. This paper will focus on two main methods of deploying exploits 1 onto a host computer in order to retrieve information from a compromised system. One method of deployment for an exploit that was tested is known as a "social engineering" exploit. This type of method requires interaction from unsuspecting user. With this user interaction, a deployed exploit may allow a malicious user to gain access to the unsuspecting user's computer as well as the network that such computer is connected to. Due to more advance security setting and antivirus protection and detection, this method is easily identified and defended against. The second method of exploit deployment is the method mainly focused upon within this paper. This method required extensive research on the best way to compromise a security enabled protected network. Once a network has been compromised, then any and all devices connected to such network has the potential to be compromised as well. With a compromised

  7. The production process for water penetrated brick

    Institute of Scientific and Technical Information of China (English)

    SunGuofeng

    2005-01-01

    Waste penetrated brick, which is a green building material with good water penetration, high strength, lower firing temperature, lower production cost, good appearance and good structure, can holding ground water lever. This article analysis the production process and related factor for water penetrated brick:proper particle size distribution, proper shaping method, proper press and proper firing can ensure to produce good quality water penetrated brick.

  8. Kali Linux wireless penetration testing essentials

    CERN Document Server

    Alamanni, Marco

    2015-01-01

    This book is targeted at information security professionals, penetration testers and network/system administrators who want to get started with wireless penetration testing. No prior experience with Kali Linux and wireless penetration testing is required, but familiarity with Linux and basic networking concepts is recommended.

  9. In vitro dermal penetration study of carbofuran, carbosulfan, and furathiocarb

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Kwang-Hyeon [Department of Pharmacology, Inje University, College of Medicine, 614-735, Busan (Korea); Kim, Jeong-Han [School of Agricultural Biotechnology, Seoul National University, 441-744, Suwon (Korea)

    2003-05-01

    In this study, the dermal penetration rate of carbofuran, carbosulfan, and furathiocarb has been measured with rat abdominal skin using the static diffusion cell. The technical grades of three compounds were applied at different doses on skin surface mounted in static diffusion cell and incubated at 32 C for 48 h with shaking. The same procedures were carried out with furathiocarb EC (emulsifiable concentrate) and WP (wettable powder). At regular intervals, the receptor fluid in cell was sampled and analyzed by HPLC. Only carbofuran was found in carbosulfan- or furathiocarb-treated samples, suggesting they converted into carbofuran while passing through the skin layer. The quantity of insecticide penetrating skin increased with time and applied dose. The skin penetration rate increased with the water solubility of insecticides. The dermal penetration rates of carbofuran, furathiocarb, and carbosulfan were determined as 1.05 {mu}g/cm{sup 2} per h (r{sup 2}=0.991), 0.46 {mu}g/cm{sup 2} per h (r{sup 2}=0.984) and 0.14 {mu}g/cm{sup 2} per h (r{sup 2}=0.967), respectively. There was no significant difference in rate of skin penetration between furathiocarb EC (1.42 {mu}g/cm{sup 2} per h, r{sup 2}=0.988) and WP (1.35 {mu}g/cm{sup 2} per h, r{sup 2}=0.982), while furathiocarb technical grade showed a lower skin penetration rate. In vitro models may be used to predict percutaneous absorption and are useful in selecting safer formulations for field application of pesticide. (orig.)

  10. Isolation, Co-Crystallization and Structure-Based Characterization of Anabaenopeptins as Highly Potent Inhibitors of Activated Thrombin Activatable Fibrinolysis Inhibitor (TAFIa).

    Science.gov (United States)

    Schreuder, Herman; Liesum, Alexander; Lönze, Petra; Stump, Heike; Hoffmann, Holger; Schiell, Matthias; Kurz, Michael; Toti, Luigi; Bauer, Armin; Kallus, Christopher; Klemke-Jahn, Christine; Czech, Jörg; Kramer, Dan; Enke, Heike; Niedermeyer, Timo H J; Morrison, Vincent; Kumar, Vasant; Brönstrup, Mark

    2016-01-01

    Mature thrombin activatable fibrinolysis inhibitor (TAFIa) is a carboxypeptidase that stabilizes fibrin clots by removing C-terminal arginines and lysines from partially degraded fibrin. Inhibition of TAFIa stimulates the degradation of fibrin clots and may help to prevent thrombosis. Applying a lead finding approach based on literature-mining, we discovered that anabaenopeptins, cyclic peptides produced by cyanobacteria, were potent inhibitors of TAFIa with IC50 values as low as 1.5 nM. We describe the isolation and structure elucidation of 20 anabaenopeptins, including 13 novel congeners, as well as their pronounced structure-activity relationships (SAR) with respect to inhibition of TAFIa. Crystal structures of the anabaenopeptins B, C and F bound to the surrogate protease carboxypeptidase B revealed the binding modes of these large (~850 Da) compounds in detail and explained the observed SAR, i.e. the strong dependence of the potency on a basic (Arg, Lys) exocyclic residue that addressed the S1' binding pocket, and a broad tolerance towards substitutions in the pentacyclic ring that acted as a plug of the active site. PMID:27604544

  11. FAA Fluorescent Penetrant Activities - An Update

    Energy Technology Data Exchange (ETDEWEB)

    Moore, D.G.

    1998-10-20

    The Federal Aviation Administration's Airworthiness Assurance NDI Validation Center (AANC) is currently characterizing low cycle fatigue specimens that will support the needs of penetrant manufacturers, commercial airline industry and the Federal Aviation Administration. The main focus of this characterization is to maintain and enhance the evaluation of penetrant inspection materials and apply resources to support the aircraft community needs. This paper discusses efforts to-date to document the Wright Laboratory penetrant evaluation process and characterize penetrant brightness readings in the initial set of sample calibration panels using Type 1 penetrant.

  12. Web penetration testing with Kali Linux

    CERN Document Server

    Muniz, Joseph

    2013-01-01

    Web Penetration Testing with Kali Linux contains various penetration testing methods using BackTrack that will be used by the reader. It contains clear step-by-step instructions with lot of screenshots. It is written in an easy to understand language which will further simplify the understanding for the user.""Web Penetration Testing with Kali Linux"" is ideal for anyone who is interested in learning how to become a penetration tester. It will also help the users who are new to Kali Linux and want to learn the features and differences in Kali versus Backtrack, and seasoned penetration testers

  13. Penetration testing with Raspberry Pi

    CERN Document Server

    Muniz, Joseph

    2015-01-01

    If you are looking for a low budget, small form-factor remotely accessible hacking tool, then the concepts in this book are ideal for you. If you are a penetration tester who wants to save on travel costs by placing a low-cost node on a target network, you will save thousands by using the methods covered in this book. You do not have to be a skilled hacker or programmer to use this book. It will be beneficial to have some networking experience; however, it is not required to follow the concepts covered in this book.

  14. An electrochemical modeling of lithium-ion battery nail penetration

    Science.gov (United States)

    Chiu, Kuan-Cheng; Lin, Chi-Hao; Yeh, Sheng-Fa; Lin, Yu-Han; Chen, Kuo-Ching

    2014-04-01

    Nail penetration into a battery pack, resulting in a state of short-circuit and thus burning, is likely to occur in electric car collisions. To demonstrate the behavior of a specific battery when subject to such incidents, a standard nail penetration test is usually performed; however, conducting such an experiment is money consuming. The purpose of this study is to propose a numerical electrochemical model that can simulate the test accurately. This simulation makes two accurate predictions. First, we are able to model short-circuited lithium-ion batteries (LIBs) via electrochemical governing equations so that the mass and charge transfer effect could be considered. Second, the temperature variation of the cell during and after nail penetration is accurately predicted with the help of simulating the temperature distribution of thermal runaway cells by thermal abuse equations. According to this nail penetration model, both the onset of battery thermal runaway and the cell temperature profile of the test are obtained, both of which are well fitted with our experimental results.

  15. Evaluation of nicotinamide microemulsion on the skin penetration enhancement.

    Science.gov (United States)

    Boonme, Prapaporn; Boonthongchuay, Chalida; Wongpoowarak, Wibul; Amnuaikit, Thanaporn

    2016-01-01

    This study purposed to evaluate a microemulsion containing nicotinamide for its characteristics, stability, and skin penetration and retention comparing with a solution of nicotinamide in 2:1 mixture of water and isopropyl alcohol (IPA). The microemulsion system was composed of 1:1 mixture of Span80 and Tween80 as a surfactant mixture, isopropyl palmitate (IPP) as an oil phase, and 2:1 mixture of water and IPA as an aqueous phase. Nicotinamide microemulsion was prepared by dissolving the active in the aqueous phase before simply mixing with the other components. It was determined for its characteristics and stability under various conditions. The skin penetration and retention studies of nicotinamide microemulsion and solution were performed by modified Franz diffusion cells, using newborn pig skin as the membrane. The results showed that nicotinamide microemulsion could be obtained as clear yellowish liquid, was water-in-oil (w/o) type, possessed Newtonian flow, and exhibited physicochemical stability when kept at 4 °C and room temperature (≈30 ± 2 °C) during 3 months. From the skin penetration data, the microemulsion could enhance the skin penetration of nicotinamide comparing with the solution. Additionally, nicotinamide microemulsion could provide much higher amount of skin retention than that of skin penetration, resulting in suitability for a cosmeceutical product.

  16. Evaluation of nicotinamide microemulsion on the skin penetration enhancement.

    Science.gov (United States)

    Boonme, Prapaporn; Boonthongchuay, Chalida; Wongpoowarak, Wibul; Amnuaikit, Thanaporn

    2016-01-01

    This study purposed to evaluate a microemulsion containing nicotinamide for its characteristics, stability, and skin penetration and retention comparing with a solution of nicotinamide in 2:1 mixture of water and isopropyl alcohol (IPA). The microemulsion system was composed of 1:1 mixture of Span80 and Tween80 as a surfactant mixture, isopropyl palmitate (IPP) as an oil phase, and 2:1 mixture of water and IPA as an aqueous phase. Nicotinamide microemulsion was prepared by dissolving the active in the aqueous phase before simply mixing with the other components. It was determined for its characteristics and stability under various conditions. The skin penetration and retention studies of nicotinamide microemulsion and solution were performed by modified Franz diffusion cells, using newborn pig skin as the membrane. The results showed that nicotinamide microemulsion could be obtained as clear yellowish liquid, was water-in-oil (w/o) type, possessed Newtonian flow, and exhibited physicochemical stability when kept at 4 °C and room temperature (≈30 ± 2 °C) during 3 months. From the skin penetration data, the microemulsion could enhance the skin penetration of nicotinamide comparing with the solution. Additionally, nicotinamide microemulsion could provide much higher amount of skin retention than that of skin penetration, resulting in suitability for a cosmeceutical product. PMID:25318786

  17. Effects of soap-water wash on human epidermal penetration.

    Science.gov (United States)

    Zhu, Hanjiang; Jung, Eui-Chang; Phuong, Christina; Hui, Xiaoying; Maibach, Howard

    2016-08-01

    Skin decontamination is a primary interventional method used to decrease dermal absorption of hazardous contaminants, including chemical warfare agents, pesticides and industrial pollutants. Soap and water wash, the most common and readily available decontamination system, may enhance percutaneous absorption through the "wash-in effect." To understand better the effect of soap-water wash on percutaneous penetration, and provide insight to improving skin decontamination methods, in vitro human epidermal penetration rates of four C(14) -labeled model chemicals (hydroquinone, clonidine, benzoic acid and paraoxon) were assayed using flow-through diffusion cells. Stratum corneum (SC) absorption rates of these chemicals at various hydration levels (0-295% of the dry SC weights) were determined and compared with the results of the epidermal penetration study to clarify the effect of SC hydration on skin permeability. Results showed accelerated penetration curves of benzoic acid and paraoxon after surface wash at 30 min postdosing. Thirty minutes after washing (60 min postdosing), penetration rates of hydroquinone and benzoic acid decreased due to reduced amounts of chemical on the skin surface and in the SC. At the end of the experiment (90 min postdosing), a soap-water wash resulted in lower hydroquinone penetration, greater paraoxon penetration and similar levels of benzoic acid and clonidine penetration compared to penetration levels in the non-wash groups. The observed wash-in effect agrees with the enhancement effect of SC hydration on the SC chemical absorption rate. These results suggest SC hydration derived from surface wash to be one cause of the wash-in effect. Further, the occurrence of a wash-in effect is dependent on chemical identity and elapsed time between exposure and onset of decontamination. By reducing chemical residue quantity on skin surface and in the SC reservoir, the soap-water wash may decrease the total quantity of chemical absorbed in the

  18. Effects of soap-water wash on human epidermal penetration.

    Science.gov (United States)

    Zhu, Hanjiang; Jung, Eui-Chang; Phuong, Christina; Hui, Xiaoying; Maibach, Howard

    2016-08-01

    Skin decontamination is a primary interventional method used to decrease dermal absorption of hazardous contaminants, including chemical warfare agents, pesticides and industrial pollutants. Soap and water wash, the most common and readily available decontamination system, may enhance percutaneous absorption through the "wash-in effect." To understand better the effect of soap-water wash on percutaneous penetration, and provide insight to improving skin decontamination methods, in vitro human epidermal penetration rates of four C(14) -labeled model chemicals (hydroquinone, clonidine, benzoic acid and paraoxon) were assayed using flow-through diffusion cells. Stratum corneum (SC) absorption rates of these chemicals at various hydration levels (0-295% of the dry SC weights) were determined and compared with the results of the epidermal penetration study to clarify the effect of SC hydration on skin permeability. Results showed accelerated penetration curves of benzoic acid and paraoxon after surface wash at 30 min postdosing. Thirty minutes after washing (60 min postdosing), penetration rates of hydroquinone and benzoic acid decreased due to reduced amounts of chemical on the skin surface and in the SC. At the end of the experiment (90 min postdosing), a soap-water wash resulted in lower hydroquinone penetration, greater paraoxon penetration and similar levels of benzoic acid and clonidine penetration compared to penetration levels in the non-wash groups. The observed wash-in effect agrees with the enhancement effect of SC hydration on the SC chemical absorption rate. These results suggest SC hydration derived from surface wash to be one cause of the wash-in effect. Further, the occurrence of a wash-in effect is dependent on chemical identity and elapsed time between exposure and onset of decontamination. By reducing chemical residue quantity on skin surface and in the SC reservoir, the soap-water wash may decrease the total quantity of chemical absorbed in the

  19. Cable Braid Electromagnetic Penetration Model.

    Energy Technology Data Exchange (ETDEWEB)

    Warne, Larry K. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Langston, William L. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Basilio, Lorena I. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States); Johnson, W. A. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)

    2015-06-01

    The model for penetration of a wire braid is rigorously formulated. Integral formulas are developed from energy principles and reciprocity for both self and transfer immittances in terms of potentials for the fields. The detailed boundary value problem for the wire braid is also setup in a very efficient manner; the braid wires act as sources for the potentials in the form of a sequence of line multipoles with unknown coefficients that are determined by means of conditions arising from the wire surface boundary conditions. Approximations are introduced to relate the local properties of the braid wires to a simplified infinite periodic planar geometry. This is used in a simplified application of reciprocity to be able to treat nonuniform coaxial geometries including eccentric interior coaxial arrangements and an exterior ground plane.

  20. Ethical hacking and penetration testing guide

    CERN Document Server

    Baloch, Rafay

    2014-01-01

    Requiring no prior hacking experience, Ethical Hacking and Penetration Testing Guide supplies a complete introduction to the steps required to complete a penetration test, or ethical hack, from beginning to end. You will learn how to properly utilize and interpret the results of modern-day hacking tools, which are required to complete a penetration test. The book covers a wide range of tools, including Backtrack Linux, Google reconnaissance, MetaGooFil, dig, Nmap, Nessus, Metasploit, Fast Track Autopwn, Netcat, and Hacker Defender rootkit. Supplying a simple and clean explanation of how to effectively utilize these tools, it details a four-step methodology for conducting an effective penetration test or hack.Providing an accessible introduction to penetration testing and hacking, the book supplies you with a fundamental understanding of offensive security. After completing the book you will be prepared to take on in-depth and advanced topics in hacking and penetration testing. The book walks you through each ...

  1. Proniosomes for Penetration Enhancement in Transdermal System

    OpenAIRE

    Samita Singla; S. L. Harikumar; Geeta Aggarwal

    2012-01-01

    Over the last few years an inclusive research has been done over provesicular approach for transdermal drug delivery. Skin has a very tough diffusion barrier inhibiting penetration of drug moiety which is rate limiting barrier for penetration of drugs. There are several approaches that deal with penetration enhancement across the skin. Vesicular and provesicular systems are promising amongst them. Vesicular systems including (niosomes, ethosomes, transfersomes and liposomes) are promising sys...

  2. A Multiple Scattering Theory for Proton Penetration

    Institute of Scientific and Technical Information of China (English)

    YANG Dai-Lun; WU Zhang-Wen; JIANG Steve-Bin; LUO Zheng-Ming

    2004-01-01

    @@ We extend the electron small-angle multiple scattering theory to proton penetration. After introducing the concept of narrow energy spectra, the proton energy loss process is included in the proton deep penetration theory. It precisely describes the whole process of proton penetration. Compared to the Monte Carlo method,this method maintains the comparable precision and possesses much higher computational efficiency. Thus, it shows the real feasibility of applying this algorithm to proton clinical radiation therapy.

  3. Site-specific conjugation of the quencher on peptide's N-terminal for the synthesis of a targeted non-spreading activatable optical probe.

    Science.gov (United States)

    Simard, Bryan; Mironov, Gleb G; Tomanek, Boguslaw; van Veggel, Frank C J M; Abulrob, Abedelnasser

    2016-06-01

    Optical imaging offers high sensitivity and portability at low cost. The design of 'smart' or 'activatable' probes can decrease the background noise and increase the specificity of the signal. By conjugating a fluorescent dye and a compatible quencher on each side of an enzyme's substrate, the signal remains in its 'off ' state until it reaches the area where a specific enzyme is expressed. However, the signal can leak from that area unless the dye is attached to a molecule able to bind to a specific target also presented in that area. The aim of this study was to (i) specifically conjugate the quencher on the α-amino group of the peptide's N-terminus, (ii) conjugate the dye on the ε-amino group of a lysine in C-terminus, and (iii) conjugate the carboxyl group of the peptide's C-terminus to an amino group present on an antibody, using carbodiimide chemistry. The use of protecting groups, such as Boc or Fmoc, to allow site-specific conjugation, presents several drawbacks including 'on beads labeling', additional steps required for deprotection and removal from the resin, decreased yield, and dye degradation. A method of preferential labeling of α-amino N-terminal group in slightly acidic solution, proposed by Selo et al. (1996) has partially solved the problem. The present study reports improvements of the method allowing to (i) avoid the homo-bilabeling, (ii) increase the yield of the N-terminal labeling by two folds, and (iii) decrease the cost by 44-fold. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. PMID:27282138

  4. Penetration depth time history measurement method

    OpenAIRE

    Liu, C; Ahrens, T. J.

    2000-01-01

    A new method for measuring the depth time history of rigid body penetration into brittle materials under a deceleration of ~10^5 g. The method includes: sabot-projectile, sabot-projectile separation and penetration depth detection systems. Relatively small intrinsic time error (3%) and depth error (0.3–0.7 mm) results. Penetration depth time history in a series of 4140 steel projectile penetrations into a mortar are measured at velocities of 100 to 500 m/sec with sufficient accuracy such that...

  5. In-place HEPA filter penetration test

    Energy Technology Data Exchange (ETDEWEB)

    Bergman, W.; Wilson, K.; Elliott, J. [Lawrence Livermore National Lab., CA (United States)] [and others

    1997-08-01

    We have demonstrated the feasibility of conducting penetration tests on high efficiency particulate air (HEPA) filters as installed in nuclear ventilation systems. The in-place penetration test, which is designed to yield equivalent penetration measurements as the standard DOP efficiency test, is based on measuring the aerosol penetration of the filter installation as a function of particle size using a portable laser particle counter. This in-place penetration test is compared to the current in-place leak test using light scattering photometers for single HEPA filter installations and for HEPA filter plenums using the shroud method. Test results show the in-place penetration test is more sensitive than the in-place leak test, has a similar operating procedure, but takes longer to conduct. Additional tests are required to confirm that the in-place penetration test yields identical results as the standard dioctyl phthalate (DOP) penetration test for HEPA filters with controlled leaks in the filter and gasket and duct by-pass leaks. Further development of the procedure is also required to reduce the test time before the in-place penetration test is practical. 14 refs., 14 figs., 3 tabs.

  6. Market penetration rates of new energy technologies

    International Nuclear Information System (INIS)

    The market penetration rates of 11 different new energy technologies were studied covering energy production and end-use technologies. The penetration rates were determined by fitting observed market data to an epidemical diffusion model. The analyses show that the exponential penetration rates of new energy technologies may vary from 4 up to over 40%/yr. The corresponding take-over times from a 1% to 50% share of the estimated market potential may vary from less than 10 to 70 years. The lower rate is often associated with larger energy impacts. Short take-over times less than 25 years seem to be mainly associated with end-use technologies. Public policies and subsides have an important effect on the penetration. Some technologies penetrate fast without major support explained by technology maturity and competitive prices, e.g. compact fluorescent lamps show a 24.2%/yr growth rate globally. The penetration rates determined exhibit some uncertainty as penetration has not always proceeded close to saturation. The study indicates a decreasing penetration rate with increasing time or market share. If the market history is short, a temporally decreasing functional form for the penetration rate coefficient could be used to anticipate the probable behavior

  7. Ultrasonic/Sonic Impacting Penetrators

    Science.gov (United States)

    Bao, Xiaoqi; Bar-Cohen, Yoseph; Chang, Zensheu; Sherrit, Stewart; Stark, Randall A.

    2008-01-01

    Ultrasonic/sonic impacting penetrators (USIPs) are recent additions to the series of apparatuses based on ultrasonic/sonic drill corers (USDCs). A USIP enables a rod probe to penetrate packed soil or another substance of similar consistency, without need to apply a large axial force that could result in buckling of the probe or in damage to some buried objects. USIPs were conceived for use in probing and analyzing soil to depths of tens of centimeters in the vicinity of buried barrels containing toxic waste, without causing rupture of the barrels. USIPs could also be used for other purposes, including, for example, searching for pipes, barrels, or other hard objects buried in soil; and detecting land mines. USDCs and other apparatuses based on USDCs have been described in numerous previous NASA Tech Briefs articles. The ones reported previously were designed, variously, for boring into, and/or acquiring samples of, rock or other hard, brittle materials of geological interest. To recapitulate: A USDC can be characterized as a lightweight, low-power, piezoelectrically driven jackhammer in which ultrasonic and sonic vibrations are generated and coupled to a tool bit. As shown in the figure, a basic USDC includes a piezoelectric stack, a backing and a horn connected to the stack, a free mass (free in the sense that it can slide axially a short distance between the horn and the shoulder of tool bit), and a tool bit, i.e., probe for USIP. The piezoelectric stack is driven at the resonance frequency of the stack/horn/backing assembly to create ultrasonic vibrations that are mechanically amplified by the horn. To prevent fracture during operation, the piezoelectric stack is held in compression by a bolt. The bouncing of the free mass between the horn and the tool bit at sonic frequencies generates hammering actions to the bit that are more effective for drilling than is the microhammering action of ultrasonic vibrations in ordinary ultrasonic drills. The hammering actions

  8. RELATIONSHIP BETWEEN PLASMA OPTIC SIGNAL AND PENETRATION DEPTH FOR PARTIAL-PENETRATION LASER WELDING

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Through sampling and analyzing of plasma optic signals of 400~600 nm emitted from partial-penetration laser welding processes, how the penetration depth is related to the welding parameter and the plasma optic signal is studied. Under the experimental conditions, the plasma optic signal has good response to variety of the weld penetration, and the signal's RMS value increases with the penetration in a quadratic curve mode. The inherent relation between the plasma optic signal and the penetration depth is also analyzed. It is also found that, between the two common parameters of laser power and welding speed, laser power has more influence on penetration while welding speed has more influence on weld width. The research results provide theoretic and practical bases for penetration real-time monitoring or predicting in partial-penetration laser welding.

  9. The equine herpesvirus 1 glycoprotein gp21/22a, the herpes simplex virus type 1 gM homolog, is involved in virus penetration and cell-to-cell spread of virions.

    OpenAIRE

    Osterrieder, N.; Neubauer, A.; Brandmuller, C; Braun, B.; Kaaden, O R; Baines, J D

    1996-01-01

    Experiments to analyze the function of the equine herpesvirus 1 (EHV-1) glycoprotein gM homolog were conducted. To this end, an Rk13 cell line (TCgM) that stably expressed EHV-1 gM was constructed. Proteins with apparent M(r)s of 46,000 to 48,000 and 50,000 to 55,000 were detected in TCgM cells with specific anti-gM antibodies, and the gM protein pattern was indistinguishable from that in cells infected with EHV-1 strain RacL11. A viral mutant (L11deltagM) bearing an Escherichia coli lacZ gen...

  10. Influence of massage and occlusion on the ex vivo skin penetration of rigid liposomes and invasomes.

    Science.gov (United States)

    Trauer, Sindy; Richter, Heike; Kuntsche, Judith; Büttemeyer, Rolf; Liebsch, Manfred; Linscheid, Michael; Fahr, Alfred; Schäfer-Korting, Monika; Lademann, Jürgen; Patzelt, Alexa

    2014-02-01

    Liposomes are frequently described as drug delivery systems for dermal and transdermal applications. Recently, it has been shown that particulate substances penetrate effectively into hair follicles and that the follicular penetration depth can be increased by massaging the skin, which simulates the in vivo movement of hairs in the hair follicles. In the present study, massage was applied to skin mounted to Franz diffusion cells. By means of confocal laser scanning microscopy, the influence of massage and occlusion on the follicular penetration depths of rigid and flexible liposomes loaded with a hydrophilic and lipophilic dye was investigated. The application of massage increased follicular penetration significantly. Occlusion resulted in an increased follicular penetration depth only for rigid liposomes, whereas invasomes did not penetrate more effectively if occlusion was applied. The results confirm that massage is an important tool for increasing follicular penetration in ex vivo studies using Franz diffusion cells. Occlusion may reduce the efficacy of follicular penetration depending on the specific liposomal preparation. Rigidity in particular appears to be a relevant parameter.

  11. Instability of a penetrating blade

    Science.gov (United States)

    Bigoni, D.; Bosi, F.; Dal Corso, F.; Misseroni, D.

    2014-03-01

    Application of a dead compressive load at the free end of an elastic rod (the ‘blade') induces its penetration into a sliding sleeve ending with a linear elastic spring. Bifurcation and stability analysis of this simple elastic system shows a variety of unexpected behaviors: (i) an increase of buckling load at decreasing of elastic stiffness; (ii) a finite number of buckling loads for a system with infinite degrees of freedom (leading to a non-standard Sturm-Liouville problem); (iii) more than one bifurcation load associated to each bifurcation mode; (iv) a restabilization of the straight configuration after the second bifurcation load associated to the first instability mode; (v) the presence of an Eshelby-like (or configurational) force, deeply influencing stability. Only the first of these behaviors was previously known, the second and third ones disprove common beliefs, the fourth highlights a sort of ‘island of instability', and the last one shows surprising phenomena and effects on stability.

  12. Stepped frequency ground penetrating radar

    Science.gov (United States)

    Vadnais, Kenneth G.; Bashforth, Michael B.; Lewallen, Tricia S.; Nammath, Sharyn R.

    1994-01-01

    A stepped frequency ground penetrating radar system is described comprising an RF signal generating section capable of producing stepped frequency signals in spaced and equal increments of time and frequency over a preselected bandwidth which serves as a common RF signal source for both a transmit portion and a receive portion of the system. In the transmit portion of the system the signal is processed into in-phase and quadrature signals which are then amplified and then transmitted toward a target. The reflected signals from the target are then received by a receive antenna and mixed with a reference signal from the common RF signal source in a mixer whose output is then fed through a low pass filter. The DC output, after amplification and demodulation, is digitized and converted into a frequency domain signal by a Fast Fourier Transform. A plot of the frequency domain signals from all of the stepped frequencies broadcast toward and received from the target yields information concerning the range (distance) and cross section (size) of the target.

  13. Quantitative penetration testing with item response theory

    NARCIS (Netherlands)

    Arnold, Florian; Pieters, Wolter; Stoelinga, Mariëlle

    2014-01-01

    Existing penetration testing approaches assess the vulnerability of a system by determining whether certain attack paths are possible in practice. Thus, penetration testing has so far been used as a qualitative research method. To enable quantitative approaches to security risk management, including

  14. Quantitative penetration testing with item response theory

    NARCIS (Netherlands)

    Pieters, W.; Arnold, F.; Stoelinga, M.I.A.

    2013-01-01

    Existing penetration testing approaches assess the vulnerability of a system by determining whether certain attack paths are possible in practice. Therefore, penetration testing has thus far been used as a qualitative research method. To enable quantitative approaches to security risk management, in

  15. FRACTIONAL PENETRATION OF PAINT OVERSPRAY ARRESTORS

    Science.gov (United States)

    The report describes the development of fractional penetration curves for liquid droplet penetration of overspray arrestors for discrete droplet diameters from 0.3 to 10 micrometers. (NOTE: Fine particulates are particles with diameters of 10 micrometers or less.) These data poin...

  16. Endoscopic biopsy: Duodenal ulcer penetrating into liver

    Institute of Scientific and Technical Information of China (English)

    Baybora Kircali; Tülay Saricam; Aysegul Ozakyol; Eser Vardareli

    2005-01-01

    @@ TO THE EDITOR We have read with interest the recent report by E Kayacetin and S Kayacetin of Gastric ulcer penetrating to liver diagnosed by endoscopic biopsy[1] since we diagnosed the duodenal ulcer which penetrated into liver similarly. This is a rather unusual case because of the fifth case in the literature and responding to medical therapy.

  17. Penetrating Moral Education in English Teaching

    Institute of Scientific and Technical Information of China (English)

    汪中跃

    2015-01-01

    Middle school students’ moral education is not ful y implemented or even ignored in the present English teaching,so in this paper,the importance of penetrating the cultivation of middle school students’ moral education in English teaching is analyzed,the strategies of penetrating moral education are discussed and the significance is also pointed out.

  18. A study of the penetration of projectiles into marine sediments

    International Nuclear Information System (INIS)

    The work described in this document consists of three main parts: - Application, after having reviewed calculation methods and known codes, of a dynamic plasticity model based on the upper bound-method (with dissipated energy calculations by plastic deformations). The soil model used for this calculation is the Cambridge Clay Model. - Carrying out a programme of tests with instrumented small scale penetrators in centrifuge on a consolidated clay-target. The trials are done under 50 g, with projectiles, fired with an airgun at high impact velocity (50 m/s). The penetrators' instrumentation consists of either measuring acceleration, or tip force with strain gauges. - The mounting of a small instrumented penetrator for shallow water depth experimentations, with an accelerometer, and a local cell for tip resistance. A rapid electronic data acquisition system has been developed for these experimentations. The preliminary tests are done in a large tank filled with clay. The geotechnical characteristics of the clay are perfectly controlled. The tests performed under these conditions have shown the following observations: the rapid electronic data acquisition system works perfectly and could be used as a basic component for a deep water instrumentated penetrator; the results, obtained in these particular test conditions, are in a perfect agreement with the predictions of the model developed in the frame of this contract

  19. Mathematical Based Calculation of Drug Penetration Depth in Solid Tumors

    Directory of Open Access Journals (Sweden)

    Hamidreza Namazi

    2016-01-01

    Full Text Available Cancer is a class of diseases characterized by out-of-control cells’ growth which affect cells and make them damaged. Many treatment options for cancer exist. Chemotherapy as an important treatment option is the use of drugs to treat cancer. The anticancer drug travels to the tumor and then diffuses in it through capillaries. The diffusion of drugs in the solid tumor is limited by penetration depth which is different in case of different drugs and cancers. The computation of this depth is important as it helps physicians to investigate about treatment of infected tissue. Although many efforts have been made on studying and measuring drug penetration depth, less works have been done on computing this length from a mathematical point of view. In this paper, first we propose phase lagging model for diffusion of drug in the tumor. Then, using this model on one side and considering the classic diffusion on the other side, we compute the drug penetration depth in the solid tumor. This computed value of drug penetration depth is corroborated by comparison with the values measured by experiments.

  20. Diesel spray penetration studied by simultaneous Mie and shadowgraphy measurements

    Energy Technology Data Exchange (ETDEWEB)

    Bougie, B.; Tulej, M.; Beaud, P.; Knopp, G.; Radi, P.; Gerber, T.

    2003-03-01

    The influence of gas density and vaporization on penetration and dispersion of Diesel sprays were investigated in a High Temperature High Pressure Cell (HTDZ) within the range of typical Diesel engine injection parameters. The temporal evolution of cold and vaporizing Diesel sprays were probed simultaneously by laser elastic scattering- and shadowgraphy-techniques. The experimental results were used to verify recently proposed Diesel spray models. (author)

  1. Transconjunctival penetration of mitomycin C

    Directory of Open Access Journals (Sweden)

    Velpandian T

    2008-01-01

    Full Text Available Aims: The study was performed to estimate transconjunctival penetration of mitomycin C (MMC to Tenon′s tissue following application over the intact conjunctiva before routine trabeculectomy. Settings and Design: Institution-based case series. Materials and Methods: In 41 eyes of 41 patients, MMC (0.4 mg/ml for 3 min was applied over the intact conjunctiva before beginning trabeculectomy. Tenon′s capsule directly beneath the site of application was excised during trabeculectomy and was homogenized, centrifuged and MMC concentrations were analyzed using high-performance liquid chromatography (HPLC. Statistical Analysis Used: Statistical analysis was performed using stata0 8.0 version software (STATA Corporation, Houston, TX, USA. In this study, P -values less than 0.05 were considered as statistically significant. Results: The average weight of the sample of Tenon′s tissue excised was 5.51 ± 4.42 mg (range: 0.9-17.1 and the average estimated MMC concentration found to be present in Tenon′s tissue using HPLC was 18.67 ± 32.36 x 10−6 moles/kg of the tissue (range: 0.38-197.05 x 10−6 . In 36 of the 41 patients (87.80%, the MMC concentration reached above 2 x 10−6 moles/kg of the tissue concentration required to inhibit human conjunctival fibroblasts. Conclusions: Mitomycin C does permeate into the subconjunctival tissue after supraconjunctival application for 3 min. Application of MMC over the conjunctiva may be a useful alternative to subconjunctival or subscleral application during routine trabeculectomy and as an adjunct for failing blebs.

  2. Development of coring, consolidating, subterrene penetrators

    Energy Technology Data Exchange (ETDEWEB)

    Murphy, H.D.; Neudecker, J.W.; Cort, G.E.; Turner, W.C.; McFarland, R.D.; Griggs, J.E.

    1976-02-01

    Coring penetrators offer two advantages over full face-melting penetrators, i.e., formation of larger boreholes with no increase in power and the production of glass-lined, structurally undisturbed cores which can be recovered with conventional core-retrieval systems. These cores are of significant value in geological exploratory drilling programs. The initial design details and fabrication features of a 114-mm-diam coring penetrator are discussed; significant factors for design optimization are also presented. Results of laboratory testing are reported and compared with performance predictions, and an initial field trial is described.

  3. Sphere impact and penetration into wet sand

    KAUST Repository

    Marston, J. O.

    2012-08-07

    We present experimental results for the penetration of a solid sphere when released onto wet sand. We show, by measuring the final penetration depth, that the cohesion induced by the water can result in either a deeper or shallower penetration for a given release height compared to dry granular material. Thus the presence of water can either lubricate or stiffen the granular material. By assuming the shear rate is proportional to the impact velocity and using the depth-averaged stopping force in calculating the shear stress, we derive effective viscosities for the wet granular materials.

  4. 穿膜肽引导的体外表达转录因子蛋白Sox2进入红鳍东方鲀精巢细胞系%Intracellular delivery of the expressed fusion protein Sox2 with cell penetrating peptides to cultured Takifugu rupies spermary cells

    Institute of Scientific and Technical Information of China (English)

    杨秀霞; 侯雪宁; 徐彬; 郝萧; 姜国建; 樊廷俊

    2014-01-01

    为了探索适用于体外培养的鱼细胞外源基因转入方法,本研究通过构建红鳍东方鲀(Takifugu rubripes)转录因子Sox2的重组表达载体 pET32a(+)-Sox2-11R-6His,诱导表达并纯化得到了 C末端连接多聚精氨酸(11R)的重组蛋白Sox2-11R-6His,以其与红鳍东方鲀精巢细胞系细胞共孵育12 h后,光镜观察结合Western Blot检测发现重组蛋白进入细胞的效率与浓度呈剂量依赖关系且最佳孵育浓度为8μg/mL,当重组蛋白质量浓度达到10μg/mL 时,表现出明显的细胞毒性。对外源蛋白进行免疫荧光标记定位,发现重组蛋白分布于细胞质,部分进入到细胞核中。证明了穿膜肽11R可以有效运载转录因子重组蛋白至红鳍东方鲀的细胞系细胞中。本研究旨在将广泛应用于哺乳动物的细胞基因递送载体穿膜肽应用于鱼类细胞系细胞。%Research on exogenous gene function in cultured cell line cells is guaranteed by two biological processes, efficient gene transduction and errorless gene expression. Fish cells are not easy for exogenous gene transduction and expression depending on the gene delivery method used and the transgene promoter. Cell penetrating peptides (CPPs), such as the CPP of the human immunodeficiency virus type 1 TAT protein, poly-arginine (6–12 residues), and the CPP derived from flock house virus (FHV), are short cationic and/or amphipathic peptides that are able to transport various biological compounds such as peptides, proteins, and oligonucleotides into mammalian cells to modulate biological activities inside cells. Although it has been shown that CPPs can mediate efficient delivery into a wide variety of mam-malian cell types, the transduction of proteins to cultured fish cells is considered more challenging. In this work, the Sox2 gene from Takifugu rupies genomic DNA was cloned and subsequent expression was carried out in Escherichia coli in the form of recombinant proteins by introducing cell

  5. A Mass Loss Penetration Model to Investigate the Dynamic Response of a Projectile Penetrating Concrete considering Mass Abrasion

    Directory of Open Access Journals (Sweden)

    NianSong Zhang

    2015-01-01

    Full Text Available A study on the dynamic response of a projectile penetrating concrete is conducted. The evolutional process of projectile mass loss and the effect of mass loss on penetration resistance are investigated using theoretical methods. A projectile penetration model considering projectile mass loss is established in three stages, namely, cratering phase, mass loss penetration phase, and remainder rigid projectile penetration phase.

  6. Increased in vivo skin penetration of quantum dots with UVR and in vitro quantum dot cytotoxicity

    Science.gov (United States)

    Mortensen, Luke; Zheng, Hong; Faulknor, Renea; De Benedetto, Anna; Beck, Lisa; DeLouise, Lisa A.

    2009-02-01

    The growing presence of quantum dots (QD) in a variety of biological, medical, and electronics applications means an increased risk of human exposure in manufacturing, research, and consumer use. However, very few studies have investigated the susceptibility of skin to penetration of QD - the most common exposure route- and the results of those that exist are conflicting. This suggests that a technique allowing determination of skin barrier status and prediction of skin permeability to QD would be of crucial interest as recent findings have provided evidence of in vitro cytotoxicity and long-term in vivo retention in the body for most QD surface chemistries. Our research focuses on barrier status of the skin (intact and with ultraviolet radiation induced barrier defect) and its impact on QD skin penetration. These model studies are particularly relevant to the common application condition of NP containing sunscreen and SPF cosmetics to UV exposed skin. Herein we present our initial efforts to develop an in vivo model of nanoparticle skin penetration using the SKH-1 hairless mouse with transepidermal water loss (TEWL) to evaluate skin barrier status and determine its ability to predict QD penetration. Our results show that ultraviolet radiation increases both TEWL and skin penetration of QD. Additionally, we demonstrate cytotoxic potential of QD to skin cells using a metastatic melanoma cell line. Our research suggests future work in specific targeting of nanoparticles, to prevent or enhance penetration. This knowledge will be used to develop powerful therapeutic agents, decreased penetration cosmetic nanoparticles, and precise skin cancer imaging modalities.

  7. Penetration testing protecting networks and systems

    CERN Document Server

    Henry, Kevin M

    2012-01-01

    This book is a preparation guide for the CPTE examination, yet is also a general reference for experienced penetration testers, ethical hackers, auditors, security personnel and anyone else involved in the security of an organization's computer systems.

  8. An Observational Investigation of Penetrative Convection

    DEFF Research Database (Denmark)

    Jensen, Niels Otto; Lenschow, D. H.

    1978-01-01

    Data taken during the Air Mass Transformation Experiment (AMTEX) by the NCAR Electra aircraft have proven useful for investigating the structure of thermals penetrating into the turbulent inversion layer which caps the convective mixed layer. Variances, covariances, spectra and cospectra...

  9. Temporary fire sealing of penetrations on TFTR

    Energy Technology Data Exchange (ETDEWEB)

    Hondorp, H.L.

    1981-02-01

    The radiation shielding provided for TFTR for D-D and D-T operation will be penetrated by numerous electrical and mechanical services. Eventually, these penetrations will have to be sealed to provide the required fire resistance, tritium sealability, pressure integrity and radiation attenuation. For the initial hydrogen operation, however, fire sealing of the penetrations in the walls and floor is the primary concern. This report provides a discussion of the required and desirable properties of a temporary seal which can be used to seal these penetrations for the hydrogen operation and then subsequently be removed and replaced as required for the D-D and D-T operations. Several candidate designs are discussed and evaluated and recommendations are made for specific applications.

  10. Prediction of weld penetration extent in GTAW

    Institute of Scientific and Technical Information of China (English)

    Zhao Ming; Zhai Lei

    2008-01-01

    The governing equations of surface-deformation in fully-penetrated GTA W weld pool are derived. The solution scheme of the Lagrange multiplier, which makes the top sutface-deformation equation be coupled with that for the bottomsurface of a single weld pool, is given out. Through numerical analysis of weld pool surface-deformation in GTA W,it is found that there is clear correlation between the penetration extent and the Lagrange multiplier, in addition to the relationship between the penetration extent and the maximum depression and hump as well as the depressed volume of the top surface of weld pool. It lays good foundation for top-side sensing based weld penetration control.

  11. Laser weld penetration estimation using temperature measurements

    Energy Technology Data Exchange (ETDEWEB)

    Lankalapalli, K.N.; Tu, J.F. [Purdue Univ., West Lafayette, IN (United States). School of Industrial Engineering; Leong, K.H. [Argonne National Lab., IL (United States); Gartner, M. [Ford Motor Co., Livonia, MI (United States)

    1997-10-01

    Penetration depth is an important factor critical to the quality of a laser weld. This paper examines the feasibility of using temperature measurements on the bottom surface of the work-piece to estimate weld penetration. A three-dimensional analytical model relating penetration depth, weld bead width and welding speed to temperature distribution at the bottom surface of the workpiece is developed. Temperatures on the bottom surface of the workpiece are measured using infrared thermocouples located behind the laser beam. Experimental results from bead-on-plate welds on low carbon steel plates of varying thickness at different levels of laser power and speeds validate the model and show that the temperature on the bottom surface is a sensitive indicator of penetration depth. The proposed model is computationally efficient and is suitable for on-line process monitoring application.

  12. Kali Linux wireless penetration testing beginner's guide

    CERN Document Server

    Ramachandran, Vivek

    2015-01-01

    If you are a security professional, pentester, or anyone interested in getting to grips with wireless penetration testing, this is the book for you. Some familiarity with Kali Linux and wireless concepts is beneficial.

  13. Temporary fire sealing of penetrations on TFTR

    International Nuclear Information System (INIS)

    The radiation shielding provided for TFTR for D-D and D-T operation will be penetrated by numerous electrical and mechanical services. Eventually, these penetrations will have to be sealed to provide the required fire resistance, tritium sealability, pressure integrity and radiation attenuation. For the initial hydrogen operation, however, fire sealing of the penetrations in the walls and floor is the primary concern. This report provides a discussion of the required and desirable properties of a temporary seal which can be used to seal these penetrations for the hydrogen operation and then subsequently be removed and replaced as required for the D-D and D-T operations. Several candidate designs are discussed and evaluated and recommendations are made for specific applications

  14. Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.

    Directory of Open Access Journals (Sweden)

    Steven J Husson

    Full Text Available Essentially any behavior in simple and complex animals depends on neuronal network function. Currently, the best-defined system to study neuronal circuits is the nematode Caenorhabditis elegans, as the connectivity of its 302 neurons is exactly known. Individual neurons can be activated by photostimulation of Channelrhodopsin-2 (ChR2 using blue light, allowing to directly probe the importance of a particular neuron for the respective behavioral output of the network under study. In analogy, other excitable cells can be inhibited by expressing Halorhodopsin from Natronomonas pharaonis (NpHR and subsequent illumination with yellow light. However, inhibiting C. elegans neurons using NpHR is difficult. Recently, proton pumps from various sources were established as valuable alternative hyperpolarizers. Here we show that archaerhodopsin-3 (Arch from Halorubrum sodomense and a proton pump from the fungus Leptosphaeria maculans (Mac can be utilized to effectively inhibit excitable cells in C. elegans. Arch is the most powerful hyperpolarizer when illuminated with yellow or green light while the action spectrum of Mac is more blue-shifted, as analyzed by light-evoked behaviors and electrophysiology. This allows these tools to be combined in various ways with ChR2 to analyze different subsets of neurons within a circuit. We exemplify this by means of the polymodal aversive sensory ASH neurons, and the downstream command interneurons to which ASH neurons signal to trigger a reversal followed by a directional turn. Photostimulating ASH and subsequently inhibiting command interneurons using two-color illumination of different body segments, allows investigating temporal aspects of signaling downstream of ASH.

  15. Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans.

    Science.gov (United States)

    Husson, Steven J; Liewald, Jana F; Schultheis, Christian; Stirman, Jeffrey N; Lu, Hang; Gottschalk, Alexander

    2012-01-01

    Essentially any behavior in simple and complex animals depends on neuronal network function. Currently, the best-defined system to study neuronal circuits is the nematode Caenorhabditis elegans, as the connectivity of its 302 neurons is exactly known. Individual neurons can be activated by photostimulation of Channelrhodopsin-2 (ChR2) using blue light, allowing to directly probe the importance of a particular neuron for the respective behavioral output of the network under study. In analogy, other excitable cells can be inhibited by expressing Halorhodopsin from Natronomonas pharaonis (NpHR) and subsequent illumination with yellow light. However, inhibiting C. elegans neurons using NpHR is difficult. Recently, proton pumps from various sources were established as valuable alternative hyperpolarizers. Here we show that archaerhodopsin-3 (Arch) from Halorubrum sodomense and a proton pump from the fungus Leptosphaeria maculans (Mac) can be utilized to effectively inhibit excitable cells in C. elegans. Arch is the most powerful hyperpolarizer when illuminated with yellow or green light while the action spectrum of Mac is more blue-shifted, as analyzed by light-evoked behaviors and electrophysiology. This allows these tools to be combined in various ways with ChR2 to analyze different subsets of neurons within a circuit. We exemplify this by means of the polymodal aversive sensory ASH neurons, and the downstream command interneurons to which ASH neurons signal to trigger a reversal followed by a directional turn. Photostimulating ASH and subsequently inhibiting command interneurons using two-color illumination of different body segments, allows investigating temporal aspects of signaling downstream of ASH. PMID:22815873

  16. Effect of penetrants on the aramid Nomex

    Indian Academy of Sciences (India)

    Anjana Jain; Kalyani Vijayan

    2000-06-01

    The response of Nomex sheets to some penetrants has been analyzed. The process of moisture uptake depends on the relative humidity (RH) of the ambient atmosphere and the initial characteristics of the polymer. In the case of common laboratory solvents, the uptake by the polymer shows an inverse dependence on the molar volume of the former. Although the calendered and the uncalendered Nomex sheets exhibit an overall similarity in their response to various penetrants, quantitative comparison reveals distinct differences.

  17. Penetration of amalgam constituents into dentine

    OpenAIRE

    Scholtanus, J.D.; Özcan, M; Huysmans, M.C.D.N.J.M.

    2009-01-01

    OBJECTIVES: Amalgam restorations are replaced by adhesively placed composite resin restorations at an increasing rate. After the removal of amalgam dentine often shows marked dark discoloration that is attributed to the penetration of corrosion products from overlying amalgams. It is questioned whether penetration of metals into dentine affects the dentine as a substrate for adhesive procedures. This study has been performed to clarify the origin of dark discoloration of dentine by metals fro...

  18. Low Force Penetration of Icy Regolith

    Science.gov (United States)

    Mantovani, J. G.; Galloway, G. M.; Zacny, K.

    2016-01-01

    A percussive cone penetrometer measures the strength of granular material by using percussion to deliver mechanical energy into the material. A percussive cone penetrometer was used in this study to penetrate a regolith ice mixture by breaking up ice and decompacting the regolith. As compared to a static cone penetrometer, percussion allows low reaction forces to push a penetrometer probe tip more easily into dry regolith in a low gravity environment from a planetary surface rover or a landed spacecraft. A percussive cone penetrates icy regolith at ice concentrations that a static cone cannot penetrate. In this study, the percussive penetrator was able to penetrate material under 65 N of down-force which could not be penetrated using a static cone under full body weight. This paper discusses using a percussive cone penetrometer to discern changes in the concentration of water-ice in a mixture of lunar regolith simulant and ice to a depth of one meter. The rate of penetration was found to be a function of the ice content and was not significantly affected by the down-force. The test results demonstrate that this method may be ideal for a small platform in a reduced gravity environment. However, there are some cases where the system may not be able to penetrate the icy regolith, and there is some risk of the probe tip becoming stuck so that it cannot be retracted. It is also shown that a percussive cone penetrometer could be used to prospect for water ice in regolith at concentrations as high as 8 by weight.

  19. DRUG VEHICLE BASED APPROACHES OF PENETRATION ENHANCEMENT

    OpenAIRE

    Swaroop R Lahoti; MOHD. HASSAN G DEHGHAN; Dinesh L Dhamecha; AMIT A RATHI; MARIA SAIFEE

    2009-01-01

    Transdermal delivery of drugs through the skin to the systemic circulation provides a convenient route of administration for a variety of clinical indications. For transdermal delivery of drugs, stratum corneum is the main barrier layer for permeation of drug. So to circumvent the stratum corneum and to increase the flux through skin membrane, different approaches of penetration enhancement are used. Many reviews had described regarding the chemical penetration enhancement but vehicle based e...

  20. Influence of massage and occlusion on the ex vivo skin penetration of rigid liposomes and invasomes

    DEFF Research Database (Denmark)

    Trauer, S.; Richter, H.; Kuntsche, Judith;

    2014-01-01

    Liposomes are frequently described as drug delivery systems for dermal and transdermal applications. Recently, it has been shown that particulate substances penetrate effectively into hair follicles and that the follicular penetration depth can be increased by massaging the skin, which simulates...... the in vivo movement of hairs in the hair follicles. In the present study, massage was applied to skin mounted to Franz diffusion cells. By means of confocal laser scanning microscopy, the influence of massage and occlusion on the follicular penetration depths of rigid and flexible liposomes loaded...

  1. Simulation of Mine Electrical Penetration Technology

    Institute of Scientific and Technical Information of China (English)

    LIU Zhi-xin; YUE Jian-hua; LIU Shu-cai

    2007-01-01

    Based on the principle of electrical penetration, the reflection characteristics of collapse columns at different locations of a working face is numerically simulated by using a 3D finite element method. The data collected by the electrical penetration is processed and interpreted using "tunnel penetration" which is similar to radio wave penetration. Reflection characteristics of collapse columns at different locations below floors of coal seams are analyzed, providing a new paradigm and a theoretical foundation for processing and interpreting electrical penetration data. The tomography analysis is made based on data simulation and calculation results and alltransmitting-receiving points are analyzed for their corresponding maximum attenuation values and maximum absorption coefficients. On the basis of this, a new method for precisely interpreting the spatial positions of geological anomalous bodies is suggested. The simulation shows that 1) the detection result of both roof and floors of the working face by electrical penetration is a volumetric effect and 2) there exists a corresponding relation between the detection depth and the working face width, with the optimal detection depth within 40% of the working face width.

  2. Giant gastric ulcer penetrating into the pancreas.

    Science.gov (United States)

    Fujihara, Shintaro; Mori, Hirohito; Nishiyama, Noriko; Kobara, Hideki; Masaki, Tsutomu

    2012-09-01

    A 57-year-old man developed haematemesis and was referred to our institution. His discomfort had begun 3 weeks earlier and localised to the upper abdomen. Abdominal CT showed a defect of gastric mucosa and gastric wall thickening. Oesophagogastroduodenoscopy showed that he had an active gastric ulcer, 40 mm in diameter, on the lesser curvature in the upper third of the corpus and the presence of a pancreatic body at the ulcer base, penetration to the pancreas. Open gastrectomy was performed with a preoperative diagnosis of localised peritonitis caused by penetration of the stomach into the pancreas. Histopathology of resected specimens showed a benign peptic ulcer, 40×40 mm in size, was found on the lesser curvature in the antrum, and this had penetrated through the pancreas. The most serious complications of PUD include haemorrhage, perforation, penetration, and gastric outlet obstruction. Approximately 7% of patients experience perforation, which occurs when an ulcer erodes through the wall and leaks air and digestive contents into the peritoneal cavity. Antral and duodenal ulcers can penetrate into the pancreas. We report a case of gastric ulcer penetrating into the pancreas.

  3. Complementary optical and nuclear imaging of caspase-3 activity using combined activatable and radio-labeled multimodality molecular probe

    Science.gov (United States)

    Lee, Hyeran; Akers, Walter J.; Cheney, Philip P.; Edwards, W. Barry; Liang, Kexian; Culver, Joseph P.; Achilefu, Samuel

    2009-07-01

    Based on the capability of modulating fluorescence intensity by specific molecular events, we report a new multimodal optical-nuclear molecular probe with complementary reporting strategies. The molecular probe (LS498) consists of tetraazacyclododecanetetraacetic acid (DOTA) for chelating a radionuclide, a near-infrared fluorescent dye, and an efficient quencher dye. The two dyes are separated by a cleavable peptide substrate for caspase-3, a diagnostic enzyme that is upregulated in dying cells. LS498 is radiolabeled with 64Cu, a radionuclide used in positron emission tomography. In the native form, LS498 fluorescence is quenched until caspase-3 cleavage of the peptide substrate. Enzyme kinetics assay shows that LS498 is readily cleaved by caspase-3, with excellent enzyme kinetic parameters kcat and KM of 0.55+/-0.01 s-1 and 1.12+/-0.06 μM, respectively. In mice, the initial fluorescence of LS498 is ten-fold less than control. Using radiolabeled 64Cu-LS498 in a controlled and localized in-vivo model of caspase-3 activation, a time-dependent five-fold NIR fluorescence enhancement is observed, but radioactivity remains identical in caspase-3 positive and negative controls. These results demonstrate the feasibility of using radionuclide imaging for localizing and quantifying the distribution of molecular probes and optical imaging for reporting the functional status of diagnostic enzymes.

  4. In vitro penetration of bleaching agents into the pulp chamber

    DEFF Research Database (Denmark)

    Benetti, Ana Raquel; Valera, M C; Mancini, M N G;

    2004-01-01

    To investigate pulp chamber penetration of bleaching agents in teeth following restorative procedures.......To investigate pulp chamber penetration of bleaching agents in teeth following restorative procedures....

  5. Rapid penetration into granular media visualizing the fundamental physics of rapid earth penetration

    CERN Document Server

    Iskander, Magued

    2015-01-01

    Rapid Penetration into Granular Media: Visualizing the Fundamental Physics of Rapid Earth Penetration introduces readers to the variety of methods and techniques used to visualize, observe, and model the rapid penetration of natural and man-made projectiles into earth materials. It provides seasoned practitioners with a standard reference that showcases the topic's most recent developments in research and application. The text compiles the findings of new research developments on the subject, outlines the fundamental physics of rapid penetration into granular media, and assembles a com

  6. Heat flux in a penetrative convection experiment in water

    Science.gov (United States)

    Corre, Yoann; Alboussière, Thierry; Labrosse, Stéphane; Odier, Philippe; Joubaud, Sylvain

    2015-11-01

    In geophysical systems, stably stratified fluids adjacent to convective regions often experience thermal plume penetration from the latter. This penetrative convection occurs in stellar interiors between radiative and convective regions and possibly in liquid envelopes of planets, such as the Earth's core. We are interested in quantifying this process experimentally as it could play a crucial role in their dynamics. A volume of water initially at ambiant temperature is cooled from below at 0 degrees Celsius. Due to the maximum density of water near 4 degrees, a convective region develops and grows below a purely conductive region. A laser sheet crosses the experimental cell, lightening both neutrally buoyant particles and a thermosensitive fluorescent dye, which allows to monitor the velocity and temperature fields respectively (PIV-LIF technique), giving access to the local convective and conductive heat flux. The apparatus is placed on a rotating table to inspect the effect of the Coriolis force on the interfacial region. We find that increasing the rotation rate deepens the penetration of vortices into the conductive region, thus changing the structure of the interfacial layer and possibly eroding the stable region.

  7. Overcoming the nail barrier: A systematic investigation of ungual chemical penetration enhancement.

    Science.gov (United States)

    Brown, M B; Khengar, R H; Turner, R B; Forbes, B; Traynor, M J; Evans, C R G; Jones, S A

    2009-03-31

    This study investigated the in vitro nail permeability of penetrants of varying lipophilicity-caffeine (CF, logP -0.07), methylparaben (MP, logP 1.96) and terbinafine (TBF, logP 3.3) and the effect of 2 novel penetration enhancers (PEs), thioglycolic acid (TA) and urea hydrogen peroxide (urea H(2)O(2)) on their permeation. Studies were conducted using full thickness human nail clippings and ChubTur((R)) diffusion cells and penetrants were applied as saturated solutions. The rank order of steady-state penetrant flux through nails without PE application (MP>CF>TBF) suggested a greater sensitivity to penetrant molecular weight rather than logP. TA increased the flux of CF and MP approximately 4- and approximately 2-fold, respectively, whilst urea H(2)O(2) proved ineffective at enhancing permeability. The sequential application of TA followed by urea H(2)O(2) increased TBF and CF flux ( approximately 19- and approximately 4-fold, respectively) but reversing the application order of the PEs was only mildly effective at increasing just MP flux ( approximately 2-fold). Both nail PEs are likely to function via disruption of keratin disulphide bonds and the associated formation of pores that provide more 'open' drug transport channels. Effects of the PEs were penetrant specific, but the use of a reducing agent (TA) followed by an oxidising agent (urea H(2)O(2)) dramatically improved human nail penetration. PMID:19071202

  8. Overcoming the nail barrier: A systematic investigation of ungual chemical penetration enhancement.

    Science.gov (United States)

    Brown, M B; Khengar, R H; Turner, R B; Forbes, B; Traynor, M J; Evans, C R G; Jones, S A

    2009-03-31

    This study investigated the in vitro nail permeability of penetrants of varying lipophilicity-caffeine (CF, logP -0.07), methylparaben (MP, logP 1.96) and terbinafine (TBF, logP 3.3) and the effect of 2 novel penetration enhancers (PEs), thioglycolic acid (TA) and urea hydrogen peroxide (urea H(2)O(2)) on their permeation. Studies were conducted using full thickness human nail clippings and ChubTur((R)) diffusion cells and penetrants were applied as saturated solutions. The rank order of steady-state penetrant flux through nails without PE application (MP>CF>TBF) suggested a greater sensitivity to penetrant molecular weight rather than logP. TA increased the flux of CF and MP approximately 4- and approximately 2-fold, respectively, whilst urea H(2)O(2) proved ineffective at enhancing permeability. The sequential application of TA followed by urea H(2)O(2) increased TBF and CF flux ( approximately 19- and approximately 4-fold, respectively) but reversing the application order of the PEs was only mildly effective at increasing just MP flux ( approximately 2-fold). Both nail PEs are likely to function via disruption of keratin disulphide bonds and the associated formation of pores that provide more 'open' drug transport channels. Effects of the PEs were penetrant specific, but the use of a reducing agent (TA) followed by an oxidising agent (urea H(2)O(2)) dramatically improved human nail penetration.

  9. Opportunities for high wind energy penetration

    DEFF Research Database (Denmark)

    Tande, J.O.; Hansen, J.C.

    1997-01-01

    Wind power is today a mature technology, which at windy locations, is economically competitive to conventional power generation technologies. This and growing global environmental concerns have led governments to encourage and plan for wind energy development, a typical aim being 10% of electricity...... consumption. The successful operation of the three major power systems of Cape Verde, with a total wind energy penetration of about 15% since December 1994, demonstrates that power systems can be operated with high penetration of wind energy by adding simple control and monitoring systems only. Thorough...... analyses conclude that expanding to even above 15% wind energy penetration in the Cape Verde power systems is economical. Worldwide, numerous locations with favorable wind conditions and power systems similar to the Capeverdean provide good opportunities for installing wind farms and achieving high wind...

  10. WAPTT - Web Application Penetration Testing Tool

    Directory of Open Access Journals (Sweden)

    DURIC, Z.

    2014-02-01

    Full Text Available Web applications vulnerabilities allow attackers to perform malicious actions that range from gaining unauthorized account access to obtaining sensitive data. The number of reported web application vulnerabilities in last decade is increasing dramatically. The most of vulnerabilities result from improper input validation and sanitization. The most important of these vulnerabilities based on improper input validation and sanitization are: SQL injection (SQLI, Cross-Site Scripting (XSS and Buffer Overflow (BOF. In order to address these vulnerabilities we designed and developed the WAPTT (Web Application Penetration Testing Tool tool - web application penetration testing tool. Unlike other web application penetration testing tools, this tool is modular, and can be easily extended by end-user. In order to improve efficiency of SQLI vulnerability detection, WAPTT uses an efficient algorithm for page similarity detection. The proposed tool showed promising results as compared to six well-known web application scanners in detecting various web application vulnerabilities.

  11. Penetration of Projectiles in Composite Laminates

    Directory of Open Access Journals (Sweden)

    B. P. Patel

    2004-04-01

    Full Text Available This paper deals with the prediction of the penetration phenomenon of a cylindro-conical impactor on the ke;labepoxy-laminated composites using C eight-nded serendipity q&drilateral finite elementbased on first-order shear deformation theory (FSDT. Local as well as global deformations during impact is considered in the evaluation of indentation, penetration, and perforation phases. Local strainsduring impact have been evaluated using the hypothesis made from the available experimental observations of bulging during penetration. A detailed parametric study, considering various projectilesand target plate variables, has been carried out to find their effect on the response of the plate, and ballistic parameters, such as ballistic limit and absorbed energy.

  12. Proniosomes for Penetration Enhancement in Transdermal System

    Directory of Open Access Journals (Sweden)

    Samita Singla

    2012-06-01

    Full Text Available Over the last few years an inclusive research has been done over provesicular approach for transdermal drug delivery. Skin has a very tough diffusion barrier inhibiting penetration of drug moiety which is rate limiting barrier for penetration of drugs. There are several approaches that deal with penetration enhancement across the skin. Vesicular and provesicular systems are promising amongst them. Vesicular systems including (niosomes, ethosomes, transfersomes and liposomes are promising systems to cross this permeation barrier. But their major drawback is their unstability, which can be overcome by using provesicular approaches like proniosomes, protransfersomes and proliposomes. Provesicular approaches have been proposed to enhance the stability of vesicles. These approaches may overcome skin barrier properties and enhance percutaneous absorption. In these systems both type of drugs, hydrophilic and lipophillic drugs can be incorporated. The focus of this review is to bring together different aspects related to mechanism of skin permeation of provesicular system, proniosomes preparation and its applications as a carrier system

  13. Diagnosis and treatment of penetrating anorectal wounds

    Institute of Scientific and Technical Information of China (English)

    LIU Xin-sheng; HUI Xi-zeng; ZHANG Yang-de; LI Kun

    2006-01-01

    Objective:To research the diagnosis and effective treatment of penetrating anorectal wounds.Methods: Retrospective analysis was done in 16 cases of penetrating anorectal wounds from 1985 to 2004.Debridement and suture of anorectal and vesical wounds,effective diversion of fecal and urinary stream and sufficient presacral drainage were performed in all cases.Results: All the 16 cases were cured. Among them, 2cases with infection in presacral space were cured by sufficient drainage after operation, one case was cured by secondary repair after anal sphincter was repaired unsuccessfully and one case with rectovesical fistula was cured with conservative treatment. None of them suffered from complications such as anal stenosis, dysuria or importence etc.Conclusions: For penetrating anorectal wound, to master early recognition of concomitant injures, to select appropriate surgical intervention and to strengthen perioperative treatment are the keys to improve the curative effects.

  14. Coagulation factor Va binds to human umbilical vein endothelial cells and accelerates protein C activation.

    OpenAIRE

    Maruyama, I.; Salem, H H; Majerus, P W

    1984-01-01

    In vitro the rate of protein C activation by thrombin is significantly accelerated by two distinct cofactors (a) the endothelial cell surface protein, thrombomodulin, and (b) human coagulation Factor Va. We have recently reported that the activity of Factor Va is contained in the 78,000-D light chain. In this study we have investigated the effects of Factor Va and its light chain on the activation of protein C in the presence of cultured endothelial cells. Thrombin-catalyzed protein C activat...

  15. Development Of The Nuclear Optical Penetration

    Science.gov (United States)

    Inoue, K.; Koike, K.; Imada, Y.

    1984-10-01

    We have developed the nuclear optical penetration to be incorporated in the wall penetration of the shell to introduce a data transmission system using optical fibers into a nuclear power plant with a pressurized water reactor. Radiation-induced coloration in optical glass seriously affects transmission characteristics of optical fibers, whereas it has been revealed that the pure-silica core optical fiber without any dopant in the core has wide applicability in radiation fields thanks to its very low radiation-induced attenuation. The wall penetration of the shell should have airtightness and resistivity to heat, vibration, and pressure, let alone radiation, excellent enough to be invariable in data transmission efficiency even when subjected to severe environmental tests. The sealing modules of this newly developed nuclear optical penetration are hermetically sealed. The gap between the optical fiber rod (100 pm in core diameter and 5 mm in rod diameter) and stainless steel tube is sealed with lamingted glass layer. As the result of He gas leakage test, high airtightness of less than 10 cc/sec was achieved. No thermal deformation of the core was caused by sealing with laminated glass layer, nor was observed transmission loss. Then the sealiing modules were subjected to the irradiation test using 60 Co gamma ray exposure of 2 x 10 rads. Though silica glass layer supporting the fiber rod and sealing glass portion turned blackish purple, transparency of the fiber was not affected. Only less than 0.5 dB of connecting loss was observed at the connecting point with the optical fiber cable. The sealing modules were also found to have resistivity to vibration and pressure as excellent as that of existing nuclear electric penetrations. We expect the nuclear optical fiber penetration will be much effective in improving reliability of data transmission systems using optical fibers in radiation fields.

  16. Ex vivo penetration of low-level laser light through equine skin and flexor tendons.

    Science.gov (United States)

    Duesterdieck-Zellmer, Katja F; Larson, Maureen K; Plant, Thomas K; Sundholm-Tepper, Andrea; Payton, Mark E

    2016-09-01

    OBJECTIVE To measure penetration efficiencies of low-level laser light energy through equine skin and to determine the fraction of laser energy absorbed by equine digital flexor tendons (superficial [SDFT] and deep [DDFT]). SAMPLE Samples of skin, SDFTs, and DDFTs from 1 metacarpal area of each of 19 equine cadavers. PROCEDURES A therapeutic laser with wavelength capabilities of 800 and 970 nm was used. The percentage of energy penetration for each wavelength was determined through skin before and after clipping and then shaving of hair, through shaved skin over SDFTs, and through shaved skin, SDFTs, and DDFTs (positioned in anatomically correct orientation). Influence of hair color; skin preparation, color, and thickness; and wavelength on energy penetration were assessed. RESULTS For haired skin, energy penetration was greatest for light-colored hair and least for dark-colored hair. Clipping or shaving of skin improved energy penetration. Light-colored skin allowed greatest energy penetration, followed by medium-colored skin and dark-colored skin. Greatest penetration of light-colored skin occurred with the 800-nm wavelength, whereas greatest penetration of medium- and dark-colored skin occurred with the 970-nm wavelength. As skin thickness increased, energy penetration of samples decreased. Only 1% to 20% and 0.1% to 4% of energy were absorbed by SDFTs and DDFTs, respectively, depending on skin color, skin thickness, and applied wavelength. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that most laser energy directed through equine skin was absorbed or scattered by the skin. To achieve delivery of energy doses known to positively affect cells in vitro to equine SDFTs and DDFTs, skin preparation, color, and thickness and applied wavelength must be considered. PMID:27580111

  17. Penetration profile of microspheres in follicular targeting of terminal hair follicles.

    Science.gov (United States)

    Toll, R; Jacobi, U; Richter, H; Lademann, J; Schaefer, H; Blume-Peytavi, U

    2004-07-01

    The transfollicular administration of pharmacologically active molecules is of current therapeutic interest, mainly with regard to delivery to specific sites of the hair follicle (HF) and the reduction of hepatic metabolism and systemic toxicity. HF are privileged pathways for specific molecules depending on formulations, which enter faster into these shunts than through the stratum corneum. The aim was to optimize the delivery of fluorescent microspheres into the HF, thereby, developing a standardized protocol for follicular targeting with microspheres. The number of HF showing penetration, as well as the depth of penetration, was determined. Freshly excised skin samples with terminal HF were divided into groups, with or without prior treatment with cyanoacrylate skin surface stripping-technique (CSSS). Thereafter microspheres at a size of 0.75-6.0 microm were applied according to the developed standardized protocol. Skin biopsies were obtained, shock-frozen, and sectioned in 5 microm slices. We demonstrated a selective penetration route of the microspheres into the HF. Optimal microsphere size proved to be approximately 1.5 microm, with a 55% rate of all HF, and with a maximum penetration depth of >2300 microm. Without previous CSSS treatment of the skin, the transfollicular microsphere penetration was below 27% with a maximum penetration depth of 1000 microm. Thus, the basis for follicular targeting of essential structures containing stem cells for keratinocytes, melanocytes, and mast cells has been laid.

  18. FREEFALL: A seabed penetrator flight code

    International Nuclear Information System (INIS)

    This report presents a one-dimensional model and computer program for predicting the motion of seabed penetrators. The program calculates the acceleration, velocity, and depth of a penetrator as a function of time from the moment of launch until the vehicle comes to rest in the sediment. The code is written in Pascal language for use on a small personal computer. Results are presented as printed tables and graphs. A comparison with experimental data is given which indicates that the accuracy of the code is perhaps as good as current techniques for measuring vehicle performance. 31 refs., 12 figs., 5 tabs

  19. Depleted Uranium Penetrators : Hazards and Safety

    OpenAIRE

    Rao, S S; T. Balakrishna Bhat

    1997-01-01

    The depleted uranium (DU) alloy is a state-of-the-art material for kinetic energy penetrators due to its superior ballistic performance. Several countries use DU penetrators in their main battle tanks. There is no gamma radiation hazard to the crew members from stowage of DO rounds. Open air firing can result in environmental contamination and associated hazards due to airborne particles containing essentially U/sub 3/0/sub 8/ and UO/sub 2/. Inhalation of polluted air only through resp...

  20. Hybrid treatment of penetrating aortic ulcer

    Energy Technology Data Exchange (ETDEWEB)

    Lara, Juan Antonio Herrero; Martins-Romeo, Daniela de Araujo; Escudero, Carlos Caparros; Falcon, Maria del Carmen Prieto; Batista, Vinicius Bianchi, E-mail: jaherrero5@hotmail.com [Unidade de Gestao Clinica (UGC) de Diagnostico por Imagem - Hosppital Universitario Virgen Macarena, Sevilha (Spain); Vazquez, Rosa Maria Lepe [Unit of Radiodiagnosis - Hospital Nuestra Senora de la Merced, Osuna, Sevilha (Spain)

    2015-05-15

    Penetrating atherosclerotic aortic ulcer is a rare entity with poor prognosis in the setting of acute aortic syndrome. In the literature, cases like the present one, located in the aortic arch, starting with chest pain and evolving with dysphonia, are even rarer. The present report emphasizes the role played by computed tomography in the diagnosis of penetrating atherosclerotic ulcer as well as in the differentiation of this condition from other acute aortic syndromes. Additionally, the authors describe a new therapeutic approach represented by a hybrid endovascular surgical procedure for treatment of the disease. (author)

  1. METHOD ON ESTIMATION OF DRUG'S PENETRATED PARAMETERS

    Institute of Scientific and Technical Information of China (English)

    刘宇红; 曾衍钧; 许景锋; 张梅

    2004-01-01

    Transdermal drug delivery system (TDDS) is a new method for drug delivery. The analysis of plenty of experiments in vitro can lead to a suitable mathematical model for the description of the process of the drug's penetration through the skin, together with the important parameters that are related to the characters of the drugs.After the research work of the experiments data,a suitable nonlinear regression model was selected. Using this model, the most important parameter-penetrated coefficient of 20 drugs was computed.In the result one can find, this work supports the theory that the skin can be regarded as singular membrane.

  2. FREEFALL: A seabed penetrator flight code

    Energy Technology Data Exchange (ETDEWEB)

    Hickerson, J.

    1988-01-01

    This report presents a one-dimensional model and computer program for predicting the motion of seabed penetrators. The program calculates the acceleration, velocity, and depth of a penetrator as a function of time from the moment of launch until the vehicle comes to rest in the sediment. The code is written in Pascal language for use on a small personal computer. Results are presented as printed tables and graphs. A comparison with experimental data is given which indicates that the accuracy of the code is perhaps as good as current techniques for measuring vehicle performance. 31 refs., 12 figs., 5 tabs.

  3. Effects of Penetrative Convection on Solar Dynamo

    CERN Document Server

    Masada, Youhei; Kageyama, Akira

    2013-01-01

    Spherical solar dynamo simulations are performed. Self-consistent, fully compressible magnetohydrodynamic system with a stably stratified layer below the convection zone are numerically solved with a newly developed simulation code based on the Yin-Yang grid. The effects of penetrative convection are studied by comparing two models with and without the stable layer. A solar-like differential rotation profile is established when the penetrative convection is taken into account without assuming any forcing. A large-scale magnetic field is also spontaneously organized in the underlying stable layer. The embedded field has a dipole symmetry about the equator and it shows polarity reversals in time.

  4. Enhancement of skin penetration of nonsteroidal anti-inflammatory drugs from extemporaneously compounded topical-gel formulations.

    Science.gov (United States)

    Goodwin, D A; Fuhrman, L C

    1999-01-01

    Ketoprofen and ibuprofen topical gels were compounded with decyl methyl sulfoxide and the terpenes d-limonene, (-)-menthone, terpinen-4-ol, and a-terpineol as penetration enhancers. Transdermal penetration profiles for both ketoprofen and ibuprofen were determined using full-thickness human skin, modified Franz diffusion cells and an isotonic (pH7.4) phosphate buffer solution. Human skin was used in these experiments to approximate the therapeutic use of these gels. Ibuprofen was found to have superior transdermal kinetics when compared to ketoprofen. Ibuprofen is a smaller and more lipophilic molecule than ketoprofen, which gives it better penetration properties. All enhancers tested significantly increased the penetration (except (-)-menthone) and skin retention (except terpinen-4-ol) of ketoprofen. None of the enhancers tested significantly increased the penetration or retention of ibuprofen. Despite the lack of enhancer activity, ibuprofen still demonstrated higher skin penetration and retention than enhanced delivery of ketoproen. The results of these studies suggest that the addition of penetration enhancers can significantly increase the amount of ketoprofen penetration, while enhancers demonstrated no significant increase (and can actually decrease) the amount of ibuprofen penetrating into and through the skin. PMID:23985822

  5. Penetration resistance according to penetration rate, cone base size and different soil conditions

    Directory of Open Access Journals (Sweden)

    Daniel Dias Valadão Junior

    2014-06-01

    Full Text Available This study aimed to evaluate the effect of penetration rate and the size of the cone base on the resistance to penetration under different soil moistures and soil bulk density. The experimental design was completely randomized in a 4x2x2x2 factorial arrangement, with the factors, soil bulk density of 1.0; 1.2; 1.4 and 1.6 Mg m-3, soil moisture at the evaluation of 0.16 and 0.22 kg kg-1, penetration rates of 0.166 and 30 mm s-1 and areas of the cone base of 10.98 and 129.28 mm² resulting in 32 treatments with 8 replicates. To ensure greater uniformity and similarity to field conditions, samples passed through cycles of wetting and drying. Only the interaction of the four factors was not significant. Resistance values varied with the density of the soil, regardless of moisture and penetration rate. Soil penetration resistance was influenced by the size of the cone base, with higher values for the smallest base independent of moisture and soil bulk density. The relationship between resistance to penetration and moisture is not always linear, once it is influenced by soil bulk density. Reduction in the area of the cone leads to an increase in the soil resistance to penetration.

  6. Limited cerebrospinal fluid penetration of docetaxel

    NARCIS (Netherlands)

    ten Tije, Albert J; Loos, Walter J; Zhao, Ming; Baker, Sharyn D; Enting, Roelien H; van der Meulen, Hans; Verweij, Jaap; Sparreboom, Alex; Enting, Roeline

    2004-01-01

    Our purpose was to investigate the cerebrospinal fluid (CSF) penetration of docetaxel in cancer patients. Docetaxel was administered as a 1-h infusion at a dose of 75 mg/m2 to two patients with metastatic breast cancer and leptomeningeal carcinomatosis. CSF samples were obtained using a lumbar punct

  7. 7 CFR 2902.14 - Penetrating lubricants.

    Science.gov (United States)

    2010-01-01

    ... recycled material as items for which Federal agencies must give preference in their purchasing programs... Items § 2902.14 Penetrating lubricants. (a) Definition. Products formulated to provide light lubrication... ingredients, re-refined oil, and/or any other recovered material, in addition to biobased ingredients,...

  8. Penetration of lipid monolayers by psychoactive drugs

    NARCIS (Netherlands)

    Demel, R.A.; Deenen, L.L.M. van

    1966-01-01

    The ability of a number of psychoactive drugs to penetrate lipid monolayers of varying composition was examined, and the following observation were made: (1) The increase in surface pressure of a monomolecular film appeared to depend on the chemical nature of the lipid as well as on the initial film

  9. The Basics of Hacking and Penetration Testing Ethical Hacking and Penetration Testing Made Easy

    CERN Document Server

    Engebretson, Patrick

    2011-01-01

    The Basics of Hacking and Penetration Testing serves as an introduction to the steps required to complete a penetration test or perform an ethical hack. You learn how to properly utilize and interpret the results of modern day hacking tools; which are required to complete a penetration test. Tool coverage will include, Backtrack Linux, Google, Whois, Nmap, Nessus, Metasploit, Netcat, Netbus, and more. A simple and clean explanation of how to utilize these tools will allow you  to gain a solid understanding of each of the four phases and prepare them to take on more in-depth texts and topi

  10. Effects of Sizes and Conformations of Fish-Scale Collagen Peptides on Facial Skin Qualities and Transdermal Penetration Efficiency

    OpenAIRE

    Huey-Jine Chai; Jing-Hua Li; Han-Ning Huang; Tsung-Lin Li; Yi-Lin Chan; Chyuan-Yuan Shiau; Chang-Jer Wu

    2010-01-01

    Fish-scale collagen peptides (FSCPs) were prepared using a given combination of proteases to hydrolyze tilapia (Oreochromis sp.) scales. FSCPs were determined to stimulate fibroblast cells proliferation and procollagen synthesis in a time- and dose-dependent manner. The transdermal penetration capabilities of the fractionationed FSCPs were evaluated using the Franz-type diffusion cell model. The heavier FSCPs, 3500 and 4500 Da, showed higher cumulative penetration capability as opposed to the...

  11. ACUPOINT TO ACUPOINT PENETRATIVE NEEDLING AND ITS CLINICAL APPLICATION

    Institute of Scientific and Technical Information of China (English)

    汪洪明

    2001-01-01

    Acupoint to acupoint penetrative needling method has a long history in traditional Chinese medicine. Chinese physicians in various dynasties hold that this type of needling method can strengthen the needling sensations and raise the therapeutic effect in clinical practice. This needling method can be divided into straight penetration, oblique penetration and transverse penetration. In clinical application, it can also be divided into external-medial meridian penetration, a single meridian penetration, neighboring meridians penetration, and neighboring aoupoints penetration, etc. in accordance with the involved acupoints and meridians. Clinical practice demonstrates that penetrative needling possesses advantages of stronger needling sensations, better therapeutic effect and fewer aoupoints selection. For this reason, it is extensively applied in clinic.

  12. Penetration and performance of isocyanate wood binders on selected wood species

    Directory of Open Access Journals (Sweden)

    Gruver, T. M., and

    2006-11-01

    Full Text Available The penetration and performance of polymeric diphenylmethane diisocyanate (pMDI wood binder was investigated according to three factors: substrate species (aspen, yellow-poplar, or southern yellow pine; anatomical bonding plane (radial or tangential; and moisture content (0%, 5%, or 12%. Compression shear block tests and fluorescence microscopy were used to examine bond performance and resin penetration. Statistically, each of the aforementioned factors impacted results. As moisture content increased, observed bond strengths and wood failure increased. Bond formation did not occur when the substrates were equilibrated to 0% moisture content, except for the radial bonding surfaces of pine, which did adhere. At 5 and 12% moisture contents, tangential bonding surfaces out-performed radial bonding surfaces. In terms of resin penetration, moisture content was clearly the most important variable. Little penetration was observed at 0% moisture content, while extensive resin penetration was observed at elevated moisture contents. Pine was the only wood species to exhibit resin flow through radial cells, possibly explaining the enhanced resin penetration depths observed in pine samples.

  13. Common gene therapy viral vectors do not efficiently penetrate sputum from cystic fibrosis patients.

    Directory of Open Access Journals (Sweden)

    Kaoru Hida

    Full Text Available Norwalk virus and human papilloma virus, two viruses that infect humans at mucosal surfaces, have been found capable of rapidly penetrating human mucus secretions. Viral vectors for gene therapy of Cystic Fibrosis (CF must similarly penetrate purulent lung airway mucus (sputum to deliver DNA to airway epithelial cells. However, surprisingly little is known about the rates at which gene delivery vehicles penetrate sputum, including viral vectors used in clinical trials for CF gene therapy. We find that sputum spontaneously expectorated by CF patients efficiently traps two viral vectors commonly used in CF gene therapy trials, adenovirus (d∼80 nm and adeno-associated virus (AAV serotype 5; d∼20 nm, leading to average effective diffusivities that are ∼3,000-fold and 12,000-fold slower than their theoretical speeds in water, respectively. Both viral vectors are slowed by adhesion, as engineered muco-inert nanoparticles with diameters as large as 200 nm penetrate the same sputum samples at rates only ∼40-fold reduced compared to in pure water. A limited fraction of AAV exhibit sufficiently fast mobility to penetrate physiologically thick sputum layers, likely because of the lower viscous drag and smaller surface area for adhesion to sputum constituents. Nevertheless, poor penetration of CF sputum is likely a major contributor to the ineffectiveness of viral vector based gene therapy in the lungs of CF patients observed to date.

  14. Lipid peroxidation and water penetration in lipid bilayers

    DEFF Research Database (Denmark)

    Conte, Elena; Megli, Francesco Maria; Khandelia, Himanshu;

    2012-01-01

    Lipid peroxidation plays a key role in the alteration of cell membrane's properties. Here we used as model systems multilamellar vesicles (MLVs) made of the first two products in the oxidative cascade of linoleoyl lecithin, namely 1-palmitoyl-2-(13-hydroperoxy-9,11-octadecanedienoyl)-lecithin (Hp......(zz) parameters revealed that OHPLPC, but mostly HpPLPC, induced a measurable increase in polarity and H-bonding propensity in the central region of the bilayer. Molecular dynamics simulation performed on 16-DSA in the PLPC-HpPLPC bilayer revealed that water molecules are statistically favored with respect to the...... hydroperoxide groups to interact with the nitroxide at the methyl-terminal, confirming that the H-bonds experimentally observed are due to increased water penetration in the bilayer. The EPR and MD data on model membranes demonstrate that cell membrane damage by oxidative stress cause alteration of water...

  15. CNS Active O-Linked Glycopeptides that Penetrate the BBB

    Science.gov (United States)

    Jones, Evan; Polt, Robin

    2015-06-01

    Naturally occurring glycopeptides and glycoproteins play important roles in biological processes. Glycosylation is one of the most common post-translational modifications in vivo. Glycopeptides are involved in cell signaling and sorting, providing cell surface markers for recognition. From the drug design and synthesis perspective, modification of a peptide through glycosylation results in increased bioavailability and bioactivity of glycopeptides in living systems with negligible toxicity of degradation products. Glycopeptide synthesis can be accomplished through incorporation of a glycosylated amino acid in solid phase peptide synthesis (SPPS) to form the desired peptide, or via incorporation of sugar-amino acid moieties. Additionally, research indicates that glycosylation increases penetration of the blood-brain barrier (BBB) by peptides, which may lead to novel therapeutics for neurological disorders. Recent applications of glycopeptides have focused on the in vivo central nervous system effects after peripheral administration of centrally active peptides modified with various carbohydrates.

  16. CNS Active O-Linked Glycopeptides that Penetrate the BBB

    Directory of Open Access Journals (Sweden)

    Evan M Jones

    2015-06-01

    Full Text Available Naturally occurring glycopeptides and glycoproteins play important roles in biological processes. Glycosylation is one of the most common post-translational modifications in vivo. Glycopeptides are involved in cell signaling and sorting, providing cell surface markers for recognition. From the drug design and synthesis perspective, modification of a peptide through glycosylation results in increased bioavailability and bioactivity of glycopeptides in living systems with negligible toxicity of degradation products. Glycopeptide synthesis can be accomplished through incorporation of a glycosylated amino acid in solid phase peptide synthesis (SPPS to form the desired peptide, or via incorporation of sugar-amino acid moieties. Additionally, research indicates that glycosylation increases penetration of the blood-brain barrier (BBB by peptides, which may lead to novel therapeutics for neurological disorders. Recent applications of glycopeptides have focused on the in vivo central nervous system effects after peripheral administration of centrally active peptides modified with various carbohydrates.

  17. Eddy current testing with high penetration

    International Nuclear Information System (INIS)

    The low-frequency eddy current testing method is used when penetration into very deep layers is required. The achievable penetration depth is determined among other parameters by the lowest testing frequency that can be realised together with the eddy current sensor. When using inductive sensors, the measuring effect declines proportional to the lowering frequency (induction effect). Further reduction of testing frequency requires other types of sensors, as e.g. the GMR (Giant Magnetic Resistance), which achieves a constant measuring sensitivity down to the steady field. The multi-frequency eddy current testing method MFEC 3 of IZFP described here can be operated using three different scanning frequencies at a time. Two variants of eddy current probes are used in this case. Both have an inductive winding at their emitters, of the type of a measuring probe. The receiver end is either also an inductive winding, or a magnetic field-responsive resistance (GMR). (orig./CB)

  18. Vacuum Radiation Pressure Fluctuations and Barrier Penetration

    CERN Document Server

    Huang, Haiyan

    2016-01-01

    We apply recent results on the probability distribution for quantum stress tensor fluctuations to the problem of barrier penetration by quantum particles. The probability for large stress tensor fluctuations decreases relatively slowly with increasing magnitude of the fluctuation, especially when the quantum stress tensor operator has been averaged over a finite time interval. This can lead to large vacuum radiation pressure fluctuations on charged or polarizable particles, which can in turn push the particle over a potential barrier. The rate for this effect depends sensitively upon the details of the time averaging of the stress tensor operator, which might be determined by factors such as the shape of the potential. We make some estimates for the rate of barrier penetration by this mechanism and argue that in some cases this rate can exceed the rate for quantum tunneling through the barrier. The possibility of observation of this effect is discussed.

  19. Dynamic behavior of novel tungsten penetrator materials

    International Nuclear Information System (INIS)

    The scope of the investigations was to find criteria which are related to the terminal ballistic performance of high strength tungsten KE-penetrator materials. The study comprised three tungsten alloys having densities above 17.1 g/cm3 and comparable static material strengths. For this purpose the authors investigated the dynamic material behavior by means of the instrumented Taylor-test. The influence of the strain rate on the yield strength and the maximum strain when cracking occurred were determined. The measured dynamic strength data were compared with the terminal ballistic efficiency of rods penetrating a semi-infinite RHA-target. Furthermore, the structural properties of the alloys were discussed with respect to their behavior in the regime of hydrodynamic flow

  20. Large scale wind power penetration in Denmark

    DEFF Research Database (Denmark)

    Karnøe, Peter

    2013-01-01

    he Danish electricity generating system prepared to adopt nuclear power in the 1970s, yet has become the world's front runner in wind power with a national plan for 50% wind power penetration by 2020. This paper deploys a sociotechnical perspective to explain the historical transformation...... of "networks of power" via the interactions of politics, the techno-physics of electrons, and the market setting. The Danish case is about how an assemblage of new agencies has reorganized and reshaped society by building a new sociotechnical network. This has rendered developments highly unpredictable...... and highly experimental. The transformation process can be followed through the way successive technical engineering reports have represented the challenges associated with the penetration of wind power. The iteration shows how novel technical phenomena emerge and are assimilated, and how new engineering...

  1. Depleted Uranium Penetrators : Hazards and Safety

    Directory of Open Access Journals (Sweden)

    S. S. Rao

    1997-01-01

    Full Text Available The depleted uranium (DU alloy is a state-of-the-art material for kinetic energy penetrators due to its superior ballistic performance. Several countries use DU penetrators in their main battle tanks. There is no gamma radiation hazard to the crew members from stowage of DO rounds. Open air firing can result in environmental contamination and associated hazards due to airborne particles containing essentially U/sub 3/0/sub 8/ and UO/sub 2/. Inhalation of polluted air only through respirators or nose masks and refraining form ingestion of water or food materials from contaminated environment are safety measures for avoiding exposure to uranium and its toxicity. Infusion of sodium bicarbonate helps in urinary excretion of uranium that may have entered the body.

  2. CRM approach penetration in a service organization

    OpenAIRE

    Mäyry, Asko

    2012-01-01

    The purpose was to develop CRM (Customer Relationship Management) system user commitment and CRM results for a service organization. The CRM approach penetration was used as an indicator to measure and explain the success of the CRM introduction. CRM related research questions and hypotheses were created. 21 respondents (95 %) answered the questionnaires before and after development activities. After both surveys, interviews were conducted to collect additional research data. A new method...

  3. Penetration of minocycline into lung tissues.

    OpenAIRE

    Naline, E.; Sanceaume, M; Toty, L; Bakdach, H; Pays, M; Advenier, C

    1991-01-01

    The penetration of minocycline into different lung tissues and bronchial mucus was studied in 17 patients undergoing pulmonary surgery for cancer. The patients received oral minocycline 100 mg at night for 3 days preceding surgery. Minocycline concentrations were measured in plasma samples collected before the operation and in tissues and mucus taken from in and around the part of the lung that was surgically removed. Mean tissue or mucus concentration to plasma concentration ratios were 3.78...

  4. Pseudo skin factor due to partial penetration

    Energy Technology Data Exchange (ETDEWEB)

    Reynolds, A.C.; Chen, J.C.; Raghavan, R.

    1983-10-01

    In this study the authors examine the pseudo skin factor due to partial penetration in a two-layer reservoir when only one layer is open to flow. They show that the pseudo skin factor can be correlated as a unique function of three reservoir parameters and in some cases can be accurately correlated as a function of two reservoir parameters. They also present graphs and procedures for estimating the pseudo skin factor.

  5. Pseudo skin factor due to partial penetration

    Energy Technology Data Exchange (ETDEWEB)

    Reynolds, A.C.; Chen, J.C.; Raghavan, R.

    1983-01-01

    This study examines the pseudo skin factor due to partial penetration in a 2-layer reservoir when only one layer is open to flow. The pseudo skin factor can be correlated as a unique function of 3 reservoir parameters and in some cases can be correlated accurately as a function of 2 reservoir parameters. Graphs and procedures for estimating the pseudo skin factor are presented. 17 references.

  6. Penetration of Candida Biofilms by Antifungal Agents

    OpenAIRE

    Al-Fattani, Mohammed A.; Douglas, L. Julia

    2004-01-01

    A filter disk assay was used to investigate the penetration of antifungal agents through biofilms containing single and mixed-species biofilms containing Candida. Fluconazole permeated all single-species Candida biofilms more rapidly than flucytosine. The rates of diffusion of either drug through biofilms of three strains of Candida albicans were similar. However, the rates of drug diffusion through biofilms of C. glabrata or C. krusei were faster than those through biofilms of C. parapsilosi...

  7. Market penetration of new energy technologies

    Energy Technology Data Exchange (ETDEWEB)

    Packey, D.J.

    1993-02-01

    This report examines the characteristics, advantages, disadvantages, and, for some, the mathematical formulas of forecasting methods that can be used to forecast the market penetration of renewable energy technologies. Among the methods studied are subjective estimation, market surveys, historical analogy models, cost models, diffusion models, time-series models, and econometric models. Some of these forecasting methods are more effective than others at different developmental stages of new technologies.

  8. Surfactant improves irrigant penetration into unoperated sinuses

    OpenAIRE

    Rohrer, Joseph W.; Dion, Greg R.; Brenner, Pryor S.; Abadie, Wesley M.; McMains, Kevin C.; Thomas, Roy F.; Weitzel, Erik K.

    2012-01-01

    Background: Saline irrigations are proving to be a valuable intervention in the treatment of chronic sinusitis. The use of surfactants is a well established additive to topical treatments known to reduce surface tension and may prove to be a simple, nonoperative intervention to improve intrasinus douching penetration. Methods: Six 30-mL, flat-bottomed medicine cups with circular holes cut through the bottom center and varying in diameter from 1 to 6 mm were created with punch biopsies. Water,...

  9. Essential fatty acids as transdermal penetration enhancers

    OpenAIRE

    Van Zyl, Lindi; du Preez, Jan; Gerber, Minja; Du Plessis, Jeanetta; Viljoen, Joe

    2015-01-01

    The aim of this study was to investigate the effect of different penetration enhancers, containing essential fatty acids (EFAs), on the transdermal delivery of flurbiprofen. Evening primrose oil (EPO), vitamin F and Pheroid™ technology all contain fatty acids and were compared using a cream-based formulation. This selection was to ascertain whether EFAs solely, or EFAs in a Pheroid™ delivery system, would have a significant increase in the transdermal delivery of a compound. Membrane release ...

  10. Aerosol penetration inside HEPA filtration media

    International Nuclear Information System (INIS)

    Attempts made to modelize changes in high-efficiency particulate air (HEPA) filter pressure drop as a function of such parameters as the amount of aerosol collected, which also take account of filtering medium characteristics, those of the filtered aerosol and the filtration velocity, have always been limited by a lack of knowledge of the distribution of particles within the medium. For the last few years, a method, developed over 25 years ago to study the distribution of Radon daughters within filter media made of cellulose fibers, has been reused and applied to radioactive and fluorescent aerosols penetration studies. This method enables determination of aerosol distribution on the surface and inside the filter medium by peeling away successive layers using an adhesive tape and measuring the specific amounts removed each time. Knowledge of aerosol penetration within the filter has revealed that, for a give aerosol, particle distribution inside the filter rapidly decreased exponentially and that fixation on the filter's front surface rapidly superseded penetration inside the medium. The deposit profiles thus measured have made it possible to propose a model for determining the rate of filter pressure drop increase that closely agrees with experimental results

  11. Inspection and repair technology for BMI penetrations

    International Nuclear Information System (INIS)

    Historically, United States (US) nuclear power plant inspections of the reactor vessel bottom-mounted-instrument (BMI) penetrations have been limited to visual verification via a combination of walk-downs and pressure tests. However in France, more than 18 ultrasonic examination (UT) and weld-surface visual test (VT) campaigns had been performed to inspect the BMI penetration nozzles and welds since 1992 with no observed failures through 2002. In April 2003, South Texas Project Unit 1 discovered apparent leakage from two nozzles during a bare-metal examination. Based on the French inspection experience, Framatome-ANP was selected for inspection and repair services to address the leaking nozzles. Inspection activities included ultrasonic examination of the tube, enhanced visual test and eddy current testing (ECT) of the J-groove weld, bobbin ECT and profile measurement of the tube inside, helium leak test, phased-array UT, borescope VT, and boat-sample removal with destructive metallurgical analysis. Repair activities included sealing the top portion of the penetration, cutting out the old nozzles, and welding in new nozzles. This presentation discusses BMI inspection and repair technology focused particularly on the South Texas Project experience. (authors)

  12. Ultrastructure and penetration of cell co-culture model of human skin equivalent%细胞共培养人体皮肤类似物超微结构观察及其渗透性测试

    Institute of Scientific and Technical Information of China (English)

    李辉; 王景雁; 林红梅; 于竞新; 章烨雯; 毛颖秋; 吴清

    2016-01-01

    目的:观察人体皮肤类似物模型形态结构,对其屏障功能进行初步评价。方法通过胶原蛋白包被成纤维细胞,制备真皮层,真皮层表面接种角质细胞,浸没培养1周后,气-液界面培养以促进角质细胞的分化形成角质层,3周后收获皮肤模型;通过肉眼观察、电镜观察皮肤类似物的宏观结构和超微结构;以吲哚美辛作为模型药物,进行体外透皮实验考察细胞共培养皮肤类似物模型的屏障功能。结果皮肤类似物形成与人类皮肤相似的真皮层和表皮层,表皮层表面形成完整的角质层,具有与人类皮肤类似的超微结构;24 h 内渗透性实验结果显示皮肤类似物屏障功能略低于大鼠腹部皮肤,但22~24 h,皮肤类似物对模型药的透过速率远小于大鼠腹部皮肤(约为其28%),且2种皮肤模型对吲哚美辛经皮渗透的稳态流速没有显著性差异(P >0.05)。结论皮肤类似物有与人体皮肤相类似的形态结构特征,与大鼠腹部皮肤相比对吲哚美辛有相似的屏障作用,有望替代动物皮肤成为经皮给药系统评价的新型皮肤模型。%Abatract:Objective To observe the structure of human skin equivalent (HSE)model in vitro,and to preliminarily assess the barrier function of HSE.Methods The HSE model was established by two steps:preparation of dermal layer and preparation of full-thickness skin.Keratinocytes were inoculated on the surface of dermal layer prepared by type 1 collagen coating fibroblasts.After submerged cultured for one week,air-liquid interface cultured for three weeks to enhance the proliferation of keratinocytes,the HSE model was obtained.Then the macroscopical structure and electon microscopical structure of HSE were studied.To investigate the barrier function of HSE model,indometacin was selected as the model drug for the penetration testing in vitro.Results The structures similar to dermal layer and

  13. Penetration Testing Curriculum Development in Practice

    Directory of Open Access Journals (Sweden)

    Chengcheng Li

    2015-04-01

    Full Text Available As both the frequency and the severity of network breaches have increased in recent years, it is essential that cybersecurity is incorporated into the core of business operations. Evidence from the U.S. Bureau of Labor Statistics (Bureau of Labor Statistics, 2012 indicates that there is, and will continue to be, a severe shortage of cybersecurity professionals nationwide throughout the next decade. To fill this job shortage we need a workforce with strong hands-on experience in the latest technologies and software tools to catch up with the rapid evolution of network technologies. It is vital that the IT professionals possess up-to-date technical skills and think and act one step ahead of the cyber criminals who are constantly probing and exploring system vulnerabilities. There is no perfect security mechanism that can defeat all the cyber-attacks; the traditional defensive security mechanism will eventually fail to the pervasive zero-day attacks. However, there are steps to follow to reduce an organization’s vulnerability to cyber-attacks and to mitigate damages. Active security tests of the network from a cyber-criminal’s perspective can identify system vulnerabilities that may lead to future breaches. “If you know yourself but not the enemy, for every victory gained you will also suffer a defeat. But if you know the enemy and know yourself, you need not fear the result of hundred battles” (Sun, 2013. Penetration testing is a discipline within cybersecurity that focuses on identifying and exploiting the vulnerabilities of a network, eventually obtaining access to the critical business information. The pentesters, the security professionals who perform penetration testing, or ethical hackers, break the triad of information security - Confidentiality, Integrity, and Accountability (CIA - as if they were a cyber-criminal. The purpose of ethical hacking or penetration testing is to know what the “enemy” can do and then generate a

  14. Penetration Testing Professional Ethics: a conceptual model and taxonomy

    OpenAIRE

    Justin Pierce; Ashley Jones; Matthew Warren

    2006-01-01

    In an environment where commercial software is continually patched to correct security flaws, penetration testing can provide organisations with a realistic assessment of their security posture. Penetration testing uses the same principles as criminal hackers to penetrate corporate networks and thereby verify the presence of software vulnerabilities. Network administrators can use the results of a penetration test to correct flaws and improve overall security. The use of hacking techniques...

  15. Hybrid Collagenase Nanocapsules for Enhanced Nanocarrier Penetration in Tumoral Tissues.

    Science.gov (United States)

    Villegas, María Rocío; Baeza, Alejandro; Vallet-Regí, María

    2015-11-01

    Poor penetration of drug delivery nanocarriers within dense extracellular matrices constitutes one of the main liabilities of current nanomedicines. The conjugation of proteolytic enzymes on the nanoparticle surface constitutes an attractive alternative. However, the scarce resistance of these enzymes against the action of proteases or other aggressive agents present in the bloodstream strongly limits their application. Herein, a novel nanodevice able to transport proteolytic enzymes coated with an engineered pH-responsive polymeric is presented. This degradable coat protects the housed enzymes against proteolytic attack at the same time that it triggers their release under mild acidic conditions, usually present in many tumoral tissues. These enzyme nanocapsules have been attached on the surface of mesoporous silica nanoparticles, as nanocarrier model, showing a significatively higher penetration of the nanoparticles within 3D collagen matrices which housed human osteosarcoma cells (HOS). This strategy can improve the therapeutic efficacy of the current nanomedicines, allowing a more homogeneous and deeper distribution of the therapeutic nanosystems in cancerous tissues.

  16. Verifying Removal Of Red Penetrant Dye From Inspected Welds

    Science.gov (United States)

    Torkelson, Jan R.

    1996-01-01

    Clean surface assured for more sensitive inspection with fluorescent penetrant dye. Simple procedure devised to ensure visible (red) penetrant dye used to identify flaws in welded surface completely removed from surface. Consists in applying reversible penetrant developer to surface to be inspected.

  17. A SNARE-protein has opposing functions in penetration resistance and defence signalling pathways

    DEFF Research Database (Denmark)

    Zhang, Ziguo; Feechan, Angela; Pedersen, Carsten;

    2007-01-01

    Penetration resistance is often the first line of defence against fungal pathogens. Subsequently induced defences are mediated by the programmed cell death (PCD) reaction pathway and the salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) signalling pathways. We previously demonstrated...

  18. Effects of Sizes and Conformations of Fish-Scale Collagen Peptides on Facial Skin Qualities and Transdermal Penetration Efficiency

    Directory of Open Access Journals (Sweden)

    Huey-Jine Chai

    2010-01-01

    Full Text Available Fish-scale collagen peptides (FSCPs were prepared using a given combination of proteases to hydrolyze tilapia (Oreochromis sp. scales. FSCPs were determined to stimulate fibroblast cells proliferation and procollagen synthesis in a time- and dose-dependent manner. The transdermal penetration capabilities of the fractionationed FSCPs were evaluated using the Franz-type diffusion cell model. The heavier FSCPs, 3500 and 4500 Da, showed higher cumulative penetration capability as opposed to the lighter FSCPs, 2000 and 1300 Da. In addition, the heavier seemed to preserve favorable coiled structures comparing to the lighter that presents mainly as linear under confocal scanning laser microscopy. FSCPs, particularly the heavier, were concluded to efficiently penetrate stratum corneum to epidermis and dermis, activate fibroblasts, and accelerate collagen synthesis. The heavier outweighs the lighter in transdermal penetration likely as a result of preserving the given desired structure feature.

  19. Effects of sizes and conformations of fish-scale collagen peptides on facial skin qualities and transdermal penetration efficiency.

    Science.gov (United States)

    Chai, Huey-Jine; Li, Jing-Hua; Huang, Han-Ning; Li, Tsung-Lin; Chan, Yi-Lin; Shiau, Chyuan-Yuan; Wu, Chang-Jer

    2010-01-01

    Fish-scale collagen peptides (FSCPs) were prepared using a given combination of proteases to hydrolyze tilapia (Oreochromis sp.) scales. FSCPs were determined to stimulate fibroblast cells proliferation and procollagen synthesis in a time- and dose-dependent manner. The transdermal penetration capabilities of the fractionationed FSCPs were evaluated using the Franz-type diffusion cell model. The heavier FSCPs, 3500 and 4500 Da, showed higher cumulative penetration capability as opposed to the lighter FSCPs, 2000 and 1300 Da. In addition, the heavier seemed to preserve favorable coiled structures comparing to the lighter that presents mainly as linear under confocal scanning laser microscopy. FSCPs, particularly the heavier, were concluded to efficiently penetrate stratum corneum to epidermis and dermis, activate fibroblasts, and accelerate collagen synthesis. The heavier outweighs the lighter in transdermal penetration likely as a result of preserving the given desired structure feature. PMID:20625414

  20. Differential penetration of targeting agents into multicellular spheroids derived from human neuroblastoma

    International Nuclear Information System (INIS)

    The authors have used a multicellular tumour spheroid model for determination of the penetration of various targeting agents of potential use in the treatment of neuroblastoma. Both the radiopharmaceutical meta-iodobenzylguanidine (mIBG) and the β subunit of nerve growth factor (β-NGF) distributed uniformly throughout spheroids, though the latter was poorly concentrated relative to mIBG. In contrast, the anti-neuroectodermal monoclonal antibody. UJ13A bound only to peripheral cell layers with little accumulation in the spheroid interior. Differential penetration of targeting agents may influence the choice of conjugated radionuclide which is likely to achieve maximum therapeutic benefit

  1. Efficacy of sunlight-activatable porphyrin formulates on larvae of Anopheles gambiae M and S molecular forms and An. arabiensis: a potential novel biolarvicide for integrated malaria vector control.

    Science.gov (United States)

    Fabris, Clara; Ouédraogo, Robert Kossivi; Coppellotti, Olimpia; Dabiré, Roch K; Diabaté, Abdoulaye; Di Martino, Piera; Guidolin, Laura; Jori, Giulio; Lucantoni, Leonardo; Lupidi, Giulio; Martena, Valentina; Sawadogo, Simon P; Soncin, Marina; Habluetzel, Annette

    2012-09-01

    Biolarvicides, such as microbial formulations based on Bacillus thuringiensis and B. sphaericus, have been found to be highly effective against mosquito larvae and are currently employed as eco-friendly alternatives to synthetic chemical insecticides for vector control. Recently, a porphyrin of natural origin has been suggested as a sunlight-activatable larvicide against the dengue vector Aedes aegypti. In order to validate the approach for the control of the malaria vector, we tested the photo-larvicidal activity of a novel porphyrin, namely meso-tri(N-methyl-pyridyl), mono(N-dodecyl-pyridyl)porphine, C12, associated with two specifically selected carriers, against Anopheles gambiae s.s. and An. arabiensis larvae, both laboratory reared and collected from malaria endemic sites in Burkina Faso. Both C12-porphyrin formulates, when administered to larvae at a 50μM porphyrin dose, were accumulated in the alimentary canal. Subsequent exposure of the porphyrin-loaded larvae to sunlight for short times (0.5-3h) led to a complete mortality. The high efficacy exhibited by a "foodstuff" porphyrin formulate also in the presence of typical larval food particles opens promising perspectives for the development of an effective photocidal larvicide.

  2. Penetration Testing Professional Ethics: a conceptual model and taxonomy

    Directory of Open Access Journals (Sweden)

    Justin Pierce

    2006-05-01

    Full Text Available In an environment where commercial software is continually patched to correct security flaws, penetration testing can provide organisations with a realistic assessment of their security posture. Penetration testing uses the same principles as criminal hackers to penetrate corporate networks and thereby verify the presence of software vulnerabilities. Network administrators can use the results of a penetration test to correct flaws and improve overall security. The use of hacking techniques, however, raises several ethical questions that centre on the integrity of the tester to maintain professional distance and uphold the profession. This paper discusses the ethics of penetration testing and presents our conceptual model and revised taxonomy.

  3. A penetrative addictive for water infusion in coal seams

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The manufacturing process, characteristics, and application results of a rod-like penetrative additive (the penetrative rod)were discussed. The components and functions of the penetrative rod were studied carefully. A large number of orthogonal combinedtests were carried out and over 100 sample molds were made. Ultimately the components were decided after careful selection amongthese molds, mainly including hygroscopic major components and auxiliary material. The results of on-site practical applicationshow that such an addictive can increase the water penetrative ability effectively and has a remarkable effect on preventing dust pro-duction during coal seam excavation. The penetrating radius, the infusing velocity and the dust-preventive effect were systematicallystudied.

  4. Premium Efficiency Motors And Market Penetration Policy

    Energy Technology Data Exchange (ETDEWEB)

    Benhaddadi, Mohamed; Olivier, Guy

    2010-09-15

    This paper illustrates the induced enormous energy saving potential, permitted by using high-efficiency motors. Furthermore, the most important barriers to larger high-efficiency motors utilization are identified, and some incentives recommendations are given to overcome identified impediments. The authors consider that there is a strong case to enhance incentives policies for larger market penetration. The US Energy Policy Act and the Canadian Energy Efficient Act have lead to North American leadership on motor efficiency implementation. North America is not on the leading edge for energy saving and conservation. Motor efficiency is an exception that should be at least maintained.

  5. Mastering Kali Linux for advanced penetration testing

    CERN Document Server

    Beggs, Robert W

    2014-01-01

    This book provides an overview of the kill chain approach to penetration testing, and then focuses on using Kali Linux to provide examples of how this methodology is applied in the real world. After describing the underlying concepts, step-by-step examples are provided that use selected tools to demonstrate the techniques. If you are an IT professional or a security consultant who wants to maximize the success of your network testing using some of the advanced features of Kali Linux, then this book is for you. This book will teach you how to become an expert in the pre-engagement, management,

  6. Kali Linux assuring security by penetration testing

    CERN Document Server

    Ali, Shakeel; Allen, Lee

    2014-01-01

    Written as an interactive tutorial, this book covers the core of Kali Linux with real-world examples and step-by-step instructions to provide professional guidelines and recommendations for you. The book is designed in a simple and intuitive manner that allows you to explore the whole Kali Linux testing process or study parts of it individually.If you are an IT security professional who has a basic knowledge of Unix/Linux operating systems, including an awareness of information security factors, and want to use Kali Linux for penetration testing, then this book is for you.

  7. Penetrating abdominal injuries in children in Nigeria.

    Science.gov (United States)

    Ameh, E A; Nmadu, P T

    1999-09-01

    This is a report of a retrospective study of 24 children managed for penetrating abdominal injury over 10 years, and it represents 34% of all abdominal injuries in children in that period. Falls onto sharp objects within and around the home were responsible for ten of the injuries, seven were injured by animal horns and four were sporting injuries. Violence and road traffic accidents were uncommon. Most patients (67%) had evisceration of omentum or intestine, and one of these was found at laparotomy to have a jejuno-jejunal intussusception. Seven children had injury to hollow viscera. There were three deaths, one each from overwhelming sepsis, tetanus and haemorrhage.

  8. Opportunities for high wind energy penetration

    DEFF Research Database (Denmark)

    Tande, J.O.; Hansen, J.C.

    1997-01-01

    Wind power is today a mature technology, which at windy locations, is economically competitive to conventional power generation technologies. This and growing global environmental concerns have led governments to encourage and plan for wind energy development, a typical aim being 10% of electricity...... analyses conclude that expanding to even above 15% wind energy penetration in the Cape Verde power systems is economical. Worldwide, numerous locations with favorable wind conditions and power systems similar to the Capeverdean provide good opportunities for installing wind farms and achieving high wind...

  9. Hacking with Kali practical penetration testing techniques

    CERN Document Server

    Broad, James

    2013-01-01

    Hacking with Kali introduces you the most current distribution of the de facto standard tool for Linux pen testing. Starting with use of the Kali live CD and progressing through installation on hard drives, thumb drives and SD cards, author James Broad walks you through creating a custom version of the Kali live distribution. You'll learn how to configure networking components, storage devices and system services such as DHCP and web services. Once you're familiar with the basic components of the software, you'll learn how to use Kali through the phases of the penetration testing lifecycle

  10. Cephradine (Velosef) penetration of mandibular bone.

    Science.gov (United States)

    Middlehurst, R J; Rood, J P

    1990-04-01

    The concentration of cephradine in serum and mandibular bone was assayed in 28 patients undergoing 3rd molar surgery following a single 1 g intravenous injection. Serum and cortical bone samples taken simultaneously, contained mean cephradine concentrations of 42.11 micrograms/ml and 2.61 micrograms/g respectively. These results, when compared with those reported for other bony sites including the femoral head and knee, show a reduced bone penetration with a bone-to-serum ratio of approximately 0.06:1. PMID:2111359

  11. [Penetrating abdominal wounds. Apropos of 330 cases].

    Science.gov (United States)

    Nejjar, M; Bennani, S; Zerouali, O N

    1991-01-01

    Penetrating abdominal wounds are frequent and serious. 330 cases have been treated in the Department of Emergencies and visceral Surgery at Averroes Hospital of Casablanca from 1980 to 1990. The predominance of male sex is noted, and these wounds are always the result of aggression by white arm. All patients have been operated, the white laparotomy rate is of 36%. The classic interventionist attitude is still recommended in spite of this high rate, because our present conditions can't permit us a rigorous watching. According to abdominal lesions, the different interventions are reviewed, and their indications are detailed.

  12. Penetrating wounds of the head and neck.

    Science.gov (United States)

    Jahrsdoerfer, R A; Johns, M E; Cantrell, R W

    1979-12-01

    Wounding capability of bullets is primarily releated to velocity. Bullet mass and shape, and specific gravity of body tissues being struck by the missile, are lesser factors. Seventy cases of penetrating wounds of the head and neck were treated during a six-year period. Vascular injuries were more common with neck wounds, while face and head injuries (extracranial) were similar to maxillofacial trauma. It is recognized that hemorrhage at the base of the skull is difficult to treat, and contemporary training in temporal bone and base of skull surgery is mandatory for the critical management of these wounds.

  13. Suicide bomb attack causing penetrating craniocerebral injury

    Institute of Scientific and Technical Information of China (English)

    Manzar Hussain; Muhammad Ehsan Bari

    2013-01-01

    Penetrating cerebral injuries caused by foreign bodies are rare in civilian neurosurgical trauma,although there are various reports of blast or gunshot injuries in warfare due to multiple foreign bodies like pellets and nails.In our case,a 30-year-old man presented to neurosurgery clinic with signs and symptoms of right-sided weakness after suicide bomb attack.The skull X-ray showed a single intracranial nail.Small craniotomy was done and the nail was removed with caution to avoid injury to surrounding normal brain tissue.At 6 months' follow-up his right-sided power improved to against gravity.

  14. Effect of sodium lauryl sulfate (SLS) on in vitro percutaneous penetration of water, hydrocortisone and nickel

    DEFF Research Database (Denmark)

    Frankild, S; Andersen, Klaus Ejner; Nielsen, Gunnar

    1995-01-01

    as membrane in static in vitro penetration cells. Simultaneous application of SLS together with 1 of the tracer compounds showed, after 48 h, a significant dose-effect relationship between SLS concentration (0.25%, 2% and 10%) and penetration of tritiated water or nickel (p ... or hydrocortisone. Pretreatment of the skin with SLS for 2 h using 3 concentrations (0.25%, 4% and 10%) showed, after 48 h, a significant dose-effect relationship between SLS treatment and penetration of tritiated water or nickel (p .../damaging effect on the skin barrier. It should be kept in mind that the model uses a dead skin membrane without the barrier repair mechanisms of live skin....

  15. Experimental Study of Bacterial Penetration into Chalk Rock: Mechanisms and Effect on Permeability

    DEFF Research Database (Denmark)

    Halim, Amalia Yunita; Shapiro, Alexander; Eliasson Lantz, Anna;

    2014-01-01

    Bacterial selective plugging is one of the mechanisms through which microorganisms can be applied for enhanced oil recovery, as bacteria can plug the water-swept zones of a reservoir, thus altering the flow paths and improving sweep efficiency. However, complete understanding of the penetration...... behavior of bacteria is lacking, especially in chalk formations where characteristic pore throat sizes are comparable with the sizes of bacterial cells. In this study, two bacterial strains, Bacillus licheniformis 421 (spore-forming) and Pseudomonas putida K12 (non-spore forming) were used to investigate...... the penetration of bacteria into chalk and its effect on permeability reduction. The core plugs were produced from Stevns Klint outcrop with low permeability (2–4 mD) and with pore sizes comparable to bacterial sizes. Both types of bacteria were able to penetrate and to be transported through the cores to some...

  16. Variant haploinsufficiency and phenotypic non-penetrance in PRPF31-associated retinitis pigmentosa.

    Science.gov (United States)

    Rose, A M; Bhattacharya, S S

    2016-08-01

    Retinitis pigmentosa (RP) is a genetically heterogenous group of inherited disorders, characterized by death of the retinal photoreceptor cells, leading to progressive visual impairment. One form of RP is caused by mutations in the ubiquitously expressed splicing factor, PRPF31, this form being known as RP11. An intriguing feature of RP11 is the presence of non-penetrance, which has been observed in the majority of PRPF31 mutation-carrying families. In contrast to variable expressivity, which is highly pervasive, true non-penetrance is a very rare phenomenon in Mendelian disorders. In this article, the molecular mechanisms underlying phenotypic non-penetrance in RP11 are explored. It is an elegant example of how our understanding of monogenic disorders has evolved from studying only the disease gene, to considering a mutation on the genetic background of the individual - the logical evolution in this genomic era. PMID:26853529

  17. Independent regulation of reovirus membrane penetration and apoptosis by the mu1 phi domain.

    Directory of Open Access Journals (Sweden)

    Pranav Danthi

    2008-12-01

    Full Text Available Apoptosis plays an important role in the pathogenesis of reovirus encephalitis. Reovirus outer-capsid protein mu1, which functions to penetrate host cell membranes during viral entry, is the primary regulator of apoptosis following reovirus infection. Ectopic expression of full-length and truncated forms of mu1 indicates that the mu1 phi domain is sufficient to elicit a cell death response. To evaluate the contribution of the mu1 phi domain to the induction of apoptosis following reovirus infection, phi mutant viruses were generated by reverse genetics and analyzed for the capacity to penetrate cell membranes and elicit apoptosis. We found that mutations in phi diminish reovirus membrane penetration efficiency by preventing conformational changes that lead to generation of key reovirus entry intermediates. Independent of effects on membrane penetration, amino acid substitutions in phi affect the apoptotic potential of reovirus, suggesting that phi initiates apoptosis subsequent to cytosolic delivery. In comparison to wild-type virus, apoptosis-defective phi mutant viruses display diminished neurovirulence following intracranial inoculation of newborn mice. These results indicate that the phi domain of mu1 plays an important regulatory role in reovirus-induced apoptosis and disease.

  18. Root cap removal increases root penetration resistance in maize (Zea mays L).

    Science.gov (United States)

    Iijima, Morio; Higuchi, Toshifumi; Barlow, Peter W; Bengough, A Glyn

    2003-09-01

    The root cap assists the passage of the root through soil by means of its slimy mucilage secretion and by the sloughing of its outer cells. The root penetration resistance of decapped primary roots of maize (Zea mays L. cv. Mephisto) was compared with that of intact roots in loose (dry bulk density 1.0 g cm-3; penetration resistance 0.06 MPa) and compact soil (1.4 g cm-3; penetration resistance 1.0 MPa), to evaluate the contribution of the cap to decreasing the impedance to root growth. Root elongation rate and diameter were the same for decapped and intact roots when the plants were grown in loose soil. In compacted soil, however, the elongation rate of decapped roots was only about half that of intact roots, whilst the diameter was 30% larger. Root penetration resistances of intact and decapped seminal axis were 0.31 and 0.52 MPa, respectively, when the roots were grown in compacted soil. These results indicated that the presence of a root cap alleviates much of the mechanical impedance to root penetration, and enables roots to grow faster in compacted soils.

  19. Ground penetrating radar for asparagus detection

    Science.gov (United States)

    Seyfried, Daniel; Schoebel, Joerg

    2016-03-01

    Ground penetrating radar is a promising technique for detection of buried objects. Recently, radar has more and more been identified to provide benefits for a plurality of applications, where it can increase efficiency of operation. One of these fields is the industrial automatic harvesting process of asparagus, which is performed so far by cutting the soil ridge at a certain height including all the asparagus spears and subsequently sieving the latter out of the soil. However, the height where the soil is cut is a critical parameter, since a wrong value leads to either damage of the roots of the asparagus plants or to a reduced crop yield as a consequence of too much biomass remaining in the soil. In this paper we present a new approach which utilizes ground penetrating radar for non-invasive sensing in order to obtain information on the optimal height for cutting the soil. Hence, asparagus spears of maximal length can be obtained, while keeping the roots at the same time undamaged. We describe our radar system as well as the subsequent digital signal processing steps utilized for extracting the information required from the recorded radar data, which then can be fed into some harvesting unit for setting up the optimal cutting height.

  20. Foliar penetration enhanced by biosurfactant rhamnolipid.

    Science.gov (United States)

    Liu, Haojing; Shao, Bing; Long, Xuwei; Yao, Yang; Meng, Qin

    2016-09-01

    With recent environmental and health concerns, biosurfactants have obtained increasing interest in replacing conventional surfactants for diverse applications. In agriculture, the use of surfactant in stimulating foliar uptake is mainly for wetting leaf surface, resisting deposition/evaporation, enhancing penetration across cuticular membrane (CM) and translocation. This paper aimed to address the improved foliar uptake by rhamnolipid (RL) in comparison with the currently used alkyl polyglucoside (APG). As found, compared with APG at 900mg/L (1×critical micellar concentration, CMC), RL at a much lower concentration of 50mg/L (1×CMC) showed much better wettability and surface activity, indicative of its high effectiveness as surfactants. Its performance on resistance to deposition and evaporation was at least as same as APG. Moreover, RL could significantly improve the penetration of herbicide glyphosate and other two small water-soluble molecules (phenol red and Fe(2+)) across CM at an equivalent efficiency as APG at 1×CMC. Finally, the greatly enhanced herbicidal actitivity of glyphosate on greenhouse plants confirmed that RL and APG could both enhance the foliar uptake including translocation. Overall, RL should be more applicable than APG in agriculture due to its more promising properties on health/environmental friendliness.

  1. Penetration of projectiles into granular targets

    Science.gov (United States)

    Ruiz-Suárez, J. C.

    2013-06-01

    Energetic collisions of subatomic particles with fixed or moving targets have been very valuable to penetrate into the mysteries of nature. But the mysteries are quite intriguing when projectiles and targets are macroscopically immense. We know that countless debris wandering in space impacted (and still do) large asteroids, moons and planets; and that millions of craters on their surfaces are traces of such collisions. By classifying and studying the morphology of such craters, geologists and astrophysicists obtain important clues to understand the origin and evolution of the Solar System. This review surveys knowledge about crater phenomena in the planetary science context, avoiding detailed descriptions already found in excellent papers on the subject. Then, it examines the most important results reported in the literature related to impact and penetration phenomena in granular targets obtained by doing simple experiments. The main goal is to discern whether both schools, one that takes into account the right ingredients (planetary bodies and very high energies) but cannot physically reproduce the collisions, and the other that easily carries out the collisions but uses laboratory ingredients (small projectiles and low energies), can arrive at a synergistic intersection point.

  2. High Penetration Photovoltaic Case Study Report

    Energy Technology Data Exchange (ETDEWEB)

    Bank, J.; Mather, B.; Keller, J.; Coddington, M.

    2013-01-01

    Technical concerns with integrating higher penetrations of photovoltaic (PV) systems include grid stability, voltage regulation, power quality (voltage rise, sags, flicker, and frequency fluctuations), and protection and coordination. The current utility grid was designed to accommodate power flows from the central generation source to the transmission system and eventually to the distribution feeders. At the distribution level, the system was designed to carry power from the substation toward the load. Renewable distributed generation, particularly solar PV, provides power at the distribution level challenging this classical paradigm. As these resources become more commonplace the nature of the distribution network and its operation is changing to handle power flow in both directions. This report is focused on large PV installations in which penetration is significantly greater than 15% of maximum daytime feeder load. These case studies are intended to demonstrate success stories with integration of large PV plants at the distribution level as well as some of the solutions used by the utility to ensure safe, reliable operation of both the PV system and the distribution network.

  3. Feature Profile Simulations and Finite Penetration Depth

    Science.gov (United States)

    Moroz, Paul; Moroz, Daniel

    2012-10-01

    In plasma materials processing, energetic ions, neutrals and UV photons typically penetrate deep inside solid materials breaking atomic bonds and displacing atoms on their paths. These important phenomena are rarely taken into consideration in processing simulation software, primarily because the proper penetration depths and the corresponding energy depositions, breaking bonds, and atom displacements are difficult and computationally expensive to compute. The FPS-3D feature profile simulator [1-2] is doing that computationally efficiently by utilizing tabulated results obtained with other methods. We discuss, compare, and present results of such simulations made with different methods, one of which is the molecular dynamics analysis. In general, molecular dynamics could be used for simulating materials processing, etching and deposition, but it is extremely computationally expensive to be used for large groups of atoms. In practice, molecular dynamics methods are too slow to be used for feature profile simulations. However, they could help in defining proper chemical reactions and corresponding rates to be used in an advanced feature profile simulator such as FPS-3D. We present results of FPS-3D simulations for Si and SiO2 etching in Ar/Cl2 and Ar/C4F6/O2 plasmas. [4pt] [1] P. Moroz, ``General Feature Profile Simulator FPS-3D,'' ECS Transactions, 35, 25 (2011). [0pt] [2] P. Moroz, ``Numerical Simulation of Feature Profile Evolution using FPS-3D,'' IEEE Transactions of Plasma Science, 39, 2804 (2011).

  4. Penetrating eye injuries from writing instruments

    Directory of Open Access Journals (Sweden)

    Kelly SP

    2011-12-01

    Full Text Available Simon P Kelly, Graham MB ReevesThe Royal Bolton Hospital, Bolton, UKPurpose: To consider the potential for ocular injury from writing implements by presenting four such cases, and to consider the incidence of such eye injuries from analysis of a national trauma database.Methods: The Home and Leisure Accident Surveillance System was searched for records of eye injuries from writing instruments to provide UK estimates of such injuries. Four patients with ocular penetrating injury from pens or pencils (especially when caused by children, and examined by the authors, are described which illustrate mechanisms of injury.Results: It is estimated that around 748 ocular pen injuries and 892 ocular pencil injuries of undetermined severity occurred annually in the UK during the database surveillance period 2000–2002. No eye injuries from swords, including toy swords and fencing foils, were reported.Conclusion: Ocular perforation sometimes occur from writing instruments that are thrown in the community, especially by children. Implications for policy and prevention are discussed. Non-specialists should have a low threshold for referring patients with eye injuries if suspicious of ocular penetration, even where caused by everyday objects, such as writing instruments.Keywords: eye injury, eye, children, mechanism, writing instruments, prevention

  5. Penetration through a wall: Is it reality?

    CERN Document Server

    Ivlev, B

    2011-01-01

    A tennis ball is not expected to penetrate through a brick wall since a motion under a barrier is impossible in classical mechanics. With quantum effects a motion of a particle through a barrier is allowed due to quantum tunneling. According to usual theories of tunneling, the particle density decays inside a classical barrier resulting in an extremely slow pentration process. However, there are no general laws forbidding fast motion through classical barriers. The problem addressed is investigation of unusual features o quantum tunneling through a classic static barrier which is at least two-dimensional. Here we show that penetration through such barrier can be not slow. When the barrier satisfies the certain conditions, a regime of quantum lens is possible with formation of caustics. De Broglie waves are reflected from the caustics, interfere, and result in a not small flux from under the barrier. This strongly contrasts to the usual scenario with a decaying under-barrier density. We construct a particular ...

  6. Visual outcome in patients undergoing penetrating keratoplasty

    Directory of Open Access Journals (Sweden)

    Gurpal Singh

    2015-01-01

    Methods: Thirty patients were selected from department of ophthalmology, Guru Gobind Singh medical college and hospital, Faridkot for penetrating keratoplasty. Results: Visual acuity of 6/12 was seen in 1 case, 6/18 in 4 cases, 6/36 in 5 cases, 6/60 in 5 cases, 3/60 in 4 cases, hand movements in 4 cases, PL+PR+ in 2 cases and no perception of light in 2 cases. Final vision at 6 months of follow up was >6/60 in 10 cases (33.33%. 5 cases (16.67% had vision between 6/60-4/60. 13 cases (43.33% had vision of 3/60 or less. While in 2 cases (6.67% there was no vision at the end of follow up. Conclusion: Penetrating keratoplasty helps patients in regaining vision which makes them able to move independently. Although in our study visual outcome of more than 3/60 was present in only 50% cases. [Int J Res Med Sci 2015; 3(1.000: 244-249

  7. Long-rod penetration:the transition zone between rigid and hydrodynamic penetration modes

    Institute of Scientific and Technical Information of China (English)

    Jian-feng LOU; Yan-geng ZHANG; Zheng WANG; Tao HONG; Xiao-li ZHANG; Shu-dao ZHANG

    2014-01-01

    Long-rod penetration in a wide range of velocity means that the initial impact velocity varies in a range from tens of meters per second to several kilometers per second. The long rods maintain rigid state when the impact velocity is low, the nose of rod deforms and even is blunted when the velocity gets higher, and the nose erodes and fails to lead to the consumption of long projectile when the velocity is very high due to instantaneous high pressure. That is, from low velocity to high velocity, the projectile undergoes rigid rods, deforming non-erosive rods, and erosive rods. Because of the complicated changes of the projectile, no well-established theoretical model and numerical simulation have been used to study the transition zone. Based on the analysis of penetration behavior in the transition zone, a phenomenological model to describe target resistance and a formula to calculate penetration depth in transition zone are proposed, and a method to obtain the boundary velocity of transition zone is determined. A combined theoretical analysis model for three response regions is built by analyzing the characteristics in these regions. The penetration depth predicted by this combined model is in good agreement with experimental result.

  8. Stylet penetration of Cacopsylla pyri; an electrical penetration graph (EPG) study

    NARCIS (Netherlands)

    Civolani, S.; Leis, M.; Grandi, G.; Garzo, E.; Pasqualini, E.; Musacchi, S.; Chicca, M.; Castaldelli, G.; Rossie, M.; Tjallingii, W.F.

    2011-01-01

    Detailed information on plant penetration activities by pear psylla Cacopsylla pyri L. (Hemiptera Psyllidae) is essential to study phytoplasma transmission of “Candidatus Phytoplasma pyri” responsible of pear decline disease (PD) and to trace and evaluate resistant traits in new pear tree selections

  9. Electromagnetic modelling of Ground Penetrating Radar responses to complex targets

    Science.gov (United States)

    Pajewski, Lara; Giannopoulos, Antonis

    2014-05-01

    This work deals with the electromagnetic modelling of composite structures for Ground Penetrating Radar (GPR) applications. It was developed within the Short-Term Scientific Mission ECOST-STSM-TU1208-211013-035660, funded by COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar". The Authors define a set of test concrete structures, hereinafter called cells. The size of each cell is 60 x 100 x 18 cm and the content varies with growing complexity, from a simple cell with few rebars of different diameters embedded in concrete at increasing depths, to a final cell with a quite complicated pattern, including a layer of tendons between two overlying meshes of rebars. Other cells, of intermediate complexity, contain pvc ducts (air filled or hosting rebars), steel objects commonly used in civil engineering (as a pipe, an angle bar, a box section and an u-channel), as well as void and honeycombing defects. One of the cells has a steel mesh embedded in it, overlying two rebars placed diagonally across the comers of the structure. Two cells include a couple of rebars bent into a right angle and placed on top of each other, with a square/round circle lying at the base of the concrete slab. Inspiration for some of these cells is taken from the very interesting experimental work presented in Ref. [1]. For each cell, a subset of models with growing complexity is defined, starting from a simple representation of the cell and ending with a more realistic one. In particular, the model's complexity increases from the geometrical point of view, as well as in terms of how the constitutive parameters of involved media and GPR antennas are described. Some cells can be simulated in both two and three dimensions; the concrete slab can be approximated as a finite-thickness layer having infinite extension on the transverse plane, thus neglecting how edges affect radargrams, or else its finite size can be fully taken into account. The permittivity of concrete can be

  10. Intraductal Papillary-Mucinous Neoplasm of the Pancreas Penetrating to the Stomach and the Common Bile Duct

    Directory of Open Access Journals (Sweden)

    Norihiro Goto

    2012-01-01

    Full Text Available Context Intraductal papillary mucinous neoplasm (IPMN of the pancreas occasionally penetrates to others organs. We present a case of IPMN penetrating to the stomach and the common bile duct. Case report A 75-year-old man was admitted to the hospital because of epigastric pain. Computed tomography (CT showed a papillary tumor protruding into the markedly dilated main pancreatic duct and splenic vein obstruction. The tumor was diagnosed as IPMN arising in the main duct, but he rejected surgery and he was followed without treatment. One year later, gastroduodenoscopy revealed gastropancreatic fistula and we were able to pass an endoscope through the fistula and directly examine the lumen of the main pancreatic duct and the papillary tumor adjacent to the fistula. Absence of malignant cells on histopathology suggested mechanical penetration rather than invasive penetration. CT showed splenic vein reperfusion due to decreased inner pressure of the main pancreatic duct. Two and a half years later, CT revealed biliopancreatic fistula formation. Endoscope biliary drainage was performed but failed. Despite jaundice, he is still ambulatory and seen in the clinic three years after the first admission. Conclusions We have experienced a case of IPMN penetrating to the stomach and the common bile duct that has taken a slow course. It represents the importance of distinguishing mechanical penetration from invasive penetration as well as mechanical splenic vein obstruction from splenic vein invasion.

  11. Design and analysis of firewall-penetrated scheme based on trusted host

    Institute of Scientific and Technical Information of China (English)

    ZHANG Deng-yin; WANG Qian-qian; XU Jian

    2007-01-01

    On the basis of various firewall-penetrated technologies, a novel firewall-penetrated technology based on trusted host, which can realize the communication between penetrating host and protected host, is proposed in this paper.Meanwhile, the firewall-penetrated evaluating model and its formalized statement is also proposed by quantizing penetrated level, penetrated quality, and penetrated hiding. Finally, the firewall-penetrated technology based on trusted host is analyzed under the firewall-penetrated evaluating model, and the experimental results show that this technology has obvious penetrating effect and high penetrating quality.

  12. Penetrating Stab Wound of the Right Ventricle

    Directory of Open Access Journals (Sweden)

    Onursal Buğra

    2010-04-01

    Full Text Available 18 years old male patient was admitted to our emergency unit with a penetrating stab wound to the right ventricle. A stab wound to the right ventricle was found to be 3 cm in diameter. The bleeding was controlled by insertion of a Foley catheter and inflation of the balloon. The stab wound had transected distal acute marginal side ofthe right coronary artery. A successful repair was performed with the use of a foley catheter and application of the Medtronic Octopus Tissue Stabilization System. The wound was closed with pledgeted mattress sutures. The distal acute marginal side of the right coronary artery was ligated. In this presentation, the surgical intervention method was reported and followed by a discussion of emergency surgical procedures of the heart.

  13. Penetration resistance of reinforced concrete containment structures

    International Nuclear Information System (INIS)

    Containment structures not only provide a leak tight barrier, but also play a role in ensuring that the structures can withstand the impact load from projectile impacts or internal plant accidents. In assessing the containment structures of nuclear power plants, predicting the characteristics of impact resistance in relation to design and safety considerations is relevant. This investigation proposes a simple but effective method of performing numerical analysis on perforation resistance of reinforced concrete containment structures. In this work, normal and oblique impacting is considered to examine the residual velocity and impact phenomena of an ogive-nose steel projectile with various impact velocities against a reinforced concrete slab. Additionally, a phase diagram is devised to describe the ballistic terminal phenomena of projectile and target. This model could assess the resistance to penetration to results in the optimum design of the containment structures in nuclear power plants

  14. Obstacle penetrating dynamic radar imaging system

    Science.gov (United States)

    Romero, Carlos E.; Zumstein, James E.; Chang, John T.; Leach, Jr.. Richard R.

    2006-12-12

    An obstacle penetrating dynamic radar imaging system for the detection, tracking, and imaging of an individual, animal, or object comprising a multiplicity of low power ultra wideband radar units that produce a set of return radar signals from the individual, animal, or object, and a processing system for said set of return radar signals for detection, tracking, and imaging of the individual, animal, or object. The system provides a radar video system for detecting and tracking an individual, animal, or object by producing a set of return radar signals from the individual, animal, or object with a multiplicity of low power ultra wideband radar units, and processing said set of return radar signals for detecting and tracking of the individual, animal, or object.

  15. Civil engineering applications of ground penetrating radar

    CERN Document Server

    Pajewski, Lara

    2015-01-01

    This book, based on Transport and Urban Development COST Action TU1208, presents the most advanced applications of ground penetrating radar (GPR) in a civil engineering context, with documentation of instrumentation, methods, and results. It explains clearly how GPR can be employed for the surveying of critical transport infrastructure, such as roads, pavements, bridges, and tunnels, and for the sensing and mapping of underground utilities and voids. Detailed attention is also devoted to use of GPR in the inspection of geological structures and of construction materials and structures, including reinforced concrete, steel reinforcing bars, and pre/post-tensioned stressing ducts. Advanced methods for solution of electromagnetic scattering problems and new data processing techniques are also presented. Readers will come to appreciate that GPR is a safe, advanced, nondestructive, and noninvasive imaging technique that can be effectively used for the inspection of composite structures and the performance of diagn...

  16. Attack Penetration System for SQL Injection

    Directory of Open Access Journals (Sweden)

    Manju Kaushik

    2014-06-01

    Full Text Available The SQL Injection attack is a popular way of attack in terms of document structure and common threats now a day. There are several ways of attack detection as per our study and also prevention methods had been discussed in several research papers. So the main motivation of our paper to penetrate the attack. For this we have proposed an efficient framework from which the central authority can control all the visited IP and restrict those IP. If the IP is in the restricted zone then the contents are restricted by using SQL update command with some status modification states. If the restricted IP wish to access the data from their credentials, it is immediately inform to the admin and the time of attack alert will be stored in the log area of that attack. Finally by comparison we will justify our results which are better in comparison to the previous test and methodology.

  17. Wide band stepped frequency ground penetrating radar

    Science.gov (United States)

    Bashforth, Michael B.; Gardner, Duane; Patrick, Douglas; Lewallen, Tricia A.; Nammath, Sharyn R.; Painter, Kelly D.; Vadnais, Kenneth G.

    1996-01-01

    A wide band ground penetrating radar system (10) embodying a method wherein a series of radio frequency signals (60) is produced by a single radio frequency source (16) and provided to a transmit antenna (26) for transmission to a target (54) and reflection therefrom to a receive antenna (28). A phase modulator (18) modulates those portion of the radio frequency signals (62) to be transmitted and the reflected modulated signal (62) is combined in a mixer (34) with the original radio frequency signal (60) to produce a resultant signal (53) which is demodulated to produce a series of direct current voltage signals (66) the envelope of which forms a cosine wave shaped plot (68) which is processed by a Fast Fourier Transform unit 44 into frequency domain data (70) wherein the position of a preponderant frequency is indicative of distance to the target (54) and magnitude is indicative of the signature of the target (54).

  18. Cephalic Tetanus from Penetrating Orbital Wound

    Directory of Open Access Journals (Sweden)

    Eloïse Guyennet

    2009-01-01

    Full Text Available Tetanus is a neurologic disorder caused by tetanospasmin, a protein toxin elaborated by Clostridium tetani. Cephalic tetanus is a localized form of the disease causing trismus and dysfunction of cranial nerves. We report the case of a man who presented with facial trauma, complete ophthalmoplegia, exophthalmos, areactive mydriasis, and periorbital hematoma. An orbital CT revealed air bubbles in the right orbital apex. The patient was given a tetanus toxoid booster and antibiotherapy. After extraction of a wooden foreign body, the patient developed right facial nerve palsy, disorders of swallowing, contralateral III cranial nerve palsy, and trismus. Only one case of cephalic tetanus from penetrating orbital wound has been reported in literature 20 years ago. When a patient presents with an orbital wound with ophthalmoplegia and signs of anaerobic infection, cephalic tetanus should be ruled out.

  19. Moxibustion Penetration and Its Clinical Application

    Institute of Scientific and Technical Information of China (English)

    GAO Xi-yan

    2008-01-01

    Moxibustion penetration is a therapy in which the moxibustion sensation promoted by moxa is felt deeply inside the tissues.When moxibustion is practiced to certain extent,the sensation conducts from the moxibustion site to the deep tissues and far ends,or muscular pulsation appears at the moxibustion site,or the patient feels comfortable,painful,heavy and itching or the patient has reaction as skin redness and even perspiration throughout the body after the moxibustion.The best effect of moxibustion is expected whenlocal site becomes slightly and evenly red or there is sweating.The observation has shown that the moxibustion dosage is the key to enhancing the therapeutic effect.

  20. Aerodynamic Jump for Long Rod Penetrators

    Directory of Open Access Journals (Sweden)

    Mark L. Bundy

    2000-04-01

    Full Text Available Aerodynamic jump for a non-spinning kinetic energy penetrator is neither a discontinuous change in the ,direction of motion at the origin of free night, nor is it the converse, i.e. a cumulativer~direc4on over a domain of infinite extent. Rather aerodynamic jump, for such a projectile, is a localised redirection of the centre of gravity motion, caused ~ the force of lift due to yaw over ther4latively short region from entry into free flight until the yaw reaches its first maximum. The primary objective of this paper is to provide answtfrs to the questions like what is aerodynamic jump, what liauses it, !lnd wh~t aspects df the flight trajectory does it refer to, or account for .

  1. The full penetration hole as a stochastic process: controlling penetration depth in keyhole laser-welding processes

    Science.gov (United States)

    Blug, A.; Abt, F.; Nicolosi, L.; Heider, A.; Weber, R.; Carl, D.; Höfler, H.; Tetzlaff, R.

    2012-07-01

    Although laser-welding processes are frequently used in industrial production the quality control of these processes is not satisfactory yet. Until recently, the "full penetration hole" was presumed as an image feature which appears when the keyhole opens at the bottom of the work piece. Therefore it was used as an indicator for full penetration only. We used a novel camera based on "cellular neural networks" which enables measurements at frame rates up to 14 kHz. The results show that the occurrence of the full penetration hole can be described as a stochastic process. The probability to observe it increases near the full penetration state. In overlap joints, a very similar image feature appears when the penetration depth reaches the gap between the sheets. This stochastic process is exploited by a closed-loop system which controls penetration depth near the bottom of the work piece ("full penetration") or near the gap in overlap joints ("partial penetration"). It guides the welding process at the minimum laser power necessary for the required penetration depth. As a result, defects like spatters are reduced considerably and the penetration depth becomes independent of process drifts such as feeding rate or pollution on protection glasses.

  2. Enhancement of Fluorescent Probe Penetration into Tumors In Vivo Using Unseeded Inertial Cavitation.

    Science.gov (United States)

    Prieur, Fabrice; Pillon, Arnaud; Mestas, Jean-Louis; Cartron, Valérie; Cèbe, Patrick; Chansard, Nathalie; Lafond, Maxime; Lafon, Cyril

    2016-07-01

    Ultrasound-induced cavitation has found many applications in the field of cancer therapy. One of its beneficial effects is the enhancement of drug intake by tumor cells. Our group has developed a device that can create and control unseeded cavitation in tissue using ultrasound. We conducted experiments on tumor-bearing mice using our device to assess the impact of sonication on the penetration of fluorescent probes into tumor cells. We studied the influence of pressure level, timing of sonication and sonication duration on treatment efficiency. Our results indicate that fluorescent probes penetrate better into tumors exposed to ultrasound. The best results revealed an increase in penetration of 61% and were obtained when sonicating the tumor in presence of the probes with a peak negative pressure at focus of 19 MPa. At this pressure level, the treatment generated only minor skin damage. Treatments could be significantly accelerated as equivalent enhanced penetration of probes was achieved when multiplying the initial raster scan speed by a factor of four. PMID:27087691

  3. Transdermal drug delivery of labetalol hydrochloride: Feasibility and effect of penetration enhancers

    Directory of Open Access Journals (Sweden)

    Saqib Zafar

    2010-01-01

    Full Text Available Objectives : The objective of this study is to investigate the feasibility of transdermal drug delivery of Labetalol Hydrochloride (LHCl and to study the effect of different penetration enhancers on the skin permeability of LHCl. Methods : The permeability experiments were conducted using a horizontal glass diffusion cell with a diffusional area of 2.37 cm-2 on albino rat skin. The effect of various penetration enhancers namely turpentine oil, dimethyl formamide (DMF, menthol, dimethyl sulfoxide, pine oil, and 2-pyrollidone, and the effect of the concentration of drug and enhancer in the donor phase on the skin permeability of LHCl was studied. Results : The apparent partition coefficient of the drug was found to be 6.95, suggesting it to be a lipophilic drug. The preliminary skin permeation studies revealed that the permeation of LHCL through albino rat skin was moderate (Kp = 6.490 Χ 10 -2 cm hr -1 from isotonic phosphate buffer of pH 7.4. An appreciable increase in the LHCl permeability coefficient was observed on using a co-solvent (ethanol 95% with the penetration enhancers in the donor phase. DMSO (10% v/v was found to be the most effective enhancer for Labetalol hydrochloride (Enhancement Factor = 1.165. An increase in the concentration of drug and enhancer in the donor cell accentuated the permeability coefficient of LHCl. Conclusions : It was concluded that LHCl could be delivered via the dermal route with the use of 10% DMSO as the penetration enhancer.

  4. Spatial resolution of MFM measurements of penetration depth

    Science.gov (United States)

    Spanton, Eric; Luan, Lan; Kirtley, John; Moler, Kathryn

    2012-02-01

    The penetration depth and its temperature dependence are key ways to characterize superconductors. Measurements of the local Meissner response of a superconductor can determine the local penetration depth. To quantify the spatial resolution of such measurements, we seek to characterize the point spread function of magnetic force microscope (MFM) measurements of the penetration depth both numerically and experimentally. Modeling various geometries of MFM tips (pyramid, dipole, and long thin cylinder) in the presence of various geometries of spatial variation in the penetration depth (point variation, columnar defects, and planar defects or twin boundaries) shows the importance of the MFM tip geometry to achieving both excellent spatial resolution and quantitatively interpretable results. We compare these models to experimental data on pnictides and cuprates to set upper limits on the sub-micron-scale variation of the penetration depth. These results demonstrate both the feasibility and the technical challenges of submicron penetration depth mapping.

  5. Penetration of tamoxifen citrate loaded ethosomes and liposomes across human skin: a comparative study with confocal laser scanning microscopy.

    Science.gov (United States)

    Sarwa, Khomendra K; Suresh, Preeti K; Rudrapal, Mithun; Verma, Vinod K

    2014-01-01

    In the present study, ethosomal and liposomal formulations containing tamoxifen citrate were prepared and evaluated for their penetration properties in human cadaver skin using Franz diffusion cell and confocal laser scanning microscope (CLSM). The results clearly revealed that ethosomal vesicles showed a better drug permeation profile than that of liposomal vesicles. In addition, low fluorescence intensity in CLSM was recorded with liposomes as compared to ethosomes, indicating lower cumulative amount of drug permeation from liposomal vesicles. Furthermore, CLSM showed uniform fluorescence intensity across the entire depth of skin in ethosomal treatment, indicating high penetrability of ethosomal vesicles through human cadaver skin. In contrast, low penetrability of conventional liposomal vesicles was recorded as penetration was limited to the 7(th) section (i.e. upper epidermis layer) of skin as evident from visualization of intact liposomal vesicles in CLSM. PMID:24428443

  6. Depletion-induced forces and crowding in polymer-nanoparticle mixtures: Role of polymer shape fluctuations and penetrability

    Science.gov (United States)

    Lim, Wei Kang; Denton, Alan R.

    2016-01-01

    Depletion forces and macromolecular crowding govern the structure and function of biopolymers in biological cells and the properties of polymer nanocomposite materials. To isolate and analyze the influence of polymer shape fluctuations and penetrability on depletion-induced interactions and crowding by nanoparticles, we model polymers as effective penetrable ellipsoids, whose shapes fluctuate according to the probability distributions of the eigenvalues of the gyration tensor of an ideal random walk. Within this model, we apply Monte Carlo simulation methods to compute the depletion-induced potential of mean force between hard nanospheres and crowding-induced shape distributions of polymers in the protein limit, in which polymer coils can be easily penetrated by smaller nanospheres. By comparing depletion potentials from simulations of ellipsoidal and spherical polymer models with predictions of polymer field theory and free-volume theory, we show that polymer depletion-induced interactions and crowding depend sensitively on polymer shapes and penetrability, with important implications for bulk thermodynamic phase behavior.

  7. Penetration of tamoxifen citrate loaded ethosomes and liposomes across human skin: a comparative study with confocal laser scanning microscopy.

    Science.gov (United States)

    Sarwa, Khomendra K; Suresh, Preeti K; Rudrapal, Mithun; Verma, Vinod K

    2014-01-01

    In the present study, ethosomal and liposomal formulations containing tamoxifen citrate were prepared and evaluated for their penetration properties in human cadaver skin using Franz diffusion cell and confocal laser scanning microscope (CLSM). The results clearly revealed that ethosomal vesicles showed a better drug permeation profile than that of liposomal vesicles. In addition, low fluorescence intensity in CLSM was recorded with liposomes as compared to ethosomes, indicating lower cumulative amount of drug permeation from liposomal vesicles. Furthermore, CLSM showed uniform fluorescence intensity across the entire depth of skin in ethosomal treatment, indicating high penetrability of ethosomal vesicles through human cadaver skin. In contrast, low penetrability of conventional liposomal vesicles was recorded as penetration was limited to the 7(th) section (i.e. upper epidermis layer) of skin as evident from visualization of intact liposomal vesicles in CLSM.

  8. Seasonal changes in radiation penetration within mustard crop canopy

    OpenAIRE

    Adak, Tarun; Chakravarty, NVK

    2012-01-01

    Green leaf area index, dry matter production and economic seed yield are significantly influenced by dynamics of radiation penetration within oilseed crop canopy. Keeping this in view, following a two years field experiment with Indian mustard cultivars in IARI research farm, it was revealed that the radiation penetration at the early crop growth stage was high and then decreased within the canopy as the leaf area developed. The study indicated significant seasonal changes in radiation penetr...

  9. Building virtual pentesting labs for advanced penetration testing

    CERN Document Server

    Cardwell, Kevin

    2014-01-01

    Written in an easy-to-follow approach using hands-on examples, this book helps you create virtual environments for advanced penetration testing, enabling you to build a multi-layered architecture to include firewalls, IDS/IPS, web application firewalls, and endpoint protection, which is essential in the penetration testing world. If you are a penetration tester, security consultant, security test engineer, or analyst who wants to practice and perfect penetration testing skills by building virtual pen testing labs in varying industry scenarios, this is the book for you. This book is ideal if yo

  10. Genetic variability and molecular responses of root penetration in cotton.

    Science.gov (United States)

    Klueva; Joshi; Joshi; Wester; Zartman; Cantrell; Nguyen

    2000-06-12

    Compacted soils restrict root penetration hindering productivity. In this paper, genetic variability of cotton (Gossipium spp.) root capacity to penetrate hard soil layers and the patterns of gene expression during penetration event were investigated. To mimic hard soil layers, wax-petrolatum mixtures were used. Genetic variability among 27 cotton genotypes for the root capacity to penetrate wax-petrolatum disks of 500-700 g wax/kg of mixture was high indicating that breeding efforts targeted to improve this trait can be successful. In the root tips of a cotton strain with high root penetrating ability (G. hirsutum HS 200) which penetrated through wax-petrolatum disks (P), quantity of four polypeptides with molecular weights 35-66 kDa increased compared to those root tips which grew in the absence of mechanical impedance (NP). Differential display showed significant differences in the sets of mRNA expressed in P and NP roots. Out of a total of 917 cDNAs scored in the differential display experiment, 118 cDNAs, or 13%, were specific to P roots and hence could be associated with the root penetration event. Further detailed study of gene expression in penetrated roots will pinpoint molecular factors involved in root penetration ability in cotton. PMID:10773338

  11. Penetrance of Hypertrophic Cardiomyopathy in Children and Adolescents

    DEFF Research Database (Denmark)

    Jensen, Morten K; Havndrup, Ole; Christiansen, Michael;

    2013-01-01

    The penetrance of hypertrophic cardiomyopathy (HCM) during childhood and adolescence has been only sparsely described. We studied the penetrance of HCM and the short- and long-term outcomes of clinical screening and predictive genetic testing of child relatives of patients with HCM.......The penetrance of hypertrophic cardiomyopathy (HCM) during childhood and adolescence has been only sparsely described. We studied the penetrance of HCM and the short- and long-term outcomes of clinical screening and predictive genetic testing of child relatives of patients with HCM....

  12. Characterization of nuclear reactor containment penetrations. Preliminary report

    International Nuclear Information System (INIS)

    This report summarizes the survey work conducted by Argonne National Laboratory on the design and details of major penetrations in 22 nuclear power plants. The survey includes all containment types and materials in current use. It also includes details of all types of penetrations (except for electrical penetration assemblies and valves) and the seals and gaskets used in them. The report provides a test matrix for testing major penetrations and for testing seals and gaskets in order to evaluate their leakage potential under severe accident conditions

  13. Numerical Simulation of Rigid Projectile's Normally Penetrating into Granite Targets

    Institute of Scientific and Technical Information of China (English)

    张德志; 张向荣; 林俊德; 唐润棣

    2004-01-01

    The process of penetrating into granite was simulated by using program LS-DYNA3D. The granite was represented by the isotropic elastic-plastic model with failure criterion and the projectile was modeled by rigid model. The depth of penetration from simulations is identical with experiments. Penetration deceleration vs striking velocity was acquired at the same time, which can assist in the design of penetration weapons with payload and fuse. Through numerical simulation, that material model is considered with straightforward physical meaning, a few parameters which can be determined easily are more practical for engineering calculation along with experiments.

  14. Hacking and penetration testing with low power devices

    CERN Document Server

    Polstra, Philip

    2014-01-01

    Hacking and Penetration Testing with Low Power Devices shows you how to perform penetration tests using small, low-powered devices that are easily hidden and may be battery-powered. It shows how to use an army of devices, costing less than you might spend on a laptop, from distances of a mile or more. Hacking and Penetration Testing with Low Power Devices shows how to use devices running a version of The Deck, a full-featured penetration testing and forensics Linux distribution, and can run for days or weeks on batteries due to their low power consumption. Author Philip Polstra shows how to

  15. Consistent constitutive modeling of metallic target penetration using empirical, analytical, and numerical penetration models

    OpenAIRE

    John (Jack) P. Riegel III; David Davison

    2016-01-01

    Historically, there has been little correlation between the material properties used in (1) empirical formulae, (2) analytical formulations, and (3) numerical models. The various regressions and models may each provide excellent agreement for the depth of penetration into semi-infinite targets. But the input parameters for the empirically based procedures may have little in common with either the analytical model or the numerical model. This paper builds on previous work by Riegel and Anderso...

  16. Evaluating and Comparing of Three Penetrability Measuring Devices : Modified Filter Pump, Modified Penetrability Meter, and Short Slot

    OpenAIRE

    Ali Akbar, Saman; Al-Naddaf, Manar

    2015-01-01

    Rock grouting is a commonly used process for sealing rocks in tunnels to reduce water ingress. In order to achieve sufficient sealing level the grout must effectively penetrate into rock fractures while the limiting factor is filtration of cement based grout. Many devices and measuring methods have been developed to study filtration and to measure the penetrability. The filter pump and the penetrability meter are two of the most commonly used instruments for measuring filtration tendency in t...

  17. Ground Penetrating Radar Technologies in Ukraine

    Science.gov (United States)

    Pochanin, Gennadiy P.; Masalov, Sergey A.

    2014-05-01

    Transient electromagnetic fields are of great interest in Ukraine. The following topics are studied by research teams, with high-level achievements all over the world: (i) Ultra-Wide Band/Short-pulse radar techniques (IRE and LLC "Transient Technologies", for more information please visit http://applied.ire.kharkov.ua/radar%20systems_their%20components%20and%20relevant%20technologies_e.html and http://viy.ua); (ii) Ground Penetrating Radar (GPR) with stepped frequency sounding signals (IRE); (iii) Continuous-Wave (CW) radar with phase-shift keying signals (IRE); and (iv) Radio-wave interference investigation (Scientific and Technical Centre of The Subsurface Investigation, http://geophysics.ua). GPR applications are mainly in search works, for example GPR is often used to search for treasures. It is also used to identify leaks and diffusion of petroleum in soil, in storage areas, as well as for fault location of pipelines. Furthermore, GPR is used for the localization of underground utilities and for diagnostics of the technical state of hydro dams. Deeper GPR probing was performed to identify landslides in Crimea. Rescue radar with CW signal was designed in IRE to search for living people trapped under the rubble of collapsed buildings. The fourth version of this radar has been recently created, showing higher stability and noise immunity. Radio-wave interference investigation allows studying the soil down to tens of meters. It is possible to identify areas with increased conductivity (moisture) of the soil. LLC "Transient Technologies" is currently working with Shevchenko Kyiv University on a cooperation program in which the construction of a test site is one of the planned tasks. In the framework of this program, a GPR with a 300 MHz antenna was handed to the geological Faculty of the University. Employees of "Transient Technologies" held introductory lectures with a practical demonstration for students majoring in geophysics. The authors participated to GPR

  18. Penetration and Toxicity of Nanomaterials in Higher Plants

    Directory of Open Access Journals (Sweden)

    Giuseppe Chichiriccò

    2015-05-01

    Full Text Available Nanomaterials (NMs comprise either inorganic particles consisting of metals, oxides, and salts that exist in nature and may be also produced in the laboratory, or organic particles originating only from the laboratory, having at least one dimension between 1 and 100 nm in size. According to shape, size, surface area, and charge, NMs have different mechanical, chemical, electrical, and optical properties that make them suitable for technological and biomedical applications and thus they are being increasingly produced and modified. Despite their beneficial potential, their use may be hazardous to health owing to the capacity to enter the animal and plant body and interact with cells. Studies on NMs involve technologists, biologists, physicists, chemists, and ecologists, so there are numerous reports that are significantly raising the level of knowledge, especially in the field of nanotechnology; however, many aspects concerning nanobiology remain undiscovered, including the interactions with plant biomolecules. In this review we examine current knowledge on the ways in which NMs penetrate plant organs and interact with cells, with the aim of shedding light on the reactivity of NMs and toxicity to plants. These points are discussed critically to adjust the balance with regard to the risk to the health of the plants as well as providing some suggestions for new studies on this topic.

  19. GSTAMIDS ground-penetrating radar: hardware description

    Science.gov (United States)

    Sower, Gary D.; Eberly, John; Christy, Ed

    2001-10-01

    The Ground Standoff Mine Detection System (GSTAMIDS) is now in the Engineering, Manufacturing and Development (EMD) Block 0 phase for USA CECOM. The Mine Detection Subsystem (MDS) presently utilizes three different sensor technologies to detect buried anti-tank (AT) land mines; Ground Penetrating Radar (GPR), Pulsed Magnetic Induction (PMI), and passive infrared (IR). The GSTAMIDS hardware and software architectures are designed so that other technologies can readily be incorporated when and if they prove viable. Each sensor suite is designed to detect the buried mines and to discriminate against various clutter and background objects. Sensor data fusion of the outputs of the individual sensor suites then enhances the detection probability while reducing the false alarm rate from clutter objects. The metal detector is an essential tool for buried mine detection, as metal land mines still account for a large percentage of land mines. Technologies such as nuclear quadrupole resonance (NQR or QR) are presently being developed to detect or confirm the presence of explosive material in buried land mines, particularly the so-called plastic mines; unfortunately, the radio frequency signals required cannot penetrate into a metal land mine. The limitation of the metal detector is not in detection of the metal mines, but in the additional detection of metal clutter. A metal detector has been developed using singular value decomposition (SVD) extraction techniques to discriminate the mines from the clutter, thereby greatly reducing false alarm rates. This mine detector is designed to characterize the impulse response function of the metal objects, based on a parametric three-pole model of the response, and to use pattern recognition to determine the match of the responses to known mines. In addition to discrimination against clutter, the system can also generally tell one mine type from another. This paper describes the PMI sensor suite hardware and its physical incorporation

  20. Ice-Penetrating Robot for Scientific Exploration

    Science.gov (United States)

    Zimmerman, Wayne; Carsey, Frank; French, Lloyd

    2007-01-01

    The cryo-hydro integrated robotic penetrator system (CHIRPS) is a partially developed instrumentation system that includes a probe designed to deeply penetrate the European ice sheet in a search for signs of life. The CHIRPS could also be used on Earth for similar exploration of the polar ice caps especially at Lake Vostok in Antarctica. The CHIRPS probe advances downward by a combination of simple melting of ice (typically for upper, non-compacted layers of an ice sheet) or by a combination of melting of ice and pumping of meltwater (typically, for deeper, compacted layers). The heat and electric power for melting, pumping, and operating all of the onboard instrumentation and electronic circuitry are supplied by radioisotope power sources (RPSs) and thermoelectric converters energized by the RPSs. The instrumentation and electronic circuitry includes miniature guidance and control sensors and an advanced autonomous control system that has fault-management capabilities. The CHIRPS probe is about 1 m long and 15 cm in diameter. The RPSs generate a total thermal power of 1.8 kW. Initially, as this power melts the surrounding ice, a meltwater jacket about 1 mm thick forms around the probe. The center of gravity of the probe is well forward (down), so that the probe is vertically stabilized like a pendulum. Heat is circulated to the nose by means of miniature pumps and heat pipes. The probe melts ice to advance in a step-wise manner: Heat is applied to the nose to open up a melt void, then heat is applied to the side to allow the probe to slip down into the melt void. The melt void behind the probe is allowed to re-freeze. Four quadrant heaters on the nose and another four quadrant heaters on the rear (upper) surface of the probe are individually controllable for steering: Turning on two adjacent nose heaters on the nose and two adjacent heaters on the opposite side at the rear causes melt voids to form on opposing sides, such that the probe descends at an angle from

  1. Occult celiac disease prevents penetrance of hemochromatosis

    Institute of Scientific and Technical Information of China (English)

    Andreas Geier; Siegfried Matern; Carsten Gartung; Igor Theurl; Guenter Weiss; Frank Lammert; Christoph G. Dietrich; Ralf Weiskirchen; Heinz Zoller; Benita Hermanns

    2005-01-01

    AIM: To report a patient with C282Y homozygocity, depleted body iron and intestinal atrophy caused by celiac disease (CD) who experienced resolution of the enteropathy with subsequent normalization of iron metabolism upon glutenfree diet.METHODS: To obtain information on the tissue distribution and quantitative expression of proteins involved in duodenal iron trafficking, we determined the expression of divalent-metal transporter 1 (DMT1), ferroportin 1 (FP1) and transferrin receptor (TfR1) by means of immunohistochemistry and real-time PCR in duodenal biopsies of this patient.RESULTS: Whereas in hereditary hemochromatosis patients without CD, DMT1 expression was up-regulated leading to excessive uptake of iron, we identified a significant reduction in protein and mRNA expression of DMT1 as acompensatory mechanism in this patient with HH and CD.CONCLUSION: Occult CD may compensate tot increased DMT1 expression in a specific subset of individuals withhomozygous C282Y mutations in the hemochromatosis(HFE) gene, thus contributing to the low penetrance of HH.

  2. Road evaluation with ground penetrating radar

    Science.gov (United States)

    Saarenketo, Timo; Scullion, Tom

    2000-03-01

    This paper provides a status report of the Ground Penetrating Radar (GPR) highway applications based on studies conducted in both Scandinavia and the USA. After several years of research local transportation agencies are now beginning to implement GPR technology for both network and project level surveys. This paper summarizes the principles of operation of both ground-coupled and air-launched GPR systems together with a discussion of both signal processing and data interpretation techniques. In the area of subgrade soil evaluation GPR techniques have been used to nondestructively identify soil type, to estimate the thickness of overburden and to evaluate the compressibility and frost susceptibility of subgrade soil. In road structure surveys, GPR has been used to measure layer thickness, to detect subsurface defects and to evaluate base course quality. In quality control surveys, GPR techniques have been used for thickness measurements, to estimate air void content of asphalt surfaces and to detect mix segregation. Future developments are described where the technique has great potential in assisting pavement engineers with their new pavement designs and in determining the optimal repair strategies for deteriorated roadways.

  3. Surgical management of penetrating pulmonary injuries.

    Science.gov (United States)

    Petrone, Patrizio; Asensio, Juan A

    2009-01-01

    Chest injuries were reported as early as 3000 BC in the Edwin Smith Surgical Papyrus. Ancient Greek chronicles reveal that they had anatomic knowledge of the thoracic structures. Even in the ancient world, most of the therapeutic modalities for chest wounds and traumatic pulmonary injuries were developed during wartime. The majority of lung injuries can be managed non-operatively, but pulmonary injuries that require operative surgical intervention can be quite challenging. Recent progress in treating severe pulmonary injuries has relied on finding shorter and simpler lung-sparing techniques. The applicability of stapled pulmonary tractotomy was confirmed as a safe and valuable procedure. Advancement in technology have revolutionized thoracic surgery and ushered in the era of video-assisted thoracoscopic surgery (VATS), providing an alternative method for accurate and direct evaluation of the lung parenchyma, mediastinum, and diaphragmatic injuries. The aim of this article is to describe the incidence of the penetrating pulmonary injuries, the ultimate techniques used in its operative management, as well as the diagnosis, complications, and morbidity and mortality. PMID:19236703

  4. Surgical management of penetrating pulmonary injuries

    Directory of Open Access Journals (Sweden)

    Asensio Juan A

    2009-02-01

    Full Text Available Abstract Chest injuries were reported as early as 3000 BC in the Edwin Smith Surgical Papyrus. Ancient Greek chronicles reveal that they had anatomic knowledge of the thoracic structures. Even in the ancient world, most of the therapeutic modalities for chest wounds and traumatic pulmonary injuries were developed during wartime. The majority of lung injuries can be managed non-operatively, but pulmonary injuries that require operative surgical intervention can be quite challenging. Recent progress in treating severe pulmonary injuries has relied on finding shorter and simpler lung-sparing techniques. The applicability of stapled pulmonary tractotomy was confirmed as a safe and valuable procedure. Advancement in technology have revolutionized thoracic surgery and ushered in the era of video-assisted thoracoscopic surgery (VATS, providing an alternative method for accurate and direct evaluation of the lung parenchyma, mediastinum, and diaphragmatic injuries. The aim of this article is to describe the incidence of the penetrating pulmonary injuries, the ultimate techniques used in its operative management, as well as the diagnosis, complications, and morbidity and mortality.

  5. Low Force Icy Regolith Penetration Technology

    Science.gov (United States)

    Metzger, P. T.; Galloway, G. M.; Mantovani, J. G.; Zacny, K.; Zacny, Kris; Craft, Jack

    2011-01-01

    Recent data from the Moon, including LCROSS data, indicate large quantities of water ice and other volatiles frozen into the soil in the permanently shadowed craters near the poles. If verified and exploited, these volatiles will revolutionize spaceflight as an inexpensive source of propellants and other consumables outside Earth's gravity well. This report discusses a preliminary investigation of a method to insert a sensor through such a soiVice mixture to verify the presence, nature, and concentration of the ice. It uses percussion to deliver mechanical energy into the frozen mixture, breaking up the ice and decompacting the soil so that only low reaction forces are required from a rover or spacecraft to push the sensor downward. The tests demonstrate that this method may be ideal for a small platform in lunar gravity. However, there are some cases where the system may not be able to penetrate the icy soil, and there is some risk ofthe sensor becoming stuck so that it cannot be retracted, so further work is needed. A companion project (ISDS for Water Detection on the Lunar Surface) has performed preliminary investigation of a dielectric/thermal sensor for use with this system.

  6. Interpretation of the lime column penetration test

    Science.gov (United States)

    Liyanapathirana, D. S.; Kelly, R. B.

    2010-06-01

    Dry soil mix (DSM) columns are used to reduce the settlement and to improve the stability of embankments constructed on soft clays. During construction the shear strength of the columns needs to be confirmed for compliance with technical assumptions. A specialized blade shaped penetrometer known as the lime column probe, has been developed for testing DSM columns. This test can be carried out as a pull out resistance test (PORT) or a push in resistance test (PIRT). The test is considered to be more representative of average column shear strength than methods that test only a limited area of the column. Both PORT and PIRT tests require empirical correlations of measured resistance to an absolute measure of shear strength, in a similar manner to the cone penetration test. In this paper, finite element method is used to assess the probe factor, N, for the PORT test. Due to the large soil deformations around the probe, an Arbitrary Lagrangian Eulerian (ALE) based finite element formulation has been used. Variation of N with rigidity index and the friction at the probe-soil interface are investigated to establish a range for the probe factor.

  7. Improving Web Application Security Using Penetration Testing

    Directory of Open Access Journals (Sweden)

    D. SriNithi

    2014-08-01

    Full Text Available The main issues of current web application is easily hacking the user information by unauthorized person. The development of entire web application depends on scripting languages that easily displays the user authentication code to the web browser. All code must be transferred through query string parameter (URL of the web application. This kind of application security fails when verifying it by penetration testing which is based on XSS languages. This study overcomes the security issues by developing a web application based on cross site scripting technique which the user codes are encrypted using RSA algorithm and cookies, cross domain verification based on encrypted use code. XSS vulnerabilities come in different forms and may be categorized into two varieties: reflected and stored. Reflected is on type of attack which can be performed against applications that employ a dynamic page error message to users. Stored XSS vulnerability appears when data submitted by one user is store in the application or in the back-end database. The user cookies of the web browser store only the encrypted key values. These techniques applied in Enterprise web application it support multiple organization for processing product purchase order, sales order and invoice details.

  8. FINITE ELEMENT METHOD ON NUMERICAL SIMULATION OF STRATUM CORNEUM'S PENETRATION PROPERTY

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    How the outer substance could penetrate through the skin lies in the stratum corneum, because it is the main barrier in the multi-layers of the skin. Supposing the keratin cell with a special geometry as tetrakaidecahedron, the penetration property of stratum corneum was the key problem which was numerically simulated with finite element method. At first the discretization of the stratum corneum region was given in two steps: first, the discretization of the keratin cell; second, the discretization of fattiness that surrounds the keratin. Then there was the work of numerical simulation. In this procedure, the finite element method and the multi-grid method were used. The former was to obtain the discretization of basic elements; the latter was to decrease the high frequency error. At last the visualization of the numerical simulation was shown.

  9. Tumor-Penetrating Peptide-Modified DNA Tetrahedron for Targeting Drug Delivery.

    Science.gov (United States)

    Xia, Zhiwei; Wang, Ping; Liu, Xunwei; Liu, Ting; Yan, Yinan; Yan, Juan; Zhong, Jian; Sun, Gang; He, Dannong

    2016-03-01

    DNA self-assembling nanostructure has been considered as a promising candidate as a drug delivery vehicle because of its compactness, mechanical stability, and noncytotoxicity. In this work, we developed functional, multiform DNA nanostructures by appending a tumor-penetrating peptide to tetrahedral DNA nanostructure (p-TDN). This functional structure is able to efficiently increase the rate of uptake of glioblastoma cell U87MG compared with the DNA tetrahedron and the double-stranded DNA structures. We found that the DNA tetrahedron plays the main role in the endocytosis of U87MG cells, whereas the tumor-penetrating peptide could also bind to transmembrane glycoprotein neuropilin-1 and mediate the endocytosis of the p-TDN nanostructure. Moreover, given the high efficiency of the growth inhibitory effect of the p-TDN loading doxorubicin hydrochloride, the p-TDN distinguishes itself as a promising candidate as an effective delivery carrier.

  10. Rock penetration : finite element sensitivity and probabilistic modeling analyses.

    Energy Technology Data Exchange (ETDEWEB)

    Fossum, Arlo Frederick

    2004-08-01

    This report summarizes numerical analyses conducted to assess the relative importance on penetration depth calculations of rock constitutive model physics features representing the presence of microscale flaws such as porosity and networks of microcracks and rock mass structural features. Three-dimensional, nonlinear, transient dynamic finite element penetration simulations are made with a realistic geomaterial constitutive model to determine which features have the most influence on penetration depth calculations. A baseline penetration calculation is made with a representative set of material parameters evaluated from measurements made from laboratory experiments conducted on a familiar sedimentary rock. Then, a sequence of perturbations of various material parameters allows an assessment to be made of the main penetration effects. A cumulative probability distribution function is calculated with the use of an advanced reliability method that makes use of this sensitivity database, probability density functions, and coefficients of variation of the key controlling parameters for penetration depth predictions. Thus the variability of the calculated penetration depth is known as a function of the variability of the input parameters. This simulation modeling capability should impact significantly the tools that are needed to design enhanced penetrator systems, support weapons effects studies, and directly address proposed HDBT defeat scenarios.

  11. Quantitative Penetration Testing with Item Response Theory (extended version)

    NARCIS (Netherlands)

    Arnold, F.; Pieters, W.; Stoelinga, M.I.A.

    2013-01-01

    Existing penetration testing approaches assess the vulnerability of a system by determining whether certain attack paths are possible in practice. Thus, penetration testing has so far been used as a qualitative research method. To enable quantitative approaches to security risk management, including

  12. Storage Possibilities for Enabling Higher Wind Energy Penetration

    DEFF Research Database (Denmark)

    Swierczynski, Maciej Jozef; Teodorescu, Remus; Rodriguez, Pedro;

    2010-01-01

    As the installed worldwide wind energy capacity increases about 30% annually and Kyoto protocol that came in force in 2005, wind penetration level in power system is considered to significantly increase in near future. Due to increased penetration and nature of the wind, especially its...

  13. A Study on Factors Affecting Airborne LiDAR Penetration

    Directory of Open Access Journals (Sweden)

    Wei-Chen Hsu

    2015-01-01

    Full Text Available This study uses data from different periods, areas and parameters of airborne LiDAR (light detection and ranging surveys to understand the factors that influence airborne LiDAR penetration rate. A discussion is presented on the relationships between these factors and LiDAR penetration rate. The results show that the flight height above ground level (AGL does not have any relationship with the penetration rate. There are some factors that should have larger influence. For example, the laser is affected by a wet ground surface by reducing the number of return echoes. The field of view (FOV has a slightly negative correlation with the penetration rate, which indicates that the laser incidence angle close to zero should achieve the best penetration. The vegetation cover rate also shows a negative correlation with the penetration rate, thus bare ground and reduced vegetation in the aftermath of a typhoon also cause high penetration rate. More return echoes could be extracted from the full-waveform system, thereby effectively improving the penetration rate. This study shows that full-waveform LiDAR is an effective method for increasing the number of surface reflected echoes. This study suggests avoiding LiDAR survey employment directly following precipitation to prevent laser echo reduction.

  14. Radionuclide transport through penetrations in nuclear waste containers

    International Nuclear Information System (INIS)

    Penetrations may result from corrosion or cracking and may be through the container material or through deposits of corrosion products. The analysis deals with the resultant radionuclide transport, but not with how these penetrations occur. We provide numerical illustrations for diffusive nuclide flux through these apertures from mathematical expressions. 2 refs., 2 figs

  15. Flux penetrations into two- and three-dimensional nanostructured superconductors

    Energy Technology Data Exchange (ETDEWEB)

    Tamegai, T., E-mail: tamegai@ap.t.u-tokyo.ac.jp [Department of Applied Physics, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan); Tsuchiya, Y.; Tada, S.; Ibuka, J.; Mine, A.; Pyon, S. [Department of Applied Physics, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan); Mawatari, Y.; Nagasawa, S.; Hidaka, M.; Maezawa, M. [National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8568 (Japan)

    2014-08-15

    Highlights: • Flux penetrations into two- and three-dimensional nanostructured superconductors are observed by magneto-optical imaging. • In two-dimensional superconducting networks with square holes on square lattice, anomalous diagonal penetrations are observed. • In three-dimensional superconducting shifted strip arrays, various forms of vortex avalanches are observed. - Abstract: We have fabricated two- and three-dimensional nanostructured superconductors, and observed vortex penetrations by magneto-optical imaging. In the case of two-dimensional superconducting networks with square holes on a square lattice, anomalous diagonal penetrations are widely observed. Two kinds of diagonal vortex penetrations at high and low temperatures have been interpreted as originating from the repulsive interaction of vortices and sharp fan-shaped vortex penetration from the corners of the square holes, respectively. In the case of three-dimensional stack of superconducting strip arrays with double and triple layers, vortex avalanches have been observed in a wide temperature and dimension ranges due to enhanced demagnetization effect. While spotlike avalanches are observed when the overlap of strips is small, anomalous linear avalanches traversing many strips in different layers are observed when the overlap is large. In triple-layer strip arrays, in addition to the spotlike and linear avalanches, vortex penetrations along the line of strips are also observed. Origins of the anomalous diagonal penetration and vortex avalanches are discussed.

  16. The Impact of IPv6 on Penetration Testing

    NARCIS (Netherlands)

    Ottow, Christiaan; Vliet, van Frank; Boer, de Pieter-Tjerk; Pras, Aiko

    2012-01-01

    In this paper we discuss the impact the use of IPv6 has on remote penetration testing of servers and web applications. Several modifications to the penetration testing process are proposed to accommodate IPv6. Among these modifications are ways of performing fragmentation attacks, host discovery and

  17. 3D Numerical Simulation of Projectile Penetration into Concrete Target

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Basing on the explicit instantaneous dynamics software MSC-Dytran and the general coupling arithmetic, the process of the projectile penetration into concrete target was simulated with the point-line-surface-body modeling method. Simulation results are in agreement with experimental results. The simulated data could provide design reference for the defense engineering construction and penetrator design.

  18. Influence of anatomical site and topical formulation on skin penetration of sunscreens.

    Science.gov (United States)

    Benson, Heather Ae; Sarveiya, Vikram; Risk, Stacey; Roberts, Michael S

    2005-09-01

    Sunscreen products are widely used to protect the skin from sun-related damage. Previous studies have shown that some sunscreen chemicals are absorbed across the skin to the systemic circulation. The current study shows that absorption into the skin of sunscreen chemicals applied to the face is up to four times greater than that of the same product applied to the back. This has implications for the way sunscreen products are formulated and may allow the use of less potent products on the face compared with the rest of the body. The effect of formulation vehicles on the release and skin penetration of the common sunscreen agent benzophenone-3 (common name oxybenzone) was also assessed. Penetration of benzophenone-3 across excised human epidermis and high-density polyethylene (HDPE) membrane was measured using in vitro Franz-type diffusion cells. Penetration and epidermal retention was measured following application of infinite and finite (epidermis only) doses of benzophenone-3 in five vehicles: liquid paraffin, coconut oil, 50:50 ethanol:coconut oil, aqueous cream BP, and oily cream BP. Highest benzophenone-3 skin retention was observed for the ethanol:coconut oil combination. Maximal and minimal benzophenone-3 fluxes were observed from liquid paraffin and coconut oil, respectively. The alcohol-based vehicle exhibited low benzophenone-3 release from the vehicle but high skin penetration and retention.

  19. Sperm penetration assay and its correlation with semen analysis parameters

    Directory of Open Access Journals (Sweden)

    Laxmi Kant Pandey

    2015-11-01

    Full Text Available Background: Aim of current study was to determine whether the Sperm Penetration Assay (SPA can be used as a test to discriminate the infertile male from fertile one. We have also correlated the SPA with semen analysis. Methods: Sperm characteristics namely Semen analysis and the sperm penetration assay were tested in 44 infertile and 10 fertile men. Sperm penetration assay was determined by using zona free hamster eggs. Results: With decreasing spermatozoa concentration in the semen there was significant decrease in percentage penetration of zona free Hamster eggs (p0.05. Conclusions: The Sperm penetration assay could discriminate the infertile group from fertile group significantly (p<0.001. The test appeared to be highly reproducible and probably identifies a truly infertile male. [Int J Res Med Sci 2015; 3(11.000: 3197-3201

  20. Standard practice for liquid penetrant examination for general industry

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2009-01-01

    1.1 This practice covers procedures for penetrant examination of materials. Penetrant testing is a nondestructive testing method for detecting discontinuities that are open to the surface such as cracks, seams, laps, cold shuts, shrinkage, laminations, through leaks, or lack of fusion and is applicable to in-process, final, and maintenance testing. It can be effectively used in the examination of nonporous, metallic materials, ferrous and nonferrous metals, and of nonmetallic materials such as nonporous glazed or fully densified ceramics, as well as certain nonporous plastics, and glass. 1.2 This practice also provides a reference: 1.2.1 By which a liquid penetrant examination process recommended or required by individual organizations can be reviewed to ascertain its applicability and completeness. 1.2.2 For use in the preparation of process specifications and procedures dealing with the liquid penetrant testing of parts and materials. Agreement by the customer requesting penetrant inspection is strongly rec...

  1. Evaporation and skin penetration characteristics of mosquito repellent formulations

    Energy Technology Data Exchange (ETDEWEB)

    Reifenrath, W.G.; Hawkins, G.S.; Kurtz, M.S.

    1989-03-01

    Formulations of the mosquito repellent N,N-diethyl-3-methylbenzamide (deet) in combination with a variety of additives were developed to control repellent evaporation and percutaneous penetration. Deet was also formulated with the repellent dimethyl phthalate to study the interaction of the two compounds on the skin. The evaporation and penetration processes were evaluated on whole and split-thickness pig skin using radiolabeled repellents with an in vitro apparatus. Under essentially still air and air flow conditions, one of the deet formulations resulted in significantly reduced total evaporation and percutaneous penetration of deet as compared to unformulated repellent. When deet and dimethyl phthalate were combined, neither repellent affected the total amount of evaporation and penetration of the other compound. However, initial percutaneous penetration and evaporation rates were slightly less and decayed less rapidly than when both chemicals were tested separately at the same dose. These results indicated a degree of competition of the two compounds for the same avenues of loss.

  2. The aerosol penetration through an electret fibrous filter

    Institute of Scientific and Technical Information of China (English)

    Jiang Wei; Cheung Chun-Shun; Chan Cheong-Ki; Zhu Chao

    2006-01-01

    The aim of this paper is to present a theoretical study of the aerosol penetration through an electret fibrous filter, using a numerical approach. The aerosol sizes considered in this study were in the submicron range, and in the numerical model, the conventional mechanical mechanisms (impaction, interception, diffusion and gravitationally settling) were taken into consideration along with the electrostatic mechanisms, including the Coulombic and dielectrophoretic effects. The aerosol penetration through an electret fibrous filter is heavily dependent on the aerosol penetration of a single fibre. The aerosol penetration through a single electret fibre under various filtration conditions was calculated. The effects of aerosol diameter, aerosol and fibre charge state, face velocity, packing density and aerosol dielectric constant on the aerosol penetration were investigated.

  3. Albumin-bound nanoparticle (nab) paclitaxel exhibits enhanced paclitaxel tissue distribution and tumor penetration

    OpenAIRE

    Chen, Nianhang; Brachmann, Carrie; Liu, Xiping; Pierce, Daniel W.; Dey, Joyoti; William S Kerwin; Li, Yan; Zhou, Simon; Hou, Shihe; Carleton, Michael; Klinghoffer, Richard A.; Palmisano, Maria; Chopra, Rajesh

    2015-01-01

    Purpose nab-paclitaxel demonstrates improved clinical efficacy compared with conventional Cremophor EL (CrEL)-paclitaxel in multiple tumor types. This study explored the distinctions in drug distribution between nab-paclitaxel and CrEL-paclitaxel and the underlying mechanisms. Methods Uptake and transcytosis of paclitaxel were analyzed by vascular permeability assay across human endothelial cell monolayers. The tissue penetration of paclitaxel within tumors was evaluated by local injections i...

  4. Irradiated survival effect of melanoma B16 cells at different penetration depths of intermediate energy heavy-ion beams%中能碳离子束对不同贯穿深度上黑色素瘤B16细胞的辐照存活效应

    Institute of Scientific and Technical Information of China (English)

    李强; 周光明; 王菊芳; 李文建; 温小琼; 党秉荣; 颉红梅; 卫增泉

    2001-01-01

    Survival fractions of melanoma B16 cells at different penetrationdepths exposed to 49.30MeV/u 12C ion beam and modulated 74.55MeV/u 12C ion beam with a ridge filter to form a spread-out Bragg peak (SOBP) were measured. The results show the survival response of B16 cells as the biological endpoint to heavy ions increases synchronously with the deposited energy along the beam traversal, that is, the survival fraction near or at the Bragg peak or SOBP is far lower than that at the beam's entrance or exit channel. Comparisons of the experimental data with calculated survival fractions at different depths predicted by the linear-quadratic survival model combined with dose-averaged LET display a good agreement within the experimental errors. It is prompted that the linear-quadratic model combined with dose-averaged LET possibly is a valid way to calculate the survival effect of B16 cells along the beam penetration under the exposure of the intermediate energy heavy-ion beam.%测量了49.30MeV/u及经脊形过滤器展宽的Bragg峰的74.55MeV/u12C离子束在不同贯穿深度上辐射后黑色素瘤B16细胞的存活率。结果显示,以存活为终点的B16细胞生物学效应与碳离子束能量沉积同步增大,即Bragg或展宽的Bragg峰区内细胞存活率远低于离子束入射或出射通道处的细胞存活率。通过实验测量与剂量平均LET结合的线性平方模型理论计算存活曲线的比较,发现在实验测量误差范围内两者符合较好。提示:剂量平均LET结合线性平方模型可能是预测中能重离子束贯穿深度上B16细胞存活效应的一种有效的方法。

  5. Investigation of the Efficacy of Transdermal Penetration Enhancers Through the Use of Human Skin and a Skin Mimic Artificial Membrane.

    Science.gov (United States)

    Balázs, Boglárka; Vizserálek, Gábor; Berkó, Szilvia; Budai-Szűcs, Mária; Kelemen, András; Sinkó, Bálint; Takács-Novák, Krisztina; Szabó-Révész, Piroska; Csányi, Erzsébet

    2016-03-01

    The aim of this study was to investigate the behavior of promising penetration enhancers through the use of 2 different skin test systems. Hydrogel-based transdermal formulations were developed with ibuprofen as a nonsteroidal anti-inflammatory drug. Transcutol and sucrose esters were used as biocompatible penetration enhancers. The permeability measurements were performed with ex vivo Franz diffusion cell methods and a newly developed Skin Parallel Artificial Membrane Permeability Assays (PAMPA) model. Franz diffusion measurement is commonly used as a research tool in studies of diffusion through synthetic membranes in vitro or penetration through ex vivo human skin, whereas Skin PAMPA involves recently published artificial membrane-based technology for the fast prediction of skin penetration. It is a 96-well plate-based model with optimized artificial membrane structure containing free fatty acid, cholesterol, and synthetic ceramide analog compounds to mimic the stratum corneum barrier function. Transdermal preparations containing 2.64% of different sucrose esters and/or Transcutol and a constant (5%) of ibuprofen were investigated to determine the effects of these penetration enhancers. The study demonstrated the good correlation of the permeability data obtained through use of human skin membrane and the in vitro Skin PAMPA system. The Skin PAMPA artificial membrane serves as quick and relatively deep tool in the early stages of transdermal delivery systems, through which the enhancing efficacy of excipients can be screened so as to facilitate the choice of effective penetration components. PMID:26886318

  6. Assessment of the percutaneous penetration of indomethacin from soybean oil microemulsion: effects of the HLB value of mixed surfactants.

    Science.gov (United States)

    Chen, Liangmei; Tan, Fengping; Wang, Jinfeng; Liu, Feng

    2012-01-01

    The objective of this study was to evaluate the influence of the ratios or the hydrophile-lipophile balance (HLB) values of Cremophor EL and Span 80 on the phase behavior of the O/W microemulsions and the percutaneous absorption and penetration of indomethacin microemulsions. The existence of microemulsion regions is investigated in quaternary systems composed of soybean oil/Cremophor EL and Span 80 (mixed surfactants)/diethylene glycol monoethyl ether (cosurfactant)/water by constructing pseudo-ternary phase diagrams at various Cremophor EL/Span 80 ratios. In addition, five microemulsion formulations with various mixed surfactants HLB values were evaluated by in vitro penetration experiments using mouse skin and Franz diffusion cells. The flux and amount of indomethacin penetration from 5 microemulsion formulations were significantly different from the control, and the enhance ratios ranged from 2.38 to 4.68 and 2.11 to 4.23, respectively. The HLB value of mixed surfactants in the formulations was a principal factor in determining the percutaneous penetration of the drug. The flux and amount of drug penetration increased gradually with increasing content of the lipophilic surfactant Span 80 and skin retention was highest for mixed surfactants with a HLB value of 7.6. Therefore, it is suggested that the presence of mixed surfactants was beneficial in the formation of O/W microemulsions and enhanced percutaneous penetration of indomethacin. PMID:22393828

  7. Investigation of the Efficacy of Transdermal Penetration Enhancers Through the Use of Human Skin and a Skin Mimic Artificial Membrane.

    Science.gov (United States)

    Balázs, Boglárka; Vizserálek, Gábor; Berkó, Szilvia; Budai-Szűcs, Mária; Kelemen, András; Sinkó, Bálint; Takács-Novák, Krisztina; Szabó-Révész, Piroska; Csányi, Erzsébet

    2016-03-01

    The aim of this study was to investigate the behavior of promising penetration enhancers through the use of 2 different skin test systems. Hydrogel-based transdermal formulations were developed with ibuprofen as a nonsteroidal anti-inflammatory drug. Transcutol and sucrose esters were used as biocompatible penetration enhancers. The permeability measurements were performed with ex vivo Franz diffusion cell methods and a newly developed Skin Parallel Artificial Membrane Permeability Assays (PAMPA) model. Franz diffusion measurement is commonly used as a research tool in studies of diffusion through synthetic membranes in vitro or penetration through ex vivo human skin, whereas Skin PAMPA involves recently published artificial membrane-based technology for the fast prediction of skin penetration. It is a 96-well plate-based model with optimized artificial membrane structure containing free fatty acid, cholesterol, and synthetic ceramide analog compounds to mimic the stratum corneum barrier function. Transdermal preparations containing 2.64% of different sucrose esters and/or Transcutol and a constant (5%) of ibuprofen were investigated to determine the effects of these penetration enhancers. The study demonstrated the good correlation of the permeability data obtained through use of human skin membrane and the in vitro Skin PAMPA system. The Skin PAMPA artificial membrane serves as quick and relatively deep tool in the early stages of transdermal delivery systems, through which the enhancing efficacy of excipients can be screened so as to facilitate the choice of effective penetration components.

  8. Consistent constitutive modeling of metallic target penetration using empirical, analytical, and numerical penetration models

    Directory of Open Access Journals (Sweden)

    John (Jack P. Riegel III

    2016-04-01

    Full Text Available Historically, there has been little correlation between the material properties used in (1 empirical formulae, (2 analytical formulations, and (3 numerical models. The various regressions and models may each provide excellent agreement for the depth of penetration into semi-infinite targets. But the input parameters for the empirically based procedures may have little in common with either the analytical model or the numerical model. This paper builds on previous work by Riegel and Anderson (2014 to show how the Effective Flow Stress (EFS strength model, based on empirical data, can be used as the average flow stress in the analytical Walker–Anderson Penetration model (WAPEN (Anderson and Walker, 1991 and how the same value may be utilized as an effective von Mises yield strength in numerical hydrocode simulations to predict the depth of penetration for eroding projectiles at impact velocities in the mechanical response regime of the materials. The method has the benefit of allowing the three techniques (empirical, analytical, and numerical to work in tandem. The empirical method can be used for many shot line calculations, but more advanced analytical or numerical models can be employed when necessary to address specific geometries such as edge effects or layering that are not treated by the simpler methods. Developing complete constitutive relationships for a material can be costly. If the only concern is depth of penetration, such a level of detail may not be required. The effective flow stress can be determined from a small set of depth of penetration experiments in many cases, especially for long penetrators such as the L/D = 10 ones considered here, making it a very practical approach. In the process of performing this effort, the authors considered numerical simulations by other researchers based on the same set of experimental data that the authors used for their empirical and analytical assessment. The goals were to establish a

  9. Consistent constitutive modeling of metallic target penetration using empirical, analytical, and numerical penetration models

    Institute of Scientific and Technical Information of China (English)

    John Jack P. RIEGEL III; David DAVISON

    2016-01-01

    Historically, there has been little correlation between the material properties used in (1) empirical formulae, (2) analytical formulations, and (3) numerical models. The various regressions and models may each provide excellent agreement for the depth of penetration into semi-infinite targets. But the input parameters for the empirically based procedures may have little in common with either the analytical model or the numerical model. This paper builds on previous work by Riegel and Anderson (2014) to show how the Effective Flow Stress (EFS) strength model, based on empirical data, can be used as the average flow stress in the analytical Walker–Anderson Penetration model (WAPEN) (Anderson and Walker, 1991) and how the same value may be utilized as an effective von Mises yield strength in numerical hydrocode simulations to predict the depth of penetration for eroding projectiles at impact velocities in the mechanical response regime of the materials. The method has the benefit of allowing the three techniques (empirical, analytical, and numerical) to work in tandem. The empirical method can be used for many shot line calculations, but more advanced analytical or numerical models can be employed when necessary to address specific geometries such as edge effects or layering that are not treated by the simpler methods. Developing complete constitutive relationships for a material can be costly. If the only concern is depth of penetration, such a level of detail may not be required. The effective flow stress can be determined from a small set of depth of penetration experiments in many cases, especially for long penetrators such as the L/D=10 ones considered here, making it a very practical approach. In the process of performing this effort, the authors considered numerical simulations by other researchers based on the same set of experimental data that the authors used for their empirical and analytical assessment. The goals were to establish a baseline with a full

  10. Bioconjugated Gold Nanoparticles Penetrate Into Spermatozoa Depending on Plasma Membrane Status.

    Science.gov (United States)

    Barchanski, Annette; Taylor, Ulrike; Sajti, Csaba L; Gamrad, Lisa; Kues, Wilfried A; Rath, Detlef; Barcikowski, Stephan

    2015-09-01

    Spermatozoa are not only essential for animal reproduction they also represent important tools for the manipulation of animal genetics. For instance, the genetic labeling and analysis of spermatozoa could provide a prospective complementation of pre-fertilization diagnosis and could help to prevent the inheritance of defective alleles during artificial insemination or to select beneficial traits in livestock. Spermatozoa feature extremely specialized membrane organization and restricted transport mechanisms making the labeling of genetically interesting DNA-sequences, e.g., with gold nanoparticles, a particular challenge. Here, we present a systematic study on the size-related internalization of ligand-free, monovalent and bivalent polydisperse gold nanoparticles, depending on spermatozoa membrane status. While monovalent conjugates were coupled solely to either negatively-charged oligonucleotides or positively-charged cell-penetrating peptides, bivalent conjugates were functionalized with both molecules simultaneously. The results clearly indicate that the cell membrane of acrosome-intact, bovine spermatozoa was neither permeable to ligand-free or oligonucleotide-conjugated nanoparticles, nor responsive to the mechanisms of cell-penetrating peptides. Interestingly, after acrosome reaction, which comprises major changes in sperm membrane composition, fluidity and charge, high numbers of monovalent and bivalent nanoparticles were found in the postequatorial segment, depicting a close and complex correlation between particle internalization and membrane organization. Additionally, depending on the applied peptide and for nanoparticle sizes < 10 nm even a successive nuclear penetration was observed, making the bivalent conjugates promising for future genetic delivery and sorting issues. PMID:26485929

  11. Diagnosis and treatment of penetrating craniocerebral injury

    Institute of Scientific and Technical Information of China (English)

    童鹰; 徐增斌; 詹仁雅; 刘凡隆

    2004-01-01

    Objective:To evaluate the clinical characteristics and present the experience in the treatment of patients with penetrating craniocerebral injury (PCCI).Methods: The data of 7 cases with PCCI by foreign body were retrospectively studied and compared with associated literatures. The strategies of diagnosis and treatment of PCCI were analyzed. In this series, 3 cases underwent emergency debridements and 4 cases underwent craniotomies. All patients received surgical intervention within 3 hours after admission.Results: Outcomes were good in 3 cases, moderate disability was in 2 cases, severe disability in 1 case and persistent vegetative state in 1 case. One case developed wound and intracranial infection, but made good recovery after treatment. During the follow-up period, one patient died one month after discharge and other six patients (range from 8 months to 3 years) recovered well and no epilepsy, leakage of cerebrospinal fluid (CSF), or traumatic vascular disease occurred.Conclusions: Early diagnosis and prompt debridement are the fundamental factors affecting the outcome of PCCI. CT scans are the mainstay in evaluating PCCI and three dimensional (3D) images reconstructed from spiral CT scans provide more information. Efficient debridement should be performed as early as possible.Minimizing the degree of surgical management of PCCI is preferred when there is no indication for aggressive operation. It is important to stress the rapid and effective management of CSF leakage in early stage of PCCI. Use of prophylactic broad-spectrum antibiotics is recommended for patients with PCCI. Traumatic vascular injury should be paid attention to after PCCI.

  12. Severe accident testing of electrical penetration assemblies

    International Nuclear Information System (INIS)

    This report describes the results of tests conducted on three different designs of full-size electrical penetration assemblies (EPAs) that are used in the containment buildings of nuclear power plants. The objective of the tests was to evaluate the behavior of the EPAs under simulated severe accident conditions using steam at elevated temperature and pressure. Leakage, temperature, and cable insulation resistance were monitored throughout the tests. Nuclear-qualified EPAs were produced from D. G. O'Brien, Westinghouse, and Conax. Severe-accident-sequence analysis was used to generate the severe accident conditions (SAC) for a large dry pressurized-water reactor (PWR), a boiling-water reactor (BWR) Mark I drywell, and a BWR Mark III wetwell. Based on a survey conducted by Sandia, each EPA was matched with the severe accident conditions for a specific reactor type. This included the type of containment that a particular EPA design was used in most frequently. Thus, the D. G. O'Brien EPA was chosen for the PWR SAC test, the Westinghouse was chosen for the Mark III test, and the Conax was chosen for the Mark I test. The EPAs were radiation and thermal aged to simulate the effects of a 40-year service life and loss-of-coolant accident (LOCA) before the SAC tests were conducted. The design, test preparations, conduct of the severe accident test, experimental results, posttest observations, and conclusions about the integrity and electrical performance of each EPA tested in this program are described in this report. In general, the leak integrity of the EPAs tested in this program was not compromised by severe accident loads. However, there was significant degradation in the insulation resistance of the cables, which could affect the electrical performance of equipment and devices inside containment at some point during the progression of a severe accident. 10 refs., 165 figs., 16 tabs

  13. Severe accident testing of electrical penetration assemblies

    Energy Technology Data Exchange (ETDEWEB)

    Clauss, D.B. (Sandia National Labs., Albuquerque, NM (USA))

    1989-11-01

    This report describes the results of tests conducted on three different designs of full-size electrical penetration assemblies (EPAs) that are used in the containment buildings of nuclear power plants. The objective of the tests was to evaluate the behavior of the EPAs under simulated severe accident conditions using steam at elevated temperature and pressure. Leakage, temperature, and cable insulation resistance were monitored throughout the tests. Nuclear-qualified EPAs were produced from D. G. O'Brien, Westinghouse, and Conax. Severe-accident-sequence analysis was used to generate the severe accident conditions (SAC) for a large dry pressurized-water reactor (PWR), a boiling-water reactor (BWR) Mark I drywell, and a BWR Mark III wetwell. Based on a survey conducted by Sandia, each EPA was matched with the severe accident conditions for a specific reactor type. This included the type of containment that a particular EPA design was used in most frequently. Thus, the D. G. O'Brien EPA was chosen for the PWR SAC test, the Westinghouse was chosen for the Mark III test, and the Conax was chosen for the Mark I test. The EPAs were radiation and thermal aged to simulate the effects of a 40-year service life and loss-of-coolant accident (LOCA) before the SAC tests were conducted. The design, test preparations, conduct of the severe accident test, experimental results, posttest observations, and conclusions about the integrity and electrical performance of each EPA tested in this program are described in this report. In general, the leak integrity of the EPAs tested in this program was not compromised by severe accident loads. However, there was significant degradation in the insulation resistance of the cables, which could affect the electrical performance of equipment and devices inside containment at some point during the progression of a severe accident. 10 refs., 165 figs., 16 tabs.

  14. Ground penetrating radar field evaluation in Angola

    Science.gov (United States)

    Walls, Richard; Brown, Todd; Clodfelter, Fred; Coors, Jeff; Laudato, Stephen; Lauziere, Steve; Patrikar, Ajay; Poole, Michael; Price, Mike

    2006-05-01

    Deminers around the globe are still using handheld metal detectors that lack the capability to distinguish mines from clutter, detect mines containing very little metal, or find mines buried at deeper depths. In the southern African country of Angola, many areas and roads are impassable due to the threat of anti-tank landmines. Some of these mines are undetectable using current metal detector technology. The US Army has funded the development of the NIITEK ground penetrating radar (GPR) for detection of anti-tank (AT) landmines. This radar detects metal and plastic mines as well as mines that are buried too deep for handheld metal detectors to find. The US Department of Defense Humanitarian Demining (HD) Research & Development Program focuses on developing, testing, demonstrating, and validating new technology for immediate use in humanitarian demining operations around the globe. The HD team provided funding and guidance to NIITEK Incorporated for development of a prototype system called Mine Stalker - a relatively light-weight, remote-controlled vehicle outfitted with the NIITEK GPR, detection algorithms, and a marking system. Individuals from the HD team, NIITEK Inc, and the non-governmental organization Meschen Gegen Minen (MgM) participated in a field evaluation of the Mine Stalker in Angola. The primary aim was to evaluate the effectiveness and reliability of the NIITEK GPR under field conditions. The Mine Stalker was extremely reliable during the evaluation with no significant maintenance issues. All AT mines used to verify GPR performance were detected, even when buried to depths as deep as 25-33cm.

  15. Groundwater contamination downstream of a contaminant penetration site. II. Horizontal penetration of the contaminant plume

    Science.gov (United States)

    Rubin, H.; Buddemeier, R.W.

    2002-01-01

    Part I of this study (Rubin, H.; Buddemeier, R.W. Groundwater Contamination Downstream of a Contaminant Penetration Site Part 1: Extension-Expansion of the Contaminant Plume. J. of Environmental Science and Health Part A (in press).) addressed cases, in which a comparatively thin contaminated region represented by boundary layers (BLs) developed within the freshwater aquifer close to contaminant penetration site. However, at some distance downstream from the penetration site, the top of the contaminant plume reaches the top or bottom of the aquifer. This is the location of the "attachment point," which comprises the entrance cross section of the domain evaluated by the present part of the study. It is shown that downstream from the entrance cross section, a set of two BLs develop in the aquifer, termed inner and outer BLs. It is assumed that the evaluated domain, in which the contaminant distribution gradually becomes uniform, can be divided into two sections, designated: (a) the restructuring section, and (b) the establishment section. In the restructuring section, the vertical concentration gradient leads to expansion of the inner BL at the expense of the outer BL, and there is almost no transfer of contaminant mass between the two layers. In the establishment section, each of the BLs occupies half of the aquifer thickness, and the vertical concentration gradient leads to transfer of contaminant mass from the inner to the outer BL. By use of BL approximations, changes of salinity distribution in the aquifer are calculated and evaluated. The establishment section ends at the uniformity point, downstream from which the contaminant concentration profile is practically uniform. The length of the restructuring section, as well as that of the establishment section, is approximately proportional to the aquifer thickness squared, and is inversely proportional to the transverse dispersivity. The study provides a convenient set of definitions and terminology that are

  16. Bone marrow-specific knock-in of a non-activatable Ikkα kinase mutant influences haematopoiesis but not atherosclerosis in Apoe-deficient mice.

    Directory of Open Access Journals (Sweden)

    Pathricia V Tilstam

    Full Text Available BACKGROUND: The Ikkα kinase, a subunit of the NF-κB-activating IKK complex, has emerged as an important regulator of inflammatory gene expression. However, the role of Ikkα-mediated phosphorylation in haematopoiesis and atherogenesis remains unexplored. In this study, we investigated the effect of a bone marrow (BM-specific activation-resistant Ikkα mutant knock-in on haematopoiesis and atherosclerosis in mice. METHODS AND RESULTS: Apolipoprotein E (Apoe-deficient mice were transplanted with BM carrying an activation-resistant Ikkα gene (Ikkα(AA/AAApoe(-/- or with Ikkα(+/+Apoe(-/- BM as control and were fed a high-cholesterol diet for 8 or 13 weeks. Interestingly, haematopoietic profiling by flow cytometry revealed a significant decrease in B-cells, regulatory T-cells and effector memory T-cells in Ikkα(AA/AAApoe(-/- BM-chimeras, whereas the naive T-cell population was increased. Surprisingly, no differences were observed in the size, stage or cellular composition of atherosclerotic lesions in the aorta and aortic root of Ikkα(AA/AAApoe(-/- vs Ikkα(+/+Apoe(-/- BM-transplanted mice, as shown by histological and immunofluorescent stainings. Necrotic core sizes, apoptosis, and intracellular lipid deposits in aortic root lesions were unaltered. In vitro, BM-derived macrophages from Ikkα(AA/AAApoe(-/- vs Ikkα(+/+Apoe(-/- mice did not show significant differences in the uptake of oxidized low-density lipoproteins (oxLDL, and, with the exception of Il-12, the secretion of inflammatory proteins in conditions of Tnf-α or oxLDL stimulation was not significantly altered. Furthermore, serum levels of inflammatory proteins as measured with a cytokine bead array were comparable. CONCLUSION: Our data reveal an important and previously unrecognized role of haematopoietic Ikkα kinase activation in the homeostasis of B-cells and regulatory T-cells. However, transplantation of Ikkα(AA mutant BM did not affect atherosclerosis in Apoe(-/- mice. This

  17. Ultrasonic enhancement of drug penetration in solid tumors

    Directory of Open Access Journals (Sweden)

    Chun-yen eLai

    2013-08-01

    Full Text Available Increasing the penetration of drugs within solid tumors can be accomplished through multiple ultrasound-mediated mechanisms. The application of ultrasound can directly change the structure or physiology of tissues or can induce changes in a drug or vehicle in order to enhance delivery and efficacy. With each ultrasonic pulse, a fraction of the energy in the propagating wave is absorbed by tissue and results in local heating. When ultrasound is applied to achieve mild hyperthermia, the thermal effects are associated with an increase in perfusion or the release of a drug from a temperature-sensitive vehicle. Higher ultrasound intensities locally ablate tissue and result in increased drug accumulation surrounding the ablated region of interest. Further, the mechanical displacement induced by the ultrasound pulse can result in the nucleation, growth and collapse of gas bubbles. As a result of such cavitation, the permeability of a vessel wall or cell membrane can be increased. Finally, the radiation pressure of the propagating pulse can translate particles or tissues. In this perspective, we will review recent progress in ultrasound-mediated tumor delivery and the opportunities for clinical translation.

  18. Investigation on penetration model of shaped charge jet in water

    Science.gov (United States)

    Shi, Jinwei; Luo, Xingbai; Li, Jinming; Jiang, Jianwei

    2016-01-01

    To analyze the process of jet penetration in water medium quantitatively, the properties of jet penetration spaced target with water interlayer were studied through test and numerical simulation. Two theoretical models of jet penetration in water were proposed. The theoretical model 1 was established considering the impact of the shock wave, combined with the shock equation Rankine-Hugoniot and the virtual origin calculation method. The theoretical model 2 was obtained by fitting theoretical analysis and numerical simulation results. The effectiveness and universality of the two theoretical models were compared through the numerical simulation results. Both the models can reflect the relationship between the penetration velocity and the penetration distance in water well, and both the deviation and stability of theoretical model 1 are better than 2, the lower penetration velocity, and the larger deviation of the theoretical model 2. Therefore, the theoretical model 1 can reflect the properties of jet penetration in water effectively, and provide the reference of model simulation and theoretical research.

  19. Proton Radiography of Shape Charge Jets Penetrating Teflon and Explosive

    Science.gov (United States)

    Ferm, Eric N.; Burkett, Michael W.; Hull, Larry M.; Marr-Lyon, Mark; McNeil, Wendy V.; Morris, Chris L.; Rightley, Paul M.; Lansce Proton Radiography Team

    2011-06-01

    We have used proton radiography at the Los Alamos Neutron Science Center to observe viper shaped charge jets penetrating inert and explosive materials. A viper jet was observed penetrating both Teflon and PBX 9501. Radiographs captured the penetration events at several times and are analyzed to determine the density of the materials imaged at each time. The interfaces and shock waves in the flow are clearly evident in the images. Multiple time images allow the determination of the velocities of the interfaces and shock waves. Comparisons are made in the Teflon case with estimates of penetration rates and densities using the quasi-steady approximation analysis used in many terminal ballistics models. The PBX 9501 clearly detonated from the impact of the shape charge jet tip traveling at 9.1 mm/s. The detonation wave is examined to see what support it obtains from the pursing jet and the jet is examined to find the influence of the explosive products on penetration velocity. This experiment gives us experimental results of in-situ penetration process that can be used to verify common modeling techniques and fluid mechanic calculations of the penetration process.

  20. USDC-based rapid penetrator of packed soil

    Science.gov (United States)

    Bao, X.; Bar-Cohen, Y.; Chang, Z.; Sherrit, S.; Badescu, M.; Du, S.; Song, T.; Peterson, T.

    2006-03-01

    Environment protection requires more testing and analysis tools. To detect buried chemical containers or other objects embedded in soil and avoid possible damage to them, a penetrator was developed for packed soil that requires low penetration force (the force needed to push rod probe into the soil). The design was based on the novel mechanism used by the ultrasonic/sonic driller/corer (USDC) that was developed jointly by scientists at the NDEAA lab at JPL and engineers at Cybersonics, Inc. [Bar-Cohen et al 2001, Bao et al 2003]. In the penetrator, a small free-flying mass is energized by a piezoelectric transducer and impacts a rod probe on its shoulder at frequencies of several hundred Hetz. The impacts help the probe to penetrate the packed soil with low pushing force. A large reduction of the penetration force was achieved. Preliminary tests show that the effects of the penetrator on plastic containers and other objectors are minimal. The details of the design of the prototype penetrator and the results of performance tests are presented.