Sample records for high cell permeability

  1. Water-permeability measurement of high performance concrete using a high-pressure triaxial cell

    El-Dieb, A.S. [Ain Shams Univ., Cairo (Egypt). Dept. of Civil Engineering; Hooton, R.D. [Univ. of Toronto, Ontario (Canada). Dept. of Civil Engineering


    Water permeability of concrete is used to indicate its durability. Accurate and reproducible measurement of water permeability is difficult and becomes more difficult as the quality of concrete increases. When high-performance concrete (HPC) is tested, these concerns become more pronounced. HPC is used widely to improve the durability and performance of structures but there are few test procedures able to evaluate its permeability-related properties. In this study the water permeabilities of concretes including HPC were measured using a high-pressure triaxial cell with a sensitive and automated measurement capability. Special analysis procedures were developed to obtain useful data from the extremely low volume of water being measured. This method was able to measure a wide range of permeability values from 10{sup {minus}12} m/s to 10{sup {minus}16} m/s, with reproducible measurements on replicates.

  2. Highly Potent Cell-Permeable and Impermeable NanoLuc Luciferase Inhibitors.

    Walker, Joel R; Hall, Mary P; Zimprich, Chad A; Robers, Matthew B; Duellman, Sarah J; Machleidt, Thomas; Rodriguez, Jacquelynn; Zhou, Wenhui


    Novel engineered NanoLuc (Nluc) luciferase being smaller, brighter, and superior to traditional firefly (Fluc) or Renilla (Rluc) provides a great opportunity for the development of numerous biological, biomedical, clinical, and food and environmental safety applications. This new platform created an urgent need for Nluc inhibitors that could allow selective bioluminescent suppression and multiplexing compatibility with existing luminescence or fluorescence assays. Starting from thienopyrrole carboxylate 1, a hit from a 42 000 PubChem compound library with a low micromolar IC50 against Nluc, we derivatized four different structural fragments to discover a family of potent, single digit nanomolar, cell permeable inhibitors. Further elaboration revealed a channel that allowed access to the external Nluc surface, resulting in a series of highly potent cell impermeable Nluc inhibitors with negatively charged groups likely extending to the protein surface. The permeability was evaluated by comparing EC50 shifts calculated from both live and lysed cells expressing Nluc cytosolically. Luminescence imaging further confirmed that cell permeable compounds inhibit both intracellular and extracellular Nluc, whereas less permeable compounds differentially inhibit extracellular Nluc and Nluc on the cell surface. The compounds displayed little to no toxicity to cells and high luciferase specificity, showing no activity against firefly luciferase or even the closely related NanoBit system. Looking forward, the structural motifs used to gain access to the Nluc surface can also be appended with other functional groups, and therefore interesting opportunities for developing assays based on relief-of-inhibition can be envisioned.

  3. A high pressure triaxial cell with improved measurement sensitivity for saturated water permeability of high performance concrete

    El-Dieb, A.S.; Hooton, R.D. (Univ. of Toronto, Ontario (Canada). Dept. of Civil Engineering)


    The measurement of the saturated water permeability of concrete is of great interest, but with the rapid improvements in properties of high performance concretes, the most common problem is the ability and accuracy of measuring the very small flow volumes. A high pressure triaxial cell with improved measurement sensitivity, capable of continuously measuring saturated water permeability of the order of < 10[sup [minus]15] m/s, is presented in this paper.

  4. Effects of serum of the rats ventilated with high tidal volume on endothelial cell permeability and therapeutic effects of ulinastatin

    HUO Guo-dong; CAI Shao-xi; CHEN Bo; CHEN Ying-hua


    Background With the widespread use of ventilators in treating critically ill patients, the morbidity of ventilator-induced lung injury (VILI) is increasing accordingly. VILI is characterized by a considerable increase in microvascular leakiness and activation of inflammatory processes. In this study we investigated the effects of inflammatory mediators in VILI rat serum on endothelial cytoskeleton and monolayer cellular permeability, as well as the therapeutic effect of ulinastatin, to explore the pathogenesis and the relationship between biotrauma and lung oedema induced by VILI.Methods Thirty healthy male Sprague-Dawley rats were randomly divided into three groups: group A (normal tidal volume ventilation), group B (high tidal volume ventilation) and group C (high tidal volume ventilation plus ulinastatin). The serum of each rat after ventilation was added to endothelial cell line ECV-304 medium for two hours to observe the effects of serum and/or ulinastatin on endothelial fibrous actin and permeability. Results Compared to rats ventilated with normal tidal volume, serum of rats ventilated with high tidal volume caused a striking reorganization of actin cytoskeleton with a weakening of fluorescent intensity at the peripheral filament bands and formation of the long and thick stress fibres in the centre resulting in endothelial contraction and higher permeability. Prior treatment with ulinastatin lessened the above changes significantly. The changes of permeability coefficient of endothelial permeability after group A, B or C rats serum stimulation were (6.95 ±1.66)%, (27.50±7.77)% and (17.71±4.66)% respectively with statistically significant differences (P<0.05)among the three groups.Conclusions The proinflammatory mediators in the serum of the rats given high tidal volume ventilation increases endothelial permeability by reorganizing actin cytoskeleton, and pretreatment with ulinastatin lessens the permeability by inhibiting of proinflammatory mediators.

  5. Non-Ionic Highly Permeable Polymer Shells for Encapsulation of Living Cells


    thermal gelation. Artif Cells Blood Substit Immobil Biotechnol 2004, 32 (2), 275-91. 2. Wilson, J. T.; Chaikof, E. L., Challenges and emerging...R.; Korsgren, O.; Nilsson, B., Islet surface heparinization prevents the instant blood -mediated inflammatory reaction in islet transplantation...R. J.; Poncelet, D., Encapsulation of brewers yeast in chitosan coated carrageenan microspheres by emulsification/thermal gelation. Artif Cells

  6. Engineered Trehalose Permeable to Mammalian Cells.

    Alireza Abazari

    Full Text Available Trehalose is a naturally occurring disaccharide which is associated with extraordinary stress-tolerance capacity in certain species of unicellular and multicellular organisms. In mammalian cells, presence of intra- and extracellular trehalose has been shown to confer improved tolerance against freezing and desiccation. Since mammalian cells do not synthesize nor import trehalose, the development of novel methods for efficient intracellular delivery of trehalose has been an ongoing investigation. Herein, we studied the membrane permeability of engineered lipophilic derivatives of trehalose. Trehalose conjugated with 6 acetyl groups (trehalose hexaacetate or 6-O-Ac-Tre demonstrated superior permeability in rat hepatocytes compared with regular trehalose, trehalose diacetate (2-O-Ac-Tre and trehalose tetraacetate (4-O-Ac-Tre. Once in the cell, intracellular esterases hydrolyzed the 6-O-Ac-Tre molecules, releasing free trehalose into the cytoplasm. The total concentration of intracellular trehalose (plus acetylated variants reached as high as 10 fold the extracellular concentration of 6-O-Ac-Tre, attaining concentrations suitable for applications in biopreservation. To describe this accumulation phenomenon, a diffusion-reaction model was proposed and the permeability and reaction kinetics of 6-O-Ac-Tre were determined by fitting to experimental data. Further studies suggested that the impact of the loading and the presence of intracellular trehalose on cellular viability and function were negligible. Engineering of trehalose chemical structure rather than manipulating the cell, is an innocuous, cell-friendly method for trehalose delivery, with demonstrated potential for trehalose loading in different types of cells and cell lines, and can facilitate the wide-spread application of trehalose as an intracellular protective agent in biopreservation studies.

  7. Intestinal permeability study of minoxidil: assessment of minoxidil as a high permeability reference drug for biopharmaceutics classification.

    Ozawa, Makoto; Tsume, Yasuhiro; Zur, Moran; Dahan, Arik; Amidon, Gordon L


    The purpose of this study was to evaluate minoxidil as a high permeability reference drug for Biopharmaceutics Classification System (BCS). The permeability of minoxidil was determined in in situ intestinal perfusion studies in rodents and permeability studies across Caco-2 cell monolayers. The permeability of minoxidil was compared with that of metoprolol, an FDA reference drug for BCS classification. In rat perfusion studies, the permeability of minoxidil was somewhat higher than that of metoprolol in the jejunum, while minoxidil showed lower permeability than metoprolol in the ileum. The permeability of minoxidil was independent of intestinal segment, while the permeability of metoprolol was region-dependent. Similarly, in mouse perfusion study, the jejunal permeability of minoxidil was 2.5-fold higher than that of metoprolol. Minoxidil and metoprolol showed similar permeability in Caco-2 study at apical pH of 6.5 and basolateral pH of 7.4. The permeability of minoxidil was independent of pH, while metoprolol showed pH-dependent transport in Caco-2 study. Minoxidil exhibited similar permeability in the absorptive direction (AP-BL) in comparison with secretory direction (BL-AP), while metoprolol had higher efflux ratio (ER > 2) at apical pH of 6.5 and basolateral pH of 7.4. No concentration-dependent transport was observed for either minoxidil or metoprolol transport in Caco-2 study. Verapamil did not alter the transport of either compounds across Caco-2 cell monolayers. The permeability of minoxidil was independent of both pH and intestinal segment in intestinal perfusion studies and Caco-2 studies. Caco-2 studies also showed no involvement of carrier mediated transport in the absorption process of minoxidil. These results suggest that minoxidil may be an acceptable reference drug for BCS high permeability classification. However, minoxidil exhibited higher jejunal permeability than metoprolol and thus to use minoxidil as a reference drug would raise the

  8. In situ cryopreservation of human embryonic stem cells in gas-permeable membrane culture cassettes for high post-thaw yield and good manufacturing practice.

    Amps, K J; Jones, M; Baker, D; Moore, H D


    The development of efficient and robust methods for the cryopreservation of human embryonic stem cells (hESCs) is important for the production of master and working cell banks for future clinical applications. Such methods must meet requirements of good manufacturing practice (GMP) and maintain genetic stability of the cell line. We investigated the culture of four Shef hESC lines in gas permeable 'culture cassettes' which met GMP compliance. hESCs adhered rapidly to the membrane and colonies displayed good proliferation and expansion. After 5-7 days of culture, hESCs were cryopreserved in situ using 10% dimethyl sulphoxide in foetal calf serum at approximately 1 degrees C/min. This method was compared with a control of standard flask culture and cryopreservation in vials. Post-thaw cassette culture displayed relative proliferation ratios (fold increase above flask/cryovial culture) of 114 (Shef 4), 8.2 (Shef 5), 195 (shef 6) and 17.5 (Shef 7). The proportion of cells expressing pluripotency markers after cryopreservation was consistently greater in cassette culture than for the control with the markers SSEA3 and SSEA4 exhibiting a significant increase (P> or =0.05). The efficiency of cell line culture in cassette was associated with the overall passage number of the cell line. The procedure enables cryopreservation of relatively large quantities of hESCs in situ, whilst returning high yields of viable, undifferentiated stem cells, thereby increasing capacity to scale up with greater efficacy.

  9. Nitrogen-doped MoS2/carbon as highly oxygen-permeable and stable catalysts for oxygen reduction reaction in microbial fuel cells

    Hao, Liang; Yu, Jia; Xu, Xin; Yang, Liu; Xing, Zipeng; Dai, Ying; Sun, Ye; Zou, Jinlong


    Developing non-noble metal catalysts with high oxygen-permeability and activity for oxygen reduction reaction (ORR) is crucial for microbial fuel cells (MFCs). In this study, nitrogen-doped molybdenum disulfide/carbon (N-MoS2/C) is prepared using melamine as nitrogen and carbon sources. Ammonium molybdate, thiourea and Pluronic F127 are used as Mo source, S source and surfactant, respectively. Mo-S-melamine complex precursor is obtained through the evaporation-induced self-assembly route, which is then carbonized at 800, 900 and 1000 °C to fabricate N-MoS2/C. Defect-rich N-MoS2/C has a large number of exposed active sites and a high oxygen permeability. N-MoS2/C (900 °C) with regular honeycomb structure shows the maximum power density of 0.815 W m-2, which is far higher than that of Pt/C (0.520 W m-2) and only has a decline of 1.23% after 1800 h operation in MFCs. Four-electron (4e-) reduction of O2 is the main ORR pathway for N-MoS2/C (900 °C), attributing to the efficient permeation, adsorption, activation and reduction of O2 on the active sites. The synergy among abundant defects, N-species (pyridinic N, graphitic N and Mo-Nx) and high conductivity contributes to the promising ORR activity. This simple synthetic route of N-doped metal sulfides/carbon composites displays a new prospect for preparation of ORR catalyst.

  10. Degradation of permeability resistance of high strength concrete after combustion

    Min LI; Hongtao KAO; Chunxiang QIAN


    To evaluate the remaining durability of con-crete materials after combustion, the permeability of high strength concrete (HSC) after combustion was studied. The transport behavior of chloride ion, water and air in concrete after combustion and the effect of temperature, strength grade, and aggregation on the permeability of HSC after combustion are investigated by chloride ion permeability coefficient (Dc), water permeability coef-ficient (Dw) and air permeability coefficient (Da). The experiment results show that all three permeability coeffi-cients commendably reflect changes of permeability. The permeability coefficient increases with the evaluation tem-perature. After the same temperature, the permeability coefficient of HSC is lower than that of normal strength concrete (NSC). However, the degree of degradation of permeability coefficient of HSC is greater than that of NSC. The permeability resistance of HSC containing limestone is better than that of HSC containing basalt. Combining changes of compressive strength and per-meability, the remaining durability of concrete materials after combustion is appropriately evaluated.

  11. High surface area, high permeability carbon monoliths

    Lagasse, R.R.; Schroeder, J.L. [Sandia National Labs., Albuquerque, NM (United States). Organic Materials Processing Dept.


    The goal of this work is to prepare carbon monoliths having precisely tailored pore size distribution. Prior studies have demonstrated that poly(acrylonitrile) can be processed into a precursor having tailored macropore structure. Since the macropores were preserved during pyrolysis, this synthetic process provided a route to porous carbon having macropores with size =0.1 to 10{mu}m. No micropores of size <2 nm could be detected in the carbon, however, by nitrogen adsorption. In the present work, the authors have processed a different polymer, poly(vinylidene chloride) into a macroporous precursor, Pyrolysis produced carbon monoliths having macropores derived from the polymer precursor as well as extensive microporosity produced during the pyrolysis of the polymer. One of these carbons had BET surface area of 1,050 m{sup 2}/g and about 1.2 cc/g total pore volume, with about 1/3 of the total pore volume in micropores and the remainder in 1{mu}m macropores. No mesopores in the intermediate size range could be detected by nitrogen adsorption. Carbon materials having high surface area as well as micron size pores have potential applications as electrodes for double layer supercapacitors containing liquid electrolyte, or as efficient media for performing chemical separations.

  12. Permeabilidade a quente de refratários para revestimento de cubas eletrolíticas Hot permeability of refractories for aluminum electrolytic cells lining at high temperatures

    D. Y. Miyaji


    Full Text Available Na indústria do alumínio primário, um dos principais responsáveis pela deterioração do revestimento refratário nas cubas de redução eletrolítica é o ataque por banho criolítico com alta concentração de NaF, que penetra e reage com o refratário podendo, em condições extremas, causar a parada prematura de operação da cuba e grandes prejuízos econômicos. Baseando-se nos mais recentes estudos de caracterização de refratários para cubas, uma boa correlação tem sido encontrada entre a resistência à corrosão e a permeabilidade, a qual é calculada pela equação de Forchheimer para a obtenção das constantes k1 Darciana (efeito viscoso e k2 não Darciana (efeito inercial. Entretanto, na maioria das situações, as medidas da permeabilidade têm sido efetuadas à temperatura ambiente, embora o refratário em uso esteja a superior temperatura. Este trabalho visa avaliar como esta permeabilidade se altera em temperaturas de até 700 ºC buscando, assim, uma melhor correlação dos resultados com as condições reais de operação. Pretende-se também, investigar essa propriedade em refratários empregados no revestimento de cadinhos para o transporte de alumínio líquido, com o intuito de verificar a aplicabilidade da permeametria como uma ferramenta de caracterização desses materiais cujo interesse de aplicação e desenvolvimento tem sido cada vez maior.In the aluminium primary industry, one of the main causes for electrolytic cells lining deterioration is the chemical attack by high NaF content cryolitic bath, that penetrates and reacts with the refractory, shortening the cell’s service life and resulting great economic losses. Based on the most recent studies on characterization of alumino-silicate refractories for aluminum cell linings, a good correlation has been found between its chemical attack by molten fluorides and the permeability, which is calculated by Forchheimer’s equation in order to obtain the Darcian

  13. Neurogenesis is enhanced by stroke in multiple new stem cell niches along the ventricular system at sites of high BBB permeability.

    Lin, Ruihe; Cai, Jingli; Nathan, Cody; Wei, Xiaotao; Schleidt, Stephanie; Rosenwasser, Robert; Iacovitti, Lorraine


    Previous studies have established the subventricular (SVZ) and subgranular (SGZ) zones as sites of neurogenesis in the adult forebrain (Doetsch et al., 1999a; Doetsch, 2003a). Work from our laboratory further indicated that midline structures known as circumventricular organs (CVOs) also serve as adult neural stem cell (NSC) niches (Bennett et al., 2009, 2010). In the quiescent rat brain, NSC proliferation remains low in all of these sites. Therefore, we recently examined whether ischemic stroke injury (MCAO) or sustained intraventricular infusion of the mitogen bFGF could trigger an up-regulation in NSC proliferation, inducing neurogenesis and gliogenesis. Our data show that both stroke and bFGF induce a dramatic and long-lasting (14day) rise in the proliferation (BrdU+) of nestin+Sox2+GFAP+ NSCs capable of differentiating into Olig2+ glial progenitors, GFAP+nestin-astrocyte progenitors and Dcx+ neurons in the SVZ and CVOs. Moreover, because of the upsurge in NSC number, it was possible to detect for the first time several novel stem cell niches along the third (3V) and fourth (4V) ventricles. Importantly, a common feature of all brain niches was a rich vasculature with a blood-brain-barrier (BBB) that was highly permeable to systemically injected sodium fluorescein. These data indicate that stem cell niches are more extensive than once believed and exist at multiple sites along the entire ventricular system, consistent with the potential for widespread neurogenesis and gliogenesis in the adult brain, particularly after injury. We further suggest that because of their leaky BBB, stem cell niches are well-positioned to respond to systemic injury-related cues which may be important for stem-cell mediated brain repair. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  14. A cell-permeable fluorescent polymeric thermometer for intracellular temperature mapping in mammalian cell lines.

    Teruyuki Hayashi

    Full Text Available Changes in intracellular temperatures reflect the activity of the cell. Thus, the tool to measure intracellular temperatures could provide valuable information about cellular status. We previously reported a method to analyze the intracellular temperature distribution using a fluorescent polymeric thermometer (FPT in combination with fluorescence lifetime imaging microscopy (FLIM. Intracellular delivery of the FPT used in the previous study required microinjection. We now report a novel FPT that is cell permeable and highly photostable, and we describe the application of this FPT to the imaging of intracellular temperature distributions in various types of mammalian cell lines. This cell-permeable FPT displayed a temperature resolution of 0.05°C to 0.54°C within the range from 28°C to 38°C in HeLa cell extracts. Using our optimized protocol, this cell-permeable FPT spontaneously diffused into HeLa cells within 10 min of incubation and exhibited minimal toxicity over several hours of observation. FLIM analysis confirmed a temperature difference between the nucleus and the cytoplasm and heat production near the mitochondria, which were also detected previously using the microinjected FPT. We also showed that this cell-permeable FPT protocol can be applied to other mammalian cell lines, COS7 and NIH/3T3 cells. Thus, this cell-permeable FPT represents a promising tool to study cellular states and functions with respect to temperature.

  15. A cell-permeable fluorescent polymeric thermometer for intracellular temperature mapping in mammalian cell lines.

    Hayashi, Teruyuki; Fukuda, Nanaho; Uchiyama, Seiichi; Inada, Noriko


    Changes in intracellular temperatures reflect the activity of the cell. Thus, the tool to measure intracellular temperatures could provide valuable information about cellular status. We previously reported a method to analyze the intracellular temperature distribution using a fluorescent polymeric thermometer (FPT) in combination with fluorescence lifetime imaging microscopy (FLIM). Intracellular delivery of the FPT used in the previous study required microinjection. We now report a novel FPT that is cell permeable and highly photostable, and we describe the application of this FPT to the imaging of intracellular temperature distributions in various types of mammalian cell lines. This cell-permeable FPT displayed a temperature resolution of 0.05°C to 0.54°C within the range from 28°C to 38°C in HeLa cell extracts. Using our optimized protocol, this cell-permeable FPT spontaneously diffused into HeLa cells within 10 min of incubation and exhibited minimal toxicity over several hours of observation. FLIM analysis confirmed a temperature difference between the nucleus and the cytoplasm and heat production near the mitochondria, which were also detected previously using the microinjected FPT. We also showed that this cell-permeable FPT protocol can be applied to other mammalian cell lines, COS7 and NIH/3T3 cells. Thus, this cell-permeable FPT represents a promising tool to study cellular states and functions with respect to temperature.

  16. Recovery of Porosity and Permeability for High Plasticity Clays

    Krogsbøll, Anette; Foged, Niels Nielsen

    Clays, which have been loaded to a high stress level, will under certain conditions keep low porosity and permeability due to the high degree of compression. In some situations it seems that porosity and permeability will recover to a very high extent when the clay is unloaded. This seems...... the clay will expand to an even higher porosity....

  17. Influence of decenylsuccinic Acid on water permeability of plant cells.

    Lee, O Y; Stadelmann, E J; Weiser, C J


    Decenylsuccinic acid altered permeability to water of epidermal cells of bulb scales of Allium cepa and of the leaf midrib of Rhoeo discolor. Water permeability, as determined by deplasmolysis time measurements, was related to the dose of undissociated decenylsuccinic acid (mm undissociated decenylsuccinic acid x minute). No relationship was found between permeability and total dose of decenylsuccinic acid, or dose of dissociated decenylsuccinic acid, suggesting that the undissociated molecule was the active factor in permeability changes and injury.At doses which did not damage cells (0.0008 to 0.6 [mm of the undissociated molecule x minute]) decenylsuccinic acid decreased water permeability. At higher doses (e.g., 4 to 8 [mm x minute]) injury to cells was common and decenylsuccinic acid increased permeability. Doses above the 10 to 20 (mm x minute) range were generally lethal. The plasmolysis form of uninjured cells was altered and protoplasmic swelling occasionally was observed. The dose-dependent reversal of water permeability changes (decreased to increased permeability) may reflect decenylsuccinic acid-induced changes in membrane structure. Reported effects of decenylsuccinic acid on temperature dependence of permeability and frost resistance were not verified.

  18. High permeability-high frequency stable MnZn ferrites

    Kalarus, J. [Ferroxcube Polska, 96-100 Skierniewice (Poland); Kogias, G., E-mail: [Centre for Research and Technology-Hellas, Chemical Process Engineering Research Institute, Laboratory of Inorganic Materials, 57001 Thessaloniki (Greece); Aristotle University of Thessaloniki, Department of Chemical Engineering, Laboratory of Materials Technology, 54124 Thessaloniki (Greece); Holz, D. [Ferroxcube Polska, 96-100 Skierniewice (Poland); Zaspalis, V.T. [Centre for Research and Technology-Hellas, Chemical Process Engineering Research Institute, Laboratory of Inorganic Materials, 57001 Thessaloniki (Greece); Aristotle University of Thessaloniki, Department of Chemical Engineering, Laboratory of Materials Technology, 54124 Thessaloniki (Greece)


    Modern MnZn ferrite applications require high magnetic initial permeability and exceptional frequency stability; the former implies large grains, while the latter high grain boundary resistivity. In this article the optimization of the final firing process is described for achieving both. The optimization is based on the homogeneous dissolution of dopants under oxidative conditions and their subsequent precipitation along grain boundaries. This was accomplished by integrating isothermal plateaus at the upper stadia of the cooling stage of the final firing process. MnZn ferrites of the basic composition [Mn{sub 0.47}Zn{sub 0.47}Fe{sub 0.06}{sup 2+}]Fe{sub 2}{sup 3+}O{sub 4} were synthesized with initial permeability (measured at f=10 kHz, B<0.1 mT, T=25 Degree-Sign C) 12,600 and losses, expressed as tan({delta})/{mu}{sub i}, of 3.1 Multiplication-Sign 10{sup -6} at 10 kHz and 20.5 Multiplication-Sign 10{sup -6} at 100 kHz (B<0.1 mT, T=25 Degree-Sign C), that reflect good frequency stability. These results could be reproduced in pilot production scale. - Highlights: Black-Right-Pointing-Pointer Optimization of sintering is described for achieving high initial permeability. Black-Right-Pointing-Pointer Optimization of sintering is described for receiving frequency stability. Black-Right-Pointing-Pointer For high permeability, high densities and large grain sizes are required. Black-Right-Pointing-Pointer The achieved initial permeability is higher than 12,500. Black-Right-Pointing-Pointer The losses, tan({delta})/{mu}{sub i}, are 3.1 Multiplication-Sign 10{sup -6} at 10 kHz and 20.5 Multiplication-Sign 10{sup -6} at 100 kHz.

  19. 21 CFR 876.5860 - High permeability hemodialysis system.


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false High permeability hemodialysis system. 876.5860... hemodialysis system. (a) Identification. A high permeability hemodialysis system is a device intended for use... toxemic conditions by performing such therapies as hemodialysis, hemofiltration, hemoconcentration,...

  20. Inhibitory effects of isoproterenol on PAF-induced endothelial cell permeability and morphological changes

    丁自强; 李少华; 吴中立


    Using a model to study vascular permeability under hydrostatically perfused bovine pulmonary artery endothelial cell (EC) monolayers and a software to automatically analyse cell morphological parameters in a computer image workstation, the effects of isoproterenol (IPN) on platelet-activating factor (PAF)-induced changes in EC monolayer permeability and cell morphological parameters were studied. Albumin has the fortifying effect on endothelial barrier function. After treatment of EC monolayer with 10-8mol/L PAF, trans-monolayer permeability increased, cell surface area decreased, and intercellular space enlarged. As pretreatment with 10-4mol/L IPN, PAF-induced EC permeability increment and morphological changes were blocked. The results suggest that EC contraction and intercellular gap expansion are important mechanisms for PAF-induced high vascular permeability. IPN inhibits the effects of PAF via stabilization of EC morphology and prevention of intercellular gap formation.

  1. Review on permeability of high-strength concrete subjected to high temperature

    Zhao, Dongfu; Han, Xiao; Liu, Yuchen


    In this paper, the research results of permeability of high-strength concrete subjected to high temperature were comprehensively reviewed, the research status of permeability of high-strength concrete at elevated temperature were discussed, and existing problems were analyzed, finally, main research directions of permeability of high-strength concrete subjected to high temperature were forecasted.

  2. Cadmium substituted high permeability lithium ferrite

    S S Bellad; S C Watawe; A M Shaikh; B K Chougule


    Polycrystalline Li0.5–/2 Cd Fe2.5–/2O4 ferrites where = 0, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6 were prepared by a double sintering ceramic technique and characterized by X-ray diffraction and scanning electron microscopy (SEM). The lattice parameter is found to increase monotonically with the cadmium content. It is explained in terms of the sizes of component ions. The grain size of the samples increases up to = 0.3 and then it decreases for higher values of . A similar trend is observed in the variation of Ms with Cd2+ content. The initial permeability () is however found to increase continuously with . The increase in is attributed to decrease of anisotropy constant K1 and higher grain size of the samples.

  3. Measurement of relative permeability of fuel cell diffusion media

    Hussaini, I.S.


    Gas diffusion layer (GDL) in PEM fuel cells plays a pivotal role in water management. Modeling of liquid water transport through the GDL relies on knowledge of relative permeability functions in the in-plane and through-plane directions. In the present work, air and water relative permeabilities are experimentally determined as functions of saturation for typical GDL materials such as Toray-060, -090, -120 carbon paper and E-Tek carbon cloth materials in their plain, untreated forms. Saturation is measured using an ex situ gravimetric method. Absolute and relative permeability functions in the two directions of interest are presented and new correlations for in-plane relative permeability of water and air are established. © 2010 Elsevier B.V. All rights reserved.

  4. Hydrogen permeability of Nb membrane annealed under ultra high vacuum

    Uchida, Helmut T.; Pundt, Astrid; Kirchheim, Reiner [Institut fuer Materialphysik, Goettingen (Germany); Yamazaki, Yoshihiro [California Institute of Technology, California (United States); Kamegawa, Atsunori; Takamura, Hitoshi; Koike, Junichi; Okada, Masuo [Department of Materials Science, School of Engineering, Tohoku University, Sendai (Japan)


    In this research, hydrogen permeability of Pd-free Nb membrane using hydrogen annealing process by ultra high vacuum equipment was investigated. Pure Nb (99.85 percent) was used as membrane. The hydrogen permeation experiment was carried out in the temperature range of 773-1173 K, and the H{sub 2} pressure range of 1-3.5 atm in the inlet side. Hydrogen flux and hydrogen pressures in the inlet side and outlet side of chamber were measured using mass flow meter and capacitance manometer, respectively. Hydrogen permeability was determined by Fick's first low. Effects of annealing conditions and heat treatment under hydrogen atmosphere on hydrogen permeability were investigated. It is supposed that the surface oxidation of Nb is reduced by the hydrogen annealing. Moreover, it is found that the value of hydrogen permeability becomes almost equivalent to the theoretical value when the sample preparation was optimized by an extension of the evacuating time. The best hydrogen permeability is obtained in metal membranes using the annealing under ultra-high vacuum and pure hydrogen atmosphere.

  5. Electrical parameters and water permeability properties of monolayers formed by T84 cells cultured on permeable supports

    Ozu M.


    Full Text Available T84 is an established cell line expressing an enterocyte phenotype whose permeability properties have been widely explored. Osmotic permeability (P OSM, hydraulic permeability (P HYDR and transport-associated net water fluxes (J W-transp, as well as short-circuit current (I SC, transepithelial resistance (R T, and potential difference (deltaV T were measured in T84 monolayers with the following results: P OSM 1.3 ± 0.1 cm.s-1 x 10-3; P HYDR 0.27 ± 0.02 cm.s-1; R T 2426 ± 109 omega.cm², and deltaV T 1.31 ± 0.38 mV. The effect of 50 µM 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one (DCEBIO, a "net Cl- secretory agent", on T84 cells was also studied. We confirm the reported important increase in I SC induced by DCEBIO which was associated here with a modest secretory deltaJ W-transp. The present results were compared with those reported using the same experimental approach applied to established cell lines originating from intestinal and renal epithelial cells (Caco-2, LLC-PK1 and RCCD-1. No clear association between P HYDR and R T could be demonstrated and high P HYDR values were observed in an electrically tight epithelium, supporting the view that a "water leaky" barrier is not necessarily an "electrically leaky" one. Furthermore, the modest secretory deltaJ W-transp was not consistent with previous results obtained with RCCD-1 cells stimulated with vasopressin (absorptive fluxes or with T84 cells secreting water under the action of Escherichia coli heat stable enterotoxin. We conclude that, while the presence of aquaporins is necessary to dissipate an external osmotic gradient, coupling between water and ion transport cannot be explained by a simple and common underlying mechanism.

  6. Polybenzimidazole-based mixed membranes with exceptional high water vapor permeability and selectivity

    Akhtar, Faheem Hassan


    Polybenzimidazole (PBI), a thermal and chemically stable polymer, is commonly used to fabricate membranes for applications like hydrogen recovery at temperatures of more than 300 °C, fuel cells working in a highly acidic environment, and nanofiltration in aggressive solvents. This report shows for the first time use of PBI dense membranes for water vapor/gas separation applications. They showed an excellent selectivity and high water vapor permeability. Incorporation of inorganic hydrophilic titanium-based nano-fillers into the PBI matrix further increased the water vapor permeability and water vapor/N2 selectivity. The most selective mixed matrix membrane with 0.5 wt% loading of TiO2 nanotubes yielded a water vapor permeability of 6.8×104 Barrer and a H2O/N2 selectivity of 3.9×106. The most permeable membrane with 1 wt% loading of carboxylated TiO2 nanoparticles had a 7.1×104 Barrer water vapor permeability and a H2O/N2 selectivity of 3.1×106. The performance of these membranes in terms of water vapor transport and selectivity is among the highest reported ones. The remarkable ability of PBI to efficiently permeate water versus other gases opens the possibility to fabricate membranes for dehumidification of streams in harsh environments. This includes the removal of water from high temperature reaction mixtures to shift the equilibrium towards products.

  7. High Guanidinium Permeability Reveals Dehydration-Dependent Ion Selectivity in the Plasmodial Surface Anion Channel

    Abdullah A. B. Bokhari


    Full Text Available Malaria parasites grow within vertebrate erythrocytes and increase host cell permeability to access nutrients from plasma. This increase is mediated by the plasmodial surface anion channel (PSAC, an unusual ion channel linked to the conserved clag gene family. Although PSAC recognizes and transports a broad range of uncharged and charged solutes, it must efficiently exclude the small Na+ ion to maintain infected cell osmotic stability. Here, we examine possible mechanisms for this remarkable solute selectivity. We identify guanidinium as an organic cation with high permeability into human erythrocytes infected with Plasmodium falciparum, but negligible uptake by uninfected cells. Transport characteristics and pharmacology indicate that this uptake is specifically mediated by PSAC. The rank order of organic and inorganic cation permeabilities suggests cation dehydration as the rate-limiting step in transport through the channel. The high guanidinium permeability of infected cells also allows rapid and stringent synchronization of parasite cultures, as required for molecular and cellular studies of this pathogen. These studies provide important insights into how nutrients and ions are transported via PSAC, an established target for antimalarial drug development.

  8. Cell Permeable Ratiometric Fluorescent Sensors for Imaging Phosphoinositides.

    Mondal, Samsuzzoha; Rakshit, Ananya; Pal, Suranjana; Datta, Ankona


    Phosphoinositides are critical cell-signal mediators present on the plasma membrane. The dynamic change of phosphoinositide concentrations on the membrane including clustering and declustering mediates signal transduction. The importance of phosphoinositides is scored by the fact that they participate in almost all cell-signaling events, and a defect in phosphoinositide metabolism is linked to multiple diseases including cancer, bipolar disorder, and type-2 diabetes. Optical sensors for visualizing phosphoinositide distribution can provide information on phosphoinositide dynamics. This exercise will ultimately afford a handle into understanding and manipulating cell-signaling processes. The major requirement in phosphoinositide sensor development is a selective, cell permeable probe that can quantify phosphoinositides. To address this requirement, we have developed short peptide-based ratiometric fluorescent sensors for imaging phosphoinositides. The sensors afford a selective response toward two crucial signaling phosphoinositides, phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) and phosphatidylinositol-4-phosphate (PI4P), over other anionic membrane phospholipids and soluble inositol phosphates. Dissociation constant values indicate up to 4 times higher probe affinity toward PI(4,5)P2 when compared to PI4P. Significantly, the sensors are readily cell-permeable and enter cells within 15 min of incubation as indicated by multiphoton excitation confocal microscopy. Furthermore, the sensors light up signaling phosphoinositides present both on the cell membrane and on organelle membranes near the perinuclear space, opening avenues for quantifying and monitoring phosphoinositide signaling.

  9. Pulsed electric field reduces the permeability of potato cell wall.

    Galindo, Federico Gómez; Vernier, P Thomas; Dejmek, Petr; Vicente, António; Gundersen, Martin A


    The effect of the application of pulsed electric fields to potato tissue on the diffusion of the fluorescent dye FM1-43 through the cell wall was studied. Potato tissue was subjected to field strengths ranging from 30 to 500 V/cm, with one 1 ms rectangular pulse, before application of FM1-43 and microscopic examination. Our results show a slower diffusion of FM1-43 in the electropulsed tissue when compared with that in the non-pulsed tissue, suggesting that the electric field decreased the cell wall permeability. This is a fast response that is already detected within 30 s after the delivery of the electric field. This response was mimicked by exogenous H2O2 and blocked by sodium azide, an inhibitor of the production of H2O2 by peroxidases. (c) 2007 Wiley-Liss, Inc.

  10. Maladaptively high and low openness: the case for experiential permeability.

    Piedmont, Ralph L; Sherman, Martin F; Sherman, Nancy C


    The domain of Openness within the Five-Factor Model (FFM) has received inconsistent support as a source for maladaptive personality functioning, at least when the latter is confined to the disorders of personality included within the American Psychiatric Association's (APA) Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR; APA, ). However, an advantage of the FFM relative to the DSM-IV-TR is that the former was developed to provide a reasonably comprehensive description of general personality structure. Rather than suggest that the FFM is inadequate because the DSM-IV-TR lacks much representation of Openness, it might be just as reasonable to suggest that the DSM-IV-TR is inadequate because it lacks an adequate representation of maladaptive variants of both high and low Openness. This article discusses the development and validation of a measure of these maladaptive variants, the Experiential Permeability Inventory.

  11. A High Gain Omnidirectional Antenna Using Negative Permeability Metamaterial

    Hangfei Tang


    Full Text Available A high gain omnidirectional antenna with low profile is proposed and is investigated numerically and experimentally. Based on the conventional center-fed circular epsilon-negative (ENG zeroth-order resonator (ZOR antenna, dendritic structure negative permeability metamaterial (NPM is used as the substrate to enhance the gain of the omnidirectional antenna. The experimental results show that the gain of a center-fed circular ENG ZOR antenna with NPM substrate is enhanced about 2.2 dB, and the efficiency is enhanced about 38%, in the whole broad operating bandwidth as compared to that of the antenna without NPM substrate, which can be used to improve the reliability of wireless communications.

  12. Fluid-loss control for high-permeability rocks in hydraulic fracturing under realistic shear conditions

    Navarrete, R.C.; Mitchell, J.P.


    A study is presented on the effectiveness of different combinations of fluid and fluid-loss additives to control fluid loss in high-permeability formations under high shear rates. The impact on matrix damage and proppant-pack damage is also studied. Borate-crosslinked guars, hydroxyethylcellulose (HEC) and a surfactant water-base gravel packing fluid were investigated. The fluid-loss additive considered was silica flour. All fluid-loss tests were run in dynamic fluid-loss cells. To properly test high-permeability cores, new long core dynamic fluid-loss cells were used. The matrix damage caused by the invasion of the fluid was determined using pressure taps along the core. Conductivity tests were also run to determine the damage to the proppant pack. Results show that the effectiveness of particulate fluid-loss additives under dynamic conditions is strongly dependent on the initial leakoff rate, which depends on the pressure gradient across the core, permeability of the core and viscosity of the invading fluid. The use of silica flour helps matrix flowback, and it has a minimal effect on proppant-pack conductivity in clean fluids (e.g., surfactant water-base gravel packing fluid). With the exception of the borate-crosslinked guar with no fluid-loss additive, the variety of fluids used in these tests (with and without silica flour) have a negligible effect on postproduction.

  13. AN69: Evolution of the world's first high permeability membrane.

    Thomas, Michel; Moriyama, Kazuhiro; Ledebo, Ingrid


    In response to a request from the French government, Rhône-Poulenc started development of a synthetic membrane for dialysis in 1969. When used for patients the new membrane, AN69, showed improved efficiency and clinical outcome. Treatment times could be reduced, peripheral neuropathy improved and patients expressed improved quality of life. This was the introduction of high-flux, volume-controlled dialysis. The polymer used for the AN69 membrane is a copolymer of acrylonitrile and sodium methallylsulfonate, specially selected for the dialysis application. Contrary to most synthetic membranes AN69 is hydrophilic, because the numerous sulfonate groups attract water and create a hydrogel structure which provides high diffusive and hydraulic permeability. In the AN69 membrane the microstructure and the chemical composition offer a unique context for bulk adsorption of low-molecular-weight proteins, the high water content of the hydrogel making the polymer chains easily accessible. The highly specific adsorptive properties - limited on the surface and favored in the membrane structure and with high specificity for basic, medium-sized proteins - distinguish AN69 from other synthetic high-flux membranes as well as from other so-called adsorptive membranes in the field of dialysis. Since its creation in the early 1970s the AN69 membrane has evolved to meet the requirements and challenges of dialysis therapy. Still the basic characteristics of high permeability to fluid and to a wide range of relevant uremic retention products combined with excellent biocompatibility, whether measured by traditional or novel indicators, have remained the hallmark of the membrane. The unique adsorptive capacity has provided a base for further development. The latest version of AN69, HeprAN, has heparin grafting on the inner surface and enhanced adsorption of bacterial products on the outer surface. Modern versions of AN69 are today used in hemodialysis and contemporary modes of convective

  14. Caco-2 cells permeability evaluation of nifuroxazide derivatives with potential activity against methicillin-resistant Staphylococcus aureus (MRSA).

    B Fernandes, Mariane; Gonçalves, José E; C Tavares, Leoberto; Storpirtis, Sílvia


    Throughout the period of evaluation and selection in drug development, the assessment of the permeability potential of a compound to achieve an efficient refinement of the molecular structure has been widely appraised by the transport of substances across cell monolayers. This study aims to develop in vitro assays through Caco-2 cells in order to analyze the permeability of 5-nitro-heterocyclic compounds analogues to nifuroxazide with antimicrobial activity, especially showing promising activity against multidrug-resistant Staphylococcus aureus (MRSA). Caco-2 cell monolayers cultivated for 21 days in Transwell® plates were used for the in vitro permeability assays. The quantification of the nifuroxazide derivatives in the basolateral chambers was performed by a validated high performance liquid chromatography with UV (HPLC-UV) method. Apparent permeability values (Papp) show that these compounds can be considered as new drug candidates with the potential to present high absorption in vivo, according to the classifications of Yee and Biganzoli. The thiophenic derivatives showed permeability values higher than the furanic ones, being AminoTIO the compound with the greatest potential for the development of a new drug against MRSA, since it showed the best cytotoxicity, permeability and solubility ratio among all the derivatives.

  15. Influence of Low-Energy Ion Irradiation on Plasma MembranePermeability of Cells

    ZHANG Dong-Mei; CUI Fu-Zhai; SUN Su-Qin; LIN You-Bo; TIAN Min-Bo; CHEN Guo-Qiang


    Effect of low-energy ion irradiation on plasma membrane permeability has been investigated by using electron spin resonance (ESR) spectroscopy of spin probe technique. The investigated system is plumule cells of wheat (Triticum aestivum L.) seeds implanted by 30keV N+ ions. ESR spectra indicated that plasmalemma permeability is sensitive to low-energyion irradiation. Ion irradiations with increasing fluences up to semi-lethal dose lead to gradual increase in plasmalemma permeability of the plumule cells. The possible factors relevant to the changes in membrane permeability are discussed in relation to the changes in the physical state and chemical nature of membranes.

  16. Influence of Decenylsuccinic Acid on Water Permeability of Plant Cells 1

    Lee, O. Y.; Stadelmann, Ed. J.; Weiser, C. J.


    Decenylsuccinic acid altered permeability to water of epidermal cells of bulb scales of Allium cepa and of the leaf midrib of Rhoeo discolor. Water permeability, as determined by deplasmolysis time measurements, was related to the dose of undissociated decenylsuccinic acid (mm undissociated decenylsuccinic acid × minute). No relationship was found between permeability and total dose of decenylsuccinic acid, or dose of dissociated decenylsuccinic acid, suggesting that the undissociated molecule was the active factor in permeability changes and injury. At doses which did not damage cells (0.0008 to 0.6 [mm of the undissociated molecule × minute]) decenylsuccinic acid decreased water permeability. At higher doses (e.g., 4 to 8 [mm × minute]) injury to cells was common and decenylsuccinic acid increased permeability. Doses above the 10 to 20 (mm × minute) range were generally lethal. The plasmolysis form of uninjured cells was altered and protoplasmic swelling occasionally was observed. The dose-dependent reversal of water permeability changes (decreased to increased permeability) may reflect decenylsuccinic acid-induced changes in membrane structure. Reported effects of decenylsuccinic acid on temperature dependence of permeability and frost resistance were not verified. Images PMID:16658227

  17. Cell-permeable nanobodies for targeted immunolabelling and antigen manipulation in living cells

    Herce, Henry D.; Schumacher, Dominik; Schneider, Anselm F. L.; Ludwig, Anne K.; Mann, Florian A.; Fillies, Marion; Kasper, Marc-André; Reinke, Stefan; Krause, Eberhard; Leonhardt, Heinrich; Cardoso, M. Cristina; Hackenberger, Christian P. R.


    Functional antibody delivery in living cells would enable the labelling and manipulation of intracellular antigens, which constitutes a long-thought goal in cell biology and medicine. Here we present a modular strategy to create functional cell-permeable nanobodies capable of targeted labelling and manipulation of intracellular antigens in living cells. The cell-permeable nanobodies are formed by the site-specific attachment of intracellularly stable (or cleavable) cyclic arginine-rich cell-penetrating peptides to camelid-derived single-chain VHH antibody fragments. We used this strategy for the non-endocytic delivery of two recombinant nanobodies into living cells, which enabled the relocalization of the polymerase clamp PCNA (proliferating cell nuclear antigen) and tumour suppressor p53 to the nucleolus, and thereby allowed the detection of protein-protein interactions that involve these two proteins in living cells. Furthermore, cell-permeable nanobodies permitted the co-transport of therapeutically relevant proteins, such as Mecp2, into the cells. This technology constitutes a major step in the labelling, delivery and targeted manipulation of intracellular antigens. Ultimately, this approach opens the door towards immunostaining in living cells and the expansion of immunotherapies to intracellular antigen targets.

  18. Structural and permeability characterization of biosynthetic PVA hydrogels designed for cell-based therapy.

    Nafea, Eman H; Poole-Warren, Laura A; Martens, Penny J


    Incorporation of extracellular matrix (ECM) components to synthetic hydrogels has been shown to be the key for successful cell encapsulation devices, by providing a biofunctional microenvironment for the encapsulated cells. However, the influence of adding ECM components into synthetic hydrogels on the permeability as well as the physical and mechanical properties of the hydrogel has had little attention. Therefore, the aim of this study was to investigate the effect of incorporated ECM analogues on the permeability performance of permselective synthetic poly(vinyl alcohol) (PVA) hydrogels in addition to examining the physico-mechanical characteristics. PVA was functionalized with a systematically increased number of methacrylate functional groups per chain (FG/c) to tailor the permselectivity of UV photopolymerized hydrogel network. Heparin and gelatin were successfully incorporated into PVA network at low percentage (1%), and co-hydrogels were characterized for network properties and permeability to bovine serum albumin (BSA) and immunoglobulin G (IgG) proteins. Incorporation of these ECM analogues did not interfere with the base PVA network characteristics, as the controlled hydrogel mesh sizes, swelling and compressive modulii remained unchanged. While the permeation profiles of both BSA and IgG were not affected by the addition of heparin and gelatin as compared with pure PVA, increasing the FG/c from 7 to 20 significantly limited the diffusion of the larger IgG. Consequently, biosynthetic hydrogels composed of PVA with high FG/c and low percent ECM analogues show promise in their ability to be permselective for various biomedical applications.

  19. Experimental Research on Permeability of Airbag Fabrics at High Pressure Differential

    WANG Xin-hou


    The air permeability of airbag fabrics was measured at high pressure differential up to 200kPa. It was found that permeability varied with pressure differential nonlinearly. The relationship between air permeability and the pressure differential was fitted well with power law curve. The study revealed that the coefficient c and exponent b in the power law equation had a strong correlation with porosity, which was chosen to characterize the airbag fabrics.

  20. Risk factor(s) related to high membrane permeability in peritoneal dialysis.

    Unal, Aydin; Sipahioglu, Murat Hayri; Kocyigit, Ismail; Tunca, Onur; Tokgoz, Bulent; Oymak, Oktay


    Peritoneal dialysis (PD) patients have different peritoneal membrane permeability (transport) characteristics. High peritoneal membrane permeability is associated with increased mortality risk in the patient population. In this study, we aimed to investigate possible risk factor(s) related to high peritoneal membrane permeability. The study included 475 PD patients (46.1 ± 14.5 years of mean age; 198 female and 277 male). The patients were divided two groups according to peritoneal equilibration test (PET) result: high-permeability group (high and high-average) and low- permeability group (low-average and low). In both the univariate and multivariate logistic regression analyses, it was found that diabetes mellitus and hypoalbuminemia was significantly associated with high peritoneal membrane permeability [relative risk (RR): 1.90, 95% confidence interval (CI): 1.26-2.86, p: 0.002 and RR: 2.14, 95% CI: 1.44-3.18, ppermeability. Diabetic patients had 1.9 times the likelihood of having high permeability. However, the relationship between hypoalbuminemia and high peritoneal permeability appears to be a result rather than cause.

  1. An experimental insight into the evolution of permeability at high temperatures and applications for shallow conduit and lava dome degassing

    Chadderton, Amy; Sammonds, Peter; Meredith, Philip; Smith, Rosanna; Tuffen, Hugh; Gaunt, Elizabeth


    Two recent eruptions in Chile, at Chaitén Volcano in 2008-10 and Cordón Caulle in 2011-12, allowed the first detailed observations of rhyolitic activity and provided insights into the evolution of highly silicic eruptions. Both events exhibited simultaneous explosive and effusive activity, with both lava and ash plumes emitted from the same vent [1]. The permeability of fracture networks that act as fluid flow pathways is key to understanding such eruptive behaviour. Here, we report results from a systematic experimental investigation of permeability in volcanic rocks at magmatic temperatures and pressures, in the presence of pore fluids using our newly-developed high-temperature permeability facility. Enhancements to the High Temperature Triaxial Deformation Cell at UCL [2] have enabled us to make permeability measurements on 25mm x 50mm cores at both elevated temperature and elevated hydrostatic pressure [3]. We present results from several suites of permeability measurements on samples of dome dacite from the 2004-08 eruption of Mount St Helens, and rhyolite collected from the lava dome formed during the 2008-10 eruption of Chaitén, Chile. Tests were conducted at temperatures up to 900oC and under an effective pressure of 5 MPa, using the steady-state flow technique. Samples were cooled to room temperature between each high temperature test, and the permeability of each sample was re-measured before heating to the next temperature increment in the series. Additional longer duration high temperature tests were also conducted to investigate the development of a permeable network at high temperatures over time. The results show a complex permeability evolution that includes a reduction in permeability by approximately 3 orders of magnitude up to 600oC. Together with thermal cracking tests, AE data and SEM/thin section analysis these new experimental permeability results are applied to enhance our understanding of the complex issue of shallow conduit and lava

  2. Glioblastoma cell-secreted interleukin-8 induces brain endothelial cell permeability via CXCR2.

    Julie Dwyer

    Full Text Available Glioblastoma constitutes the most aggressive and deadly of brain tumors. As yet, both conventional and molecular-based therapies have met with limited success in treatment of this cancer. Among other explanations, the heterogeneity of glioblastoma and the associated microenvironment contribute to its development, as well as resistance and recurrence in response to treatments. Increased vascularity suggests that tumor angiogenesis plays an important role in glioblastoma progression. However, the molecular crosstalk between endothelial and glioblastoma cells requires further investigation. To examine the effects of glioblastoma-derived signals on endothelial homeostasis, glioblastoma cell secretions were collected and used to treat brain endothelial cells. Here, we present evidence that the glioblastoma secretome provides pro-angiogenic signals sufficient to disrupt VE-cadherin-mediated cell-cell junctions and promote endothelial permeability in brain microvascular endothelial cells. An unbiased angiogenesis-specific antibody array screen identified the chemokine, interleukin-8, which was further demonstrated to function as a key factor involved in glioblastoma-induced permeability, mediated through its receptor CXCR2 on brain endothelia. This underappreciated interface between glioblastoma cells and associated endothelium may inspire the development of novel therapeutic strategies to induce tumor regression by preventing vascular permeability and inhibiting angiogenesis.

  3. Study of the effect of membrane thickness on microcapsule strength, permeability, and cell proliferation

    Ma, Ying; Zhang, Ying; Wang, Yu;


    Cell microencapsulation is one of the promising strategies for in vitro production of proteins or in vivo delivery of therapeutic products. Membrane thickness controls microcapsule strength and permeability, which may in return affect cell growth and metabolism. In this study, the strength......, permeability, and encapsulated Chinese hamster ovary cell proliferation and metabolism of four groups of microcapsules with different membrane thicknesses were investigated. It was found that increasing membrane thickness increases microcapsule strength, whereas decreases membrane permeability. During...... the first 6 days, cells within microcapsules with 10 μm thickness membrane proliferated fast and could reach a cell density of 1.9 × 10(7) cells/mL microcapsule with 92% cell density. A cell density of 5.5 × 10(7) cells/mL microcapsule with >85% cell density was achieved within microcapsules with 15 μm...

  4. A comparative evaluation of corneal epithelial cell cultures for assessing ocular permeability.

    Becker, Ulrich; Ehrhardt, Carsten; Schneider, Marc; Muys, Leon; Gross, Dorothea; Eschmann, Klaus; Schaefer, Ulrich F; Lehr, Claus-Michael


    The purpose of this study was to evaluate the potential value of different epithelial cell culture systems as in vitro models for studying corneal permeability. Transformed human corneal epithelial (HCE-T) cells and Statens Serum Institut rabbit corneal (SIRC) cells were cultured on permeable filters. SkinEthic human corneal epithelium (S-HCE) and Clonetics human corneal epithelium (C-HCE) were received as ready-to-use systems. Excised rabbit corneas (ERCs) and human corneas (EHCs) were mounted in Ussing chambers, and used as references. Barrier properties were assessed by measuring transepithelial electrical resistance, and by determining the apparent permeability of markers with different physico-chemical properties, namely, fluorescein, sodium salt; propranolol hydrochloride; moxaverine hydrochloride; timolol hydrogenmaleate; and rhodamine 123. SIRC cells and the S-HCE failed to develop epithelial barrier properties, and hence were unable to distinguish between the permeation markers. Barrier function and the power to differentiate compound permeabilities were evident with HCE-T cells, and were even more pronounced in the case of C-HCE, corresponding very well with data from ERCs and EHCs. A net secretion of rhodamine 123 was not observed with any of the models, suggesting that P-glycoprotein or similar efflux systems have no significant effects on corneal permeability. Currently available corneal epithelial cell culture systems show differences in epithelial barrier function. Systems lacking functional cell-cell contacts are of limited value for assessing corneal permeability, and should be critically evaluated for other purposes.

  5. Reservoir characteristics and genesis of high-porosity and high-permeability reservoirs in Tarim Basin


    Based on detailed studies, this paper proposes that in the Tarim Basin, hydrocarbon reservoirs widespread either in vertical sequences or in plane and high-porosity and high-permeability reservoirs are developed all over the basin. However, obvious difference and heterogeneity exist among different kinds of reservoirs. The lithologic characteristics, reservoir space types and reservoir properties in various strata have been probed. The result indicates that although the Paleozoic carbonates have been deeply buried for a long period, high-quality reservoirs with the porosity of up to 5%-8% (12% as the maximum) and the permeability of 10×10?3-100×10?3 ?m2 (1000×10?3 ?m2 as the maximum) can be found in certain areas. These include the area with the development of reefs and carbonate beaches, the weathered-crust buried-hill belts that have undergone the long-term exposure, weathering and leaching, the area with the development of dolomitization, and those areas that have experienced the resolution of carbonic acid and organic acid generated by the maturity of the organic matter. Finally, the genesis of the high-porosity and high-permeability reservoirs in deep-buried conditions (with the depth more than 3500 m) have been investigated thoroughly.

  6. Assembly of MOF Microcapsules with Size-Selective Permeability on Cell Walls.

    Li, Wanbin; Zhang, Yufan; Xu, Zehai; Meng, Qin; Fan, Zheng; Ye, Shuaiju; Zhang, Guoliang


    The assembly of metal-organic frameworks (MOFs) into microcapsules has attracted great interest because of their unique properties. However, it remains a challenge to obtain MOF microcapsules with size selectivity at the molecular scale. In this report, we used cell walls from natural biomaterials as non-toxic, stable, and inexpensive support materials to assemble MOF/cell wall (CW) microcapsules with size-selective permeability. By making use of the hollow structure, small pores, and high density of heterogeneous nucleation sites of the cell walls, uniform and continuous MOF layers could be easily obtained by inside/outside interfacial crystallization. The prepared MOF/CW microcapsules have excellent stability and enable the steady, slow, and size-selective release of small molecules. Moreover, the size selectivity of the microcapsules can be adjusted by changing the type of deposited MOF.

  7. Perm-Fit: a new program to estimate permeability at high P-T conditions

    Moulas, Evangelos; Madonna, Claudio


    Several geological processes are controlled by porous fluid flow. The circulation of porous fluids influences many physical phenomena and in turn it depends on the rock permeability. The permeability of rocks is a physical property that needs to be measured since it depends on many factors such as secondary porosity (fractures etc). We present a numerical approach to estimate permeability using the transient step method (Brace et al., 1968). When a non-reacting, compressible fluid is considered in a relative incompressible solid matrix, the only unknown parameter in the equations of porous flow is permeability. Porosity is assumed to be known and the physical properties of the fluid (compressibility, density, viscosity) are taken from the NIST database. Forward numerical calculations for different values of permeability are used and the results are compared to experimental measurements. The extracted permeability value is the one that minimizes the misfit between experimental and numerical results. The uncertainty on the value of permeability is estimated using a Monte Carlo method. REFERENCES Brace, W.F., Walsh J.B., & Frangos, W.T. 1968: Permeability of Granite under High Pressure, Journal of Geophysical Research, 73, 6, 2225-2236

  8. Investigation on the Permeability Evolution of Gypsum Interlayer Under High Temperature and Triaxial Pressure

    Tao, Meng; Yechao, You; Jie, Chen; Yaoqing, Hu


    The permeability of the surrounding rock is a critical parameter for the designing and assessment of radioactive waste disposal repositories in the rock salt. Generally, in the locations that are chosen for radioactive waste storage, the bedded rock salt is a sedimentary rock that contains NaCl and Na2SO4. Most likely, there are also layers of gypsum ( {CaSO}_{ 4} \\cdot 2 {H}_{ 2} {O)} present in the salt deposit. Radioactive wastes emit a large amount of heat and hydrogen during the process of disposal, which may result in thermal damage of the surrounding rocks and cause a great change in their permeability and tightness. Therefore, it is necessary to investigate the permeability evolution of the gypsum interlayer under high temperature and high pressure in order to evaluate the tightness and security of the nuclear waste repositories in bedded rock salt. In this study, a self-designed rock triaxial testing system by which high temperature and pressure can be applied is used; the μCT225kVFCB micro-CT system is also employed to investigate the permeability and microstructure of gypsum specimens under a constant hydrostatic pressure of 25 MPa, an increasing temperature (ranging from 20 to 650 °C), and a variable inlet gas pressure (1, 2, 4, 6 MPa). The experimental results show: (a) the maximum permeability measured during the whole experiment is less than 10-17 m2, which indicates that the gypsum interlayer has low permeability under high temperature and pressure that meet the requirements for radioactive waste repository. (b) Under the same temperature, the permeability of the gypsum specimen decreases at the beginning and then increases as the pore pressure elevates. When the inlet gas pressure is between 0 and 2 MPa, the Klinkenberg effect is very pronounced. Then, as the pore pressure increases, the movement behavior of gas molecules gradually changes from free motion to forced directional motion. So the role of free movement of gas molecules gradually

  9. Intracellular protease activation in apoptosis and cell-mediated cytotoxicity characterized by cell-permeable fluorogenic protease substrates

    Beverly Z Packard; Akira Komoriya


    Over the past decade the importance of signaling from reporter molecules inside live cells and tissues has been clearly established. Biochemical events related to inflammation, tumor metastasis and proliferation, and viral infectivity and replication are examples of processes being further defined as more molecular tools for live cell measurements become available. Moreover, in addition to quantitating parameters related to physiologic processes, real-time imaging of molecular interactions that compose basic cellular activities are providing insights into understanding disease mechanisms as well as extending clinical efficacy of therapeutic regimens. In this review the use of highly cell-permeable fluorogenic substrates that report protease activities inside live cells is described; applications to defining the molecular events of two cellular processes, i.e., apoptosis and cell-mediated cytotoxicity, are then illustrated.

  10. Research on Permeability of Multiphase Medium of Middle to High-Rank Coals

    傅雪海; 秦勇; 姜波; 王文峰; 周世宁


    The permeability of coal of middle to high ranks were tested using He,CH4 and H2O in single phase medium and using CH4 and H2O in double phase medium. The relation between adsorption and permeability of those media was discussed, and the seepage flow characteristics of methane-water medium in coals were analyzed. The result shows that the coalbed methane resource of high-rank coal reservoirs in China is still recoverable.

  11. Polycations increase the permeability of Mycobacterium vaccae cell envelopes to hydrophobic compounds.

    Korycka-Machala, M; Ziółkowski, A; Rumijowska-Galewicz, A; Lisowska, K; Sedlaczek, L


    Polycations [protamine, polymyxin B nonapeptide (PMBN) and polyethyleneimine (PEI)] have been shown to increase the cell wall permeability of Mycobacterium vaccae to highly hydrophobic compounds, as manifested in enhanced intracellular bioconversion of beta-sitosterol to 4-androsten-3,17-dione (AD) and 1,4-androstadien-3,17-dione (ADD), and cell sensitization to erythromycin and rifampicin. The quantity of AD(D) formed per biomass unit was twice as high in the presence of PMBN and PEI, and three times higher with protamine. The sensitization factor, i.e. the MIC(50) ratio of the control bacteria to those exposed to polycations, ranged from 4 to 16, depending on the polycation/antibiotic combination. Non-covalently bound free lipids were extracted from the control and polycation-treated cells and fractionated with the use of chloroform, acetone and methanol. Chloroform- and acetone-eluted fractions (mainly neutral lipids and glycolipids, respectively) showed significant polycation-induced alterations in their quantitative and qualitative composition. The fatty acid profile of neutral lipids was reduced in comparison to control, whereas acetone-derived lipids were characterized by a much higher level of octadecenoic acid (C(18:1)) and a considerably lower content of docosanoic acid (C(22:0)), the marker compound of mycolate-containing glycolipids. Methanol-eluted fractions remained unaltered. Cell-wall-linked mycolates obtained from delipidated cells were apparently unaffected by the action of polycations, as judged from the TLC pattern of mycolic acid subclasses, the mean weight of mycolate preparations and the C(22:0) acid content in the mycolates, determined by GC/MS and pyrolysis GC. The results suggest the involvement of the components of non-covalently bound lipids in the outer layer in the M. vaccae permeability barrier.

  12. Use of novel permeable membrane and air cathodes in acetate microbial fuel cells

    Pant, Deepak, E-mail: deepak.pant@vito.b [Separation and Conversion Technology, VITO - Flemish Institute for Technological Research, Boeretang 200, Mol 2400 (Belgium); Van Bogaert, Gilbert; De Smet, Mark; Diels, Ludo; Vanbroekhoven, Karolien [Separation and Conversion Technology, VITO - Flemish Institute for Technological Research, Boeretang 200, Mol 2400 (Belgium)


    In the existing microbial fuel cells (MFCs), the use of platinized electrodes and Nafion as proton exchange membrane (PEM) leads to high costs leading to a burden for wastewater treatment. In the present study, two different novel electrode materials are reported which can replace conventional platinized electrodes and can be used as very efficient oxygen reducing cathodes. Further, a novel membrane which can be used as an ion permeable membrane (Zirfon) can replace Nafion as the membrane of choice in MFCs. The above mentioned gas porous electrodes were first tested in an electrochemical half cell configuration for their ability to reduce oxygen and later in a full MFC set up. It was observed that these non-platinized air electrodes perform very well in the presence of acetate under MFC conditions (pH 7, room temperature) for oxygen reduction. Current densities of -0.43 mA cm{sup -2} for a non-platinized graphite electrode and -0.6 mA cm{sup -2} for a non-platinized activated charcoal electrode at -200 mV vs. Ag/AgCl of applied potential were obtained. The proposed ion permeable membrane, Zirfonwas tested for its oxygen mass transfer coefficient, K{sub 0} which was compared with Nafion. The K{sub 0} for Zirfon was calculated as 1.9 x 10{sup -3} cm s{sup -1}.

  13. The low/high BCS permeability class boundary: physicochemical comparison of metoprolol and labetalol.

    Zur, Moran; Gasparini, Marisa; Wolk, Omri; Amidon, Gordon L; Dahan, Arik


    Although recognized as overly conservative, metoprolol is currently the common low/high BCS permeability class boundary reference compound, while labetalol was suggested as a potential alternative. The purpose of this study was to identify the various characteristics that the optimal marker should exhibit, and to investigate the suitability of labetalol as the permeability class reference drug. Labetalol's BCS solubility class was determined, and its physicochemical properties and intestinal permeability were thoroughly investigated, both in vitro and in vivo in rats, considering the complexity of the whole of the small intestine. Labetalol was found to be unequivocally a high-solubility compound. In the pH range throughout the small intestine (6.5-7.5), labetalol exhibited pH-dependent permeability, with higher permeability at higher pH values. While in vitro octanol-buffer partitioning (Log D) values of labetalol were significantly higher than those of metoprolol, the opposite was evident in the in vitro PAMPA permeability assay. The results of the in vivo perfusion studies in rats lay between the two contradictory in vitro studies; metoprolol was shown to have moderately higher rat intestinal permeability than labetalol. Theoretical distribution of the ionic species of the drugs was in corroboration with the experimental in vitro and the in vivo data. We propose three characteristics that the optimal permeability class reference drug should exhibit: (1) fraction dose absorbed in the range of 90%; (2) the optimal marker drug should be absorbed largely via passive transcellular permeability, with no/negligible carrier-mediated active intestinal transport (influx or efflux); and (3) the optimal marker drug should preferably be nonionizable. The data presented in this paper demonstrate that neither metoprolol nor labetalol can be regarded as optimal low/high-permeability class boundary standard. While metoprolol is too conservative due to its complete absorption

  14. Effect of Ce3+ on membrane permeability of Escherichia coli cell

    CHEN Aimei; SHI Qingshan; OUYANG Yousheng; CHEN Yiben


    This study aimed to delineate the antibacterial mechanism of rare-earth ion Ce3+ to the target organism Escherichia coli cell,and the most important purpose was to identify its biological effect of increasing the E.coli cell membrane permeability.The antibacterial activities of Ce3+ to E.coli cells were tested,and then the permeability of outer membrane (OM) and inner membrane (IM) were studied by N-phenyl-1-naphthylamine (NPN) and o-nitrophenyl-β-D-galactopyranoside (ONPG) methods separately.Through these experiments we concluded that the E.coli cells grown to log phage were more sensitive to Ce3+ than the ones not at this stage; the structure of membrane was destroyed and the permeability of both OM and IM was obviously increased by Ce3+; there should be certain interactions between Ce3+ and some proteins inside the cell,which impeded the physiological activities of bacteria.

  15. Cell-permeable intrinsic cellular inhibitors of apoptosis protect and rescue intestinal epithelial cells from radiation-induced cell death.

    Matsuzaki-Horibuchi, Shiori; Yasuda, Takeshi; Sakaguchi, Nagako; Yamaguchi, Yoshihiro; Akashi, Makoto


    One of the important mechanisms for gastrointestinal (GI) injury following high-dose radiation exposure is apoptosis of epithelial cells. X-linked inhibitor of apoptosis (XIAP) and cellular IAP2 (cIAP2) are intrinsic cellular inhibitors of apoptosis. In order to study the effects of exogenously added IAPs on apoptosis in intestinal epithelial cells, we constructed bacterial expression plasmids containing genes of XIAP (full-length, BIR2 domain and BIR3-RING domain with and without mutations of auto-ubiquitylation sites) and cIAP2 proteins fused to a protein-transduction domain (PTD) derived from HIV-1 Tat protein (TAT) and purified these cell-permeable recombinant proteins. When the TAT-conjugated IAPs were added to rat intestinal epithelial cells IEC6, these proteins were effectively delivered into the cells and inhibited apoptosis, even when added after irradiation. Our results suggest that PTD-mediated delivery of IAPs may have clinical potential, not only for radioprotection but also for rescuing the GI system from radiation injuries.

  16. The inhibitors of antioxidant cell enzymes induce permeability transition in yeast mitochondria.

    Deryabina, Yulia; Isakova, Elena; Antipov, Alexey; Saris, Nils-Erik L


    In this study we investigated the effects of exogenous and endogenous oxidative stress on mitochondrial membrane permeability transition in yeast cells. E. magnusii yeast was used in the study as it is the only yeast strain possessing a natural high-capacity Са²⁺ transport system. The key reactive oxygen species (ROS) detoxifying enzymes in the yeast cells--catalases (CATs) and superoxide dismutases (SODs)--were fully characterized. At least five isoforms of SODs and only one isoform of CATs were found in the E. magnusii mitochondria. The assessment of the main properties of mitochondrial non-specific permeability under physiological conditions such as dynamics of the membrane potential (∆Ψ) and swelling in mitochondria showed that under physiological conditions classical inhibitors of CATs (ATZ--3-amino-1, 2, 4-triazole) and of SODs (DDC--diethyldithiocarbamate) caused irreversible decline in ∆Ψ in the yeast mitochondria. This decline was accelerated in the presence of 500 μM Са²⁺. The combined action of the inhibitors (ATZ + DDC) promoted moderate swelling in the isotonic medium, which was confirmed by transmission electron microscopy. Mitochondrial swelling in the cells exposed to antioxidant system inhibitors was accompanied by typical signs of early apoptosis, namely by chromatin margination and condensation, vacuolization of the cytosol, and damage of the plasma membrane. Here we showed, at both cellular and mitochondrial levels, that the deregulation of oxidant-scavenging enzymes directly leads to the opening of the mPTP, followed by induction of apoptotic markers in the whole yeast cells. Our studies are the first to clarify the highly contradictory data in the literature on mPTP in yeast mitochondria.

  17. Changes in the permeability of blood brain barrier and endothelial cell damage after cerebral ischemia

    Ke Liu; Jiansheng Li


    OBJECTIVE: To investigate the effect of endothelial cells on the permeability of blood brain barrier (BBB) after brain injury and its effect mechanism.DATA SOURCES: We searched for the articles of permeability of BBB and endothelial cell injury after brain ischemia, which were published between January 1982 and December 2005, with the key words of "cerebral ischemia damage,blood brain barrier ( BBB),permeability,effect of endothelial cell (EC) and its variation mechanism"in English.STUDY SELECTION: The materials were primarily selected. The articles related to the changes in the permeability of BBB and the effect of endothelial cells as well as the change mechanism after cerebral ischemia damage were chosen. Repetitive studies or review articles were excluded.DATA EXTRACTION: Totally 55 related articles were collected, and 35 were excluded due to repetitive or review articles, finally 20 articles were involved.DATA SYNTHESIS: The content or viewpoints of involved literatures were analyzed. Cerebral ischemia had damage for endothelial cells, such as the inflow of a lot of Ca2+, the production of nitrogen monoxide and oxygen free radical, and aggravated destruction of BBB. After acceptors of inflammatory mediators on cerebrovascular endothelial cell membrane, such as histamine, bradykinin , 5-hydroxytryptamine and so on are activated, endothelial cells shrink and the permeability of BBB increases. Its mechanism involves in the inflow of extracellular Ca2+and the release of intracellular Ca2+ in the cells. Glycocalyx molecule on the surface of endothelial cell, having structural polytropy, is the determinative factor of the permeability of BBB. VEGF, intensively increasing the vasopermeability and mainly effecting on postcapillary vein and veinlet, is the strongest known blood vessel permeation reagent. Its chronic overexpression in the brain can lead the destruction of BBB.CONCLUSION: The injury of endothelial cell participants in the pathological mechanism of BBB

  18. Graphene oxide membranes with high permeability and selectivity for dehumidification of air

    Shin, Yongsoon; Liu, Wei; Schwenzer, Birgit; Manandhar, Sandeep; Chase-Woods, Dylan G.; Engelhard, Mark H.; Devanathan, Ram; Fifield, Leonard S.; Bennett, Wendy D.; Ginovska-Pangovska, Bojana; Gotthold, David W.


    Hierarchically stacked 2D graphene oxide (GO) membranes are a fascinating and promising new class of materials with the potential for radically improved water vapor/gas separation with excellent selectivity and high permeability. This paper details dehumidification results from flowing gas mixtures through free-standing GO membrane samples prepared by a casting method. The first demonstrated use of free-standing GO membranes for water vapor separation reveals outstanding water vapor permeability and H2O/N2 selectivity. Free-standing GO membranes exhibit extremely high water vapor permeability of 1.82 x 105 Barrer and a water vapor permeance of 1.01 x 10-5 mol/m2sPa, while the nitrogen permeability was below the system’s detection limit, yielding a selectivity >104 in 80% relative humidity (RH) air at 30.8 °C. The results show great potential for a range of energy conversion and environmental applications

  19. A laminate of ferromagnetic films with high effective permeability at high frequencies

    I. T. Iakubov


    Full Text Available The paper reports on development of magnetodielectric material with high microwave permeability. The material is a laminate of multi-layer permalloy films deposited onto a thin mylar substrate by magnetron sputtering. The deposited films are arranged into a stack and glued together under pressure to obtain the laminate. With the content of ferromagnetic component in the laminate being 22 % vol., its measured quasistatic permeability is 60. The peak value of imaginary permeability attains 50 and the peak is located near 1 GHz. As compared with the multi-layer films, which the laminate is made of, it exhibits lower magnetic loss tangent at frequencies below the magnetic loss peak and may therefore be useful for many technical applications. Lower low-frequency loss may be attributed to pressing of the glued sample. This rectifies wrinkling appearing due to sputtering of rigid multi-layer film onto flexible mylar substrate and, therefore, makes the magnetic structure of the film more uniform.

  20. Vacuum Infusion Molding Process Part 1:VIMP Based on a High-Permeable Medium

    ZHU Ying-dan; WANG Ji-hui; YANG Zui; TAN Hua


    The visualization experiments were carried out to investigate the permeability of the high-permeable medium (HPM) and the roles of the peel ply and the HPM in the mold filling.The influence of process parameters on mold filling is discussed.Furthermore,the whole vacuum infusion molding process (VIMP) procedure is introduced in detail taking the manufacture of a model boat for example.

  1. Effect of Aβ1-40 on membrane permeability and intracellular free Ca2+ of nerve cells


    The effects of soluble and fibrillar Aβ1-40 on membrane permeability and intracellular free Ca2+ of nerve cells were investigated by the laser confocal microscopy. Results indicate that: i) Effects of soluble and fibrillar Aβ1-40 on cell membrane permeability are both concentration-de- pendent. Soluble Aβ1-40 increases membrane permeability only at concentration of 3 ?mol/L, while the toxic effect of fibrillar Aβ1-40 is much stronger, its evident effect begins from 1 ?mol/L. When its concentration rose to 3 ?mol/L, not only the membrane permeability increased, but also the nuclear membrane broke seriously. ii) Both soluble and fibrillar Aβ1-40 at high concentrations increased the intracellular free Ca2+, and the increased amplitudes are concentration-dependent. However, the fibrillar one induces the increase of intracellular Ca2+ much quicker and synchronously. These results indicate that some correlation exists between the neurotoxicity of high concentration soluble and fibrillar Aβ1-40 and the change of physico-chemical properties and intracellular Ca ion imbalance.

  2. Numerical simulation and experiment analysis of improving permeability by deep-hole presplitting explosion in high gassy and low permeability coal seam

    CAI Feng; LIU Ze-gong; LIN Bai-quan; LI Wei


    Created a new damage model for explosive for LS-DYNA3D, taking advantage of the Taylor method aimed at the high gassy and low permeability coal seam, and nu-merically simulated and analyzed the deep-hole presplitting explosion. The entire process of explosion was represented, including cracks caused by dynamic pressure, transmission and vibration superposition of stress waves, as well as cracks growth driven by gas gen-erated by explosion. The influence of the cracks generated in the process of explosion and the performance of improving permeability caused by the difference of interval between explosive holes were analyzed. A reasonable interval between explosive holes of deep-hole presplitting explosions in high gassy and low permeability coal seams was proposed, and the resolution of gas drainage in high gassy and low permeability coal seam was put forward.

  3. Negative-permittivity plasma generation in negative-permeability space with high-energy metamaterials

    Sakai, Osamu; Nakamura, Yoshihiro; Iwai, Akinori; Iio, Satoshi


    Plasma generation by electromagnetic waves in negative-permeability space is analyzed using experimental results and theoretical models. Installation of negative-permeability metamaterials triggers drastic changes to the propagation of electromagnetic waves. Unlike usual cases in which permeability is  +1, negative permeability induces evanescent modes in a space without plasma. However, if permittivity becomes negative due to high-electron-density or overdense plasma, electromagnetic waves can propagate because negative-refractive-index states emerge. In this study, reviewing our previous experimental data, we study the underlying physical processes in plasma generation in terms of wave propagation and parameters of wave media. We confirm nonlinear (transition) processes in the phase of density evolution up to the negative permittivity state and negative-refractive-index states in the quasi-steady phase. We also note that such energetic metamaterials are built up when we use plasma, unlike conventional metamaterials composed of solid-state materials.

  4. Avidin-biotin-based approach to forming heterotypic cell clusters and cell sheets on a gas-permeable membrane

    Hamon, M; Ozawa, T; Montagne, K; Kojima, N; Ishii, R; Sakai, Y [Institute of Industrial Science (IIS), University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505 (Japan); Yamaguchi, S; Nagamune, T [Department of Bioengineering, Graduate School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan); Ushida, T, E-mail: [Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)


    Implantation of sheet-like liver tissues is a promising method in hepatocyte-based therapies, because angiogenesis is expected to occur upon implantation from the surrounding tissues. In this context, we introduce here a new methodology for the formation of a functional thick hepatic tissue usable for cell sheet technology. First, we report the formation of composite tissue elements in suspension culture. Composite elements were composed of human hepatoma Hep G2 cells and mouse NIH/3T3 fibroblasts which are important modulators for thick-tissue formation. To overcome the very low attachment and organization capability between different cells in suspension, we synthesized a new cell-to-cell binding molecule based on the avidin-biotin binding system that we previously applied to attach hepatocytes on artificial substrata. This newly synthesized biotin-conjugated biocompatible anchoring molecule was inserted in the plasma membrane of both cell types. NIH/3T3 cells were further conjugated with avidin and incubated with biotin-presenting Hep G2 cells to form highly composite tissue elements. Then, we seeded those elements on highly gas-permeable membranes at their closest packing density to induce the formation of a thick, composite, functional hepatic tissue without any perfusion. This methodology could open a new way to engineer implantable thick liver tissue sheets where different cell types are spatially organized and well supplied with oxygen.

  5. Study of the effect of membrane thickness on microcapsule strength, permeability, and cell proliferation.

    Ma, Ying; Zhang, Ying; Wang, Yu; Wang, Qiuyan; Tan, Mingqian; Liu, Yang; Chen, Li; Li, Na; Yu, Weiting; Ma, Xiaojun


    Cell microencapsulation is one of the promising strategies for in vitro production of proteins or in vivo delivery of therapeutic products. Membrane thickness controls microcapsule strength and permeability, which may in return affect cell growth and metabolism. In this study, the strength, permeability, and encapsulated Chinese hamster ovary cell proliferation and metabolism of four groups of microcapsules with different membrane thicknesses were investigated. It was found that increasing membrane thickness increases microcapsule strength, whereas decreases membrane permeability. During the first 6 days, cells within microcapsules with 10 μm thickness membrane proliferated fast and could reach a cell density of 1.9 × 10(7) cells/mL microcapsule with 92% cell density. A cell density of 5.5 × 10(7) cells/mL microcapsule with >85% cell density was achieved within microcapsules with 15 μm membrane thickness and these microcapsules kept over 88% integrity ratio after 11 days, which was much higher than that of microcapsules with 10 μm membrane thickness. Membrane with more than 20 μm thickness was not suited for encapsulated cell culture owing to low-protein diffusion rate. These results indicated that cells survived shortly within the thinnest membrane thickness. There was a specific membrane thickness more suitable for cell growth for a long-time culture. These findings will be useful for preparing microcapsules with the desired membrane thickness for microencapsulated cell culture dependent on various purposes.

  6. Ca2+-permeable and Ca2+-impermeable AMPA receptors coexist on horizontal cells.

    Huang, Shi-Yong; Liang, Pei-Ji


    Fura-2 fluorescent calcium imaging was used for analyzing the subtype of AMPA receptors in freshly dissociated horizontal cells of carp retina. Exogenous application of AMPA induced an increase of intracellular concentration of free Ca2+ ([Ca2+]i) in horizontal cells, while the [Ca2+]i increase was partly inhibited by nifedipine. The residual [Ca2+]i increase was completely eliminated by joro spider toxin-3, a blocker of Ca2+-permeable AMPA receptors. On the other hand, the application of pentobarbital, which blocked Ca2+-impermeable AMPA receptors, could also partly inhibit the increase of [Ca2+]i, implying that the application of AMPA induced the activation of both Ca2+-permeable and Ca2+-impermeable AMPA receptors and the consequent activation of voltage-gated Ca2+ channels. Taken together, these results suggested that Ca2+-permeable and Ca2+-impermeable AMPA receptors were coexpressed on horizontal cells.

  7. Hepatitis C virus NS3 protease inhibitors: large, flexible molecules of peptide origin show satisfactory permeability across Caco-2 cells.

    Bergström, Christel A S; Bolin, Sara; Artursson, Per; Rönn, Robert; Sandström, Anja


    The purpose of this study was to investigate the intestinal absorption of tripeptide-based compounds intended for treatment of hepatitis C virus (HCV) infection. The intestinal permeability of 11 HCV NS3 protease inhibitors (Mw 687-841, ClogD(pH 7.4) 1.2-7.3 and 10-13 hydrogen bond donors/acceptors) was measured using Caco-2 cells. Each compound was investigated in the apical to basolateral (a-b) and basolateral to apical (b-a) direction at pH 7.4. For compounds displaying efflux the experiment was repeated in the presence of 1 microM GF120918 to investigate possible involvement of P-glycoprotein (Pgp; ABCB1). All compounds displayed intermediate to high permeability. Seven of them showed extensive efflux, with 31-114-fold higher permeability in the b-a direction than the a-b direction. Addition of the Pgp inhibitor GF120918 reduced the b-a transport rate for the effluxed compounds. However, for inhibitors with a C-terminal carboxylic acid and the acidic bioisosteres thereof the efflux was still significant. Hence, the negative charge resulted in efflux by other ABC-transporters than Pgp. From this study it can be concluded that small changes in the overall structure can lead to a large variation in permeability and efflux as shown by the inhibitors herein, properties that also may influence the resulting inhibition potency of the compounds when performing cell-based pharmacological assays.

  8. The Water Permeability Reduction After Successive Hypo-Osmotic Shocks in Kidney Principal Cells is Apically Regulated

    Liubov E. Katkova


    Full Text Available Background/Aims: Renal principal cells maintain their intracellular water and electrolyte content despite significant fluctuations of the extracellular water and salt concentrations. Their water permeability decreases rapidly (within a few seconds after successive hypo-osmotic shocks. Our aim was to investigate the contribution of the apical and basolateral surface to this effect and the potential influence of fast reduction in AQP-2, -3 or -4 plasma membrane content. Methods: Rat principal cells of kidney collecting duct fragments underwent hypo-osmotic challenge applied apically or basolaterally and the regulatory volume decrease (RVD was measured by the calcein quenching method. The AQP -2, -3 and -4 content of the plasma membrane fraction was quantified by Western blotting. Results: The hypo-osmotic shock applied apically causes rapid swelling with high apparent water permeability and fast RVD. An identical successive shock after 15-20 sec causes significantly lower swelling rate with 3-fold reduction in apparent water permeability. This reaction is accompanied by AQP2 decrease in the plasma membrane while AQP3 and AQP4 are unaffected. The contribution of the basolateral cell surface to RVD is significantly lower than the apical. Conclusion: These results indicate that in principal cells the effective mechanism of RVD is mainly regulated by the apical cell plasma membrane.

  9. The influence of naphthenic acids and their fractions onto cell membrane permeability

    Pavlović Ksenija


    Full Text Available The influence of naphthenic acids (NAs mixture and their narrow fractions (called NA pH 4, pH 8 and pH 10 onto permeability of beetroot cell membrane is examined. The results showed that the effect depends on treatment duration, concentration and NAs structure. Longer treatment of plant cell membranes with sodium naphthenate (Na-naph resulted in the increase of membrane permeability (e.g. 4-hour treatment with Na-naph (C=100 μmol L-1 increased membrane permeability about 3 times, while prolongation of treatment to 24 hour resulted in the 18 times increasing of the effect. NAs in the concentration range from 0.1 to 10 μmol L-1 does not change membrane permeability, while membrane permeability is increasing linearly with concentration increasing from 10-100 μmol L-1. The strongest effect expressed fraction pH 8, where bi- and tricyclic carboxylic acids are the most abundant. These structures are predominant in the total NAs mixture as well. Thereby could be explained their closest, but a little bit weaker effect, comparing to NAs present in fraction pH 8. The effect of NAs onto beetroot cell membrane is between the effects of anionic (SDS and LS and non-ionic surfactants (Triton X-100. [Projekat Ministarstva nauke Republike Srbije, br. 172006. i br. TR31036

  10. Acyl-homoserine lactones suppresses IEC-6 cell proliferation and increase permeability of isolated rat colon.

    Joe, Ga-Hyun; Andoh, Midori; Nomura, Mikako; Iwaya, Hitoshi; Lee, Jae-Sung; Shimizu, Hidehisa; Tsuji, Youhei; Maseda, Hideaki; Miyazaki, Hitoshi; Hara, Hiroshi; Ishizuka, Satoshi


    We investigated to determine whether a variety of acyl-homoserine lactones (AHLs) influences epithelial cell proliferation and mucosal permeability. 3-Oxo-C12-homoserine lactone (HSL) and 3-oxo-C14-HSL significantly suppressed IEC-6 cell proliferation. A significant increase in mucosal permeability was observed in isolated rat colon tissue exposed to C12-HSL, 3-oxo-C12-HSL, and 3-oxo-C14-HSL. These data indicate that AHLs suppress epithelial proliferation and disrupt barrier function in intestinal mucosa.

  11. A PCNA-derived cell permeable peptide selectively inhibits neuroblastoma cell growth.

    Long Gu

    Full Text Available Proliferating cell nuclear antigen (PCNA, through its interaction with various proteins involved in DNA synthesis, cell cycle regulation, and DNA repair, plays a central role in maintaining genome stability. We previously reported a novel cancer associated PCNA isoform (dubbed caPCNA, which was significantly expressed in a broad range of cancer cells and tumor tissues, but not in non-malignant cells. We found that the caPCNA-specific antigenic site lies between L126 and Y133, a region within the interconnector domain of PCNA that is known to be a major binding site for many of PCNA's interacting proteins. We hypothesized that therapeutic agents targeting protein-protein interactions mediated through this region may confer differential toxicity to normal and malignant cells. To test this hypothesis, we designed a cell permeable peptide containing the PCNA L126-Y133 sequence. Here, we report that this peptide selectively kills human neuroblastoma cells, especially those with MYCN gene amplification, with much less toxicity to non-malignant human cells. Mechanistically, the peptide is able to block PCNA interactions in cancer cells. It interferes with DNA synthesis and homologous recombination-mediated double-stranded DNA break repair, resulting in S-phase arrest, accumulation of DNA damage, and enhanced sensitivity to cisplatin. These results demonstrate conceptually the utility of this peptide for treating neuroblastomas, particularly, the unfavorable MYCN-amplified tumors.

  12. Aquaporin 1 Facilitated Hepatocellular Carcinoma SMMC7221 Cell Migration Associated with Water Permeability

    ZHANG Ai-li; LI Jiang; WANG Yan-qing; ZAKNROU Zohra; MA Tong-hui; LI Xiao-meng


    The authors investigated the regulation of human aquaporin l(hAQPl) and the involvement of aquaporin l(AQPl) in the migration of human hepatocellular carcinoma SMMC-7221 cells using RNA intereference technology.Firstly, two short hairpin RNA(shRNA) constructs in PBSU6 vector were reconstructed and their knockdown effects were identified in SMMC-7221 cells. Next, the involvement of endogenous hAQPl in regulating the migration of SMMC-7221 cells was investigated via siRNA technology. HAQPl-shRNA can specifically inhibit AQPl dependent osmotic water permeability. Meanwhile the migration of SMMC-7221 cells was inhibited remarkably after silencing AQPl by performing transwell cell migration assay and in vitro wound healing assay. Furthermore, in the presence of an inhibitor HgCl2, the water permeability of the cell membrane was remarkably decreased, the expression of AQPl was upregulated after HgCl2 treatment and the cell movement was decreased at the moment. Increased AQPl cannot attenuate cell migration ability when cell membrane loses its water permeability function. This demonstrates that the cell migration was remarkably related to the transporting water function of cell membrane.

  13. Electrical and optical characterization of thrombin-induced permeability of cultured endothelial cell monolayers on semiconductor electrode arrays

    Hillebrandt, H.; Abdelghani, A.; Abdelghani-Jacquin, C.; Aepfelbacher, M.; Sackmann, E.

    Impedance spectroscopy and phase-contrast microscopy are combined to monitor the electrical and morphological properties of human umbilical vein endothelial cell monolayers. The cells were cultured on optically transparent indium-tin-oxide (ITO) semiconductor electrode arrays coated with collagen IV, and the effect of the inflammatory mediator thrombin on monolayer permeability was monitored in real time. ITO electrodes provide several advantages for these kinds of experiments, because they are optically transparent, polarizable and highly sensitive due to the absence of insulating oxide layers. A qualitative correlation between the thrombin-induced gap formation and the electrical parameters of the cell layer is established.

  14. Method for the preparation of high surface area high permeability carbons

    Lagasse, R.R.; Schroeder, J.L.


    A method for preparing carbon materials having high surface area and high macropore volume to provide high permeability. These carbon materials are prepared by dissolving a carbonizable polymer precursor, in a solvent. The solution is cooled to form a gel. The solvent is extracted from the gel by employing a non-solvent for the polymer. The non-solvent is removed by critical point drying in CO{sub 2} at an elevated pressure and temperature or evaporation in a vacuum oven. The dried product is heated in an inert atmosphere in a first heating step to a first temperature and maintained there for a time sufficient to substantially cross-link the polymer material. The cross-linked polymer material is then carbonized in an inert atmosphere. 3 figs.

  15. Method for the preparation of high surface area high permeability carbons

    Lagasse, Robert R. (Albuquerque, NM); Schroeder, John L. (Albuquerque, NM)


    A method for preparing carbon materials having high surface area and high macropore volume to provide high permeability. These carbon materials are prepared by dissolving a carbonizable polymer precursor, in a solvent. The solution is cooled to form a gel. The solvent is extracted from the gel by employing a non-solvent for the polymer. The non-solvent is removed by critical point drying in CO.sub.2 at an elevated pressure and temperature or evaporation in a vacuum oven. The dried product is heated in an inert atmosphere in a first heating step to a first temperature and maintained there for a time sufficient to substantially cross-link the polymer material. The cross-linked polymer material is then carbonized in an inert atmosphere.

  16. Slit2-Robo4 receptor responses inhibit ANDV directed permeability of human lung microvascular endothelial cells.

    Gorbunova, Elena E; Gavrilovskaya, Irina N; Mackow, Erich R


    Hantaviruses nonlytically infect human endothelial cells (ECs) and cause edematous and hemorrhagic diseases. Andes virus (ANDV) causes hantavirus pulmonary syndrome (HPS), and Hantaan virus (HTNV) causes hemorrhagic fever with renal syndrome (HFRS). Hantaviruses enhance vascular endothelial growth factor directed EC permeability resulting in the disassembly of inter-endothelial cell adherens junctions (AJs). Recent studies demonstrate that Slit2 binding to Robo1/Robo4 receptors on ECs has opposing effects on AJ disassembly and vascular fluid barrier functions. Here we demonstrate that Slit2 inhibits ANDV and HTNV induced permeability and AJ disassembly of pulmonary microvascular ECs (PMECs) by interactions with Robo4. In contrast, Slit2 had no effect on the permeability of ANDV infected human umbilical vein ECs (HUVECs). Analysis of Robo1/Robo4 expression determined that PMECs express Robo4, but not Robo1, while HUVECs expressed both Robo4 and Robo1 receptors. SiRNA knockdown of Robo4 in PMECs prevented Slit2 inhibition of ANDV induced permeability demonstrating that Robo4 receptors determine PMEC responsiveness to Slit2. Collectively, this data demonstrates a selective role for Slit2/Robo4 responses within PMECs that inhibits ANDV induced permeability and AJ disassembly. These findings suggest Slit2s utility as a potential HPS therapeutic that stabilizes the pulmonary endothelium and antagonizes ANDV induced pulmonary edema.

  17. Diethylentriaminepenta acetic acid glucose conjugates as a cell permeable iron chelator

    Mona Mosayebnia


    Full Text Available Objective: To find out whether DTPA-DG complex can enhance clearance of intracellular free iron. Materials and Methods: Diethylenetriaminepentaacetic acid-D-deoxy-glucosamine (DTPA-DG was synthesized and examined for its activity as a cell-permeable iron chelator in human hepatocellular carcinoma (HEPG2 cell line exposed to high concentration of iron sulfate and compared with deferoxamine (DFO, a prototype iron chelator. The effect of DTPA-DG on cell viability was monitored using the 3-(4,5-dimethythiazol-2-yl-2,5-diphenyl tetrazolium bromide MTT assay as well. Results: There was a significant increase of iron level after iron overload induction in HEPG2 cell culture. DTPA-DG presented a remarkable capacity to iron burden reducing with estimated 50% inhibitory concentration value of 65.77 nM. In fact, glycosyl moiety was gained access of DTPA to intracellular iron deposits through glucose transporter systems. Conclusion: DTPA-DG, more potent than DFO to sequester deposits of free iron with no profound toxic effect. The results suggest the potential of DTPA-DG in chelating iron and permitting its excretion from primary organ storage.

  18. Permittivity and permeability determination for high index specimens using partially filled shorted rectangular waveguides

    Scott, Mark M; Bean, Jeffrey A; Allen, Kenneth W


    A method for determining the permittivity and permeability for specimens with high refractive index and variable shape is investigated. The method extracts the permeability and permittivity tensor elements from reflection measurements made with a partially-filled shorted rectangular waveguide on an electrically small specimen. Measurements are performed for two isotropic, heavily loaded coaxial magnetic composites. Supporting measurements from a stripline cavity and coaxial airline are used to validate the method. The results demonstrate the methods ability to handle frequency dispersive and high index materials.

  19. High-pressure homogenization lowers water vapor permeability of soybean protein isolate-beeswax films.

    Zhang, Chao; Ma, Yue; Guo, Kuan; Zhao, Xiaoyan


    Soybean-protein isolate (SPI) has excellent film-forming capacity. However, the water vapor permeability of SPI film is high, which will cause the moisture lose of packaged products. The effect of high-pressure homogenization (HPH) on the water vapor permeability of SPI-beeswax films was evaluated. The HPH was effective at lowering the water vapor permeability of SPI-beeswax films to about 50% of the control. The HPH reduced the particle size of films and made their matrix more compact. The HPH improved the hydrophobicity of SPI-beeswax films. For the first time, we proved that the HPH improved the bound-beeswax content in SPI-beeswax films. The bound beeswax was effective at lowering the water vapor permeability of films rather than the free beeswax in the film matrix. In summary, the HPH lowered water vapor permeability of SPI-beeswax films by reducing their particle size and raising their hydrophobicity and bound-beeswax content.

  20. Development of implantable hemodialysis system using PES membranes with high water-permeability.

    To, N; Sanada, I; Ito, H; Morita, S; Kanno, Y; Miki, N


    This paper presents development of high water-permeable dialysis membranes. We proposed the system that does not use dialysis fluid for the implantable micro dialysis treatment and development of such membranes is crucial. We developed micro dialysis system composed by nanoporous membranes and microfluidic channels in our prior work. The membranes were made of nanoporous polyethersulfone (PES), which was not water-permeable. By not using dialysate, our device can be simplified because the pumps and storage tanks for the dialysis fluid are not necessary. This treatment is termed as hemofiltration. We measured the water permeability of PES membrane with respect to the concentrations of the PES, the additives, and the solvents in the casting solution. We could find the membranes with sufficiently high water permeability through in vitro experiments using a syringe pomp and whole cow blood, and the membrane had enough mechanical strength. We conducted experiments with multi-layered device in in vitro and in vivo using rats, where the system was connected to the vein and artery. We successfully collected the filtrate beyond target line, which was set by a medical doctor, without any leakage of blood from the device. The results verified that the filtration device can be scaled-up by increasing a number of the layer. We connected the device to a rat for 5h. It was verified the device maintained almost constant water permeability beyond our target line.

  1. Experimental Measurement of Relative Permeability Functions for Fuel Cell GDL Materials

    Hussaini, Irfan


    Gas diffusion layer in PEM fuel cells plays a pivotal role in water management. Modeling of liquid water transport through the GDL relies on knowledge of relative permeability functions in the in-plane and through-plane directions. In the present work, air and water relative permeabilities are experimentally determined as functions of saturation for typical GDL materials such as Toray-060, -090, -120 carbon paper and E-Tek carbon cloth materials in their plain, untreated forms. Saturation is measured using an ex-situ gravimetric method. Absolute and relative permeability functions in the two directions of interest are presented. Significant departure from the generally assumed cubic function of saturation is observed. ©The Electrochemical Society.

  2. Resistin impairs glucose permeability in EA.hy926 cells by down-regulating GLUT1 expression.

    Li, Qiang; Cai, Yuxi; Huang, Jing; Yu, Xiaolan; Sun, Jun; Yang, Zaiqing; Zhou, Lei


    Type 2 diabetes mellitus (T2DM) is a chronic disease which is now affecting the health of more and more people in the world. Resistin, discovered in 2001, is considered to be closely related to metabolic dysfunction and obesity. Previous study showed that hyperglycemia is always accompanied by a high serum resistin concentration. We therefore investigated whether resistin can mediate glucose transfer across the blood-tissue barrier. Here, we employed a transwell system to analyze glucose permeability in EA.hy926 human endothelial cells treated without or with human resistin. In EA.hy926 cells treated with resistin, the permeability to glucose was heavily impaired. This was due to the down-regulation of GLUT1 expression as a result of the treatment, rather than regulation of tight junctions. In addition, overexpression of GLUT1 in EA.hy926 cells was able to recover the blocking effect of resistin on glucose permeability. We further found that resistin could inhibit the expression of peroxisome proliferator-activated receptor gamma (PPARγ) and consequently impede the transcription of GLUT1. The results of the present study suggested that resistin could cause glucose retention in serum and thus result in hyperglycemia. This provides a novel explanation for hyperglycemia and a potential new way of treating type 2 diabetes mellitus. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  3. Influence of pore roughness on high-frequency permeability

    Cortis, A.; Smeulders, D.M.J.; Guermond, J.L.; Lafarge, D.


    The high-frequency behavior of the fluid velocity patterns for smooth and corrugated pore channels is studied. The classical approach of Johnson et al. [J. Fluid Mech. 176, 379 (1987)] for smooth geometries is obtained in different manners, thus clarifying differences with Sheng and Zhou [Phys. Rev.

  4. Calculation of High Frequency Complex Permeability of Carbonyl Iron Flakes in a Nomagnetic Matrix

    WEN Fu-Sheng; QIAO Liang; YI Hai-Bo; ZHOU Dong; LI Fa-Shen


    The carbonyl iron flakes are fabricated by high-energy ball milling.The effective permeability is measured and calculated for the composite consisting of flakes embedded in a nonmagnetic matrix.The magnetic flakes with a shape anisotropy and random spatial distribution of normal direction are considered to calculate the complex permeability of magnetic flake materials.Its analytical model is derived from the Landau-Lifshitz- Gilbert equation and Bruggeman's effective medium theory.The calculated results agree well with the experiment.

  5. Ultra-low percolation threshold in ferrite-metal cofired ceramics brings both high permeability and high permittivity.

    Wang, Liang; Bai, Yang; Lu, Xuefei; Cao, Jiang-Li; Qiao, Li-Jie


    High permeability and high permittivity are hard to be achieved simultaneously, either in single-phased materials or in composite materials, such as ferrite-ferroelectric ceramic composites and ferrite-metal percolative composites. In this work, ultra-low percolation threshold is achieved in NiZnCu ferrite-Ag cofired ceramics, which endows the composite with both high permeability and high permittivity by minimizing the negative effect of nonmagnetic conductive fillers on magnetic properties. The percolation threshold is controlled by the temperature matching between ferrite densification and Ag melting. A thin and long percolative net forms between large ferrite grains under a proper cofiring process, which brings a low percolation threshold of 1.21vol%, more than one order of magnitude lower than the theoretical value of 16vol%. Near the ultra-low threshold, the composite exhibits a high permeability of 585 and a high permittivity of 78.

  6. Enzymatically active high-flux selectively gas-permeable membranes

    Jiang, Ying-Bing; Cecchi, Joseph L.; Rempe, Susan; FU, Yaqin; Brinker, C. Jeffrey


    An ultra-thin, catalyzed liquid transport medium-based membrane structure fabricated with a porous supporting substrate may be used for separating an object species such as a carbon dioxide object species. Carbon dioxide flux through this membrane structures may be several orders of magnitude higher than traditional polymer membranes with a high selectivity to carbon dioxide. Other gases such as molecular oxygen, molecular hydrogen, and other species including non-gaseous species, for example ionic materials, may be separated using variations to the membrane discussed.

  7. A biomimic thermal fabric with high moisture permeability

    Fan Jie


    Full Text Available Moisture comfort is an essential factor for functional property of thermal cloth, especially for thick thermal cloth, since thick cloth may hinder effective moisture permeation, and high moisture concentration in the micro-climate between skin and fabric would cause cold feeling. Here, we report a biomimic thermal fabric with excellent warm retention and moisture management properties. In this fabric, the warp yarn system constructs many tree-shaped channel nets in the thickness direction of the fabric. Experimental result indicates that the special hierarchic configuration of warp yarns endows the biomimic thermal fabric with a better warm retention and water vapor management properties compared with the traditional fabrics.

  8. A plate reader-based method for cell water permeability measurement

    Fenton, Robert A.; Moeller, H B; Nielsen, S


    Cell volume and water permeability measurements in cultured mammalian cells are typically conducted under a light microscope. Many of the employed approaches are time consuming and not applicable to a study of confluent epithelial cell monolayers. We present here an adaptation of a calcein-quenching-based......-mannitol concentrations. Similarly, according average cell volumes have been measured in suspension in a Coulter counter (particle-sizing device). Based on these measurements, we have derived an equation that facilitates the modeling of cell volume changes based on fluorescence intensity changes. We have utilized...

  9. Effects of low-intensity ultrasound on the growth, cell membrane permeability and ethanol tolerance of Saccharomyces cerevisiae.

    Dai, Chunhua; Xiong, Feng; He, Ronghai; Zhang, Weiwei; Ma, Haile


    Effects of low-intensity ultrasound (at different frequency, treatment time and power) on Saccharomyces cerevisiae in different growth phase were evaluated by the biomass in the paper. In addition, the cell membrane permeability and ethanol tolerance of sonicated Saccharomyces cerevisiae were also researched. The results revealed that the biomass of Saccharomyces cerevisiae increased by 127.03% under the optimum ultrasonic conditions such as frequency 28kHz, power 140W/L and ultrasonic time 1h when Saccharomyces cerevisiae cultured to the latent anaphase. And the membrane permeability of Saccharomyces cerevisiae in latent anaphase enhanced by ultrasound, resulting in the augment of extracellular protein, nucleic acid and fructose-1,6-diphosphate (FDP) contents. In addition, sonication could accelerate the damage of high concentration alcohol to Saccharomyces cerevisiae although the ethanol tolerance of Saccharomyces cerevisiae was not affected significantly by ultrasound. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Endothelial cell permeability during hantavirus infection involves factor XII-dependent increased activation of the kallikrein-kinin system.

    Shannon L Taylor

    Full Text Available Hemorrhagic fever with renal syndrome (HFRS and hantavirus pulmonary syndrome (HPS are diseases caused by hantavirus infections and are characterized by vascular leakage due to alterations of the endothelial barrier. Hantavirus-infected endothelial cells (EC display no overt cytopathology; consequently, pathogenesis models have focused either on the influx of immune cells and release of cytokines or on increased degradation of the adherens junction protein, vascular endothelial (VE-cadherin, due to hantavirus-mediated hypersensitization of EC to vascular endothelial growth factor (VEGF. To examine endothelial leakage in a relevant in vitro system, we co-cultured endothelial and vascular smooth muscle cells (vSMC to generate capillary blood vessel-like structures. In contrast to results obtained in monolayers of cultured EC, we found that despite viral replication in both cell types as well as the presence of VEGF, infected in vitro vessels neither lost integrity nor displayed evidence of VE-cadherin degradation. Here, we present evidence for a novel mechanism of hantavirus-induced vascular leakage involving activation of the plasma kallikrein-kinin system (KKS. We show that incubation of factor XII (FXII, prekallikrein (PK, and high molecular weight kininogen (HK plasma proteins with hantavirus-infected EC results in increased cleavage of HK, higher enzymatic activities of FXIIa/kallikrein (KAL and increased liberation of bradykinin (BK. Measuring cell permeability in real-time using electric cell-substrate impedance sensing (ECIS, we identified dramatic increases in endothelial cell permeability after KKS activation and liberation of BK. Furthermore, the alterations in permeability could be prevented using inhibitors that directly block BK binding, the activity of FXIIa, or the activity of KAL. Lastly, FXII binding and autoactivation is increased on the surface of hantavirus-infected EC. These data are the first to demonstrate KKS activation

  11. CCR7 and IRF4-dependent dendritic cells regulate lymphatic collecting vessel permeability.

    Ivanov, Stoyan; Scallan, Joshua P; Kim, Ki-Wook; Werth, Kathrin; Johnson, Michael W; Saunders, Brian T; Wang, Peter L; Kuan, Emma L; Straub, Adam C; Ouhachi, Melissa; Weinstein, Erica G; Williams, Jesse W; Briseño, Carlos; Colonna, Marco; Isakson, Brant E; Gautier, Emmanuel L; Förster, Reinhold; Davis, Michael J; Zinselmeyer, Bernd H; Randolph, Gwendalyn J


    Lymphatic collecting vessels direct lymph into and from lymph nodes (LNs) and can become hyperpermeable as the result of a previous infection. Enhanced permeability has been implicated in compromised immunity due to reduced flow of lymph and immune cells to LNs, which are the primary site of antigen presentation to T cells. Presently, very little is known about the molecular signals that affect lymphatic collecting vessel permeability. Here, we have shown that lymphatic collecting vessel permeability is controlled by CCR7 and that the chronic hyperpermeability of collecting vessels observed in Ccr7-/- mice is followed by vessel fibrosis. Reexpression of CCR7 in DCs, however, was sufficient to reverse the development of such fibrosis. IFN regulatory factor 4-positive (IRF4+) DCs constitutively interacted with collecting lymphatics, and selective ablation of this DC subset in Cd11c-Cre Irf4fl/fl mice also rendered lymphatic collecting vessels hyperpermeable and fibrotic. Together, our data reveal that CCR7 plays multifaceted roles in regulating collecting vessel permeability and fibrosis, with one of the key players being IRF4-dependent DCs.

  12. Electrochemical impedance spectroscopic measurements of FCCP-induced change in membrane permeability of MDCK cells.

    Zhao, Lingzhi; Li, Xianchan; Lin, Yuqing; Yang, Lifen; Yu, Ping; Mao, Lanqun


    This study demonstrates a new electrochemical impedance spectroscopic (EIS) method for measurements of the changes in membrane permeability during the process of cell anoxia. Madin-Darby canine kidney (MDCK) cells were employed as the model cells and were cultured onto gelatin-modified glassy carbon (GC) electrodes. EIS measurements were conducted at the MDCK/gelatin-modified GC electrodes with Fe(CN)(6)(3-/4-) as the redox probe. The anoxia of the cells grown onto electrode surface was induced by the addition of carbonycyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) into the cell culture, in which the MDCK/gelatin-modified GC electrodes were immersed for different times. The EIS results show that the presence of FCCP in the cell culture clearly decreases the charge-transfer resistance of the Fe(CN)(6)(3-/4-) redox probe at the MDCK/gelatin-modified GC electrodes, and the charge-transfer resistance decreases with increasing time employed for immersing the MDCK/gelatin-modified GC electrodes into the cell culture containing FCCP. These results demonstrate that the EIS method could be used to monitor the changes in the cell membrane permeability during the FCCP-induced cell anoxia. To simulate the EIS system, a rational equivalent circuit was proposed and the values of ohmic resistance of the electrolyte, charge-transfer resistance and constant phase elements for both the gelatin and the cell layers are given with the fitting error in an acceptable value. This study actually offers a new and simple approach to measuring the dynamic process of cell death induced by anoxia through monitoring the changes in the cell membrane permeability.

  13. A low-frequency asymptotic model of seismic reflection from a high-permeability layer

    Silin, Dmitriy; Goloshubin, Gennady


    Analysis of compression wave propagation through a high-permeability layer in a homogeneous poroelastic medium predicts a peak of reflection in the low-frequency end of the spectrum. An explicit formula expresses the resonant frequency through the elastic moduli of the solid skeleton, the permeability of the reservoir rock, the fluid viscosity and compressibility, and the reservoir thickness. This result is obtained through a low-frequency asymptotic analysis of the Biot's model of poroelasticity. A new physical interpretation of some coefficients of the classical poroelasticity is a result of the derivation of the main equations from the Hooke's law, momentum and mass balance equations, and the Darcy's law. The velocity of wave propagation, the attenuation factor, and the wave number, are expressed in the form of power series with respect to a small dimensionless parameter. The latter is equal to the product of the kinematic reservoir fluid mobility, an imaginary unit, and the frequency of the signal. Retaining only the leading terms of the series leads to explicit and relatively simple expressions for the reflection and transmission coefficients for a planar wave crossing an interface between two permeable media, as well as wave reflection from a thin highly-permeable layer (a lens). The practical implications of the theory developed here are seismic modeling, inversion, and attribute analysis.

  14. Measurement of the permeability and resealing time constant of the electroporated mammalian cell membranes

    Shirakashi, Ryo [Tokyo Univ., Inst. of Industrial Science, Tokyo (Japan); Sukhorukov, Vladimir L.; Zimmermann, Ulrich [Wuerzburg Univ. Biozentrum, Lehrstuhl fuer Biotechnologie, Wuerzburg (Germany); Tanasawa, Ichiro [Nihon Univ., Dept. of Mechanical Engineering, Koriyama (Japan)


    In this study a new method is presented for measuring the transient permeability of mammalian cell membranes to sugar and electrolyte molecules based on the volumetric response of cells subjected to electroporation. The time constant of membrane resealing was determined independently by flow cytometry using a fluorescent dye as the reporter molecule. The volumetric and dye uptake data were analyzed with a model relating the cell volume changes to the solute transport across the reversibly permeabilized cell membrane. The experimental approach developed here might be useful for estimating the amount of electroinjected molecules, which are difficult to measure directly. (Author)

  15. Enhancement of beta-sitosterol transformation in Mycobacterium vaccae with increased cell wall permeability.

    Korycka-Machała, M; Rumijowska-Galewicz, A; Lisowska, K; Ziolkowskit, A; Sedlacze, L


    Mycobacterium vaccae exposed to compounds which are known to disorganise the cell wall composition and architecture (protamine, glycine) showed increased specific activity in beta-sitosterol biotransformation to androstene derivatives, intennediates in the production of most medical steroids. GC/MS analysis of free lipid fatty acids revealed higher content of unsaturated compounds, mainly C16:1 and C18:1 in protamine- and glycine-treated cells than that in control cells, which seems to change the permeability features of the cell wall barrier, facilitating hydrophobic beta-sitosterol diffusion.

  16. High gastrointestinal permeability and local metabolism of naringenin: influence of antibiotic treatment on absorption and metabolism.

    Orrego-Lagarón, Naiara; Martínez-Huélamo, Miriam; Vallverdú-Queralt, Anna; Lamuela-Raventos, Rosa M; Escribano-Ferrer, Elvira


    The present study aims to determine the permeability of naringenin in the stomach, small intestine and colon, to evaluate intestinal and hepatic first-pass metabolism, and to study the influence of the microbiota on the absorption and disposition of naringenin (3.5 μg/ml). A single-pass intestinal perfusion model in mice (n 4-6) was used. Perfusate (every 10 min), blood (at 60 min) and bile samples were taken and analysed to evaluate the presence of naringenin and its metabolites by an HPLC-MS/MS method. To study the influence of the microbiota on the bioavailability of naringenin, a group of animals received the antibiotic rifaximin (50 mg/kg per d) for 5 d, and naringenin permeability was determined in the colon. Naringenin was absorbed well throughout the gastrointestinal tract but mainly in the small intestine and colon (mean permeability coefficient 7.80 (SD 1.54) × 10(-4) cm/s and 5.49 (SD 1.86) × 10(-4) cm/s, respectively), at a level similar to the highly permeable compound, naproxen (6.39 (SD 1.23) × 10(-4) cm/s). According to the high amounts of metabolites found in the perfusate compared to the bile and plasma, naringenin underwent extensive intestinal first-pass metabolism, and the main metabolites excreted were sulfates (84.00 (SD 12.14)%), followed by glucuronides (8.40 (SD 5.67)%). Phase II metabolites were found in all perfusates from 5 min of sampling. Mice treated with rifaximin showed a decrease in naringenin permeability and in the amounts of 4-hydroxyhippuric acid and hippuric acid in the lumen. Naringenin was well absorbed throughout the gastrointestinal tract and its poor bioavailability was due mainly to high intestinal metabolism.

  17. Regulation of endothelial permeability and transendothelial migration of cancer cells by tropomyosin-1 phosphorylation

    Simoneau Bryan


    Full Text Available Abstract Background Loss of endothelial cell integrity and selective permeability barrier is an early event in the sequence of oxidant-mediated injury and may result in atherosclerosis, hypertension and facilitation of transendothelial migration of cancer cells during metastasis. We already reported that endothelial cell integrity is tightly regulated by the balanced co-activation of p38 and ERK pathways. In particular, we showed that phosphorylation of tropomyosin-1 (tropomyosin alpha-1 chain = Tm1 at Ser283 by DAP kinase, downstream of the ERK pathway might be a key event required to maintain the integrity and normal functions of the endothelium in response to oxidative stress. Methods Endothelial permeability was assayed by monitoring the passage of Dextran-FITC through a tight monolayer of HUVECs grown to confluence in Boyden chambers. Actin and Tm1 dynamics and distribution were evaluated by immunofluorescence. We modulated the expression of Tm1 by siRNA and lentiviral-mediated expression of wild type and mutated forms of Tm1 insensitive to the siRNA. Transendothelial migration of HT-29 colon cancer cells was monitored in Boyden chambers similarly as for permeability. Results We provide evidence indicating that Tm1 phosphorylation at Ser283 is essential to regulate endothelial permeability under oxidative stress by modulating actin dynamics. Moreover, the transendothelial migration of colon cancer cells is also regulated by the phosphorylation of Tm1 at Ser283. Conclusion Our finding strongly support the role for the phosphorylation of endothelial Tm1 at Ser283 to prevent endothelial barrier dysfunction associated with oxidative stress injury.

  18. Studies on bleomycin-induced repair DNA synthesis in permeable mouse ascites sarcoma cells.



    Full Text Available To study the mechanism of DNA excision repair, a DNA repair system employing permeable mouse sarcoma (SR-C3H/He cells was established and characterized. SR-C3H/He cells were permeabilized with a 0.0175% Triton X-100 solution. The permeable cells were treated with 1 mM ATP and 0.11 mM bleomycin, and then washed thoroughly to remove ATP and bleomycin. Repair DNA synthesis occurred in the bleomycin-damaged, permeable SR-C3H/He cells when incubated with ATP and four deoxyribonucleoside triphosphates. The repair nature of the DNA synthesis was confirmed by the BrdUMP density shift technique, and by the reduced sensitivity of the newly synthesized DNA to Escherichia coli exonuclease III. The DNA synthesis was optimally enhanced by addition of 0.08 M NaCl. Studies using selective inhibitors of DNA synthesis showed that aphidicolin-sensitive DNA polymerase (DNA polymerase alpha and/or delta and DNA polymerase beta were involved in the repair process. The present DNA repair system is thought to be useful to study nuclear DNA damage by bleomycin, removal of the damaged ends by an exonuclease, repair DNA synthesis by DNA polymerases and repair patch ligation by DNA ligase(s.

  19. Glycosylated hemoglobin in human and animal red cells. Role of glucose permeability.

    Higgins, P J; Garlick, R L; Bunn, H F


    We have compared red cells from man and selected animals in order to determine the effect of glucose permeability on nonenzymatic glycosylation of hemoglobin. Glucose permeability was highest in the primates (human, baboon, rhesus monkey), lower in dogs and rabbits, and nearly zero in pigs. Glycosylation of hemoglobin was measured by three independent methods: cation-exchange chromatography on Bio-Rex 70 (Bio-Rad, Inc., Richmond, California), agar gel electrophoresis, and affinity chromatography. The colorimetric thiobarbituric acid test did not provide valid data on animal hemolysates. However, this test was useful for identifying glycosylated hemoglobin (HbA1c) components isolated on Bio-Rex chromatography. In all animals tested, levels of HbA1c (from Bio-Rex chromatography) and total glycosylated hemoglobin (from affinity chromatography) correlated well with glucose exposure, the product of intracellular glucose concentration, and red cell life span. These results indicate that nonenzymatic glycosylation of hemoglobin in mammals is determined by three major variables: mean plasma glucose concentration, red cell life span, and red cell glucose permeability.

  20. Permeability of Rosmarinic acid in Prunella vulgaris and Ursolic acid in Salvia officinalis Extracts across Caco-2 Cell Monolayers

    Qiang, Zhiyi; Ye, Zhong; Hauck, Cathy; Murphy, Patricia A.; McCoy, Joe-Ann; Widrlechner, Mark P.; Reddy, Manju B.; Hendrich, Suzanne


    Ethnopharmacological relevance Rosmarinic acid (RA), a caffeic acid-related compound found in high concentrations in Prunella vulgaris (self-heal), and ursolic acid (UA), a pentacyclic triterpene acid concentrated in Salvia officinalis (sage), have been traditionally used to treat inflammation in the mouth, and may also be beneficial for gastrointestinal health in general. Aim of the study To investigate the permeabilities of RA and UA as pure compounds and in P. vulgaris and S. officinalis ethanol extracts across human intestinal epithelial Caco-2 cell monolayers. Materials and methods The permeabilities and Phase II biotransformation of RA and UA as pure compounds and in herbal extracts were compared using Caco-2 cells with HPLC detection. Results The apparent permeability coefficient (Papp) for RA and RA in P. vulgaris extracts was 0.2 ± 0.05 × 10−6 cm/s, significantly increased to 0.9 ± 0.2 × 10−6 cm/s after β-glucuronidase/sulfatase treatment. Papp for UA and UA in S. officinalis extract was 2.7 ± 0.3 × 10−6 cm/s and 2.3 ± 0.5 × 10−6 cm/s before and after β-glucuronidase/sulfatase treatment, respectively. Neither compound was affected in permeability by the herbal extract matrix. Conclusion RA and UA in herbal extracts had similar uptake as that found using the pure compounds, which may simplify the prediction of compound efficacy, but the apparent lack of intestinal glucuronidation/sulfation of UA is likely to further enhance the bioavailability of that compound compared with RA. PMID:21798330

  1. Analysis of heterogeneous characteristics in a geothermal area with low permeability and high temperature

    Alfonso Aragón-Aguilar


    Full Text Available An analytical methodology for reservoir characterization was applied in the central and southwestern zones of Los Humeros geothermal field (LHGF. This study involves analysis of temperature, pressure, enthalpy and permeability in wells and their distribution along the area. The wells located in the central western side of the geothermal field are productive, whereas those located at the central-eastern side are non-productive. Through temperature profiles, determined at steady state in the analyzed wells, it was observed that at bottom conditions (approximately 2300 m depth, temperatures vary between 280 and 360 °C. The temperatures are higher at the eastern side of central zone of LHGF. A review of transient pressure tests, laboratory measurements of core samples, and correlation of circulation losses during drilling suggest that permeability of the formation is low. The enthalpy behavior in productive wells shows a tendency of increase in the steam fraction. It was found that productivity behavior has inverse relation with permeability of rock formation. Further, it is observed that an imbalance exists between exploitation and recharge. It is concluded from the results that the wells located at central-eastern area have low permeability and high temperature, which indicates possibility of heat storage.

  2. Direct Calculation of Permeability by High-Accurate Finite Difference and Numerical Integration Methods

    Wang, Yi


    Velocity of fluid flow in underground porous media is 6~12 orders of magnitudes lower than that in pipelines. If numerical errors are not carefully controlled in this kind of simulations, high distortion of the final results may occur [1-4]. To fit the high accuracy demands of fluid flow simulations in porous media, traditional finite difference methods and numerical integration methods are discussed and corresponding high-accurate methods are developed. When applied to the direct calculation of full-tensor permeability for underground flow, the high-accurate finite difference method is confirmed to have numerical error as low as 10-5% while the high-accurate numerical integration method has numerical error around 0%. Thus, the approach combining the high-accurate finite difference and numerical integration methods is a reliable way to efficiently determine the characteristics of general full-tensor permeability such as maximum and minimum permeability components, principal direction and anisotropic ratio. Copyright © Global-Science Press 2016.

  3. Endothelial Cell Permeability and Adherens Junction Disruption Induced by Junín Virus Infection

    Lander, Heather M.; Grant, Ashley M.; Albrecht, Thomas; Hill, Terence; Peters, Clarence J.


    Junín virus (JUNV) is endemic to the fertile Pampas of Argentina, maintained in nature by the rodent host Calomys musculinus, and the causative agent of Argentine hemorrhagic fever (AHF), which is characterized by vascular dysfunction and fluid distribution abnormalities. Clinical as well as experimental studies implicate involvement of the endothelium in the pathogenesis of AHF, although little is known of its role. JUNV has been shown to result in productive infection of endothelial cells (ECs) in vitro with no visible cytopathic effects. In this study, we show that direct JUNV infection of primary human ECs results in increased vascular permeability as measured by electric cell substrate impedance sensing and transwell permeability assays. We also show that EC adherens junctions are disrupted during virus infection, which may provide insight into the role of the endothelium in the pathogenesis of AHF and possibly, other viral hemorrhagic fevers. PMID:24710609

  4. Mast cells and histamine alter intestinal permeability during malaria parasite infection.

    Potts, Rashaun A; Tiffany, Caitlin M; Pakpour, Nazzy; Lokken, Kristen L; Tiffany, Connor R; Cheung, Kong; Tsolis, Renée M; Luckhart, Shirley


    Co-infections with malaria and non-typhoidal Salmonella serotypes (NTS) can present as life-threatening bacteremia, in contrast to self-resolving NTS diarrhea in healthy individuals. In previous work with our mouse model of malaria/NTS co-infection, we showed increased gut mastocytosis and increased ileal and plasma histamine levels that were temporally associated with increased gut permeability and bacterial translocation. Here, we report that gut mastocytosis and elevated plasma histamine are also associated with malaria in an animal model of falciparum malaria, suggesting a broader host distribution of this biology. In support of mast cell function in this phenotype, malaria/NTS co-infection in mast cell-deficient mice was associated with a reduction in gut permeability and bacteremia. Further, antihistamine treatment reduced bacterial translocation and gut permeability in mice with malaria, suggesting a contribution of mast cell-derived histamine to GI pathology and enhanced risk of bacteremia during malaria/NTS co-infection.


    王晓峰; 由广旭; 皮绍文; 秦永文


    To investigate the effects of angiotensin converting enzyme inhibitor captopril, calcium channel blocker diltiazem and β-adrenoceptor antagonist dobutamine on the permeability of rat aortic endothelial monolayers.Methods Trauma-free isolation by Chen et al was adopted in the culture of rat aortic endothelial cells. Rat aortic endothelial cells were seeded on the nitrocellulose microporous filters. Eight days after seeding, the monolayers could be used for measuring the permeability. Before being perfused, monolayers were treated with captopril, diltiazem and dobutamine for 4 hours successively. The prepared filters were mounted on the Boydon chambers and perfused with hyperlipemia containing FITC-labeled albumin. The fluid filtering through the monolayers and the filter was collected and the albumin concentration was measured. At the same time, cholesterol, triglyceride, lipoprotein A and lipoprotein B concentrations of the collected fluid were also measured by ELISA.Results The above three drugs decreased the permeability of aortic endothelial cell monolayers to water, cholesterol, triglyceride lipoprotein A and lipoprotein B significantly. Dobutamine had more significant effects than the other two drugs. But diltiazem worked well in the clearance of albumin, while the other two drugs had no obvious effect.Conclusion Captopril, diltiazem and dobutamine may decrease the infiltration of lipids and lipoproteins into the subendothelial space, thus they can be used to prevent and ameliorate atherosclerosis.

  6. 信息动态%Evaluation of Mitochondrial Damage of lsletβCells by Mitochondrial Permeability Transition Pore


    Objective To evaluate the mitochondrial damage of islet β cells under glucolipotoxicity by investigating the mitochondrial permeability transition pore (mPTP). Methods Pancreatic β cell lines INS-1 cells were treated with 0. 4 mmol/L palmitic acid and different concentrations of glucose (5.6 mmol/L or 25 mmol/L). The mitochondrial membrane potential, mPTP and reactive oxygen species (ROS) were measured by flow cytometry and fluorescence staining technique to assess the mitochon drial damage. Cell proliferation was measured by 5-bromodeoxyuridine incorporation and cell apoptosis was detected by Annexin V method. Results Compared with the low glucose concentration, the high glucose concentration resulted in decreased mPTP activity (P<0.05), increased mitochondrial membrane potential (P<0.05) and increased cell proliferation rate (P<0.05). There was no significant change in ROS generation. When cells were exposed to high glucose concentration and palmitic acid, both mPTP activity and mitochonhdrial membrane potential reduced (P<0.05), with increased cell apoptosis rate (P <0.05) and increased ROS generation. Conclusion The high glucose concentration decreases mPTP and increases mitochondrial membrane potential, suggesting that cells may remain in an unstable high metabolic state. Evaluation of mPTP may contribute to a more comprehensive understanding of mitochondrial dysfunction under glucotoxictiy.

  7. Ubisol-Q10 Prevents Glutamate-Induced Cell Death by Blocking Mitochondrial Fragmentation and Permeability Transition Pore Opening

    Kumari, Santosh; Mehta, Suresh L.; Milledge, Gaolin Z.; Huang, Xinyu; Li, Haining; Li, P. Andy


    Mitochondrial dysfunction and oxidative stress are the major events that lead to the formation of mitochondrial permeability transition pore (mPTP) during glutamate-induced cytotoxicity and cell death. Coenzyme Q10 (CoQ10) has widely been used for the treatment of mitochondrial disorders and neurodegenerative diseases. Comparing to traditional lipid-soluble CoQ10, water soluble CoQ10 (Ubisol-Q10) has high intracellular and intra-mitochondrial distribution. The aims of the present study are to...

  8. Dry aging of beef in a bag highly permeable to water vapour.

    Ahnström, Maria Lundesjö; Seyfert, Mark; Hunt, Melvin C; Johnson, Dallas E


    The objective of this experiment was to compare traditional dry aging of beef with a novel technique of dry aging in a highly moisture-permeable bag. Four equal-sized sections from paired beef strip loins were dry aged traditionally, unpackaged, or packaged in the experimental bag for 14 or 21d at 3°C. No differences (P>0.05) were noted for pH, moisture, fat, total plate counts, cook loss, shear force, or any measured sensory attribute between the two aging treatments after either aging period. After 21d, however, dry aging in the bag (versus traditional dry aging) decreased (PDry aging in a highly moisture-permeable bag is feasible, will positively impact yields and reduce microbial spoilage, and will have no negative impact on product quality.

  9. Rectification of the water permeability in COS-7 cells at 22, 10 and 0°C.

    Diana B Peckys

    Full Text Available The osmotic and permeability parameters of a cell membrane are essential physico-chemical properties of a cell and particularly important with respect to cell volume changes and the regulation thereof. Here, we report the hydraulic conductivity, L(p, the non-osmotic volume, V(b, and the Arrhenius activation energy, E(a, of mammalian COS-7 cells. The ratio of V(b to the isotonic cell volume, V(c iso, was 0.29. E(a, the activation energy required for the permeation of water through the cell membrane, was 10,700, and 12,000 cal/mol under hyper- and hypotonic conditions, respectively. Average values for L(p were calculated from swell/shrink curves by using an integrated equation for L(p. The curves represented the volume changes of 358 individually measured cells, placed into solutions of nonpermeating solutes of 157 or 602 mOsm/kg (at 0, 10 or 22°C and imaged over time. L(p estimates for all six combinations of osmolality and temperature were calculated, resulting in values of 0.11, 0.21, and 0.10 µm/min/atm for exosmotic flow and 0.79, 1.73 and 1.87 µm/min/atm for endosmotic flow (at 0, 10 and 22°C, respectively. The unexpected finding of several fold higher L(p values for endosmotic flow indicates highly asymmetric membrane permeability for water in COS-7. This phenomenon is known as rectification and has mainly been reported for plant cell, but only rarely for animal cells. Although the mechanism underlying the strong rectification found in COS-7 cells is yet unknown, it is a phenomenon of biological interest and has important practical consequences, for instance, in the development of optimal cryopreservation.

  10. Rectification of the water permeability in COS-7 cells at 22, 10 and 0°C.

    Peckys, Diana B; Kleinhans, F W; Mazur, Peter


    The osmotic and permeability parameters of a cell membrane are essential physico-chemical properties of a cell and particularly important with respect to cell volume changes and the regulation thereof. Here, we report the hydraulic conductivity, L(p), the non-osmotic volume, V(b), and the Arrhenius activation energy, E(a), of mammalian COS-7 cells. The ratio of V(b) to the isotonic cell volume, V(c iso), was 0.29. E(a), the activation energy required for the permeation of water through the cell membrane, was 10,700, and 12,000 cal/mol under hyper- and hypotonic conditions, respectively. Average values for L(p) were calculated from swell/shrink curves by using an integrated equation for L(p). The curves represented the volume changes of 358 individually measured cells, placed into solutions of nonpermeating solutes of 157 or 602 mOsm/kg (at 0, 10 or 22°C) and imaged over time. L(p) estimates for all six combinations of osmolality and temperature were calculated, resulting in values of 0.11, 0.21, and 0.10 µm/min/atm for exosmotic flow and 0.79, 1.73 and 1.87 µm/min/atm for endosmotic flow (at 0, 10 and 22°C, respectively). The unexpected finding of several fold higher L(p) values for endosmotic flow indicates highly asymmetric membrane permeability for water in COS-7. This phenomenon is known as rectification and has mainly been reported for plant cell, but only rarely for animal cells. Although the mechanism underlying the strong rectification found in COS-7 cells is yet unknown, it is a phenomenon of biological interest and has important practical consequences, for instance, in the development of optimal cryopreservation.

  11. Negative correlation of CD34+ cells with blood-brain barrier permeability following traumatic brain injury in a rat model.

    Jin, Xuelong; Wang, Feifei; Liu, Xingju; Liang, Bin; Chen, Zequn; He, Junfeng; Zhang, Hong; Zhang, Jianning


    TBI causes localized cerebral ischemia that, in turn, is accompanied by both changes in BBB permeability and recruitment of CD34(+) cells to the injured tissue. However, it remains unknown whether CD34(+) cell recruitment is linked to BBB permeability. This study is a preliminary investigation into possible correlations between CD34(+) cell recruitment and BBB permeability following TBI in a rat model. Male SD rats were subjected to mild fluid percussion injury. BBB permeability was assessed by measuring extrinsic EB dye extravasation and endogenous EBA expression at days 1, 3, 5, 7, and 12 post injury. The number of CD34(+) cells in the damaged tissue was analyzed by immunohistochemistry at each time point. EB dye extravasation reached a peak at day 3 following TBI, while EBA expression displayed the reverse profile. Accumulation of CD34(+) cells in injured brain tissue was evident at five days post injury. It revealed a negative linear correlation between CD34(+) cell and BBB permeability. The negative linear correlation between CD34(+) cell recruitment and BBB permeability following TBI provides a support for further study of CD34(+) cell transplantation for BBB repair after TBI. © 2014 John Wiley & Sons Ltd.

  12. Establishment of HEK293 cell line expressing GFPAQP1 to determine water osmotic permeability

    Jun-weiGAO; He-mingYU; Qian-liuSONG; Shu-xinLI; Xue-junLI


    AIM: To develop an osmotic water permeability assay.METHODS: We subcloned the rat AQP1 cDNA into pEGFPC3 vector. HEK293 cells were transfected with pEGFP-C3/AQP 1 or pEGFP-C3 and selected by G418. The expression of AQP1was detected by RT-PCR and Western blot. Confocal laser fluorescence microscopy was used to record the change of fluorescent density corresponding to the volume change of the cells

  13. Claudin-binder C-CPE mutants enhance permeability of insulin across human nasal epithelial cells.

    Kojima, Takashi; Kondoh, Masuo; Keira, Takashi; Takano, Ken-Ichi; Kakuki, Takuya; Kaneko, Yakuto; Miyata, Ryo; Nomura, Kazuaki; Obata, Kazufumi; Kohno, Takayuki; Konno, Takumi; Sawada, Norimasa; Himi, Tetsuo


    Intranasal insulin administration has therapeutic potential for Alzheimer's disease and in intranasal administration across the nasal mucosa, the paracellular pathway regulated by tight junctions is important. The C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE) binds the tight junction protein claudin and disrupts the tight junctional barrier without a cytotoxic effect. The C-CPE mutant called C-CPE 194 binds only to claudin-4, whereas the C-CPE 194 mutant called C-CPE m19 binds not only to claudin-4 but also to claudin-1. In the present study, to investigate the effects of C-CPE mutants on the tight junctional functions of human nasal epithelial cells (HNECs) and on the permeability of human recombinant insulin across the cells, HNECs were treated with C-CPE 194 and C-CPE m19. C-CPE 194 and C-CPE m19 disrupted the barrier and fence functions without changes in expression of claudin-1, -4, -7, and occludin or cytotoxicity, whereas they transiently increased the activity of ERK1/2 phosphorylation. The disruption of the barrier function caused by C-CPE 194 and C-CPE m19 was prevented by pretreatment with the MAPKK inhibitor U0126. Furthermore, C-CPE 194 and C-CPE m19 significantly enhanced the permeability of human recombinant insulin across HNECs and the permeability was also inhibited by U0126. These findings suggest that C-CPE mutants 194 and m19 can regulate the permeability of insulin across HNECs via the MAPK pathway and may play a crucial role in therapy for the diseases such as Alzheimer's disease via the direct intranasal insulin administration.

  14. Modelling of the magnetic and magnetostrictive properties of high permeability Mn–Zn ferrites

    R Szewczyk


    This paper presents the results of modelling of both magnetic and magnetostrictive properties of high permeability Mn0.51Zn0.44Fe2.05O4 ferrites. The parameters of energy-based Jiles–Atherton–Sablik (J–A–S) model were calculated for each experimental hysteresis loop on the basis of evolutionary strategies and Hook–Jevis optimization method. Finally, high conformity between experimental and modelling results was achieved. This high conformity indicates that the presented results create new opportunity of modelling of the properties of inductive components based on ferrites as well as quantitative description of magnetization process.

  15. Discovery of a Highly Potent, Cell-Permeable Macrocyclic Peptidomimetic (MM-589) Targeting the WD Repeat Domain 5 Protein (WDR5)–Mixed Lineage Leukemia (MLL) Protein–Protein Interaction

    Karatas, Hacer; Li, Yangbing; Liu, Liu; Ji, Jiao; Lee, Shirley; Chen, Yong; Yang, Jiuling; Huang, Liyue; Bernard, Denzil; Xu, Jing; Townsend, Elizabeth C.; Cao, Fang; Ran, Xu; Li, Xiaoqin; Wen, Bo; Sun, Duxin; Stuckey, Jeanne A; Lei, Ming; Dou, Yali; Wang, Shaomeng (Michigan)


    We report herein the design, synthesis, and evaluation of macrocyclic peptidomimetics that bind to WD repeat domain 5 (WDR5) and block the WDR5–mixed lineage leukemia (MLL) protein–protein interaction. Compound 18 (MM-589) binds to WDR5 with an IC50 value of 0.90 nM (Ki value <1 nM) and inhibits the MLL H3K4 methyltransferase (HMT) activity with an IC50 value of 12.7 nM. Compound 18 potently and selectively inhibits cell growth in human leukemia cell lines harboring MLL translocations and is >40 times better than the previously reported compound MM-401. Cocrystal structures of 16 and 18 complexed with WDR5 provide structural basis for their high affinity binding to WDR5. Additionally, we have developed and optimized a new AlphaLISA-based MLL HMT functional assay to facilitate the functional evaluation of these designed compounds. Compound 18 represents the most potent inhibitor of the WDR5–MLL interaction reported to date, and further optimization of 18 may yield a new therapy for acute leukemia.

  16. System for exposing cultured cells to intermittent hypoxia utilizing gas permeable cultureware.

    Polak, Jan; Studer-Rabeler, Karen; McHugh, Holly; Hussain, Mehboob A; Shimoda, Larissa A


    Tissue intermittent hypoxia (IH) occurs in obstructive sleep apnea, sickle cell anemia, physical exercise and other conditions. Poor gas solubility and slow diffusion through culture media hampers mimicking IH-induced transitions of O(2) in vitro. We aimed to develop a system enabling exposure of cultured cells to IH and to validate such exposure by real-time O(2) measurements and cellular responses. Standard 24-well culture plates and plates with bottoms made from a gas permeable film were placed in a heated cabinet. Desired cycling of O(2) levels was induced using programmable solenoids to purge mixtures of 95% N(2) + 5% CO(2) or 95% O(2) + 5% CO(2). Dissolved oxygen, gas pressure, temperature, and water evaporation were measured during cycling. IH-induced cellular effects were evaluated by hypoxia inducible factor (HIF) and NF-κB luciferase reporters in HEK296 cells and by insulin secretion in rat insulinoma cells. Oxygen cycling in the cabinet was translated into identical changes of O(2) at the well bottom in gas permeable, but not in standard cultureware. Twenty-four hours of IH exposure increased HIF (112%), NF-κB (111%) and insulin secretion (44%). Described system enables reproducible and prolonged IH exposure in cultured cells while controlling for important environmental factors.

  17. [Investigation of permeability of intranasal formulations using Side-Bi-Side horizontal diffusion cell].

    Horváth Tamás; Ambrus, Rita; Szabóné, Révész Piroska


    Nowadays the nasal route has received a great attention as a reliable administration for the systemic administration. In the Department of Pharmaceutical Technology, University of Szeged, the main research work is the design and development of innovative nasal formulations, which can open new possibilities for some well-known agents and may also help some drug-candidates delivery problems. The aim of this work was to present some reliable models for investigation of permeability, such as Spectra/Por Dialisys Membran, ZelluTrans/Roth Mini Dialyzer, μFLUX diffusion Cell, Navicyte Vertical and Horizontal Diffusion Chamber System and In-line Cell. In addition, the horizontal membrane diffusion model (Side-Bi-Side) was used to investigate in vitro and ex vivo studies of permeability of meloxicam in comparison with the vertical diffusion cell (Franz). The present study investigated the meloxicam in different dosage forms (powder, spray, gel). It was found that the Side-Bi-Side cell is suitable to test the nasal formulations, but the uniform distribution of the active substance cannot be ensured in donor place by increasing the viscosity of the compositions, therefore the Franz cell is recommended for investigation of nasal gel. Previous measurement cannot be found related to this topic.

  18. Dynamic effects of high-pressure pulsed water jet in low-permeability coal seams

    LI Xiao-hong; ZHOU Dong-ping; LU Yi-yu; KANG Yong; ZHAO Yu; WANG Xiao-chuan


    Mine gas extraction in China is difficult due to the characteristics such as mi-cro-porosity, low-permeability and high adsorption of coal seams. The pulsed mechanism of a high-pressure pulsed water jet was studied through theoretical analysis, experiment and field measurement. The results show that high-pressure pulsed water jet has three dynamic properties. What's more, the three dynamic effects can be found in low-perme-ability coal seams. A new pulsed water jet with 200-1 000 Hz oscillation frequency and peak pressure 2.5 times than average pressure was introduced. During bubble collapsing, sound vibration and instantaneous high pressures over 100 MPa enhanced the cutting ability of the high-pressure jet. Through high-pressure pulsed water jet drilling and slotting, the exposure area of coal bodies was greatly enlarged and pressure of the coal seams rapidly decreased. Therefore, the permeability of coal seams was improved and gas ab-sorption rate also decreased. Application results show that gas adsorption rate decreased by 30%-40% and the penetrability coefficient increased 100 times. This proves that high-pressure pulsed water is more efficient than other conventional methods.

  19. High-resolution nanopatterning of biodegradable polylactide by thermal nanoimprint lithography using gas permeable mold

    Satoshi Takei


    Full Text Available We report high-resolution (150 nm nanopatterning of biodegradable polylactide by thermal nanoimprint lithography using dichloromethane as a volatile solvent for improving the liquidity and a porous cyclodextrin-based gas-permeable mold. This study demonstrates the high-resolution patterning of polylactic acid and other non-liquid functional materials with poor fluidity by thermal nanoimprinting. Such a patterning is expected to expand the utility of thermal nanoimprint lithography and fabricate non-liquid functional materials suitable for eco-friendly and biomedical applications.

  20. High-resolution nanopatterning of biodegradable polylactide by thermal nanoimprint lithography using gas permeable mold

    Takei, Satoshi; Hanabata, Makoto


    We report high-resolution (150 nm) nanopatterning of biodegradable polylactide by thermal nanoimprint lithography using dichloromethane as a volatile solvent for improving the liquidity and a porous cyclodextrin-based gas-permeable mold. This study demonstrates the high-resolution patterning of polylactic acid and other non-liquid functional materials with poor fluidity by thermal nanoimprinting. Such a patterning is expected to expand the utility of thermal nanoimprint lithography and fabricate non-liquid functional materials suitable for eco-friendly and biomedical applications.

  1. Mapping Cd²⁺-induced membrane permeability changes of single live cells by means of scanning electrochemical microscopy.

    Filice, Fraser P; Li, Michelle S M; Henderson, Jeffrey D; Ding, Zhifeng


    Scanning Electrochemical Microscopy (SECM) is a powerful, non-invasive, analytical methodology that can be used to investigate live cell membrane permeability. Depth scan SECM imaging allowed for the generation of 2D current maps of live cells relative to electrode position in the x-z or y-z plane. Depending on resolution, one depth scan image can contain hundreds of probe approach curves (PACs). Individual PACs were obtained by simply extracting vertical cross-sections from the 2D image. These experimental PACs were overlaid onto theoretically generated PACs simulated at specific geometry conditions. Simulations were carried out using 3D models in COMSOL Multiphysics to determine the cell membrane permeability coefficients at different locations on the surface of the cells. Common in literature, theoretical PACs are generated using a 2D axially symmetric geometry. This saves on both compute time and memory utilization. However, due to symmetry limitations of the model, only one experimental PAC right above the cell can be matched with simulated PAC data. Full 3D models in this article were developed for the SECM system of live cells, allowing all experimental PACs over the entire cell to become usable. Cd(2+)-induced membrane permeability changes of single human bladder (T24) cells were investigated at several positions above the cell, displaced from the central axis. The experimental T24 cells under study were incubated with Cd(2+) in varying concentrations. It is experimentally observed that 50 and 100 μM Cd(2+) caused a decrease in membrane permeability, which was uniform across all locations over the cell regardless of Cd(2+) concentration. The Cd(2+) was found to have detrimental effects on the cell, with cells shrinking in size and volume, and the membrane permeability decreasing. A mapping technique for the analysis of the cell membrane permeability under the Cd(2+) stress is realized by the methodology presented.

  2. Bioinspired Tuning of Hydrogel Permeability-Rigidity Dependency for 3D Cell Culture

    Lee, Min Kyung; Rich, Max H.; Baek, Kwanghyun; Lee, Jonghwi; Kong, Hyunjoon


    Hydrogels are being extensively used for three-dimensional immobilization and culture of cells in fundamental biological studies, biochemical processes, and clinical treatments. However, it is still a challenge to support viability and regulate phenotypic activities of cells in a structurally stable gel, because the gel becomes less permeable with increasing rigidity. To resolve this challenge, this study demonstrates a unique method to enhance the permeability of a cell-laden hydrogel while avoiding a significant change in rigidity of the gel. Inspired by the grooved skin textures of marine organisms, a hydrogel is assembled to present computationally optimized micro-sized grooves on the surface. Separately, a gel is engineered to preset aligned microchannels similar to a plant's vascular bundles through a uniaxial freeze-drying process. The resulting gel displays significantly increased water diffusivity with reduced changes of gel stiffness, exclusively when the microgrooves and microchannels are aligned together. No significant enhancement of rehydration is achieved when the microgrooves and microchannels are not aligned. Such material design greatly enhances viability and neural differentiation of stem cells and 3D neural network formation within the gel.

  3. Short-term control of maize cell and root water permeability through plasma membrane aquaporin isoforms.

    Hachez, Charles; Veselov, Dmitry; Ye, Qing; Reinhardt, Hagen; Knipfer, Thorsten; Fricke, Wieland; Chaumont, François


    Although it is widely accepted that aquaporins are involved in the regulation of root water uptake, the role of specific isoforms in this process is poorly understood. The mRNA expression and protein level of specific plasma membrane intrinsic proteins (PIPs) were analysed in Zea mays in relation to cell and root hydraulic conductivity. Plants were analysed during the day/night period, under different growth conditions (aeroponics/hydroponics) and in response to short-term osmotic stress applied through polyethylene glycol (PEG). Higher protein levels of ZmPIP1;2, ZmPIP2;1/2;2, ZmPIP2;5 and ZmPIP2;6 during the day coincided with a higher water permeability of root cortex cells during the day compared with night period. Similarly, plants which were grown under aeroponic conditions and which developed a hypodermis ('exodermis') with Casparian bands, effectively forcing more water along a membranous uptake path across roots, showed increased levels of ZmPIP2;5 and ZmPIP1;2 in the rhizodermis and exodermis. When PEG was added to the root medium (2-8 h), expression of PIPs and cell water permeability in roots increased. These data support a role of specific PIP isoforms, in particular ZmPIP1;2 and ZmPIP2;5, in regulating root water uptake and cortex cell hydraulic conductivity in maize.

  4. The integrin antagonist cilengitide activates alphaVbeta3, disrupts VE-cadherin localization at cell junctions and enhances permeability in endothelial cells.

    Gian Carlo Alghisi

    Full Text Available Cilengitide is a high-affinity cyclic pentapeptdic alphaV integrin antagonist previously reported to suppress angiogenesis by inducing anoikis of endothelial cells adhering through alphaVbeta3/alphaVbeta5 integrins. Angiogenic endothelial cells express multiple integrins, in particular those of the beta1 family, and little is known on the effect of cilengitide on endothelial cells expressing alphaVbeta3 but adhering through beta1 integrins. Through morphological, biochemical, pharmacological and functional approaches we investigated the effect of cilengitide on alphaVbeta3-expressing human umbilical vein endothelial cells (HUVEC cultured on the beta1 ligands fibronectin and collagen I. We show that cilengitide activated cell surface alphaVbeta3, stimulated phosphorylation of FAK (Y(397 and Y(576/577, Src (S(418 and VE-cadherin (Y(658 and Y(731, redistributed alphaVbeta3 at the cell periphery, caused disappearance of VE-cadherin from cellular junctions, increased the permeability of HUVEC monolayers and detached HUVEC adhering on low-density beta1 integrin ligands. Pharmacological inhibition of Src kinase activity fully prevented cilengitide-induced phosphorylation of Src, FAK and VE-cadherin, and redistribution of alphaVbeta3 and VE-cadherin and partially prevented increased permeability, but did not prevent HUVEC detachment from low-density matrices. Taken together, these observations reveal a previously unreported effect of cilengitide on endothelial cells namely its ability to elicit signaling events disrupting VE-cadherin localization at cellular contacts and to increase endothelial monolayer permeability. These effects are potentially relevant to the clinical use of cilengitide as anticancer agent.

  5. Bleomycin-induced DNA synthesis in a cell-free system using a permeable mouse sarcoma cell Extract.



    Full Text Available To investigate factors involved in excision repair DNA synthesis, a soluble extract was prepared from permeable mouse sarcoma (SR-C3H/He cells by homogenization and ultracentrifugation. DNA synthesis measured by using native calf thymus DNA as the template-primer and the extract as the polymerase source showed low activity. The DNA synthesis was enhanced more than ten-fold by the addition of an appropriate concentration of bleomycin, a radiomimetic DNA-damaging drug. Using selective inhibitors of DNA polymerases, it was shown that the DNA polymerase involved in the bleomycin-induced DNA synthesis was DNA polymerase beta. In addition to DNA polymerase beta, an exonuclease which converts bleomycin-damaged DNA into suitable template-primers for repair DNA synthesis appeared to be present in the permeable cell extract.

  6. Nicotine permeability across the buccal TR146 cell culture model and porcine buccal mucosa in vitro

    Nielsen, Hanne Mørck; Rassing, Margrethe Rømer


    The present study was conducted to investigate and compare the effect of pH and drug concentration on nicotine permeability across the TR146 cell culture model and porcine buccal mucosa in vitro. As a further characterization of the TR146 cell culture model, it was explored whether the results were...... comparable for bi-directional and uni-directional transport in the presence of a transmembrane pH gradient. Nicotine concentrations between 10(-5) and 10(-2) M were applied to the apical side of the TR146 cell culture model or the mucosal side of porcine buccal mucosa. Buffers with pH values of 5.5, 7.......4 and 8.1 were used to obtain different fractions of non- and mono-ionized nicotine. The apparent permeability (P(app)) of nicotine across both models increased significantly with increasing pH, and the P(app) values obtained with the two models could be correlated in a linear manner. With increasing...

  7. Homoharringtonine increases intestinal epithelial permeability by modulating specific claudin isoforms in Caco-2 cell monolayers.

    Watari, Akihiro; Hashegawa, Maki; Yagi, Kiyohito; Kondoh, Masuo


    Homoharringtonine (HHT), a natural alkaloid produced by various Cephalotaxus species, has antileukemic activity in acute and chronic myelogenous leukemia. However, HHT can also induce unanticipated effects in the gastrointestinal tract, such as diarrhea and nausea/vomiting, but the mechanism behind these adverse effects has not been clarified. In the present study, we show that HHT affects the epithelial permeability of intestinal Caco-2 cell monolayers. HHT reduced the transepithelial electrical resistance (TER) of Caco-2 cells in a dose- and time-dependent manner. The HHT effect was reversible and no cytotoxicity was observed at the concentrations used. HHT simultaneously increased the paracellular flux of the 4 kDa and 40 kDa FITC-dextrans associated with the TER reduction. Immunoblotting analysis revealed that HHT decreased the protein expression of TJ components such as claudin-3, -5, and -7. However, the transcription levels of these claudins were not repressed by HHT treatment. HHT also disturbed the cellular localization of claudin-1 and -4. These changes coincided with the reduced barrier function. Our findings suggest that HHT enhances the paracellular permeability of Caco-2 cell monolayers by modulating the protein expression and localization of claudin isoforms; these actions might be responsible for the gastrointestinal effects of HHT. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. 4-Quinolone-3-carboxylic acids as cell-permeable inhibitors of protein tyrosine phosphatase 1B.

    Zhi, Ying; Gao, Li-Xin; Jin, Yi; Tang, Chun-Lan; Li, Jing-Ya; Li, Jia; Long, Ya-Qiu


    Protein tyrosine phosphatase 1B is a negative regulator in the insulin and leptin signaling pathways, and has emerged as an attractive target for the treatment of type 2 diabetes and obesity. However, the essential pharmacophore of charged phosphotyrosine or its mimetic confer low selectivity and poor cell permeability. Starting from our previously reported aryl diketoacid-based PTP1B inhibitors, a drug-like scaffold of 4-quinolone-3-carboxylic acid was introduced for the first time as a novel surrogate of phosphotyrosine. An optimal combination of hydrophobic groups installed at C-6, N-1 and C-3 positions of the quinolone motif afforded potent PTP1B inhibitors with low micromolar IC50 values. These 4-quinolone-3-carboxylate based PTP1B inhibitors displayed a 2-10 fold selectivity over a panel of PTP's. Furthermore, the bidentate inhibitors of 4-quinolone-3-carboxylic acids conjugated with aryl diketoacid or salicylic acid were cell permeable and enhanced insulin signaling in CHO/hIR cells. The kinetic studies and molecular modeling suggest that the 4-quinolone-3-carboxylates act as competitive inhibitors by binding to the PTP1B active site in the WPD loop closed conformation. Taken together, our study shows that the 4-quinolone-3-carboxylic acid derivatives exhibit improved pharmacological properties over previously described PTB1B inhibitors and warrant further preclinical studies.

  9. Expression of Ca²⁺-permeable two-pore channels rescues NAADP signalling in TPC-deficient cells.

    Ruas, Margarida; Davis, Lianne C; Chen, Cheng-Chang; Morgan, Anthony J; Chuang, Kai-Ting; Walseth, Timothy F; Grimm, Christian; Garnham, Clive; Powell, Trevor; Platt, Nick; Platt, Frances M; Biel, Martin; Wahl-Schott, Christian; Parrington, John; Galione, Antony


    The second messenger NAADP triggers Ca(2+) release from endo-lysosomes. Although two-pore channels (TPCs) have been proposed to be regulated by NAADP, recent studies have challenged this. By generating the first mouse line with demonstrable absence of both Tpcn1 and Tpcn2 expression (Tpcn1/2(-/-)), we show that the loss of endogenous TPCs abolished NAADP-dependent Ca(2+) responses as assessed by single-cell Ca(2+) imaging or patch-clamp of single endo-lysosomes. In contrast, currents stimulated by PI(3,5)P2 were only partially dependent on TPCs. In Tpcn1/2(-/-) cells, NAADP sensitivity was restored by re-expressing wild-type TPCs, but not by mutant versions with impaired Ca(2+)-permeability, nor by TRPML1. Another mouse line formerly reported as TPC-null likely expresses truncated TPCs, but we now show that these truncated proteins still support NAADP-induced Ca(2+) release. High-affinity [(32)P]NAADP binding still occurs in Tpcn1/2(-/-) tissue, suggesting that NAADP regulation is conferred by an accessory protein. Altogether, our data establish TPCs as Ca(2+)-permeable channels indispensable for NAADP signalling.

  10. Rescue From Lethal Shiga Toxin 2-induced Renal Failure With A Cell-Permeable Peptide

    Stearns-Kurosawa, Deborah J.; Collins, Valta; Freeman, Scott; Debord, Diann; Nishikawa, Kiyotaka; Oh, Sun-Young; Leibowitz, Caitlin S.; Kurosawa, Shinichiro


    Background Intestinal infection with Shiga toxin (Stx)-producing E.coli is a leading cause of hemolytic uremic syndrome and acute renal injury in otherwise healthy children in the US. Antibiotics are contraindicated and a therapeutic priority is agents that act intracellularly against the bacterial toxins which drive kidney injury. Our aim was to evaluate whether intravenous administration of a cell-permeable peptide (TVP) that binds to Stx2 will reduce disease severity and rescue juvenile baboons from a lethal Stx2 dose (50ng/kg). Methods TVP (5mg/kg) was delivered i.v. simultaneously with toxin (prevention protocol) or at 6 or 24 hours after toxin with daily 1mg/kg supplements up to Day 4 (rescue protocols). Biomarkers were monitored in blood and urine up to 28 days. Results and Conclusions TVP therapy resulted in either absence of clinical signs of acute kidney injury and normal urine output (prevention), or delayed and reduced BUN and creatinine levels (rescue) with concomitant survival. Delayed peptide administration significantly reduced thrombocytopenia, but surprisingly did not alter anemia even when monitored for 28 days in rescued survivors. This is the first successful cell-permeable therapeutic that counteracts Stx2 lethality in an animal model which recapitulates many of the human responses to enteric infection. PMID:21603905

  11. Metaxin deficiency alters mitochondrial membrane permeability and leads to resistance to TNF-induced cell killing.

    Ono, Koh; Wang, Xiaofei; Kim, Sung Ouk; Armstrong, Lucas C; Bornstein, Paul; Han, Jiahuai


    Metaxin, a mitochondrial outer membrane protein, is critical for TNF-induced cell death in L929 cells. Its deficiency, caused by retroviral insertion-mediated mutagenesis, renders L929 cells resistance to TNF killing. In this study, we further characterized metaxin deficiency-caused TNF resistance in parallel with Bcl-X(L) overexpression-mediated death resistance. We did not find obvious change in mitochondria membrane potential in metaxin-deficient (Met(mut)) and Bcl-X(L)-overexpressing cells, but we did find an increase in the release rate of the mitochondrial membrane potential probe rhodamine 123 (Rh123) that was preloaded into mitochondria. In addition, overexpression of a function-interfering mutant of metaxin (MetaΔTM/C) or Bcl-X(L) in MCF-7.3.28 cells also resulted in an acquired resistance to TNF killing and a faster rate of Rh123 release, indicating a close correlation between TNF resistance and higher rates of the dye release from the mitochondria. The release of Rh123 can be controlled by the mitochondrial membrane permeability transition (PT) pore, as targeting an inner membrane component of the PT pore by cyclosporin A (CsA) inhibited Rh123 release. However, metaxin deficiency and Bcl-X(L) overexpression apparently affect Rh123 release from a site(s) different from that of CsA, as CsA can overcome their effect. Though both metaxin and Bcl-X(L) appear to function on the outer mitochondrial membrane, they do not interact with each other. They may use different mechanisms to increase the permeability of Rh123, since previous studies have suggested that metaxin may influence certain outer membrane porins while Bcl-X(L) may form pores on the outer membrane. The alteration of the mitochondrial outer membrane properties by metaxin deficiency and Bcl-X(L) overexpression, as indicated by a quicker Rh123 release, may be helpful in maintaining mitochondrial integrity.

  12. Iron oxide nanoparticles induce human microvascular endothelial cell permeability through reactive oxygen species production and microtubule remodeling

    Shi Xianglin


    Full Text Available Abstract Background Engineered iron nanoparticles are being explored for the development of biomedical applications and many other industry purposes. However, to date little is known concerning the precise mechanisms of translocation of iron nanoparticles into targeted tissues and organs from blood circulation, as well as the underlying implications of potential harmful health effects in human. Results The confocal microscopy imaging analysis demonstrates that exposure to engineered iron nanoparticles induces an increase in cell permeability in human microvascular endothelial cells. Our studies further reveal iron nanoparticles enhance the permeability through the production of reactive oxygen species (ROS and the stabilization of microtubules. We also showed Akt/GSK-3β signaling pathways are involved in iron nanoparticle-induced cell permeability. The inhibition of ROS demonstrate ROS play a major role in regulating Akt/GSK-3β – mediated cell permeability upon iron nanoparticle exposure. These results provide new insights into the bioreactivity of engineered iron nanoparticles which can inform potential applications in medical imaging or drug delivery. Conclusion Our results indicate that exposure to iron nanoparticles induces an increase in endothelial cell permeability through ROS oxidative stress-modulated microtubule remodeling. The findings from this study provide new understandings on the effects of nanoparticles on vascular transport of macromolecules and drugs.

  13. Hard magnetization direction and its relation with magnetic permeability of highly grain-oriented electrical steel

    Hao Wang; Chang-sheng Li; Tao Zhu


    The magnetic properties of highly grain-oriented electrical steel vary along different directions. In order to investigate these prop-erties, standard Epstein samples were cut at different angles to the rolling direction. The hard magnetization direction was found at an angle of 60° to the rolling direction. To compare the measured and fitting curves, when the magnetic field intensity is higher than 7000 A/m, it is appropriate to simulate the relation of magnetic permeability and magnetization angle using the conventional elliptical model. When the magnetic field intensity is less than 3000 A/m, parabolic fitting models should be used; but when the magnetic field intensity is between 3000 and 7000 A/m, hybrid models with high accuracy, as proposed in this paper, should be applied. Piecewise relation models of magnetic per-meability and magnetization angle are significant for improving the accuracy of electromagnetic engineering calculations of electrical steel, and these new models could be applied in further industrial applications.

  14. Influence of a high-fat diet on gut microbiota, intestinal permeability and metabolic endotoxaemia.

    Moreira, Ana Paula Boroni; Texeira, Tatiana Fiche Salles; Ferreira, Alessandra Barbosa; Peluzio, Maria do Carmo Gouveia; Alfenas, Rita de Cássia Gonçalves


    Lipopolysaccharide (LPS) may play an important role in chronic diseases through the activation of inflammatory responses. The type of diet consumed is of major concern for the prevention and treatment of these diseases. Evidence from animal and human studies has shown that LPS can diffuse from the gut to the circulatory system in response to the intake of high amounts of fat. The method by which LPS move into the circulatory system is either through direct diffusion due to intestinal paracellular permeability or through absorption by enterocytes during chylomicron secretion. Considering the impact of metabolic diseases on public health and the association between these diseases and the levels of LPS in the circulatory system, this review will mainly discuss the current knowledge about high-fat diets and subclinical inflammation. It will also describe the new evidence that correlates gut microbiota, intestinal permeability and alkaline phosphatase activity with increased blood LPS levels and the biological effects of this increase, such as insulin resistance. Although the majority of the studies published so far have assessed the effects of dietary fat, additional studies are necessary to deepen the understanding of how the amount, the quality and the structure of the fat may affect endotoxaemia. The potential of food combinations to reduce the negative effects of fat intake should also be considered in future studies. In these studies, the effects of flavonoids, prebiotics and probiotics on endotoxaemia should be investigated. Thus, it is essential to identify dietetic strategies capable of minimising endotoxaemia and its postprandial inflammatory effects.

  15. Design and characterization of genetically engineered zebrafish aquaporin-3 mutants highly permeable to the cryoprotectant ethylene glycol

    Lubzens Esther


    Full Text Available Abstract Background Increasing cell membrane permeability to water and cryoprotectants is critical for the successful cryopreservation of cells with large volumes. Artificial expression of water-selective aquaporins or aquaglyceroporins (GLPs, such as mammalian aquaporin-3 (AQP3, enhances cell permeability to water and cryoprotectants, but it is known that AQP3-mediated water and solute permeation is limited and pH dependent. To exploit further the possibilities of using aquaporins in cryobiology, we investigated the functional properties of zebrafish (Danio rerio GLPs. Results Water, glycerol, propylene glycol and ethylene glycol permeability of zebrafish Aqp3a, -3b, -7, -9a, -9b, -10a and -10b, and human AQP3, was examined. Expression in Xenopus laevis oocytes indicated that the permeability of DrAqp3a and -3b to ethylene glycol was higher than for glycerol or propylene glycol under isotonic conditions, unlike other zebrafish GLPs and human AQP3, which were more permeable to glycerol. In addition, dose-response experiments and radiolabeled ethylene glycol uptake assays suggested that oocytes expressing DrAqp3b were permeated by this cryoprotectant more efficiently than those expressing AQP3. Water and ethylene glycol transport through DrAqp3a and -3b were, however, highest at pH 8.5 and completely abolished at pH 6.0. Point mutations in the DrAqp3b amino acid sequence rendered two constructs, DrAqp3b-T85A showing higher water and ethylene glycol permeability at neutral and alkaline pH, and DrAqp3b-H53A/G54H/T85A, no longer inhibited at acidic pH but less permeable than the wild type. Finally, calculation of permeability coefficients for ethylene glycol under concentration gradients confirmed that the two DrAqp3b mutants were more permeable than wild-type DrAqp3b and/or AQP3 at neutral pH, resulting in a 2.6- to 4-fold increase in the oocyte intracellular concentration of ethylene glycol. Conclusion By single or triple point mutations in the Dr

  16. Isoflavones in food supplements: chemical profile, label accordance and permeability study in Caco-2 cells.

    Almeida, I M C; Rodrigues, F; Sarmento, B; Alves, R C; Oliveira, M B P P


    Consumers nowadays are playing an active role in their health-care. A special case is the increasing number of women, who are reluctant to use exogenous hormone therapy for the treatment of menopausal symptoms and are looking for complementary therapies. However, food supplements are not clearly regulated in Europe. The EFSA has only recently begun to address the issues of botanical safety and purity regulation, leading to a variability of content, standardization, dosage, and purity of available products. In this study, isoflavones (puerarin, daidzin, genistin, daidzein, glycitein, genistein, formononetin, prunetin, and biochanin A) from food supplements (n = 15) for menopausal symptoms relief are evaluated and compared with the labelled information. Only four supplements complied with the recommendations made by the EC on the tolerable thresholds. The intestinal bioavailability of these compounds was investigated using Caco-2 cells. The apparent permeability coefficients of the selected isoflavonoids across the Caco-2 cells were affected by the isoflavone concentration and product matrix.

  17. Structural analysis and Caco-2 cell permeability of the celiac-toxic A-gliadin peptide 31-55.

    Iacomino, Giuseppe; Fierro, Olga; D'Auria, Sabato; Picariello, Gianluca; Ferranti, Pasquale; Liguori, Claudia; Addeo, Francesco; Mamone, Gianfranco


    Celiac disease is a chronic enteropathy caused by the ingestion of wheat gliadin and other cereal prolamines. The synthetic peptides 31-43 (P31-43) and 31-49 (P31-49) from A-gliadin are considered to be model peptides for studying innate immunity in celiac disease. Our previous study demonstrated that P31-43 and P31-49 are encrypted within peptide 31-55 (P31-55), which is naturally released from gastropancreatic digestion and is not susceptible to hydrolysis by brush border membrane enzymes. Here, we analyzed the permeability of P31-55 through the epithelial cell layer of confluent Caco-2 cells using high-performance liquid chromatography, mass spectrometry, and fluorescence-activated cell sorting. Twenty-three percent of the P31-55 added to the apical chamber was transported to the basolateral chamber after 4 h of incubation without being degraded by hydrolysis. Treatment of Caco-2 cells with whole gliadin digests extracted from a common wheat cultivar increased the epithelial P31-55 translocation by approximately 35%. Moreover, we observed an atypical chromatographic profile consisting of a double peak. Chromatography using different column temperatures and circular dichroism highlighted the presence of more conformational structures around the amide bond of the two adjacent prolines 38 and 39. These findings confirm that P31-55 is gastrointestinally resistant and is permeable across a Caco-2 monolayer. Moreover, we hypothesize that the various conformations of P31-55 may play a role in the activation of innate immunity.

  18. The Blood-Brain Barrier Permeability of Lignans and Malabaricones from the Seeds of Myristica fragrans in the MDCK-pHaMDR Cell Monolayer Model

    Ni Wu


    Full Text Available The blood-brain barrier (BBB permeability of twelve lignans and three phenolic malabaricones from the seeds of Myristica fragrans (nutmeg were studied with the MDCK-pHaMDR cell monolayer model. The samples were measured by high-performance liquid chromatography and the apparent permeability coefficients (Papp were calculated. Among the fifteen test compounds, benzonfuran-type, dibenzylbutane-type and arylnaphthalene-type lignans showed poor to moderate permeabilities with Papp values at 10−8–10−6 cm/s; those of 8-O-4′-neolignan and tetrahydrofuran-lignan were at 10−6–10−5 cm/s, meaning that their permeabilities are moderate to high; the permeabilities of malabaricones were poor as their Papp values were at 10−8–10−7 cm/s. To 5-methoxy-dehydrodiisoeugenol (2, erythro-2-(4-allyl-2,6-dimethoxyphenoxy-1-(3,4-dimethoxyphenyl-propan-1-ol acetate (6, verrucosin (8, and nectandrin B (9, an efflux way was involved and the main transporter for 6, 8 and 9 was demonstrated to be P-glycoprotein. The time and concentration dependency experiments indicated the main transport mechanism for neolignans dehydrodiisoeugenol (1, myrislignan (7 and 8 was passive diffusion. This study summarized the relationship between the BBB permeability and structure parameters of the test compounds, which could be used to preliminarily predict the transport of a compound through BBB. The results provide a significant molecular basis for better understanding the potential central nervous system effects of nutmeg.

  19. Human Lysozyme Synergistically Enhances Bactericidal Dynamics and Lowers the Resistant Mutant Prevention Concentration for Metronidazole to Helicobacter pylori by Increasing Cell Permeability

    Xiaolin Zhang


    Full Text Available Metronidazole (MNZ is an effective agent that has been employed to eradicate Helicobacter pylori (H. pylori. The emergence of broad MNZ resistance in H. pylori has affected the efficacy of this therapeutic agent. The concentration of MNZ, especially the mutant prevention concentration (MPC, plays an important role in selecting or enriching resistant mutants and regulating therapeutic effects. A strategy to reduce the MPC that can not only effectively treat H. pylori but also prevent resistance mutations is needed. H. pylori is highly resistant to lysozyme. Lysozyme possesses a hydrolytic bacterial cell wall peptidoglycan and a cationic dependent mode. These effects can increase the permeability of bacterial cells and promote antibiotic absorption into bacterial cells. In this study, human lysozyme (hLYS was used to probe its effects on the integrity of the H. pylori outer and inner membranes using as fluorescent probe hydrophobic 1-N-phenyl-naphthylamine (NPN and the release of aspartate aminotransferase. Further studies using a propidium iodide staining method assessed whether hLYS could increase cell permeability and promote cell absorption. Finally, we determined the effects of hLYS on the bactericidal dynamics and MPC of MNZ in H. pylori. Our findings indicate that hLYS could dramatically increase cell permeability, reduce the MPC of MNZ for H. pylori, and enhance its bactericidal dynamic activity, demonstrating that hLYS could reduce the probability of MNZ inducing resistance mutations.

  20. Influence of high permeability disks in an axisymmetric model of the Cadarache dynamo experiment

    Giesecke, A; Stefani, F; Gerbeth, G; Léorat, J; Herreman, W; Luddens, F; Guermond, J -L


    Numerical simulations of the kinematic induction equation are performed on a model configuration of the Cadarache von-K\\'arm\\'an-Sodium dynamo experiment. The effect of a localized axisymmetric distribution of relative permeability {\\mu} that represents soft iron material within the conducting fluid flow is investigated. The critical magnetic Reynolds number Rm^c for dynamo action of the first non-axisymmetric mode roughly scales like Rm^c({\\mu})-Rm^c({\\mu}->infinity) ~ {\\mu}^(-1/2) i.e. the threshold decreases as {\\mu} increases. This scaling law suggests a skin effect mechanism in the soft iron disks. More important with regard to the Cadarache dynamo experiment, we observe a purely toroidal axisymmetric mode localized in the high permeability disks which becomes dominant for large {\\mu}. In this limit, the toroidal mode is close to the onset of dynamo action with a (negative) growth-rate that is rather independent of the magnetic Reynolds number. We qualitatively explain this effect by paramagnetic pumping...

  1. Microbial communities in low permeability, high pH uranium mine tailings: characterization and potential effects.

    Bondici, V F; Lawrence, J R; Khan, N H; Hill, J E; Yergeau, E; Wolfaardt, G M; Warner, J; Korber, D R


    To describe the diversity and metabolic potential of microbial communities in uranium mine tailings characterized by high pH, high metal concentration and low permeability. To assess microbial diversity and their potential to influence the geochemistry of uranium mine tailings using aerobic and anaerobic culture-based methods, in conjunction with next generation sequencing and clone library sequencing targeting two universal bacterial markers (the 16S rRNA and cpn60 genes). Growth assays revealed that 69% of the 59 distinct culturable isolates evaluated were multiple-metal resistant, with 15% exhibiting dual-metal hypertolerance. There was a moderately positive correlation coefficient (R = 0·43, P iron, 22 reduced molybdate and seven oxidized arsenite. Based on next generation sequencing, tailings depth was shown to influence bacterial community composition, with the difference in the microbial diversity of the upper (0-20 m) and middle (20-40 m) tailings zones being highly significant (P iron-reducing bacteria were identified with low abundance, yet relatively high diversity. The presence of a population of metabolically-diverse, metal-resistant micro-organisms within the tailings environment, along with their demonstrated capacity for transforming metal elements, suggests that these organisms have the potential to influence the long-term geochemistry of the tailings. This study is the first investigation of the diversity and functional potential of micro-organisms present in low permeability, high pH uranium mine tailings. © 2013 The Society for Applied Microbiology.

  2. Vascular Permeability Drives Susceptibility to Influenza Infection in a Murine Model of Sickle Cell Disease

    Karlsson, Erik A.; Oguin, Thomas H.; Meliopoulos, Victoria; Iverson, Amy; Broadnax, Alexandria; Yoon, Sun-Woo; Pestina, Tamara; Thomas, Paul; Webby, Richard; Schultz-Cherry, Stacey; Rosch, Jason W.


    Sickle cell disease (SCD) is a major global health concern. Patients with SCD experience disproportionately greater morbidity and mortality in response to influenza infection than do others. Viral infection is one contributing factor for the development of Acute Chest Syndrome (ACS), a major cause of morbidity and mortality in SCD patients. We determined whether the heightened sensitivity to influenza infection could be reproduced in the two different SCD murine models to ascertain the underlying mechanisms of increased disease severity. In agreement with clinical observations, we found that both genetic and bone marrow-transplanted SCD mice had greater mortality in response to influenza infection than did wild-type animals. Despite similar initial viral titers and inflammatory responses between wild-type and SCD animals during infection, SCD mice continued to deteriorate and failed to resolve the infection, resulting in increased mortality. Histopathology of the lung tissues revealed extensive pulmonary edema and vascular damage following infection, a finding confirmed by heightened vascular permeability following virus challenge. These findings implicate the development of exacerbated pulmonary permeability following influenza challenge as the primary factor underlying heightened mortality. These studies highlight the need to focus on prevention and control strategies against influenza infection in the SCD population. PMID:28256526

  3. Proteomics-Based Metabolic Modeling Reveals That Fatty Acid Oxidation (FAO) Controls Endothelial Cell (EC) Permeability*

    Patella, Francesca; Schug, Zachary T.; Persi, Erez; Neilson, Lisa J.; Erami, Zahra; Avanzato, Daniele; Maione, Federica; Hernandez-Fernaud, Juan R.; Mackay, Gillian; Zheng, Liang; Reid, Steven; Frezza, Christian; Giraudo, Enrico; Fiorio Pla, Alessandra; Anderson, Kurt; Ruppin, Eytan; Gottlieb, Eyal; Zanivan, Sara


    Endothelial cells (ECs) play a key role to maintain the functionality of blood vessels. Altered EC permeability causes severe impairment in vessel stability and is a hallmark of pathologies such as cancer and thrombosis. Integrating label-free quantitative proteomics data into genome-wide metabolic modeling, we built up a model that predicts the metabolic fluxes in ECs when cultured on a tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the fatty acid oxidation rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle. Remarkably, global phosphoproteomic changes measured upon acute CPT1A inhibition pinpointed altered calcium signaling. Indeed, CPT1A inhibition increases intracellular calcium oscillations. Finally, inhibiting CPT1A induces hyperpermeability in vitro and leakage of blood vessel in vivo, which were restored blocking calcium influx or replenishing the tricarboxylic acid cycle. Fatty acid oxidation emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability. PMID:25573745

  4. Proteomics-based metabolic modeling reveals that fatty acid oxidation (FAO) controls endothelial cell (EC) permeability.

    Patella, Francesca; Schug, Zachary T; Persi, Erez; Neilson, Lisa J; Erami, Zahra; Avanzato, Daniele; Maione, Federica; Hernandez-Fernaud, Juan R; Mackay, Gillian; Zheng, Liang; Reid, Steven; Frezza, Christian; Giraudo, Enrico; Fiorio Pla, Alessandra; Anderson, Kurt; Ruppin, Eytan; Gottlieb, Eyal; Zanivan, Sara


    Endothelial cells (ECs) play a key role to maintain the functionality of blood vessels. Altered EC permeability causes severe impairment in vessel stability and is a hallmark of pathologies such as cancer and thrombosis. Integrating label-free quantitative proteomics data into genome-wide metabolic modeling, we built up a model that predicts the metabolic fluxes in ECs when cultured on a tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the fatty acid oxidation rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle. Remarkably, global phosphoproteomic changes measured upon acute CPT1A inhibition pinpointed altered calcium signaling. Indeed, CPT1A inhibition increases intracellular calcium oscillations. Finally, inhibiting CPT1A induces hyperpermeability in vitro and leakage of blood vessel in vivo, which were restored blocking calcium influx or replenishing the tricarboxylic acid cycle. Fatty acid oxidation emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability.

  5. Science Letters:Preparation of high-permeability NiCuZn ferrite

    HU Jun; YAN Mi


    Appropriate addition of CuO/V2O5 and the reduction of the granularity of the raw materials particle decrease the sintering temperature ofNiZn ferrite from 1200 ℃ to 930 ℃. Furthermore, the magnetic properties of the NiZn ferrite prepared at low temperature of 930 ℃ is superior to that of the NiZn ferrite prepared by sintering at high temperature of 1200 ℃ because the microstructure of the NiZn ferrite sintered at 930 ℃ is more uniform and compact than that of the NiZn ferrite sintered at 1200 ℃.The high permeability of 1700 and relative loss coefficient tanδ//μi of 9.0×10-6 at 100 kHz was achieved in the (Ni0.17Zn0.63Cu0.20)Fe1.915O4 ferrite.

  6. Suppression of cell membrane permeability by suramin: involvement of its inhibitory actions on connexin 43 hemichannels.

    Chi, Yuan; Gao, Kun; Zhang, Hui; Takeda, Masayuki; Yao, Jian


    Suramin is a clinically prescribed drug for treatment of human African trypanosomiasis, cancer and infection. It is also a well-known pharmacological antagonist of P2 purinoceptors. Despite its clinical use and use in research, the biological actions of this molecule are still incompletely understood. Here, we investigated the effects of suramin on membrane channels, as exemplified by its actions on non-junctional connexin43 (Cx43) hemichannels, pore-forming α-haemolysin and channels involved in ATP release under hypotonic conditions. Hemichannels were activated by removing extracellular Ca(2+) . The influences of suramin on hemichannel activities were evaluated by its effects on influx of fluorescent dyes and efflux of ATP. The membrane permeability and integrity were assessed through cellular retention of preloaded calcein and LDH release. Suramin blocked Cx43 hemichannel permeability induced by removal of extracellular Ca(2+) without much effect on Cx43 expression and gap junctional intercellular communication. This action of suramin was mimicked by its analogue NF023 and NF449 but not by another P2 purinoceptor antagonist PPADS. Besides hemichannels, suramin also significantly blocked intracellular and extracellular exchanges of small molecules caused by α-haemolysin from Staphylococcus aureus and by exposure of cells to hypotonic solution. Furthermore, it prevented α-haemolysin- and hypotonic stress-elicited cell injury. Suramin blocked membrane channels and protected cells against toxin- and hypotonic stress-elicited injury. Our finding provides novel mechanistic insights into the pharmacological actions of suramin. Suramin might be therapeutically exploited to protect membrane integrity under certain pathological situations. © 2014 The British Pharmacological Society.

  7. High permeability dialysis membrane allows effective removal of myoglobin in acute kidney injury resulting from rhabdomyolysis.

    Sorrentino, Sajoscha A; Kielstein, Jan T; Lukasz, Alexander; Sorrentino, Janine-Nicole; Gohrbandt, Bernhard; Haller, Hermann; Schmidt, Bernhard M W


    The objective of this study was to test the ability of myoglobin removal of a novel, high-permeability polysulphone dialyzer in acute kidney injury as a result of rhabdomyolysis. Intensive care unit of a tertiary care hospital. Six patients (one female; aged 24, 36, 41, 55, 63, and 65 yrs) with oligoanuric acute kidney injury resulting from rhabdomyolysis. Extended dialysis was performed using a single-pass batch dialysis system and a novel polysulphone high-flux dialyzer (effective surface area 1.8 m; inner lumen 220 μm; wall thickness 35 μm; allowing elimination of substances with a molecular weight of up to 30 kDa). Samples were collected at prefilter and postfilter sites as well as from the collected spent dialysate. The dialyzer clearance was calculated from concentrations before and directly after the dialysis membrane, the blood flow, and the ultrafiltration rate. The total amount of the myoglobin removed was measured directly as the whole dialysate was preserved. A median myoglobin clearance of 90.5 mL/min (range, 52.4-126.3 mL/min) was achieved, resulting in a median myoglobin removal per treatment hour of 0.54 g (range, 0.15-2.21 g). Extended dialysis with a high-flux, high-permeability membrane allowed effective elimination of myoglobin with a clearance of myoglobin that surpassed all previously reported dialysis techniques. This membrane may be advantageous in preventing acute kidney injury or avoiding complete loss of kidney function in patients with rhabdomyolysis. Further studies are needed to determine whether improving renal recovery or mortality in patients with acute kidney injury resulting from rhabdomyolysis is possible.

  8. High Resolution Additive Patterning of Nanoparticles and Polymers Enabled by Vapor Permeable Polymer Templates

    Demko, Michael Thomas

    The structure and chemistry of nanoparticles and polymers are interesting for applications in electronics and sensors. However, because they are outside of the standard material set typically used for these applications, widespread use of these materials has not yet been realized. This is due in part to the limited ability of traditional manufacturing processes to adapt to these unique materials. As a result, several alternative manufacturing methods have been developed, including nanoimprint lithography, gravure printing, inkjet printing, and screen printing, among many others. However, these current processes are not able to simultaneously produce patterns with high resolution and high dimensional fidelity, rapidly, over large areas, and in a completely additive manner. In this work, high-resolution patterns of nanoparticles and polymers are created on a variety of substrates in a completely additive manner using a template-based microfluidic process. Permeation of solvent through a vapor-permeable polymer template is used to both drive the flow of ink and concentrate the solute inside of template features. This fluidic process is shown to allow gradual packing of solute inside the template features, enabling creation of three-dimensional features with low defect densities. Additionally, because the mechanical properties of the template material are found to significantly impact patterning resolution and fidelity, and a process for creating rigid, vapor permeable templates from poly(4-methyl-2-pentyne) is developed. These templates are used for creating patterning of nanoparticles and polymers with a minimum line width of smaller than 350 nm. The process is then applied to the creation of low temperature metallization for polymer electronics using metallic nanoparticles and a highly-sensitive ultraviolet light sensor from zinc oxide nanoparticles.

  9. High-density expression of Ca2+-permeable ASIC1a channels in NG2 glia of rat hippocampus.

    Yen-Chu Lin

    Full Text Available NG2 cells, a fourth type of glial cell in the mammalian CNS, undergo reactive changes in response to a wide variety of brain insults. Recent studies have demonstrated that neuronally expressed acid-sensing ion channels (ASICs are implicated in various neurological disorders including brain ischemia and seizures. Acidosis is a common feature of acute neurological conditions. It is postulated that a drop in pH may be the link between the pathological process and activation of NG2 cells. Such postulate immediately prompts the following questions: Do NG2 cells express ASICs? If so, what are their functional properties and subunit composition? Here, using a combination of electrophysiology, Ca2+ imaging and immunocytochemistry, we present evidence to demonstrate that NG2 cells of the rat hippocampus express high density of Ca2+-permeable ASIC1a channels compared with several types of hippocampal neurons. First, nucleated patch recordings from NG2 cells revealed high density of proton-activated currents. The magnitude of proton-activated current was pH dependent, with a pH for half-maximal activation of 6.3. Second, the current-voltage relationship showed a reversal close to the equilibrium potential for Na+. Third, psalmotoxin 1, a blocker specific for the ASIC1a channel, largely inhibited proton-activated currents. Fourth, Ca2+ imaging showed that activation of proton-activated channels led to an increase of [Ca2+]i. Finally, immunocytochemistry showed co-localization of ASIC1a and NG2 proteins in the hippocampus. Thus the acid chemosensor, the ASIC1a channel, may serve for inducing membrane depolarization and Ca2+ influx, thereby playing a crucial role in the NG2 cell response to injury following ischemia.

  10. Cell-permeable p38 MAP kinase promotes migration of adult neural stem/progenitor cells

    Hamanoue, Makoto; Morioka, Kazuhito; Ohsawa, Ikuroh; Ohsawa, Keiko; Kobayashi, Masaaki; Tsuburaya, Kayo; Akasaka, Yoshikiyo; Mikami, Tetsuo; Ogata, Toru; Takamatsu, Ken


    Endogenous neural stem/progenitor cells (NPCs) can migrate toward sites of injury, but the migration activity of NPCs is insufficient to regenerate damaged brain tissue. In this study, we showed that p38 MAP kinase (p38) is expressed in doublecortin-positive adult NPCs. Experiments using the p38 inhibitor SB203580 revealed that endogenous p38 participates in NPC migration. To enhance NPC migration, we generated a cell-permeable wild-type p38 protein (PTD-p38WT) in which the HIV protein transduction domain (PTD) was fused to the N-terminus of p38. Treatment with PTD-p38WT significantly promoted the random migration of adult NPCs without affecting cell survival or differentiation; this effect depended on the cell permeability and kinase activity of the fusion protein. These findings indicate that PTD-p38WT is a novel and useful tool for unraveling the roles of p38, and that this protein provides a reasonable approach for regenerating the injured brain by enhancing NPC migration. PMID:27067799

  11. Pericyte abundance affects sucrose permeability in cultures of rat brain microvascular endothelial cells.

    Parkinson, Fiona E; Hacking, Cindy


    The blood-brain barrier is a physical and metabolic barrier that restricts diffusion of blood-borne substances into brain. In vitro models of the blood-brain barrier are used to characterize this structure, examine mechanisms of damage and repair and measure permeability of test substances. The core component of in vitro models of the blood-brain barrier is brain microvascular endothelial cells. We cultured rat brain microvascular endothelial cells (RBMEC) from isolated rat cortex microvessels. After 2-14 days in vitro (DIV), immunohistochemistry of these cells showed strong labeling for zona occludens 1 (ZO-1), a tight junction protein expressed in endothelial cells. Pericytes were also present in these cultures, as determined by expression of alpha-actin. The present study was performed to test different cell isolation methods and to compare the resulting cell cultures for abundance of pericytes and for blood-brain barrier function, as assessed by 14C-sucrose flux. Two purification strategies were used. First, microvessels were preabsorbed onto uncoated plastic for 4 h, then unattached microvessels were transferred to coated culture ware. Second, microvessels were incubated with an antibody to platelet-endothelial cell adhesion molecule 1 (PECAM-1; CD31) precoupled to magnetic beads, and a magnetic separation procedure was performed. Our results indicate that immunopurification, but not preadsorption, was an effective method to purify microvessels and reduce pericyte abundance in the resulting cultures. This purification significantly reduced 14C-sucrose fluxes across cell monolayers. These data indicate that pericytes can interfere with the development of blood-brain barrier properties in in vitro models that utilize primary cultures of RBMECs.

  12. Stimulating in situ denitrification in an aerobic, highly permeable municipal drinking water aquifer

    Critchley, K.; Rudolph, D. L.; Devlin, J. F.; Schillig, P. C.


    A preliminary trial of a cross-injection system (CIS) was designed to stimulate in situ denitrification in an aquifer servicing an urban community in southern Ontario. It was hypothesized that this remedial strategy could be used to reduce groundwater nitrate in the aquifer such that it could remain in use as a municipal supply until the beneficial effects of local reduced nutrient loadings lead to long-term water quality improvement at the wellfield. The CIS application involved injecting a carbon source (acetate) into the subsurface using an injection-extraction well pair positioned perpendicular to the regional flow direction, up-gradient of the water supply wells, with the objective of stimulating native denitrifying bacteria. The pilot remedial strategy was targeted in a high nitrate flux zone within an aerobic and heterogeneous section of the glacial sand and gravel aquifer. Acetate injections were performed at intervals ranging from daily to bi-daily. The carbon additions led to general declines in dissolved oxygen concentrations; decreases in nitrate concentration were localized in aquifer layers where velocities were estimated to be less than 0.5 m/day. NO3-15N and NO3-18O isotope data indicated the nitrate losses were due to denitrification. Relatively little nitrate was removed from groundwater in the more permeable strata, where velocities were estimated to be on the order of 18 m/day or greater. Overall, about 11 percent of the nitrate mass passing through the treatment zone was removed. This work demonstrates that stimulating in situ denitrification in an aerobic, highly conductive aquifer is challenging but achievable. Further work is needed to increase rates of denitrification in the most permeable units of the aquifer.

  13. Measurement of Transient Permeability of Sp2/0 Myeloma Cells: Flow Cytometric Study

    Novickij Vitalij


    Full Text Available Electroporation is an electric field induced phenomenon occurring when the permeability of the cell membrane is increased due to the excess of critical transmembrane potential. Fluorescent dye assays are frequently used for evaluation of the permeabilization rate, however, the protocols vary, which negatively affects the repeatability of the results. In this work we have designed experiments to investigate the protocols and threshold concentrations of the Propidium Iodide (PI and YO-PRO-1 (YP fluorescent dyes for evaluation of mammalian cell permeabilization induced by electroporation. The Sp2/0 mouse myeloma cells were used and the bursts of 100 μs × 8 electrical pulses of 0.8-2 kV/cm were applied. It has been shown that the dye concentration has an influence on the detectable permeabilization, and the concentrations below 30 μM for PI and 1 μM for YP should be avoided for measurement of electropermeabilization efficacy due to unreliable fluorescence signals. Further, based on the experimental data, the permeabilization curve for the Sp2/0 myeloma cells in the 0.8-2 kV/cm range has been presented.

  14. Evaluation of Various Pulse-Decay Laboratory Permeability Measurement Techniques for Highly Stressed Coals

    Feng, Ruimin; Harpalani, Satya; Pandey, Rohit


    The transient technique for laboratory permeability measurement, proposed by Brace et al. (J Geophys Res 73:2225-2236, 1968) and widely used for conventional gas reservoir rocks, is the preferred method when testing low-permeability rocks in the laboratory. However, Brace et al.'s solution leads to considerable errors since it does not take into account compressive storage and sorption effect when applied to sorptive rocks, such as, coals and shales. To verify the applicability of this solution when used to characterize fluid flow behavior of coal, an in-depth investigation of permeability evolution for flow of helium and methane depletion was conducted for San Juan coals using the pressure pulse-decay method under best replicated in situ conditions. Three permeability solutions, Brace et al.'s (1968), Dicker and Smits's (International meeting on petroleum engineering, Society of Petroleum Engineers, 1988) and Cui et al.'s (Geofluids 9:208-223, 2009), were utilized to establish the permeability trends. Both helium and methane permeability results exhibited very small difference between the Brace et al.'s solution and Dicker and Smits's solution, indicating that the effect of compressive storage is negligible. However, methane permeability enhancement at low pressures due to coal matrix shrinkage resulting from gas desorption can be significant and this was observed in pressure response plots and the estimated permeability values using Cui et al.'s solution only. Therefore, it is recommended that Cui et al.'s solution be employed to correctly include the sorption effect when testing coal permeability using the transient technique. A series of experiments were also carried out to establish the stress-dependent permeability trend under constant effective stress condition, and then quantify the sole contribution of the sorption effect on permeability variation. By comparison with the laboratory data obtained under in situ stress/strain condition, it was verified that

  15. Puerarin transport across a Calu-3 cell monolayer – an in vitro model of nasal mucosa permeability and the influence of paeoniflorin and menthol

    Zhang L


    Full Text Available Lin Zhang, Shou-Ying Du, Yang Lu, Chang Liu, Zhi-Hao Tian, Chang Yang, Hui-Chao Wu, Zhen Wang School of Chinese Materia Medica, Beijing University of Chinese Medicine, Chaoyang District, Beijing, People’s Republic of China Abstract: Nasal administration is a high-potential delivery system, particularly because it can provide a pathway from the nose to the brain. The objective of this research is to characterize puerarin transport across a Calu-3 cell monolayer used as a model of the nasal mucosa and to evaluate the influence of puerarin in combination with paeoniflorin and menthol to explore the enhanced mechanism of the permeability at the cell level. The apparent permeability coefficients (Papp of puerarin bidirectional transport were both <1.5×10-6 cm/s, and the efflux ratio was <1.5, indicating that puerarin alone exhibited poor absorption and that its transport primarily occurred by passive diffusion through the cell monolayer. When puerarin was coad­ministered with paeoniflorin, the Papp was not changed (P>0.05. However, the addition of menthol significantly (P<0.05 improved the Papp of puerarin in both directions. Moreover, based on immunofluorescence experiments and transepithelial electrical resistance measurements, the data indicated that the drug compatibility opened tight junctions and weakened the barrier capabilities of epithelial cells, thereby promoting the permeability of puerarin. Keywords: puerarin, paeoniflorin, menthol, nasal administration, transport, tight junction

  16. A highly permeable and enhanced surface area carbon-cloth electrode for vanadium redox flow batteries

    Zhou, X. L.; Zhao, T. S.; Zeng, Y. K.; An, L.; Wei, L.


    In this work, a high-performance porous electrode, made of KOH-activated carbon-cloth, is developed for vanadium redox flow batteries (VRFBs). The macro-scale porous structure in the carbon cloth formed by weaving the carbon fibers in an ordered manner offers a low tortuosity (∼1.1) and a broad pore distribution from 5 μm to 100 μm, rendering the electrode a high hydraulic permeability and high effective ionic conductivity, which are beneficial for the electrolyte flow and ion transport through the porous electrode. The use of KOH activation method to create nano-scale pores on the carbon-fiber surfaces leads to a significant increase in the surface area for redox reactions from 2.39 m2 g-1 to 15.4 m2 g-1. The battery assembled with the present electrode delivers an energy efficiency of 80.1% and an electrolyte utilization of 74.6% at a current density of 400 mA cm-2, as opposed to an electrolyte utilization of 61.1% achieved by using a conventional carbon-paper electrode. Such a high performance is mainly attributed to the combination of the excellent mass/ion transport properties and the high surface area rendered by the present electrode. It is suggested that the KOH-activated carbon-cloth electrode is a promising candidate in redox flow batteries.

  17. Toward high permeability, selectivity and controllability of water desalination with FePc nanopores.

    Deng, Qingming; Pan, Jun; Yin, Xiaohui; Wang, Xiaofeng; Zhao, Lina; Kang, Seung-gu; Jimenez-Cruz, Camilo A; Zhou, Ruhong; Li, Jingyuan


    Nanoporous materials exhibit promising potential in water transportation applications, especially in ocean water desalination. It is highly desired to have great permeability, selectivity and controllability in the desalination performance of these nanopores. However, it is still a challenge to achieve all three features in one material or device. Here, we demonstrate efficient and controllable water desalination with a nanoporous 2D Fe phthalocyanine (FePc) membrane using molecular dynamics simulations. We find the FePc membrane not only conducts fast water flow, but it also suppresses ion permeation. The selectivity is attributed to a mechanism distinct from the traditional steric exclusion: cations are excluded due to electrostatic repulsion, whereas anions can be trapped in the nanopore and induce the reorganization of ions in the vicinity of the nanopore, which in turn creates a tendency for the trapped anions to move back into the saline reservoir. More interestingly, we find such mechanism is largely due to the sufficiently strong electrostatic interaction of the charged nanopore region with ions and is not restricted to the FePc nanopore. In addition, the number of protonated nitrogen atoms in FePc pores can be modulated by adjusting the pH value of the solution. The extent of the anion occupancy can thus be regulated, giving rise to control of the water flow. Taken together, great permeability, selectivity and controllability can be achieved with this nanosheet system. Moreover, our study suggests there is an alternative mechanism of water desalination which may be realized by intrinsically nanoporous materials such as FePc membranes.

  18. Membrane-Permeable Calpain Inhibitors Promote Rat Oral Mucosal Epithelial Cell Proliferation by Inhibiting IL-1α Signaling.

    Makoto Kondo

    Full Text Available To standardise regenerative medicine using cultured cells, the use of serum-free, chemically defined media will be necessary. We have reported that IL-1α inhibits the growth of epithelial cells in culture and that recombinant IL-1 receptor antagonist (IL-1RA significantly promotes epithelial cell growth in no feeder layer condition. In this study, we examined inhibitors of calpain, a cysteine proteinase that plays crucial roles in various cellular functions, including IL-1α maturation and secretion. The culturing of epithelial cells in serum-free media supplemented with a membrane-permeable calpain inhibitor significantly promoted growth while suppressing IL-1α maturation and secretion. By contrast, non-membrane-permeable calpain inhibitor treatment did not have these effects. Interestingly, immunoblotting analysis revealed that immature, untruncated, IL-1α expression was also downregulated by cell-permeable calpain inhibitor treatment, and the difference in IL-1α gene expression increased from day 2 to day 6. Although IL-1RA has been reported to promote epithelial cell growth, we detected no synergistic promotion of epithelial cell growth using a calpain inhibitor and IL-1RA. These findings indicate that calpain inhibitors promote epithelial cell proliferation by inhibiting IL-1α maturation at an early phase of epithelial cell culture and by suppressing the positive feedback-mediated amplification of IL-1α signalling.

  19. Epithelial Cell Damage Activates Bactericidal/Permeability Increasing-Protein (BPI Expression in Intestinal Epithelium

    Arjun Balakrishnan


    Full Text Available As the first line of defense against invading pathogen, intestinal epithelium produces various antimicrobial proteins (AMP that help in clearance of pathogen. Bactericidal/permeability-increasing protein (BPI is a 55 kDa AMP that is expressed in intestinal epithelium. Dysregulation of BPI in intestinal epithelium is associated with various inflammatory diseases like Crohn’s Disease, Ulcerative colitis, and Infectious enteritis’s. In this paper, we report a direct correlation between intestinal damage and BPI expression. In Caco-2 cells, we see a significant increase in BPI levels upon membrane damage mediated by S. aureus infection and pore-forming toxins (Streptolysin and Listeriolysin. Cells detect changes in potassium level as a Danger-associated molecular pattern associated with cell damage and induce BPI expression in a p38 dependent manner. These results are further supported by in vivo findings that the BPI expression in murine intestinal epithelium is induced upon infection with bacteria which cause intestinal damage (Salmonella Typhimurium and Shigella flexneri whereas mutants that do not cause intestinal damage (STM ΔfliC and STM ΔinvC did not induce BPI expression. Our results suggest that epithelial damage associated with infection act as a signal to induce BPI expression.

  20. Error Analysis of High Frequency Core Loss Measurement for Low-Permeability Low-Loss Magnetic Cores

    Niroumand, Farideh Javidi; Nymand, Morten


    in magnetic cores is B-H loop measurement where two windings are placed on the core under test. However, this method is highly vulnerable to phase shift error, especially for low-permeability, low-loss cores. Due to soft saturation and very low core loss, low-permeability low-loss magnetic cores are favorable....... The analysis has been validated by experimental measurements for relatively low-loss magnetic cores with different permeability values.......Magnetic components significantly contribute to the dissipated loss in power electronic converters. Measuring the true value of dissipated power in these components is highly desirable, since it can be used to verify the optimum design of these components. The common approach for measuring the loss...

  1. Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide

    Mehrdad Moosazadeh Moghaddam


    Full Text Available Introduction: Some of bacterial species are able to uptake DNA molecule from environment, the yield of this process depends on some conditions such as plasmid size and host type. In the case of Bacillus subtilis, DNA uptake has low efficacy. Using Spizizen minimal medium is common method in plasmid transformation into B. subtilis, but rate of this process is not suitable and noteworthy. The aim of this study was investigation of novel method for improvement of DNA transformation into B. subtilis based on CM11 cationic peptide as a membrane permeable agent.Materials and methods: In this study, for optimization of pWB980 plasmid transformation into B. subtilis, the CM11 cationic peptide was used. For this purpose, B. subtilis competent cell preparation in the present of different concentration of peptide was implemented by two methods. In the first method, after treatment of bacteria with different amount of peptide for 14h, plasmid was added. In the second method, several concentration of peptide with plasmid was exposed to bacteria simultaneously. Bacteria that uptake DNA were screened on LB agar medium containing kanamycin. The total transformed bacteria per microgram of DNA was calculated and compared with the control.Results: Plasmid transformation in best conditions was 6.5 folds higher than the control. This result was statistically significant (P value <0.001.Discussion and conclusion: This study showed that CM11 cationic peptide as a membrane permeable agent was able to increase plasmid transformation rate into B. subtilis. This property was useful for resolution of low transformation efficacy.

  2. Evaluation of the intestinal permeability of rosemary (Rosmarinus officinalis L.) extract polyphenols and terpenoids in Caco-2 cell monolayers

    Arráez-Román, David; González-Álvarez, Isabel; Ibáñez, Elena; Segura-Carretero, Antonio; Bermejo, Marival; Micol, Vicente


    Rosemary (Rosmarinus officinalis) is grown throughout the world and is widely used as a medicinal herb and to season and preserve food. Rosemary polyphenols and terpenoids have attracted great interest due to their potential health benefits. However, complete information regarding their absorption and bioavailability in Caco-2 cell model is scarce. The permeation properties of the bioactive compounds (flavonoids, diterpenes, triterpenes and phenylpropanoids) of a rosemary extract (RE), obtained by supercritical fluid extraction, was studied in Caco-2 cell monolayer model, both in a free form or liposomed. Compounds were identified and quantitated by liquid chromatography coupled to quadrupole time-of-flight with electrospray ionization mass spectrometry analysis (HPLC-ESI-QTOF-MS), and the apparent permeability values (Papp) were determined, for the first time in the extract, for 24 compounds in both directions across cell monolayer. For some compounds, such as triterpenoids and some flavonoids, Papp values found were reported for the first time in Caco-2 cells.Our results indicate that most compounds are scarcely absorbed, and passive diffusion is suggested to be the primary mechanism of absorption. The use of liposomes to vehiculize the extract resulted in reduced permeability for most compounds. Finally, the biopharmaceutical classification (BCS) of all the compounds was achieved according to their permeability and solubility data for bioequivalence purposes. BCS study reveal that most of the RE compounds could be classified as classes III and IV (low permeability); therefore, RE itself should also be classified into this category. PMID:28234919




    Seedlings of a salt sensitive line of Triticum aestivum were grown in Hoagland solution supplemented with 100 mM NaCl following a pretreatment with choline chloride (ChCl). Changes in growth, mineral content of roots and shoots, and passive permeability of the cell membrane were measured. Relative g

  4. The Effect of Capsaicin Derivatives on Tight-Junction Integrity and Permeability of Madin-Darby Canine Kidney Cells.

    Kaiser, Mathias; Chalapala, Sudharani; Gorzelanny, Christian; Perali, Ramu Sridhar; Goycoolea, Francisco Martin


    Capsaicin is known to interfere with tight junctions (TJs) of epithelial cells and therefore to enhance paracellular permeability of poorly absorbable drugs. However, due to its low water solubility, pungency, and cytotoxicity, its pharmacologic use is limited. In this study, we investigated the effect of capsaicin derivatives of synthetic (e.g., 10-hydroxy-N-(4-hydroxy-3-methoxybenzyl)decanamide, etc.) and natural (olvanil and dihydrocapsaicin) origin on Madin-Darby Canine Kidney-C7 cells. Impedance spectroscopy was used to determine the transepithelial electrical resistance and the capacitance. Permeability assays with fluorescein isothiocyanate-dextran were carried out to evaluate the impact on cell permeability. The results show that lipophilicity could play an important role for the interference with TJ and that the mechanism is independent from the ion channel TRPV-1 and hence on the flux of calcium into the cells. In summary, we synthesized 4 derivatives of capsaicin of lower lipophilicity and compared their properties with other well-known vanilloids. We show that these compounds are able to enhance the permeability of a hydrophilic macromolecule, by opening the TJ for a shorter time than capsaicin. This behavior is dependent on the lipophilicity of the molecule. Understanding of these phenomena may lead to better control of administration of therapeutic molecules.

  5. Modeling placental transport: correlation of in vitro BeWo cell permeability and ex vivo human placental perfusion

    Poulsen, Marie Sønnegaard; Rytting, Erik; Mose, Tina


    across the BeWo cells was observed in the rank order of caffeine>antipyrine>benzoic acid>glyphosate in terms of both the apparent permeability coefficient and the initial slope, defined as the linear rate of substance transferred to the fetal compartment as percent per time, a parameter used to compare...

  6. The effect of pore-scale geometry and wettability on two-phase relative permeabilities within elementary cells

    Bianchi Janetti, Emanuela; Riva, Monica; Guadagnini, Alberto


    We study the relative role of the complex pore space geometry and wettability of the solid matrix on the quantification of relative permeabilities characterizing steady state immiscible two-phase flow in porous media. We do so by considering elementary cells, which are typically employed in upscaling frameworks based on, e.g., homogenization or volume averaging. In this context one typically relies on the solution of pore-scale physics at a scale which is much smaller than that of an investigated porous system. Pressure-driven two-phase flow following simultaneous co-current injection of water and oil is numerically solved for a suite of regular and stochastically generated two-dimensional explicit elementary cells with fixed porosity and sharing main topological/morphological features. We show that relative permeabilities of the randomly generated elementary cells are significantly influenced by the formation of preferential percolation paths (principal pathways), giving rise to a strongly nonuniform distribution of fluid fluxes. These pathways are a result of the spatially variable resistance that the random pore structures exert on the fluid. The overall effect on relative permeabilities of the diverse organization of principal pathways, as driven by a given random realization at the scale of the unit cell, is significantly larger than that of the wettability of the host rock. In contrast to what can be observed for the random cells analyzed, relative permeabilities of regular cells display a clear trend with contact angle at the investigated scale. Our findings suggest the need to perform systematic upscaling studies in a stochastic context, to propagate the effects of uncertain pore space geometries to a probabilistic description of relative permeability curves at the continuum scale.

  7. Cell-permeable Ln(III) chelate-functionalized InP quantum dots as multimodal imaging agents.

    Stasiuk, Graeme J; Tamang, Sudarsan; Imbert, Daniel; Poillot, Cathy; Giardiello, Marco; Tisseyre, Céline; Barbier, Emmanuel L; Fries, Pascal Henry; de Waard, Michel; Reiss, Peter; Mazzanti, Marinella


    Quantum dots (QDs) are ideal scaffolds for the development of multimodal imaging agents, but their application in clinical diagnostics is limited by the toxicity of classical CdSe QDs. A new bimodal MRI/optical nanosized contrast agent with high gadolinium payload has been prepared through direct covalent attachment of up to 80 Gd(III) chelates on fluorescent nontoxic InP/ZnS QDs. It shows a high relaxivity of 900 mM(-1) s(-1) (13 mM(-1 )s(-1) per Gd ion) at 35 MHz (0.81 T) and 298 K, while the bright luminescence of the QDs is preserved. Eu(III) and Tb(III) chelates were also successfully grafted to the InP/ZnS QDs. The absence of energy transfer between the QD and lanthanide emitting centers results in a multicolor system. Using this convenient direct grafting strategy additional targeting ligands can be included on the QD. Here a cell-penetrating peptide has been co-grafted in a one-pot reaction to afford a cell-permeable multimodal multimeric MRI contrast agent that reports cellular localization by fluorescence and provides high relaxivity and increased tissue retention with respect to commercial contrast agents.

  8. [Effect of toluene diisocyanate on reactive oxygen species production and permeability of human bronchial epithelial cells in vitro].

    Mo, Guan-wen; Cai, Shao-xi; Zhao, Hai-jin; Li, Wen-jun; Tong, Wan-cheng; Liu, Lai-yu


    To investigate the effect of toluene diisocyanate (TDI) on the production of reactive oxygen species (ROS) and the permeability of human bronchial epithelial (HBE) cells. TDI-human serum albumin (TDI-HSA) conjugate was prepared using a modified Son's method. MTT assay was used to assess HBE cell viability after exposure to different concentrations of TDI-HSA. The level of intracellular ROS of HBE cells was detected by flow cytometry with an oxidation-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) uploading, and the permeability of cell monolayer was assessed by detecting the transepithelial electrical resistance (TEER). The exposure to 120 µg/ml TDI-HSA did not obviously affect the cell viability. Compared with the control group, the intracellular fluorescent intensity increased significantly in the cells exposed to 20, 60, and 100 µg/ml TDI-HSA (Pproduction increased significantly after 100 µg/ml TDI-HSA treatment (Pproduction was significantly suppressed by pretreatment of the cells with N-acetylcysteine (NAC) (P<0.05), which also enhanced the TEER decreased by TDI-HSA treatment (P<0.05). TDI enhances the permeability of HBE cell monolayer partially through a ROS-mediated pathway, suggesting the importance of oxidative stress in TDI-induced pulmonary diseases.

  9. Alterations in voltage-sensing of the mitochondrial permeability transition pore in ANT1-deficient cells.

    Doczi, Judit; Torocsik, Beata; Echaniz-Laguna, Andoni; Mousson de Camaret, Bénédicte; Starkov, Anatoly; Starkova, Natalia; Gál, Aniko; Molnár, Mária J; Kawamata, Hibiki; Manfredi, Giovanni; Adam-Vizi, Vera; Chinopoulos, Christos


    The probability of mitochondrial permeability transition (mPT) pore opening is inversely related to the magnitude of the proton electrochemical gradient. The module conferring sensitivity of the pore to this gradient has not been identified. We investigated mPT's voltage-sensing properties elicited by calcimycin or H2O2 in human fibroblasts exhibiting partial or complete lack of ANT1 and in C2C12 myotubes with knocked-down ANT1 expression. mPT onset was assessed by measuring in situ mitochondrial volume using the 'thinness ratio' and the 'cobalt-calcein' technique. De-energization hastened calcimycin-induced swelling in control and partially-expressing ANT1 fibroblasts, but not in cells lacking ANT1, despite greater losses of mitochondrial membrane potential. Matrix Ca(2+) levels measured by X-rhod-1 or mitochondrially-targeted ratiometric biosensor 4mtD3cpv, or ADP-ATP exchange rates did not differ among cell types. ANT1-null fibroblasts were also resistant to H2O2-induced mitochondrial swelling. Permeabilized C2C12 myotubes with knocked-down ANT1 exhibited higher calcium uptake capacity and voltage-thresholds of mPT opening inferred from cytochrome c release, but intact cells showed no differences in calcimycin-induced onset of mPT, irrespective of energization and ANT1 expression, albeit the number of cells undergoing mPT increased less significantly upon chemically-induced hypoxia than control cells. We conclude that ANT1 confers sensitivity of the pore to the electrochemical gradient.

  10. On the boundary layer structure near a highly permeable porous interface

    Dalwadi, Mohit P; Waters, Sarah L; Oliver, James M


    The method of matched asymptotic expansions is used to study the canonical problem of steady laminar flow through a narrow two-dimensional channel blocked by a tight-fitting finite-length highly permeable porous obstacle. We investigate the behaviour of the local flow close to the interface between the single-phase and porous regions (governed by the incompressible Navier--Stokes and Darcy flow equations, respectively). We solve for the local flow in the limits of low and high Reynolds number, facilitating an understanding of the nature of the transition from Poiseuille to plug to Poiseuille flow in each of these limits. Significant analytic progress is made in the high-Reynolds-number limit, as we are able to explore in detail the rich boundary layer structure that occurs. We derive general results for the interfacial stress and for the conditions that couple the flow in the regions away from the interface. We consider the three-dimensional generalization to unsteady laminar flow through and around a tight-f...

  11. Silver nanoparticles interact with the cell membrane and increase endothelial permeability by promoting VE-cadherin internalization

    Sun, Xia; Shi, Junpeng; Zou, Xiaoyan; Wang, Chengcheng; Yang, Yi; Zhang, Hongwu, E-mail:


    Highlights: • Short-term exposure to AgNPs at low doses induces increase HUVECs monolayer permeability. • AgNPs interact with the cell membrane and increase endothelial permeability by promoting VE-Cadherin internalization. • Particle effect is a major factor leading to endothelial dysfunction. - Abstract: The toxicological risks of silver nanoparticles (AgNPs) have attracted widespread attention, and many studies have been published that have contributed to understanding AgNPs-induced toxicity. However, little attention has been paid to the low-dose effects of AgNPs and the related toxicological mechanism is still unclear. Here, we show that short-term exposure to AgNPs at low doses induces a substantial increase in human umbilical vein endothelial cells (HUVECs) monolayer permeability, whereas Ag ions at low doses do not induce an observable increase in monolayer permeability. This effect is independent of oxidative stress and apoptosis. Scanning electron microscopy confirms that AgNPs adhere to the cell membrane after 1 h exposure. Furthermore, adhesion of AgNPs to the cell membrane can trigger vascular endothelial (VE)-cadherin phosphorylation at Y658 followed by VE-cadherin internalization, which lead to the increase in endothelial monolayer permeability. Our data show that surface interactions of AgNPs with the cell membrane, in other words, the particle effect, is a major factor leading to endothelial dysfunction following low-dose and short-term exposure. Our findings will contribute to understanding the health effects and the toxicological mechanisms of AgNPs.

  12. Honeycomb porous films as permeable scaffold materials for human embryonic stem cell-derived retinal pigment epithelium.

    Calejo, Maria Teresa; Ilmarinen, Tanja; Jongprasitkul, Hatai; Skottman, Heli; Kellomäki, Minna


    Age-related macular degeneration (AMD) is a leading cause of blindness in developed countries, characterised by the degeneration of the retinal pigment epithelium (RPE), a pigmented cell monolayer that closely interacts with the photoreceptors. RPE transplantation is thus considered a very promising therapeutic option to treat this disease. In this work, porous honeycomb-like films are for the first time investigated as scaffold materials for human embryonic stem cell-derived retinal pigment epithelium (hESC-RPE). By changing the conditions during film preparation, it was possible to produce films with homogeneous pore distribution and adequate pore size (∼3-5 µm), that is large enough to ensure high permeability but small enough to enable cell adherence and spreading. A brief dip-coating procedure with collagen type IV enabled the homogeneous adsorption of the protein to the walls and bottom of pores, increasing the hydrophilicity of the surface. hESC-RPE adhered and proliferated on all the collagen-coated materials, regardless of small differences in pore size. The differentiation of hESC-RPE was confirmed by the detection of specific RPE protein markers. These results suggest that the porous honeycomb films can be promising candidates for hESC-RPE tissue engineering, importantly enabling the free flow of ions and molecules across the material. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1646-1656, 2016.

  13. Passive permeability and effective pore size of HeLa cell nuclear membranes.

    Samudram, Arunkarthick; Mangalassery, Bijeesh M; Kowshik, Meenal; Patincharath, Nandakumar; Varier, Geetha K


    Nuclear pore complexes in the nuclear membrane act as the sole gateway of transport of molecules from the cytoplasm to the nucleus and vice versa. Studies on biomolecular transport through nuclear membranes provide vital data on the nuclear pore complexes. In this work, we use fluorescein isothiocyanate-labeled dextran molecules as a model system and study the passive nuclear import of biomolecules through nuclear pore complexes in digitonin-permeabilized HeLa cells. Experiments are carried out under transient conditions in the time lapse imaging scheme using an in-house constructed confocal laser scanning microscope. Transport rates of dextran molecules having molecular weights of 4-70 kDa corresponding to Stokes radius of 1.4-6 nm are determined. Analyzing the permeability of the nuclear membrane for different sizes the effective pore radius of HeLa cell nuclear membrane is determined to be 5.3 nm, much larger than the value reported earlier using proteins as probe molecules. The range of values reported for the nuclear pore radius suggest that they may not be rigid structures and it is quite probable that the effective pore size of nuclear pore complexes is critically dependent on the probe molecules and on the environmental factors.

  14. Measuring the osmotic water permeability coefficient (Pf) of spherical cells: isolated plant protoplasts as an example.

    Shatil-Cohen, Arava; Sibony, Hadas; Draye, Xavier; Chaumont, François; Moran, Nava; Moshelion, Menachem


    Studying AQP regulation mechanisms is crucial for the understanding of water relations at both the cellular and the whole plant levels. Presented here is a simple and very efficient method for the determination of the osmotic water permeability coefficient (P(f)) in plant protoplasts, applicable in principle also to other spherical cells such as frog oocytes. The first step of the assay is the isolation of protoplasts from the plant tissue of interest by enzymatic digestion into a chamber with an appropriate isotonic solution. The second step consists of an osmotic challenge assay: protoplasts immobilized on the bottom of the chamber are submitted to a constant perfusion starting with an isotonic solution and followed by a hypotonic solution. The cell swelling is video recorded. In the third step, the images are processed offline to yield volume changes, and the time course of the volume changes is correlated with the time course of the change in osmolarity of the chamber perfusion medium, using a curve fitting procedure written in Matlab (the 'PfFit'), to yield P(f).

  15. Anti-chloride permeability and anti-chloride corrodibility of a green high performance concrete admixture in concrete

    ZHENG Yongbao; LIU Xinrong; LIU Dongyan; CHEN Jiangong; HUANG Xiongjun


    The effects of green high performance concrete (GHPC) admixture on the anti-chloride permeability and anti- chloride corrodibility of concrete are studied by a series of experiments designed on the basis of the diffusion principle and electrochemical principle. The GHPC admixture consists of fly ash, gangue, slag, red mud, etc., of which the mass fraction of industrial residues is over 96 %. The anti-permeabilities and anti-corrodibilities of the tested GHPC and normal concrete (NC) are evaluated by the Diffusion Coefficients of chloride which was obtained by measuring the concentration of chloride in the tested systems by the voltage difference method. It is found that the adoption of GHPC admixture greatly improves the anti-chloride permeability and anti-chloride corrodibility by modifying the inner structure and contracting the porosity of concrete to the reduce considerably the diffusion rate of chloride. The admixture is desirable regarding its engineering performances as well as economical and environmental interests.

  16. High permeability and low loss of Ni-Zn-Fe ferrite/metal composite cores in high frequency region

    Mori, Satoshi; Mitsuoka, Takeshi; Sonehara, Makoto; Sato, Toshiro; Matsushita, Nobuhiro


    A Ni-Zn-Fe ferrite was deposited on the surface of Fe-Si and three kinds of Fe-Si-Cr powders as an insulating material using an ultrasonic-enhanced ferrite plating method. All ferrite-coated powders exhibited a high magnetization of about 180 emu/g derived from their soft magnetic metal and ferrite composition. The three different ferrite-coated Fe-Si-Cr cores annealed at 973 K exhibited a constant permeability μ' of 55-60 in the frequency range up to 50 MHz owing to the insulating effect of the ferrite layer. In contrast, the permeability μ' of the ferrite-coated Fe-3.5Si mass % core decreased in the MHz range. The Cr component acted as a passive layer that made it possible to maintain the Ni-Zn-Fe ferrite layer after annealing. The core loss of the ferrite-coated Fe-3.5Si-4.5Cr mass % core was 448 kW/m3 at 100 kHz and 50 mT, which was half of that observed for the Fe-3.5Si-4.5Cr core without ferrite layer. Additionally, the eddy-current loss of the ferrite-coated Fe-3.5Si-4.5Cr core was strongly decreased compared with the non-coated Fe-3.5Si-4.5Cr core owing to the insulating properties of the ferrite layer.

  17. Amoxicillin separation from pharmaceutical wastewater by high permeability polysulfone nanofiltration membrane.

    Derakhsheshpoor, Reza; Homayoonfal, Maryam; Akbari, Ahmad; Mehrnia, Mohammad Reza


    In this study, high permeability flat sheet polysulfone nanofiltration membranes were prepared for amoxicillin (AMX) recovery from pharmaceutical wastewater. Membrane fabrication includes two steps: raw ultrafiltration membrane synthesis by phase inversion method and nanaofiltration membrane synthesis by surface photopolymerization. Raw ultrafiltration membranes were synthesized using different molecular weights of polyethylene glycol (PEG) as pore former and different coagulation bath temperatures (CBTs). The synthesized ultrafiltration membranes were modified using UV-assisted polymerization technique and their performance in the separation of AMX at different pHs, were studied. The results showed that the more irradiation time, the smaller surface pore size. Moreover, the membranes made with higher molecular weight of PEG and coagulation bath temperatures were more susceptible for UV-modification at these conditions; fabricated membranes had higher flux as well as relatively high AMX separation. Moreover, pH enhancement increased AMX rejection by 85%. The effect of irradiation on membrane surface morphology was studied by SEM surface images and the morphological effects of pore former and coagulation bath temperatures on membrane structure were confirmed by SEM cross section images. A fairly comprehensive discussion about the effects of PEG, coagulation bath temperature and irradiation time on membrane structure and AMX recovery performance was represented in this study.

  18. Towards high water permeability in triazine-framework-based microporous membranes for dehydration of ethanol.

    Tang, Yu Pan; Wang, Huan; Chung, Tai Shung


    The microstructural evolution of a series of triazine framework-based microporous (TFM) membranes under different conditions has been explored in this work. The pristine TFM membrane is in situ fabricated in the course of polymer synthesis via a facile Brønsted-acid-catalyzed cyclotrimerizaiton reaction. The as-synthesized polymer exhibits a microporous network with high thermal stability. The free volume size of the TFM membranes gradually evolved from a unimodal distribution to a bimodal distribution under annealing, as analyzed by positron annihilation lifetime spectroscopy (PALS). The emergence of the bimodal distribution is probably ascribed to the synergetic effect of quenching and thermal cyclization reaction. In addition, the fractional free volume (FFV) of the membranes presents a concave trend with increasing annealing temperature. Vapor sorption tests reveal that the mass transport properties are closely associated with the free volume evolution, which provides an optimal condition for dehydration of biofuels. A promising separation performance with extremely high water permeability has been attained for dehydration of an 85 wt % ethanol aqueous solution at 45 °C. The study on the free volume evolution of the TFM membranes may provide useful insights about the microstructure and mass transport behavior of the microporous polymeric materials.

  19. A mixed ionic and electronic conducting dual-phase membrane with high oxygen permeability.

    Fang, Wei; Liang, Fangyi; Cao, Zhengwen; Steinbach, Frank; Feldhoff, Armin


    To combine good chemical stability and high oxygen permeability, a mixed ionic-electronic conducting (MIEC) 75 wt% Ce(0.85)Gd(0.1)Cu(0.05)O(2-δ)-25 wt% La(0.6)Ca(0.4)FeO(3-δ)(CGCO-LCF) dual-phase membrane based on a MIEC-MIEC composite has been developed. Copper doping into Ce(0.9)Gd(0.1)O(2-δ) (CGO) oxide enhances both ionic and electronic conductivity, which then leads to a change from ionic conduction to mixed conduction at elevated temperatures. For the first time we demonstrate that an intergranular film with 2-10 nm thickness containing Ce, Ca, Gd, La, and Fe has been formed between the CGCO grains in the CGCO-LCF one-pot dual-phase membrane. A high oxygen permeation flux of 0.70 mL min(-1) cm(-2) is obtained by the CGCO-LCF one-pot dual-phase membrane with 0.5 mm thickness at 950 °C using pure CO2 as the sweep gas, and the membrane shows excellent stability in the presence of CO2 even at lower temperatures (800 °C) during long-term operation.

  20. Amoxicillin Separation from Pharmaceutical Wastewater by High Permeability Polysulfone Nanofiltration Membrane

    Reza Derakhsheshpoor


    Full Text Available In this study, high permeability flat sheet polysulfone nanofiltration membranes were prepared for amoxicillin (AMX recovery from pharmaceutical wastewater. Membrane fabrication includes two steps: raw ultrafiltration membrane synthesis by phase inversion method and nanaofiltration membrane synthesis by surface photopolymerization. Raw ultrafiltration membranes were synthesized using different molecular weights of polyethylene glycol (PEG as pore former and different coagulation bath temperatures (CBTs. The synthesized ultrafiltration membranes were modified using UV-assisted polymerization technique and their performance in the separation of AMX at different pHs, were studied. The results showed that the more irradiation time, the smaller surface pore size. Moreover, the membranes made with higher molecular weight of PEG and coagulation bath temperatures were more susceptible for UV-modification at these conditions; fabricated membranes had higher flux as well as relatively high AMX separation. Moreover, pH enhancement increased AMX rejection by 85%. The effect of irradiation on membrane surface morphology was studied by SEM surface images and the morphological effects of pore former and coagulation bath temperatures on membrane structure were confirmed by SEM cross section images. A fairly comprehensive discussion about the effects of PEG, coagulation bath temperature and irradiation time on membrane structure and AMX recovery performance was represented in this study.

  1. Experimental measurements of the permeability of fibrous carbon at high temperature

    Panerai, Francesco; Cochell, Thomas J; Schroeder, Olivia M; Mansour, Nagi N; Wright, Michael J; Martin, Alexandre


    A series of experiments was performed to obtain permeability data on FiberForm(R), a commercial carbon preform used for manufacturing thermal protection systems. A porous sample was placed in a quartz flow-tube heated by an isothermal furnace. The setup was instrumented to measure mass flow through and pressure drop across the sample. The intrinsic permeability and the Klinkenberg correction, which accounts for rarefied effects, were computed from the experimental data. The role of the gas temperature and pressure on the effective permeability is shown, and it is demonstrated that with proper data reduction, the intrinsic permeability is strictly a function of the micro-structure of the material. A function for the effective permeability of FiberForm, dependent on temperature, pressure, pore geometry, and type of gas is proposed. The intrinsic permeability was evaluated at $K_0 = 5.57 \\times 10^{-11}$ m$^2$, with a Klinkenberg parameter of $8c/d_p = 2.51 \\times 10^5$ m$^{-1}$ and a reference porosity of $\\phi...

  2. Plasmolysis and cell shape depend on solute outer-membrane permeability during hyperosmotic shock in E. coli.

    Pilizota, Teuta; Shaevitz, Joshua W


    The concentration of chemicals inside the bacterial cytoplasm generates an osmotic pressure, termed turgor, which inflates the cell and is necessary for cell growth and survival. In Escherichia coli, a sudden increase in external concentration causes a pressure drop across the cell envelope that drives changes in cell shape, such as plasmolysis, where the inner and outer membranes separate. Here, we use fluorescence imaging of single cells during hyperosmotic shock with a time resolution on the order of seconds to examine the response of cells to a range of different conditions. We show that shock using an outer-membrane impermeable solute results in total cell volume reduction with no plasmolysis, whereas a shock caused by outer-membrane permeable ions causes plasmolysis immediately upon shock. Slowly permeable solutes, such as sucrose, which cross the membrane in minutes, cause plasmolysis to occur gradually as the chemical potential equilibrates. In addition, we quantify the detailed morphological changes to cell shape during osmotic shock. Nonplasmolyzed cells shrink in length with an additional lateral size reduction as the magnitude of the shock increases. Quickly plasmolyzing cells shrink largely at the poles, whereas gradually plasmolyzing cells invaginate along the cell cylinder. Our results give a comprehensive picture of the initial response of E. coli to hyperosmotic shock and offer explanations for seemingly opposing results that have been reported previously.

  3. The inner mitochondrial membrane has aquaporin-8 water channels and is highly permeable to water.

    Calamita, Giuseppe; Ferri, Domenico; Gena, Patrizia; Liquori, Giuseppa E; Cavalier, Annie; Thomas, Daniel; Svelto, Maria


    Mitochondria are remarkably plastic organelles constantly changing their shape to fulfil their various functional activities. Although the osmotic movement of water into and out of the mitochondrion is central for its morphology and activity, the molecular mechanisms and the pathways for water transport across the inner mitochondrial membrane (IMM), the main barrier for molecules moving into and out of the organelle, are completely unknown. Here, we show the presence of a member of the aquaporin family of water channels, AQP8, and demonstrate the strikingly high water permeability (Pf) characterizing the rat liver IMM. Immunoblotting, electron microscopy, and biophysical studies show that the largest mitochondria feature the highest AQP8 expression and IMM Pf. AQP8 was also found in the mitochondria of other organs, whereas no other known aquaporins were seen. The osmotic water transport of liver IMM was partially inhibited by the aquaporin blocker Hg2+, while the related activation energy remained low, suggesting the presence of a Hg2+-insensitive facilitated pathway in addition to AQP8. It is suggested that AQP8-mediated water transport may be particularly important for rapid expansions of mitochondrial volume such as those occurring during active oxidative phosphorylation and those following apoptotic signals.

  4. High permeability cores to optimize the stimulation of deeply located brain regions using transcranial magnetic stimulation

    Salvador, R; Miranda, P C [Institute of Biophysics and Biomedical Engineering, Faculty of Sciences, University of Lisbon, 1749-016 Lisbon (Portugal); Roth, Y [Advanced Technology Center, Sheba Medical Center, Tel-Hashomer (Israel); Zangen, A [Neurobiology Department, Weizmann Institute of Science, Rehovot 76100 (Israel)], E-mail:


    Efficient stimulation of deeply located brain regions with transcranial magnetic stimulation (TMS) poses many challenges, arising from the fact that the induced field decays rapidly and becomes less focal with depth. We propose a new method to improve the efficiency of TMS of deep brain regions that combines high permeability cores, to increase focality and field intensity, with a coil specifically designed to induce a field that decays slowly with increasing depth. The performance of the proposed design was investigated using the finite element method to determine the total electric field induced by this coil/core arrangement on a realistically shaped homogeneous head model. The calculations show that the inclusion of the cores increases the field's magnitude by as much as 25% while also decreasing the field's decay with depth along specific directions. The focality, as measured by the area where the field's norm is greater than 1/{radical}2 of its maximum value, is also improved by as much as 15% with some core arrangements. The coil's inductance is not significantly increased by the cores. These results show that the presence of the cores might make this specially designed coil even more suited for the effective stimulation of deep brain regions.

  5. The attenuation of microorganisms in on-site wastewater effluent discharged into highly permeable subsoils

    O'Luanaigh, N. D.; Gill, L. W.; Misstear, B. D. R.; Johnston, P. M.


    An extensive field study on percolation areas receiving both septic tank and secondary treated on-site effluents from single houses in Ireland was carried out to investigate the attenuation capacity of highly permeable subsoils with respect to E. coli bacteria and spiked bacteriophages (MS2, ΦX174 and PR772). The development of biomats across the percolation areas receiving the secondary effluent was restricted compared to the percolation area receiving septic tank effluent, promoting a much higher areal hydraulic loading which created significant differences in the potential microbiological loading to groundwater. Greatest E. coli removal in the subsoil occurred within the first 0.35 m of unsaturated subsoil for all effluent types. Analysis showed, however, that more evidence of faecal contamination occurred at depth in the subsoils receiving secondary treated effluents than that receiving septic tank effluent, despite the lower bacterial influent load. All three bacteriophages were reduced to their minimum detection limit (septic tank effluent, although isolated incidences of ΦX174 and PR772 were measured below one trench. However again, slightly higher breakthroughs of MS2 and PR772 contamination were detected at the same depth under the trenches receiving secondary treated effluent.

  6. Actin Dynamics Regulates Voltage-Dependent Calcium-Permeable Channels of the Vicia faba Guard Cell Plasma Membrane

    Wei Zhang; Liu-Min Fan


    Free cytosolic Ca~(2+) ([Ca~(2+)]_(cyt)) is an ubiquitous second messenger in plant cell signaling, and [Ca~(2+)]_(cyt) elevation is associated with Ca~(2+)-permeable channels in the plasma membrane and endomembranes regulated by a wide range of stimuli. However, knowledge regarding Ca~(2+) channels and their regulation remains limited in planta. A type of voltage-dependent Ca~(2+)-permeable channel was identified and characterized for the Vicia faba L. guard cell plasma membrane by using patch-clamp techniques. These channels are permeable to both Ba~(2+) and Ca~(2+), and their activities can be inhibited by micromolar Gd~(3+). The unitary conductance and the reversal potential of the channels depend on the Ca~(2+) or Ba~(2+) gradients across the plasma membrane. The inward whole-cell Ca~(2+) (Ba~(2+)) current, as well as the unitary current amplitude and NP. of the single Ca~(2+) channel, increase along with the membrane hyperpolarization. Pharmacological experiments suggest that actin dynamics may serve as an upstream regulator of this type of calcium channel of the guard cell plasma membrane. Cytochalasin D, an actin polymerization blocker, activated the NP_o of these channels at the single channel level and increased the current amplitude at the whole-cell level. But these channel activations and current increments could be restrained by pretreatment with an F-actin stabilizer, phalloidin. The potential physiological significance of this regulatory mechanism is also discussed.

  7. Morphology and water permeability of red blood cells from green sea turtle (Chelonia mydas).

    Benga, Gheorghe; Chapman, Bogdan E; Romeo, Tony; Cox, Guy C; Kuchel, Philip W


    The morphology and diffusional water permeability (P d) of red blood cells (RBCs) from green sea turtle (GST) (Chelonia mydas) are presented for the first time. The RBCs had an ellipsoidal shape with full-axis lengths (diameters): D = 14.4 μm; d = 10.2 μm; h = 2.8 μm. The values of P d (cm s(-1)) were 5.1 × 10(-3) at 15 °C, 5.7 × 10(-3) at 20 °C, 6.3 × 10(-3) at 25 °C, 6.8 × 10(-3) at 30 °C, and 7.9 × 10(-3) at 37 °C (i.e., significantly higher than in human RBCs in which it was measured to be 4.2 × 10(-3) at 25 °C, 5.0 × 10(-3) at 30 °C, and 6.2 × 10(-3) at 37 °C). There was a lack of inhibition of P d of GST RBCs by p-chloromercuribenzoate (PCMB), a well-known inhibitor of the RBC water channel proteins (WCPs). The activation energy of water diffusion (E a,d) in GST RBCs was 15.0 ± 1.6 kJ mol(-1) which is lower than the E a,d for human RBCs (~25 kJ mol(-1)). These results indicate that in the membrane of GST RBCs, there were no WCPs that were inhibited by the mercurial reagent, while the lipid bilayer of this membrane is unusually permeable to water. This is likely to be a phylogenetically old trait, like that found in amphibians and even the later birds, all of which have nucleated erythrocytes; and it is also likely to be a result of the animal's adaptation to a herbivorous diet (algae and seagrasses).

  8. Towards the characterization of an in vitro triple co-culture intestine cell model for permeability studies.

    Araújo, Francisca; Sarmento, Bruno


    Caco-2 based cell models have been the gold standard in vitro method to study intestinal drug permeability, despite the absence of many important features with major influence in the drug absorption mechanism. In the present work, a triple co-culture comprising Caco-2, HT29-MTX and Raji B cells was established to mimic in a closely way the human intestinal epithelium, presenting the main components in the process of drug absorption, namely the absorptive cells that resemble enterocytes, mucus producers cells and cells able to induce M-cell phenotype on Caco-2 cells. All the three cell lines maintained their function when cultured together with each other being, thus, an asset to new orally administrated drugs development. The seeding ratio of 90:10 between Caco-2 and HT29-MTX showed to be the best to achieve physiological proportions after cells maturation and differentiation in culture. The formation of M-cells phenotype from enterocytes was identified for the first time in a co-culture system comprising Caco-2 and HT29-MTX cells through immunocytochemical techniques. Thus, the triple co-culture model presented in the herein work is a good and reliable alternative to the in vitro methods already existents for the study of drugs permeability. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. On the impact of spatial heterogeneous permeability distributions on the development of free convection cells in the Perth Basin, Australia.

    Niederau, Jan; Ebigbo, Anozie; Freitag, Sebastian; Marquart, Gabriele; Clauser, Christoph


    Recent increase in exploration of the geothermal energy potential of the Perth Metropolitan Area (PMA) results in the need for reliable and robust reservoir models in order to explore rock properties and temperature distributions in the subsurface, where free convection in the main reservoir (Yarragadee Aquifer) is likely to occur [1]. While the structure of the Perth Basin has been refined recently, the heterogeneity and spatial complexity of permeability was up till now mainly neglected. An integrated, three dimensional tectonostratigraphic model of the PMA is constructed, using the modeling software '3D GeoModeller' and data of numerous artesian and petroleum wells. Comprising the region around the city of Perth, the model covers an area of about 5000 km2 up to a depth of 4.5 km, with focus on adequate representation of the main reservoir. We further construct a numerical model for fluid flow and heat transport in the Yarragadee Aquifer. Porosity distributions are deduced from well logs and linked to permeability by a calibrated correlation, based on a fractal approach. Three different cases are simulated using the FD code SHEMAT-Suite, in order to assess the influence of spatial heterogeneity of porosity and permeability on the development of free convection cells. constant porosity and permeability for the entire aquifer porosity and permeability decreasing with depth, thus reflecting compaction a conditional random permeability field within prescribed limits and for given correlation length In order to improve understanding of model correctness, as well as identification and comparison of convection cells in different simulations, we are developing a specialized visualization tool tailored to this purpose. The three different scenarios show distinctions in the distribution of convection cells. Where the Yarragadee Aquifer is in contact with overlying aquifers, regions of downflow develop. These in turn have a strong impact on the regional flow field and

  10. Influence of permeability on hydrothermal circulation in the sediment-buried oceanic crust

    WANG Xingtao; ZHAI Shikui; MENG Fanshun; LI Huaiming; YU Zenghui; SUN Ge; XUE Gang


    Hydrothermal convection in the upper oceanic crust has been inferred to be a common and important process. Under the simplified conditions of planar boundaries, permeability provides a strong constraint on the pattern of circulation, the dimensions of convective cells and flow field of hydrothermal circulation. By applying an advanced numerical modeling method, to our knowledge, it is the first time to investigate convection as it is influenced by different strata permeability structures,formational anisotropy, fracture zone and cooling intrusion. The simplified geological model is composed of 3 layers, sedimentary layer, high permeable basement layer and low permeable basement layer from top to bottom. When permeability in high permeable layer is 10 times larger than that in sedimentary layer, convection occurs in high permeable layer. The pattern of hydrothermal circulation and flow velocity of hydrothermal fluid are strongly influenced by strata permeability structures,changes of permeability in high permeable basement layer, fracture zone and cooling intrusion.However, formational anisotropy relatively exerts weak influence on hydrothermal circulation, with the ratio up to 1.5 of vertical permeability to lateral permeability in high permeable layer. Fracture zone existing in basement is the most important factor affecting the circulation field. The effects of a local intrusion are limited to convection intensity above the intrusion and have little impact on the fluid flow on a regional scale. As the result of numerical modelling, key factors affecting the hydrothermal circulation are good permeable zone and long-term heat source, not including fluid source.

  11. Determination of the Membrane Permeability to Water of Human Vaginal Mucosal Immune Cells at Subzero Temperatures Using Differential Scanning Calorimetry.

    Shu, Zhiquan; Hughes, Sean M; Fang, Cifeng; Hou, Zhiyuan; Zhao, Gang; Fialkow, Michael; Lentz, Gretchen; Hladik, Florian; Gao, Dayong


    To study mucosal immunity and conduct HIV vaccine trials, it is important to be able to cryopreserve mucosal specimens and recover them in functional viable form. Obtaining a good recovery depends, in part, on cooling the cells at the appropriate rate, which is determined by the rate of water transport across the cell membrane during the cooling process. In this study, the cell membrane permeabilities to water at subzero temperatures of human vaginal mucosal T cells and macrophages were measured using the differential scanning calorimetry method proposed by Devireddy et al. in 1998. Thermal histograms were measured before and after cell lysis using a Slow-Fast-Fast-Slow cooling program. The difference between the thermal histograms of the live intact cells and the dead lysed cells was used to calculate the temperature-dependent cell membrane permeability at subzero temperatures, which was assumed to follow the Arrhenius relationship, [Formula: see text], where Lpg is the permeability to water at the reference temperature (273.15 K). The results showed that Lpg = 0.0209 ± 0.0108 μm/atm/min and Ea = 41.5 ± 11.4 kcal/mol for T cells and Lpg = 0.0198 ± 0.0102 μm/atm/min and Ea = 38.2 ± 10.4 kcal/mol for macrophages, respectively, in the range 0°C to -40°C (mean ± standard deviation). Theoretical simulations predicted that the optimal cooling rate for both T cells and macrophages was about -3°C/min, which was proven by preliminary immune cell cryopreservation experiments.

  12. Infection of human monocyte-derived dendritic cells by ANDES Hantavirus enhances pro-inflammatory state, the secretion of active MMP-9 and indirectly enhances endothelial permeability

    Lopez-Lastra Marcelo


    Full Text Available Abstract Background Andes virus (ANDV, a rodent-borne Hantavirus, is the major etiological agent of Hantavirus cardiopulmonary syndrome (HCPS in South America, which is mainly characterized by a vascular leakage with high rate of fatal outcomes for infected patients. Currently, neither specific therapy nor vaccines are available against this pathogen. ANDV infects both dendritic and epithelial cells, but in despite that the severity of the disease directly correlates with the viral RNA load, considerable evidence suggests that immune mechanisms rather than direct viral cytopathology are responsible for plasma leakage in HCPS. Here, we assessed the possible effect of soluble factors, induced in viral-activated DCs, on endothelial permeability. Activated immune cells, including DC, secrete gelatinolytic matrix metalloproteases (gMMP-2 and -9 that modulate the vascular permeability for their trafficking. Methods A clinical ANDES isolate was used to infect DC derived from primary PBMC. Maturation and pro-inflammatory phenotypes of ANDES-infected DC were assessed by studying the expression of receptors, cytokines and active gMMP-9, as well as some of their functional status. The ANDES-infected DC supernatants were assessed for their capacity to enhance a monolayer endothelial permeability using primary human vascular endothelial cells (HUVEC. Results Here, we show that in vitro primary DCs infected by a clinical isolate of ANDV shed virus RNA and proteins, suggesting a competent viral replication in these cells. Moreover, this infection induces an enhanced expression of soluble pro-inflammatory factors, including TNF-α and the active gMMP-9, as well as a decreased expression of anti-inflammatory cytokines, such as IL-10 and TGF-β. These viral activated cells are less sensitive to apoptosis. Moreover, supernatants from ANDV-infected DCs were able to indirectly enhance the permeability of a monolayer of primary HUVEC. Conclusions Primary human DCs

  13. Effect of Light-Activated Hypocrellin B on the Growth and Membrane Permeability of Gram-Negative Escherichia coli Cells

    Yuan Jiang


    Full Text Available Aim. To investigate the effect of light-activated hypocrellin B on the growth and membrane permeability of Gram-negative bacteria. Methods. Escherichia coli (E. coli as a model bacterium of Gram-negative bacteria was incubated with various concentrations of hypocrellin B for 60 min and was subsequently irradiated by blue light with wavelength of 470 nm at the dose of 12 J/cm2. Colony forming units were counted and the growth inhibition rate of E. coli cells was calculated after light-activated hypocrellin B. Membrane permeability was measured using flow cytometry and confocal laser scanning microscopy (CLSM with propidium iodide (PI staining. Bacterial morphology was observed using transmission electron microscopy (TEM. Reactive oxygen species in bacterial cells were measured using flow cytometry with DCFH-DA staining. Results. Significant growth inhibition rate of E. coli cells was observed after photodynamic action of hypocrellin B. Remarkable damage to the ultrastructure of E. coli was also observed by TEM. Flow cytometry and CLSM observation showed that light-activated hypocrellin B markedly increased membrane permeability of E. coli. Flow cytometry showed the intracellular ROS increase in E. coli treated by photodynamic action of hypocrellin B. Conclusion. Light-activated hypocrellin B caused intracellular ROS increase and structural damages and inhibited the growth of Gram-negative E. coli cells.

  14. Evaluation of layers of the rat airway epithelial cell line RL-65 for permeability screening of inhaled drug candidates.

    Hutter, V; Hilgendorf, C; Cooper, A; Zann, V; Pritchard, D I; Bosquillon, C


    A rat respiratory epithelial cell culture system for in vitro prediction of drug pulmonary absorption is currently lacking. Such a model may however enhance the understanding of interspecies differences in inhaled drug pharmacokinetics by filling the gap between human in vitro and rat in/ex vivo drug permeability screens. The rat airway epithelial cell line RL-65 was cultured on Transwell inserts for up to 21 days at an air-liquid (AL) interface and cell layers were evaluated for their suitability as a drug permeability measurement tool. These layers were found to be morphologically representative of the bronchial/bronchiolar epithelium when cultured for 8 days in a defined serum-free medium. In addition, RL-65 layers developed epithelial barrier properties with a transepithelial electrical resistance (TEER) >300 Ω cm(2) and apparent (14)C-mannitol permeability (P(app)) values between 0.5-3.0 × 10(-6)cm/s; i.e., in the same range as established in vitro human bronchial epithelial absorption models. Expression of P-glycoprotein was confirmed by gene analysis and immunohistochemistry. Nevertheless, no vectorial transport of the established substrates (3)H-digoxin and Rhodamine123 was observed across the layers. Although preliminary, this study shows RL-65 cell layers have the potential to become a useful in vitro screening tool in the pre-clinical development of inhaled drug candidates.

  15. Development of a membrane impregnated with a poly(dimethylsiloxane)/poly(ethylene glycol) copolymer for a high-throughput screening of the permeability of drugs, cosmetics, and other chemicals across the human skin.

    Miki, Ryotaro; Ichitsuka, Yasuna; Yamada, Takumi; Kimura, Soichiro; Egawa, Yuya; Seki, Toshinobu; Juni, Kazuhiko; Ueda, Hideo; Morimoto, Yasunori


    We aimed to develop a high-throughput screening (HTS) system for preliminary predictions of human skin permeability by using an artificial membrane that can mimic the permeation behaviour of lipophilic and hydrophilic compounds across the human skin. In this study, we synthesized a copolymer containing poly(dimethylsiloxane) (PDMS) and poly(ethylene glycol) (PEG) 6000 and impregnated it onto a supportive membrane filter to prepare a PDMS/PEG 6000 copolymer-impregnated membrane. In addition, we synthesized another polymer without PEG units and used it to prepare an impregnated membrane for determining the role of PEG 6000 units in the PDMS/PEG 6000 copolymer-impregnated membrane. The permeation characteristics of the impregnated membranes were evaluated on the basis of the permeability coefficients of 12 model compounds with different lipophilicities, by using a 2-chamber diffusion cell, and these permeability coefficients were compared with those across the human skin. We obtained a good correlation between the permeability coefficients across the PDMS/PEG 6000 copolymer-impregnated membrane and human skin. Further, we evaluated the permeation characteristics of a 96-well plate model of the PDMS/PEG 6000 copolymer by using 6 model compounds. We obtained an ideal correlation between the permeability coefficients across the PDMS/PEG 6000 copolymer using a 96-well plate and those across the human skin. Thus, the PDMS/PEG 6000 copolymer would be a good candidate for preliminary evaluation of the permeability of lipophilic and hydrophilic compounds across the human skin.

  16. Cell-permeable gomesin peptide promotes cell death by intracellular Ca(2+) overload.

    Paredes-Gamero, Edgar J; Casaes-Rodrigues, Rafael L; Moura, Gioconda E D D; Domingues, Tatiana M; Buri, Marcus V; Ferreira, Victor H C; Trindade, Edvaldo S; Moreno-Ortega, Ana J; Cano-Abad, María F; Nader, Helena B; Ferreira, Alice T; Miranda, Antonio; Justo, Giselle Z; Tersariol, Ivarne L S


    In recent years, the antitumoral activity of antimicrobial peptides (AMPs) has been the goal of many research studies. Among AMPs, gomesin (Gm) displays antitumor activity by unknown mechanisms. Herein, we studied the cytotoxicity of Gm in the Chinese hamster ovary (CHO) cell line. Furthermore, we investigated the temporal ordering of organelle changes and the dynamics of Ca(2+) signaling during Gm-induced cell death. The results indicated that Gm binds to the plasma membrane and rapidly translocates into the cytoplasm. Moreover, 20 μM Gm increases the cytosolic Ca(2+) and induces membrane permeabilization after 30 min of treatment. Direct Ca(2+) measurements in CHO cells transfected with the genetically encoded D1-cameleon to the endoplasmic reticulum (ER) revealed that Gm induces ER Ca(2+) depletion, which in turn resulted in oscillatory mitochondrial Ca(2+) signal, as measured in cells expressing the genetically encoded probe to the mitochondrial matrix (mit)Pericam. This leads to mitochondria disruption, loss of mitochondrial membrane potential and increased reactive oxygen species prior to membrane permeabilization. Gm-induced membrane permeabilization by a Ca(2+)-dependent pathway involving Gm translocation into the cell, ER Ca(2+) depletion and disruption, mitochondrial Ca(2+) overload and oxidative stress.

  17. Colon-targeted cell-permeable NFκB inhibitory peptide is orally active against experimental colitis.

    Hong, Sungchae; Yum, Soohwan; Yoo, Hyun-Jung; Kang, Sookjin; Yoon, Jeong-Hyun; Min, Dosik; Kim, Young Mi; Jung, Yunjin


    For the purpose of development of orally active peptide therapeutics targeting NFκB for treatment of inflammatory bowel disease (IBD), two major barriers in oral delivery of therapeutic peptides, metabolic lability and tissue impermeability, were circumvented by introduction of a colon-targeted delivery system and cell permeable peptides (CPP) to NFκB inhibitory peptides (NIP). Suppression of NFκB activation was compared following treatment with various CPP conjugated NIPs (CPP-NIP). The most potent CPP-NIP was loaded in a capsule coated with a colon specific polymer, which was administered orally to colitic rats. The anti-inflammatory activity of the colon-targeted CPP-NIP was evaluated by measuring inflammatory indices in the inflamed colonic tissue. For confirmation of the local action of the CPP-NIP, the same experiment was done after rectal administration. Tissue permeability of the CPP-NIP was examined microscopically and spectrophotometrically using FITC-labeled CPP-NIP (CPP-NIP-FITC). NEMO binding domain peptide (NBD, TALDWSWLQTE) fused with a cell permeable peptide CTP (YGRRARRRARR), CTP-NBD, was most potent in inhibiting NFκB activity in cells. Colon-targeted CTP-NBD, but not colon-targeted NBD and CTP-NBD in an enteric capsule, ameliorated the colonic injury, which was in parallel with decrease in MPO activity and the levels of inflammatory mediators. Intracolonic treatment with CTP-NBD alleviated rat colitis and improved all the inflammatory indicators. CTP-NBD-FITC was detected at much greater level in the inflamed tissue than was NBD-FITC. Taken together, introduction of cell permeability and colon targetability to NIP may be a feasible strategy for an orally active peptide therapy for treatment of IBD.

  18. Metamaterials and the Landau-Lifshitz permeability argument: large permittivity begets high-frequency magnetism.

    Merlin, Roberto


    Homogeneous composites, or metamaterials, made of dielectric or metallic particles are known to show magnetic properties that contradict arguments by Landau and Lifshitz [Landau LD, Lifshitz EM (1960) Electrodynamics of Continuous Media (Pergamon, Oxford, UK), p 251], indicating that the magnetization and, thus, the permeability, loses its meaning at relatively low frequencies. Here, we show that these arguments do not apply to composites made of substances with Im square root(epsilon(S)) > lambda/l or Re square root(epsilon(S)) approximately lambda/l (epsilon(S) and l are the complex permittivity and the characteristic length of the particles, and lambda > l is the vacuum wavelength). Our general analysis is supported by studies of split rings, one of the most common constituents of electromagnetic metamaterials, and spherical inclusions. An analytical solution is given to the problem of scattering by a small and thin split ring of arbitrary permittivity. Results reveal a close relationship between epsilon(S) and the dynamic magnetic properties of metamaterials. For |square root(epsilon(S))| magnetic activity, consistent with the Landau-Lifshitz argument and similar to that of molecular crystals. In contrast, large values of the permittivity lead to strong diamagnetic or paramagnetic behavior characterized by susceptibilities whose magnitude is significantly larger than that of natural substances. We compiled from the literature a list of materials that show high permittivity at wavelengths in the range 0.3-3000 microm. Calculations for a system of spherical inclusions made of these materials, using the magnetic counterpart to Lorentz-Lorenz formula, uncover large magnetic effects the strength of which diminishes with decreasing wavelength.

  19. Extracellular K+ activates a K(+)- and H(+)-permeable conductance in frog taste receptor cells.

    Kolesnikov, S S; Margolskee, R F


    1. The effect of extracellular K+ on membrane currents of bull frog (Rana catesbeiana) taste receptor cells (TRCs) was investigated by the patch clamp and fast perfusion techniques. Extracellular K+ (2.5-90 mM) increased a TRC resting conductance and enhanced both inward and outward whole-cell currents. 2. To isolate the inward current activated by external potassium (PA current), TRCs were dialysed with 110 mM NMGCl while extracellular NaCl was replaced with NMGCl. Under these conditions, the PA current displayed an S-shaped current-voltage (I-V) curve in the -100 to 100 mV range. Extracellular Rb+ and NH4+, but not Li+, Na+ or Cs+, evoked similar currents. 3. The PA current reversal potential (Vr) did not follow the equilibrium K+ potential under experimental conditions. Therefore, K+ ions were not the only current carriers. The influence of other ions on the PA current Vr indicated that the channels involved are permeable to K+ and H+ and much less so to Na+, Ca2+ and Mg2+. Relative permeabilities were estimated on the basis of the Goldman-Hodgkin-Katz equation as follows: PH:PK:PNa = 4000:1:0.04. 4. All I-V curves of the PA current were nearly linear at low negative potentials. The slope conductance at these voltages was used to characterize the dependence of the PA current on external K+ and H+. The slope conductance versus K+ concentration was fitted by the Hill equation. The data yielded a half-maximal concentration, K1/2 = 19 +/- 3 mM and a Hill coefficient, nH = 1.53 +/- 0.36 (means +/- S.E.M.). 5. The dependence of the mean PA current and the current variance on the K+ concentration indicated a rise in the open probability of the corresponding channels as extracellular K+ was increased. With 110 mM KCl in the bath, the single channel conductance was estimated at about 6 pS. Taken together, the data suggest that extracellular K+ may serve as a ligand to activate specific small-conductance cation channels (PA channels). The mean number of the PA channels

  20. Prediction of in vivo atenolol removal by high-permeability hemodialysis based on an in vitro model.

    Daheb, Kahina; Lecours, Jean-Philippe; Lipman, Mark L; Hildgen, Patrice; Roy, Julie J


    In order to update our data on drug dialyzability using the high-permeability dialysis membranes, atenolol elimination by an in vitro dialysis model was compared to that observed in six patients during high-permeability hemodialysis (HD), and the predictive value of the model was evaluated. Atenolol clearance was evaluated in six patients undergoing chronic HD. They were considered as eligible candidates if they were between 18 and 80 years of age, had a body mass index between 19 and 30 kg/m2, underwent HD and were taking atenolol on a regular basis in oral tablet form for at least 1 month before the study started. Atenolol clearance was also evaluated in three in vitro dialysis sessions with high-permeability polysulfone membrane. Atenolol was dissolved in 6 L of Krebs-Henseleit buffer with bovine serum albumin. Dialysis parameters were set to mirror as much as possible the patients' parameters (flow rate: 300 mL/min, dialyzate flow: 500 mL/min). After sample collection, drug concentrations were measured with high performance liquid chromatography. The comparison between in vivo and in vitro atenolol elimination kinetics was performed by drawing the curve fittings of concentrations vs. time on SigmaPlot 12, and adding a 95% prediction interval to each elimination curve fitting. Mean dialysis clearance of atenolol in vitro and in vivo was 198 ± 4 and 235 ± 53 mL/min, respectively. Atenolol was significantly removed within the study time period in both in vitro and in vivo experiments. By the end of in vitro dialysis, atenolol remaining in the drug reservoir was less than 2% of initial arterial concentration. Our study has indicated that atenolol is almost entirely cleared during high-permeability hemodialysis. Furthermore, the in vitro prediction interval of the drug elimination curve fitting could forecast its in vivo elimination especially at the end of dialysis.

  1. Adult human dental pulp stem cells promote blood-brain barrier permeability through vascular endothelial growth factor-a expression.

    Winderlich, Joshua N; Kremer, Karlea L; Koblar, Simon A


    Stem cell therapy is a promising new treatment option for stroke. Intravascular administration of stem cells is a valid approach as stem cells have been shown to transmigrate the blood-brain barrier. The mechanism that causes this effect has not yet been elucidated. We hypothesized that stem cells would mediate localized discontinuities in the blood-brain barrier, which would allow passage into the brain parenchyma. Here, we demonstrate that adult human dental pulp stem cells express a soluble factor that increases permeability across an in vitro model of the blood-brain barrier. This effect was shown to be the result of vascular endothelial growth factor-a. The effect could be amplified by exposing dental pulp stem cell to stromal-derived factor 1, which stimulates vascular endothelial growth factor-a expression. These findings support the use of dental pulp stem cell in therapy for stroke. © The Author(s) 2015.

  2. Design and synthesis of a cell-permeable, drug-like small molecule inhibitor targeting the polo-box domain of polo-like kinase 1.

    Ganipisetti Srinivasrao

    Full Text Available Polo-like kinase-1 (Plk1 plays a crucial role in cell proliferation and the inhibition of Plk1 has been considered as a potential target for specific inhibitory drugs in anti-cancer therapy. Several research groups have identified peptide-based inhibitors that target the polo-box domain (PBD of Plk1 and bind to the protein with high affinity in in vitro assays. However, inadequate proteolytic resistance and cell permeability of the peptides hinder the development of these peptide-based inhibitors into novel therapeutic compounds.In order to overcome the shortcomings of peptide-based inhibitors, we designed and synthesized small molecule inhibitors. Among these molecules, bg-34 exhibited a high binding affinity for Plk1-PBD and it could cross the cell membrane in its unmodified form. Furthermore, bg-34-dependent inhibition of Plk1-PBD was sufficient for inducing apoptosis in HeLa cells. Moreover, modeling studies performed on Plk1-PBD in complex with bg-34 revealed that bg-34 can interact effectively with Plk1-PBD.We demonstrated that the molecule bg-34 is a potential drug candidate that exhibits anti-Plk1-PBD activity and possesses the favorable characteristics of high cell permeability and stability. We also determined that bg-34 induced apoptotic cell death by inhibiting Plk1-PBD in HeLa cells at the same concentration as PEGylated 4j peptide, which can inhibit Plk1-PBD activity 1000 times more effectively than bg-34 can in in vitro assays. This study may help to design and develop drug-like small molecule as Plk1-PBD inhibitor for better therapeutic activity.

  3. Cell-permeable Tat-NBD peptide attenuates rat pancreatitis and acinus cell inflammation response

    You-Ming Long; Ken Chen; Xue-Jin Liu; Wen-Rui Xie; Hui Wang


    AIM: To investigate the effects of Tat-NEMO-binding domain (NBD) peptide on taurocholate-induced pancreatitis and lipopolysaccharide (LPS)-stimulated AR42J acinus cells inflammatory response in rats. METHODS: Sodium taurocholate (5%) was used to induce the pancreatitis model. Forty-eight rats from the taurocholate group aeceuved an intravenous bolus of 13 mg/kg Tat-NBD (wild-type, WT) peptide, Tat-NBD (mutant-type, MT) peptide, NBD peptide or Tat peptide. The pancreatic histopathology was analyzed by hematoxylin staining. LPS was added to the culture medium to stimulate the AR42J cells. For pretreatment, cells were incubated with different peptides for 2 h before LPS stimulation. Expression of IL-1β and TNF-α mRNA was analyzed using a semi-quantitative reverse-transcript polymerase chain reaction (RT-PCR) method. IL-1β and TNF-α protein in culture medium were detected by enzyme linked immunosorbent assay (ELISA). NF-κB DNA-binding in pancreas was examined by electrophoretic mobility shift assays. P65 expression of AR42J was determined by Strept Actividin-Biotin Complex (SABC) method. RESULTS: Pretreatment with Tat-NBD (WT) peptide at a concentration of 13 mg/kg body wt showed beneficial effect in pancreaitis model. LPS (10 mg/L) resulted in an increase of IL-1β mRNA, IL-1β protein, TNF-α mRNA and TNF-α protein, whereas significantly inhibitory effects were observed when cells were incubated with Tat-NBD (WT). Consisting with p65 expression decrease analyzed by SABC method, NF-κB DNA-binding activity significantly decreased in Tat-NBD (WT) pretreatment group, especially at the largest dose. No significant changes were found in the control peptide group. CONCLUSION: Our result supports that active NF-κB participates in the pathogenesis of STC-induced acute pancreatitis in rats. Tat-NBD (WT) peptide has antiinflammatory effects in this model and inhibits the inflammation of acinus simulated by LPS.

  4. High frequency permeability and permittivity spectra of BiFeO{sub 3}/(CoTi)-BaM ferrite composites

    Peng, Yun; Wu, Xiaohan; Li, Qifan; Yu, Ting; Feng, Zekun, E-mail: [School of Optical and Electronic Information, Huazhong University of Science and Technology, Wuhan 430074 (China); Chen, Zhongyan [School of Optical and Electronic Information, Huazhong University of Science and Technology, Wuhan 430074 (China); Jiangmen Magsource New Material CO., LTD., 529000 Guangdong (China); Su, Zhijuan; Chen, Yajie; Harris, Vincent G. [Center for Microwave Magnetic Materials and Integrated Circuits, and Department of Electrical and Computer Engineering, Northeastern University, Boston, Massachusetts 02115 (United States)


    Low magnetic loss ferrite composites consisting of Ba(CoTi){sub 1.2}Fe{sub 9.6}O{sub 19} and BiFeO{sub 3} (BFO) ferrite were investigated for permeability, permittivity, and high frequency losses at 10 MHz–1 GHz. The phase fraction of BiFeO{sub 3} was quantitatively analyzed by X-ray diffraction measurements. An effective medium approach was employed to predict the effective permeability and permittivity for the ferrite composites, which was found to be in good agreement with experimental data. The experiment demonstrated low magnetic losses (<0.128), modified by BFO phase fraction, while retaining high permeability (∼10.86) at 300 MHz. More importantly, the BFO phase resulted in a reduction of magnetic loss by 32%, as BFO phase increased from 2.7 vol. % to 12.6 vol. %. The effect of BFO phase on magnetic and dielectric properties revealed great potential for use in the miniaturization of high efficiency antennas.

  5. Notional Permeability

    Kik, R.; Van den Bos, J.P.; Maertens, J.; Verhagen, H.J.; Van der Meer, J.W.


    Different layer design of a rock slope and under layers has a large effect on the strengths on the rock slope itself. In the stability formula developed of VAN DER MEER [1988] this effect is represented by the term Notional Permeability with symbol P. A more open, or permeable, structure underneath

  6. Recombinant Treponema pallidum protein Tp0965 activates endothelial cells and increases the permeability of endothelial cell monolayer.

    Rui-Li Zhang

    Full Text Available The recombinant Treponema pallidum protein Tp0965 (rTp0965, one of the many proteins derived from the genome of T. pallidum subsp. pallidum, shows strong immunogenicity and immunoreactivity. In this study, we investigated the effects of rTp0965 on the endothelial barrier. Treatment of human umbilical vein endothelial cells (HUVECs with rTp0965 resulted in increased levels of ICAM-1, E-selectin, and MCP-1 mRNA and protein expression. These increases contributed to the adhesion and chemataxis of monocytes (THP-1 cells to HUVECs preincubated with rTp0965. In addition, rTp0965 induced reorganization of F-actin and decreased expression of claudin-1 in HUVECs. Interestingly, inhibition of the RhoA/ROCK signal pathway protected against rTp0965-induced higher endothelial permeability as well as transendothelial migration of monocytes. These data indicate that Tp0965 protein may play an important role in the immunopathogenesis of syphilis.

  7. 高浓度胆汁改变肠上皮细胞氯通道蛋白-2和通透性%High concentration bile changes the chloride channel protein-2 and permeability of intestinal epithelium cell line

    陈振勇; 张劲; 杨鹏; 黄文广; 冯贤松


    Objective To investigate the effects of bile on chloride channel protein-2 (CLC-2)and permeability of intestinal epithelium cell.Methods Rat intestinal epithelium cell line IEC-6 was cultured.They were exposed to 5.0%,1.0%,0.1% rat bile and chloride channel agonist Lubiprostone.After 20 h cultured,transepithelial electrical resistance (TER) of the monostratal cells was measured.The change of CLC-2 and zonula occludens-1 (ZO-1) were examined by Western blotting,and the images were analysed quantitatively.Results The TEER of 5% group was most lower.The average relative gradations of Western blotting images in 5.0% group and 1.0% were lower obviously than those in control group (0.30 ± 0.05,0.37 ± 0.08 vs.0.56 ± 0.08) ( all P < 0.05 ).The relative gradations of ZO-1 were decreaseded only in 5.0% group.After Lubiprostone added,the TER in 5.0% group was upgraded (451.3 ± 60.5 )Ω·cm2,and the average relative gradations of ZO-1 (0.32 ± 0.04).Conclusion High concentration of bile destroied enterocyte chloride channel and tight junction protein,increase epithelium permeabihty.%目的 观察胆汁对肠上皮细胞氯离子通道和通透性的影响.方法 鼠肠上皮细胞株IEC-6分别与5.0%、1.0%、0.1%胆汁及氯通道激动剂接触.20h后检测跨膜电阻,Western blot分析氯离子通道蛋白-2(CLC-2)和紧密连接闭锁小带-1(ZO-1)表达的变化及各条带的相对灰度值.结果 5.0%浓度组降低跨膜电阻作用最强.5.0%和1.0%组的CLC-2蛋白相对灰度值(0.30±0.05和0.37±0.08)低于对照组(P均<0.05).5.0%组的ZO-1相对表达量下降.添加氯通道激动剂后,5.0%浓度组的跨膜电阻(451.3±60.5)Ω.cm2及ZO-1相对表达量(0.32±0.04)较对照组上升明显(P均<0.05).结论 高浓度胆汁破坏肠上皮细胞氯离子通道和紧密连接蛋白,增加上皮细胞通透性.

  8. Influence of zinc concentration on structure, complex permittivity and permeability of Ni–Zn ferrites at high frequency

    Jiang, Nan-Nan; Yang, Yang; Zhang, Yu-Xiang [School of Physics and Information Technology, Shaanxi Normal University, Xi' an 710119 (China); Zhou, Jian-Ping, E-mail: [School of Physics and Information Technology, Shaanxi Normal University, Xi' an 710119 (China); Liu, Peng [School of Physics and Information Technology, Shaanxi Normal University, Xi' an 710119 (China); Deng, Chao-Yong [Department of Electronic Science, Key Laboratory of Functional Composite Materials of Guizhou Province, Guizhou University, Guiyang 550025 (China)


    Polycrystalline soft magnetic nickel–zinc ferrites with chemical composition Ni{sub 1−x}Zn{sub x}Fe{sub 2}O{sub 4}, where x=0, 0.2, 0.4, 0.6, 0.65, 0.7, 0.75, and 0.8, were prepared by solid state reaction method. We researched the effect of zinc concentration on the lattice parameter, crystal morphology and electromagnetic properties at high frequency. Results show that ε′ and ε″ decline with increasing frequency until they reach almost constants over 3 MHz to 1 GHz. The dielectric constant achieves a maximum when the Zn concentration is 0.8. The value of ε′ slightly declines with increasing frequency in the range of 2–18 GHz. The spectra of the permeability displays a relaxation resonance for the ferrites with x=0, 0.2, and 0.4 in 3 MHz to 1 GHz frequency range. The permeability is ruled by Snoek's law, which results in the values of μ′ decreased fast below 2 GHz and smaller than 1 above 2 GHz. The value of μ′ reaches maximum and μ″ shows minimum for the samples around x=0.75 in 2–18 GHz range. The magnetic permeability μ′ decreases in an external magnetic field, and shows two resonance peaks corresponding to domain wall and spin rotation resonance. The resonance peaks shift to higher frequency with increasing the external magnetic field. But the permeability has no clear response for magnetic field when zinc concentration is much higher. - Graphical abstract: (a) Resonance frequencies for the domain wall and spin components as a function of external magnetic field and (b) illustration of magnetization process. - Highlights: • A detailed study on permittivity and permeability of Ni–Zn ferrites in 3 MHz to 18 GHz. • Magnetic field inspires two resonance peaks in permeability. • The resonance peaks shift to higher frequency with the magnetic field.

  9. Cardiotoxic drugs Herceptin and doxorubicin inhibit cardiac microvascular endothelial cell barrier formation resulting in increased drug permeability

    Emma L. Wilkinson


    Full Text Available Cardiotoxicity induced by anti-cancer therapeutics is a severe, and potentially fatal, adverse reaction of the heart in response to certain drugs. Current in vitro approaches to assess cardiotoxicity have focused on analysing cardiomyocytes. More recently it has become apparent that non-cardiomyocyte cells of the heart can potentially contribute to cardiotoxicity. Herceptin and doxorubicin are known to induce cardiotoxicity in the clinic. The effect of these drugs on the endothelial tight junction barrier was tested by analysing tight junction formation and zona occludens-1 (ZO-1 levels, revealing that Herceptin and doxorubicin are able to induce barrier perturbment and decrease barrier function in human cardiac microvascular endothelial cells (HCMECs leading to increased permeability. Herceptin treatment had no effect on the tight junction barrier function in human dermal and human brain microvascular endothelial cells. HCMECs showed detectable levels of HER2 compared with the other endothelial cells suggesting that Herceptin binding to HER2 in these cells may interfere with tight junction formation. Our data suggests that doxorubicin and Herceptin can affect tight junction formation in the cardiac microvasculature leading to increased drug permeability and adverse effects on the cardiac myocytes.

  10. Adjusting the Ion Permeability of Polyelectrolyte Multilayers through Layer-by-Layer Assembly under a High Gravity Field.

    Jiang, Chao; Luo, Caijun; Liu, Xiaolin; Shao, Lei; Dong, Youqing; Zhang, Yingwei; Shi, Feng


    The layer-by-layer (LbL) assembled multilayer has been widely used as good barrier film or capsule due to the advantages of its flexible tailoring of film permeability and compactness. Although many specific systems have been proposed for film design, developing a versatile strategy to control film compactness remains a challenge. We introduced the simple mechanical energy of a high gravity field to the LbL assembly process to tailor the multilayer permeability through adjusting film compactness. By taking poly(diallyldimethylammonium chloride) (PDDA) and poly{1-4[4-(3-carboxy-4-hydroxyphenylazo)benzenesulfonamido]-1,2-ethanediyl sodium salt} (PAzo) as a model system, we investigated the LbL assembly process under a high gravity field. The results showed that the high gravity field introduced effectively accelerated the multilayer deposition process by 20-fold compared with conventional dipping assembly; the adsorption rate was positively dependent on the rotating speed of the high gravity equipment and the concentration of the building block solutions. More interestingly, the film compactness of the PDDA/PAzo multilayer prepared under the high gravity field increased remarkably with the growing rotational speed of the high gravity equipment, as demonstrated through comparisons of surface morphology, cyclic voltammetry curves, and photoisomerization kinetics of PDDA/PAzo multilayers fabricated through the conventional dipping method and through LbL assembly under a high gravity field, respectively. In this way, we have introduced a simple and versatile external form of mechanical energy into the LbL assembling process to improve film compactness, which should be useful for further applications in controlled ion permeability, anticorrosion, and drug loading.

  11. Retinoic acid and hydrocortisone strengthen the barrier function of human RPMI 2650 cells, a model for nasal epithelial permeability.

    Kürti, Levente; Veszelka, Szilvia; Bocsik, Alexandra; Ozsvári, Béla; Puskás, László G; Kittel, Agnes; Szabó-Révész, Piroska; Deli, Mária A


    The nasal pathway represents an alternative route for non-invasive systemic administration of drugs. The main advantages of nasal drug delivery are the rapid onset of action, the avoidance of the first-pass metabolism in the liver and the easy applicability. In vitro cell culture systems offer an opportunity to model biological barriers. Our aim was to develop and characterize an in vitro model based on confluent layers of the human RPMI 2650 cell line. Retinoic acid, hydrocortisone and cyclic adenosine monophosphate, which influence cell attachment, growth and differentiation have been investigated on the barrier formation and function of the nasal epithelial cell layers. Real-time cell microelectronic sensing, a novel label-free technique was used for dynamic monitoring of cell growth and barrier properties of RPMI 2650 cells. Treatments enhanced the formation of adherens and tight intercellular junctions visualized by electron microscopy, the presence and localization of junctional proteins ZO-1 and β-catenin demonstrated by fluorescent immunohistochemistry, and the barrier function of nasal epithelial cell layers. The transepithelial resistance of the RPMI 2650 cell model reached 50 to 200 Ω × cm(2), the permeability coefficient for 4.4 kDa FITC-dextran was 9.3 to 17 × 10(-6) cm/s, in agreement with values measured on nasal mucosa from in vivo and ex vivo experiments. Based on these results human RPMI 2650 cells seem to be a suitable nasal epithelial model to test different pharmaceutical excipients and various novel formulations, such as nanoparticles for toxicity and permeability.

  12. On Blastocystis secreted cysteine proteases: a legumain-activated cathepsin B increases paracellular permeability of intestinal Caco-2 cell monolayers.

    Nourrisson, C; Wawrzyniak, I; Cian, A; Livrelli, V; Viscogliosi, E; Delbac, F; Poirier, P


    Blastocystis spp. pathogenic potential remains unclear as these anaerobic parasitic protozoa are frequently isolated from stools of both symptomatic and asymptomatic subjects. In silico analysis of the whole genome sequence of Blastocystis subtype 7 revealed the presence of numerous proteolytic enzymes including cysteine proteases predicted to be secreted. To assess the potential impact of proteases on intestinal cells and gut function, we focused our study on two cysteine proteases, a legumain and a cathepsin B, which were previously identified in Blastocystis subtype 7 culture supernatants. Both cysteine proteases were produced as active recombinant proteins. Activation of the recombinant legumain was shown to be autocatalytic and triggered by acidic pH, whereas proteolytic activity of the recombinant cathepsin B was only recorded after co-incubation with the legumain. We then measured the diffusion of 4-kDa FITC-labelled dextran across Caco-2 cell monolayers following exposition to either Blastocystis culture supernatants or each recombinant protease. Both Blastocystis culture supernatants and recombinant activated cathepsin B induced an increase of Caco-2 cell monolayer permeability, and this effect was significantly inhibited by E-64, a specific cysteine protease inhibitor. Our results suggest that cathepsin B might play a role in pathogenesis of Blastocystis by increasing intestinal cell permeability.

  13. Morphine induces expression of platelet-derived growth factor in human brain microvascular endothelial cells: implication for vascular permeability.

    Hongxiu Wen

    Full Text Available Despite the advent of antiretroviral therapy, complications of HIV-1 infection with concurrent drug abuse are an emerging problem. Morphine, often abused by HIV-infected patients, is known to accelerate neuroinflammation associated with HIV-1 infection. Detailed molecular mechanisms of morphine action however, remain poorly understood. Platelet-derived growth factor (PDGF has been implicated in a number of pathological conditions, primarily due to its potent mitogenic and permeability effects. Whether morphine exposure results in enhanced vascular permeability in brain endothelial cells, likely via induction of PDGF, remains to be established. In the present study, we demonstrated morphine-mediated induction of PDGF-BB in human brain microvascular endothelial cells, an effect that was abrogated by the opioid receptor antagonist-naltrexone. Pharmacological blockade (cell signaling and loss-of-function (Egr-1 approaches demonstrated the role of mitogen-activated protein kinases (MAPKs, PI3K/Akt and the downstream transcription factor Egr-1 respectively, in morphine-mediated induction of PDGF-BB. Functional significance of increased PDGF-BB manifested as increased breach of the endothelial barrier as evidenced by decreased expression of the tight junction protein ZO-1 in an in vitro model system. Understanding the regulation of PDGF expression may provide insights into the development of potential therapeutic targets for intervention of morphine-mediated neuroinflammation.

  14. Glucose and calcium ions may modulate the efficiency of bovine β-casomorphin-7 permeability through a monolayer of Caco-2 cells.

    Jarmołowska, Beata; Teodorowicz, Małgorzata; Fiedorowicz, Ewa; Sienkiewicz-Szłapka, Edyta; Matysiewicz, Michał; Kostyra, Elżbieta


    Milk and dairy products provide a lot of valuable nutritive elements. They are also sources of biologically active peptides, including β-casomorphins that manifest the properties of morphine. An activity of DPPIV seems to be most crucial factor decreasing the efficiency of the β-casomorphin-7 (BCM7) transport. The increase of BCM7 concentration in blood may intensify symptoms of apparent life threatening events (ALTE), autism, schizophrenia, and allergy. This study aimed at identifying the influence of several selected substances on a transport efficiency of bovine BCM7 through an intestinal monolayer in a Caco-2 cell model system. Applying the ELISA method, the permeability coefficient of BCM7 through the Caco-2 monolayer was calculated. TEER values were used to evaluate the integrity of Caco-2 cell monolayers. An increase of glucose and Ca(2+) concentrations in the culture medium was accompanied by an increase of the BCM7 transport efficiency. The lowest permeability coefficients of BCM7 were observed for the membranes with high electrical resistances. The transport was enhanced in the presence of milk infant formulas, whereas no changes were observed when using μ-opioid receptor antagonist (casoxin-6). The results may be useful in understanding the pathogenesis of inflammation and food allergy in infants.

  15. Polymer coated liposomes for use in the oral cavity - A study of the in vitro toxicity, effect on cell permeability and interaction with mucin

    Klemetsrud, Therese; Kjøniksen, Anna-Lena; Hiorth, Marianne


    (NIPAAM-co-MAA)), hydrophobically modified hydroxyethyl cellulose (HM-HEC), and hydrophobically modified ethyl hydroxyethyl cellulose (HM-EHEC). With chitosan as an exception, all the systems exhibited no significant effect on cell viability and permeability at the considered concentrations. Additionally, all the formulations...... formulations promising for oromucosal administration. Although the chitosan coated liposomes affected the cell viability, this formulation also influenced the cell permeability, which makes it an interesting candidate for systemic drug delivery from the oral cavity.......In this study we investigated the in vitro toxicity, impact on cell permeability and mucoadhesive potential of polymer coated liposomes intended for use in the oral cavity. A TR146 cell line was used as a model. The overall aim was to end up with a selection of safe polymer coated liposomes...

  16. Homogenization of two-phase flow: high contrast of phase permeability; Homogeneisation d'ecoulement diphasique: grand contraste de permeabilite d'une phase

    Panasenko, G.P. [Saint-Etienne Universite, Equipe d' Analyse Numerique, UPRES EA 3058, 42 (France); Universite Pierre et Marie Curie, Lab. de Modelisation en Numerique, CNRS UMR 7607, 75 - Paris (France); Virnovsky, G. [R.F. - Rogaland Research, Stavanger (Norway)


    The steady-state two-phase flow non-linear equation is considered in the case when one of phases has low effective permeability in some periodic set, while on the complementary set it is high; the second phase has no contrast of permeabilities in different zones. A homogenization procedure gives the homogenized model with macroscopic effective permeability of the second phase depending on the gradient and on the second order derivatives of the macroscopic pressure of the first phase. This effect cannot be obtained by classical (one small parameter) homogenization. (author)

  17. Cell permeability, sodium transport, and the hypertensive process in the rat.

    Friedman, S M; Friedman, C L


    The tail artery of the spontaneously hypertensive rat (SHR) (Carworth Farms), excised rapidly and immersed immediately in cold (2 degrees C) Li-substituted physiologic salt solution (LiPSS), continues to exchange cell Na+ and K+ for Li+, this exchange is negligible to the control (Carworth Farms normotensive) CFN). In the incubated artery at 37 degrees C, when the vascular smooth muscle cell is slack, the leakiness of the cell membrane in the SHR is more than offset by increased Na+ pumping activity, so that cell Na+ is subnormal. A high precision technique with ion-specific electrodes was developed to follow the passive downhill and active uphill phases of Na+-K+ exchange in the perfused artery exposed to K+-free physiologic salt solution (K+-free PSS) followed by physiological salt solution (PSS). The exchange was found to be fully reversible and sufficiently equimolar to be definable in terms of movements of K+ alone. The rates of ionic movement across the vascular smooth muscle cell were found to be about 6 times faster for the vessel perfused at low pressure (less than 3 mm Hg) than for the slack incubated artery. The rate of passive downhill movement was significantly accelerated in the mature SHR compared with CFN, and the net active transport activity much enhanced. Similar changes were seen as early as 3 weeks after treatment with DOCA and were pronounced at 8 weeks. It is proposed that conditions favoring a sustained accumulation of Na+ in the vascular smooth muscle cell are countered by an enhanced synthesis of transport protein, of contractile protein, and of paracellular matrix protein which progressively restructure the wall.

  18. The permeability of red blood cells to chloride, urea and water

    Brahm, Jesper


    -1000 mmol l(-1), Michaelis-Menten-like kinetics; K½=127, 173 and 345 mmol l(-1)), and values at [urea]=1 mmol l(-1) are 29.5×10(-6), 467×10(-6) and 260×10(-6) cm s(-1), respectively. Diffusional water permeability, Pd, was 0.84×10(-3) (chick), 5.95×10(-3) (duck), 0.39×10(-3) (Amphiuma), 3.13×10(-3) (dog...

  19. A modified parallel artificial membrane permeability assay for evaluating the bioconcentration of highly hydrophobic chemicals in fish.

    Kwon, Jung-Hwan; Escher, Beate I


    Low cost in vitro tools are needed at the screening stage of assessment of bioaccumulation potential of new and existing chemicals because the number of chemical substances that needs to be tested highly exceeds the capacity of in vivo bioconcentration tests. Thus, the parallel artificial membrane permeability assay (PAMPA) system was modified to predict passive uptake/ elimination rate in fish. To overcome the difficulties associated with low aqueous solubility and high membrane affinity of highly hydrophobic chemicals, we measured the rate of permeation from the donor poly(dimethylsiloxane)(PDMS) disk to the acceptor PDMS disk through aqueous and PDMS membrane boundary layers and term the modified PAMPA system "PDMS-PAMPA". Twenty chemicals were selected for validation of PDMS-PAMPA. The measured permeability is proportional to the passive elimination rate constant in fish and was used to predict the "minimum" in vivo elimination rate constant. The in vivo data were very close to predicted values except for a few polar chemicals and metabolically active chemicals, such as pyrene and benzo[a]pyrene. Thus, PDMS-PAMPA can be an appropriate in vitro system for nonmetabolizable chemicals. Combination with metabolic clearance rates using a battery of metabolic degradation assays would enhance the applicability for metabolizable chemicals.

  20. Impulse noise transiently increased the permeability of nerve and glial cell membranes, an effect accentuated by a recent brain injury.

    Säljö, Annette; Huang, Ying-Lai; Hansson, Hans-Arne


    A single exposure to intense impulse noise may cause diffuse brain injury, revealed by increased expression of immediate early gene products, transiently altered distribution of neurofilaments, accumulation of beta-amyloid precursor protein, apoptosis, and gliosis. Neither hemorrage nor any gross structural damage are seen. The present study focused on whether impulse noise exposure increased the permeability of nerve and glial cell membranes to proteins. Also, we investigated whether a preceding, minor focal surgical brain lesion accentuated the leakage of cytosolic proteins. Anaesthetized rats were exposed to a single impulse noise at either 199 or 202 dB for 2 milliseconds. Transiently elevated levels of the cellular protein neuron specific enolase (NSE) and the glial cytoplasmic protein S-100 were recorded in the cerebrospinal fluid (CSF) during the first hours after the exposure to 202 dB. A surgical brain injury, induced the day before the exposure to the impulse noise, was associated with significantly increased concentrations of both markers in the CSF. It is concluded that intense impulse noise damages both nerve and glial cells, an effect aggravated by a preexisting surgical lesion. The impulse of the shock wave, i.e. the pressure integrated over time, is likely to be the injurious mechanism. The abnormal membrane permeability and the associated cytoskeletal changes may initiate events, which eventually result in a progressive diffuse brain injury.

  1. Notional Permeability

    Kik, R.; Van den Bos, J.P.; Maertens, J.; Verhagen, H.J.; van der Meer, J W


    Different layer design of a rock slope and under layers has a large effect on the strengths on the rock slope itself. In the stability formula developed of VAN DER MEER [1988] this effect is represented by the term Notional Permeability with symbol P. A more open, or permeable, structure underneath the armour layer has the ability to dissipate more wave energy and therefore requires less weight of the armour layer. The influence of this parameter is thus very important in economic sense. Up u...

  2. Permeability of edible coatings.

    Mishra, B; Khatkar, B S; Garg, M K; Wilson, L A


    The permeabilities of water vapour, O2 and CO2 were determined for 18 coating formulations. Water vapour transmission rate ranged from 98.8 g/m(2).day (6% beeswax) to 758.0 g/m(2).day (1.5% carboxymethyl cellulose with glycerol). O2 permeability at 14 ± 1°C and 55 ± 5% RH ranged from 1.50 to 7.95 cm(3)cm cm(-2)s(-1)Pa(-1), with CO2 permeability 2 to 6 times as high. Permeability to noncondensable gases (O2 and CO2) was higher for hydrophobic (peanut oil followed by beeswax) coatings as compared to hydrophilic (whey protein concentrate and carboxymethyl cellulose).

  3. Permeability of edible coatings

    B Mishra; Khatkar, B. S.; Garg, M. K.; Wilson, L.A.


    The permeabilities of water vapour, O2 and CO2 were determined for 18 coating formulations. Water vapour transmission rate ranged from 98.8 g/ (6% beeswax) to 758.0 g/ (1.5% carboxymethyl cellulose with glycerol). O2 permeability at 14 ± 1°C and 55 ± 5% RH ranged from 1.50 to 7.95 cm3cm cm−2s−1Pa−1, with CO2 permeability 2 to 6 times as high. Permeability to noncondensable gases (O2 and CO2) was higher for hydrophobic (peanut oil followed by beeswax) coatings as compared to hydrop...

  4. Prediction of oral absorption of cinnarizine--a highly supersaturating poorly soluble weak base with borderline permeability.

    Berlin, Mark; Przyklenk, Karl-Heinz; Richtberg, Annette; Baumann, Wolfgang; Dressman, Jennifer B


    Two important driving forces for oral absorption of active pharmaceutical ingredients are drug dissolution and permeability in the gastrointestinal tract. Poorly soluble weak bases typically exhibit high solubility under fasted gastric conditions. However, the solubility of such drugs usually decreases drastically in the fasted small intestine, constraining drug absorption. Since there is a discrepancy in solubility between the fasted state stomach and intestine, it is crucial to examine the influence of dissolution, supersaturation and precipitation on the oral absorption of poorly soluble weak bases during and after fasted state gastric emptying. Cinnarizine is a poorly soluble weak base with borderline permeability, exhibiting supersaturation and precipitation under simulated fasted state gastric emptying conditions. Interestingly, supersaturation and precipitation of cinnarizine under fed state conditions is not expected to occur, since the drug shows good solubility in fed state biorelevant media and exhibits a positive food effect in pharmacokinetic studies. The present work is aimed at investigating the dissolution, supersaturation and precipitation behavior of marketed cinnarizine tablets under fasted and fed state conditions using biorelevant dissolution and transfer methods. In order to predict the in vivo performance of these cinnarizine formulations, the in vitro results were then coupled with different physiologically based pharmacokinetic (PBPK) models, which considered either only dissolution or a combination of dissolution, supersaturation and precipitation kinetics. The results of the in silico predictions were then compared with in vivo observations. The study revealed that under fasting conditions, plasma profiles could be accurately predicted only when supersaturation and precipitation as well as dissolution were taken into account. It was concluded that for poorly soluble weak bases with moderate permeability, supersaturation and precipitation

  5. High strength, flexible and transparent nanofibrillated cellulose-nanoclay biohybrid films with tunable oxygen and water vapor permeability.

    Aulin, Christian; Salazar-Alvarez, German; Lindström, Tom


    A novel, technically and economically benign procedure to combine vermiculite nanoplatelets with nanocellulose fibre dispersions into functional biohybrid films is presented. Nanocellulose fibres of 20 nm diameters and several micrometers in length are mixed with high aspect ratio exfoliated vermiculite nanoplatelets through high-pressure homogenization. The resulting hybrid films obtained after solvent evaporation are stiff (tensile modulus of 17.3 GPa), strong (strength up to 257 MPa), and transparent. Scanning electron microscopy (SEM) shows that the hybrid films consist of stratified nacre-like layers with a homogenous distribution of nanoplatelets within the nanocellulose matrix. The oxygen barrier properties of the biohybrid films outperform commercial packaging materials and pure nanocellulose films showing an oxygen permeability of 0.07 cm(3) μm m(-2) d(-1) kPa(-1) at 50% relative humidity. The oxygen permeability of the hybrid films can be tuned by adjusting the composition of the films. Furthermore, the water vapor barrier properties of the biohybrid films were also significantly improved by the addition of nanoclay. The unique combination of excellent oxygen barrier behavior and optical transparency suggests the potential of these biohybrid materials as an alternative in flexible packaging of oxygen sensitive devices such as thin-film transistors or organic light-emitting diode displays, gas storage applications and as barrier coatings/laminations in large volume packaging applications.

  6. Cell-based in vitro blood-brain barrier model can rapidly evaluate nanoparticles' brain permeability in association with particle size and surface modification.

    Hanada, Sanshiro; Fujioka, Kouki; Inoue, Yuriko; Kanaya, Fumihide; Manome, Yoshinobu; Yamamoto, Kenji


    The possibility of nanoparticle (NP) uptake to the human central nervous system is a major concern. Recent reports showed that in animal models, nanoparticles (NPs) passed through the blood-brain barrier (BBB). For the safe use of NPs, it is imperative to evaluate the permeability of NPs through the BBB. Here we used a commercially available in vitro BBB model to evaluate the permeability of NPs for a rapid, easy and reproducible assay. The model is reconstructed by culturing both primary rat brain endothelial cells and pericytes to support the tight junctions of endothelial cells. We used the permeability coefficient (P(app)) to determine the permeability of NPs. The size dependency results, using fluorescent silica NPs (30, 100, and 400 nm), revealed that the Papp for the 30 nm NPs was higher than those of the larger silica. The surface charge dependency results using Qdots® (amino-, carboxyl-, and PEGylated-Qdots), showed that more amino-Qdots passed through the model than the other Qdots. Usage of serum-containing buffer in the model resulted in an overall reduction of permeability. In conclusion, although additional developments are desired to elucidate the NPs transportation, we showed that the BBB model could be useful as a tool to test the permeability of nanoparticles.

  7. Cell-Based in Vitro Blood–Brain Barrier Model Can Rapidly Evaluate Nanoparticles’ Brain Permeability in Association with Particle Size and Surface Modification

    Sanshiro Hanada


    Full Text Available The possibility of nanoparticle (NP uptake to the human central nervous system is a major concern. Recent reports showed that in animal models, nanoparticles (NPs passed through the blood–brain barrier (BBB. For the safe use of NPs, it is imperative to evaluate the permeability of NPs through the BBB. Here we used a commercially available in vitro BBB model to evaluate the permeability of NPs for a rapid, easy and reproducible assay. The model is reconstructed by culturing both primary rat brain endothelial cells and pericytes to support the tight junctions of endothelial cells. We used the permeability coefficient (Papp to determine the permeability of NPs. The size dependency results, using fluorescent silica NPs (30, 100, and 400 nm, revealed that the Papp for the 30 nm NPs was higher than those of the larger silica. The surface charge dependency results using Qdots® (amino-, carboxyl-, and PEGylated-Qdots, showed that more amino-Qdots passed through the model than the other Qdots. Usage of serum-containing buffer in the model resulted in an overall reduction of permeability. In conclusion, although additional developments are desired to elucidate the NPs transportation, we showed that the BBB model could be useful as a tool to test the permeability of nanoparticles.

  8. Apomorphine and its esters: Differences in Caco-2 cell permeability and chylomicron affinity.

    Borkar, Nrupa; Chen, Zhizhong; Saaby, Lasse; Müllertz, Anette; Håkansson, Anders E; Schönbeck, Christian; Yang, Mingshi; Holm, René; Mu, Huiling


    Oral delivery of apomorphine via prodrug principle may be a potential treatment for Parkinson's disease. The purpose of this study was to investigate the transport and stability of apomorphine and its esters across Caco-2 cell monolayer and their affinity towards chylomicrons. Apomorphine, monolauroyl apomorphine (MLA) and dilauroyl apomorphine (DLA) were subjected to apical to basolateral (A-B) and basolateral to apical (B-A) transport across Caco-2 cell monolayer. The stability of these compounds was also assessed by incubation at intestinal pH and physiological pH with and without Caco-2 cells. Molecular dynamics (MD) simulations were performed to understand the stability of the esters on a molecular level. The affinity of the compounds towards plasma derived chylomicrons was assessed. The A-B transport of intact DLA was about 150 times lower than the transport of apomorphine. In contrast, MLA was highly unstable in the aqueous media leading to apomorphine appearance basolaterally. MD simulations possibly explained the differences in hydrolysis susceptibilities of DLA and MLA. The affinity of apomorphine diesters towards plasma derived chylomicrons provided an understanding of their potential lymphatic transport. The intact DLA transport is not favorable; therefore, the conversion of DLA to MLA is an important step for intestinal apomorphine absorption.

  9. Hyperthermia Differently Affects Connexin43 Expression and Gap Junction Permeability in Skeletal Myoblasts and HeLa Cells

    Ieva Antanavičiūtė


    Full Text Available Stress kinases can be activated by hyperthermia and modify the expression level and properties of membranous and intercellular channels. We examined the role of c-Jun NH2-terminal kinase (JNK in hyperthermia-induced changes of connexin43 (Cx43 expression and permeability of Cx43 gap junctions (GJs in the rabbit skeletal myoblasts (SkMs and Cx43-EGFP transfected HeLa cells. Hyperthermia (42°C for 6 h enhanced the activity of JNK and its target, the transcription factor c-Jun, in both SkMs and HeLa cells. In SkMs, hyperthermia caused a 3.2-fold increase in the total Cx43 protein level and enhanced the efficacy of GJ intercellular communication (GJIC. In striking contrast, hyperthermia reduced the total amount of Cx43 protein, the number of Cx43 channels in GJ plaques, the density of hemichannels in the cell membranes, and the efficiency of GJIC in HeLa cells. Both in SkMs and HeLa cells, these changes could be prevented by XG-102, a JNK inhibitor. In HeLa cells, the changes in Cx43 expression and GJIC under hyperthermic conditions were accompanied by JNK-dependent disorganization of actin cytoskeleton stress fibers while in SkMs, the actin cytoskeleton remained intact. These findings provide an attractive model to identify the regulatory players within signalosomes, which determine the cell-dependent outcomes of hyperthermia.

  10. Determination of the temperature-dependent cell membrane permeabilities using microfluidics with integrated flow and temperature control.

    Fang, Cifeng; Ji, Fujun; Shu, Zhiquan; Gao, Dayong


    We developed an integrated microfluidic platform for instantaneous flow and localized temperature control. The platform consisted of a flow-focusing region for sample delivery and a cross-junction region embedded with a microheater for cell trapping and localized temperature control by using an active feedback control system. We further used it to measure the membrane transport properties of Jurkat cells, including the osmotically inactive cell volume (Vb) and cell membrane permeabilities to water (Lp) and to cryoprotective agent (CPA) solutions (dimethyl sulfoxide (DMSO) in this study) (PS) at various temperatures (room temperature, 30 °C, and 37 °C). Such characteristics of cells are of great importance in many applications, especially in optimal cryopreservation. With the results, the corresponding activation energy for water and CPA transport was calculated. The comparison of the results from the current study with reference data indicates that the developed platform is a reliable tool for temperature-dependent cell behavior study, which provides valuable tools for general cell manipulation applications with precise temperature control.

  11. Hyperthermia differently affects connexin43 expression and gap junction permeability in skeletal myoblasts and HeLa cells.

    Antanavičiūtė, Ieva; Mildažienė, Vida; Stankevičius, Edgaras; Herdegen, Thomas; Skeberdis, Vytenis Arvydas


    Stress kinases can be activated by hyperthermia and modify the expression level and properties of membranous and intercellular channels. We examined the role of c-Jun NH2-terminal kinase (JNK) in hyperthermia-induced changes of connexin43 (Cx43) expression and permeability of Cx43 gap junctions (GJs) in the rabbit skeletal myoblasts (SkMs) and Cx43-EGFP transfected HeLa cells. Hyperthermia (42°C for 6 h) enhanced the activity of JNK and its target, the transcription factor c-Jun, in both SkMs and HeLa cells. In SkMs, hyperthermia caused a 3.2-fold increase in the total Cx43 protein level and enhanced the efficacy of GJ intercellular communication (GJIC). In striking contrast, hyperthermia reduced the total amount of Cx43 protein, the number of Cx43 channels in GJ plaques, the density of hemichannels in the cell membranes, and the efficiency of GJIC in HeLa cells. Both in SkMs and HeLa cells, these changes could be prevented by XG-102, a JNK inhibitor. In HeLa cells, the changes in Cx43 expression and GJIC under hyperthermic conditions were accompanied by JNK-dependent disorganization of actin cytoskeleton stress fibers while in SkMs, the actin cytoskeleton remained intact. These findings provide an attractive model to identify the regulatory players within signalosomes, which determine the cell-dependent outcomes of hyperthermia.

  12. High Red Blood Cell Count

    Symptoms High red blood cell count By Mayo Clinic Staff A high red blood cell count is an increase in oxygen-carrying cells in your bloodstream. Red blood cells transport oxygen from your lungs to tissues throughout ...

  13. Cytotoxic Effect and Permeability Activities of Curcumin Analogue; 2, 6-Bis (2, 5-dimethoxybenzy-lidene cyclohexanone (BDMC33 in Caco-2 Cell Model

    N. Yakubu


    Full Text Available Previously, curcumin analogue, 2, 6-bis (2, 5-dimethoxybenzylidene cyclohexanone (BDMC33 with high anti-inflammatory activity was chemically synthesized in our laboratory to enhance the biological activity of curcumin. In this study, the toxicity and permeability activities of 2,6-bis(2,5-dimethoxybenzy-lidenecyclohexanone (BDMC33 in Caco-2 cells was investigated. Toxicity effects using MTT assay and apparent permeability coefficient (Papp, uptake (UR and efflux (ER ratios, and mass balance of BDMC33 after permeation in Caco-2 cells for 180 min were evaluated in apical (A to basolateral (B and basolateral (B to apical (A directions. The similar analyses on 3-(2-fluoro-benzylidene-5-(2-fluorocyclohexylmethylene-piperidin-4-one; (EF-24 (check control were also conducted. The 24 hr LC50 value for BDMC33 and EF-24 on Caco-2 cells were both 50 µM. The Papp value in A→B direction was 3.37 ± 0.47 cm/s (BDMC33 and 2.47 ± 0.15 cm/s (EF-24. Whereas in B→A direction, it was 1.9 ± 0.36 cm/s (BDMC33 and 1.8 ± 0.15 cm/s (EF-24 upon 120 min incubation. The UR and ER ratios calculated were 1.77% and 0.56%, respectively, and the mass balance calculated were 41-44% (BDMC33 and 31-34% (EF-24 in A→B and B→A direction. This study has suggested BDMC33 to be more absorbable than EF-24 in Caco-2 cells. Therefore, BDMC33 could be a leading feature, the anti-inflammatory agent, as it biological activities would be expected outside the intestine.

  14. Synaptic contribution of Ca2+-permeable and Ca2+-impermeable AMPA receptors on isolated carp retinal horizontal cells and their modulation by Zn2+.

    Sun, Yan; Jiang, Xiao-Dong; Liu, Xue; Gong, Hai-Qing; Liang, Pei-Ji


    Ca(2+)-permeable and Ca(2+)-impermeable AMPA receptors are co-expressed on carp retinal horizontal cells. In the present study, we examined the synaptic contribution and Zn(2+) modulatory effect of these two AMPA receptor subtypes using whole-cell patch clamp technique. Specific Ca(2+)-permeable AMPA receptor antagonist (1-naphthyl acetyl spermine, NAS) and selective Ca(2+)-impermeable AMPA receptor blocker (pentobarbital, PB) were used to separate the glutamate-response in isolated H1 horizontal cell mediated by these two subtypes of AMPA receptors respectively. Application of 100 microM NAS substantially suppressed the current elicited by 3 mM glutamate and the remaining NAS-insensitive component was completely blocked by application of 100 microM PB. In addition, Zn(2+) had dual effects on Ca(2+)-permeable AMPA receptor-mediated current: at low concentration (10 microM), Zn(2+) potentiated the current, but at higher concentrations (100 and 1000 microM), Zn(2+) reduced the current in a dose-dependent manner. However, Zn(2+) (10, 100 and 1000 microM) failed to modulate the NAS-insensitive current mediated by Ca(2+)-impermeable AMPA receptors. Overall, our results suggest that Ca(2+)-permeable AMPA receptors contribute more to the cell's glutamate-response than Ca(2+)-impermeable AMPA receptors. Furthermore, Zn(2+) has dual effects on the Ca(2+)-permeable AMPA receptor activity without affecting Ca(2+)-impermeable AMPA receptors. 2009 Elsevier B.V. All rights reserved.

  15. Hydraulic Flushing Technology and Its Practice in Outburst Coal Seam with High Gas and Low Permeability

    Qing Ye


    Full Text Available Hydraulic flushing technology can rapidly and effectively eliminate coal and gas outbursts and improve the permeability of a coal seam. Its effect mainly depends on the technical parameters of hydraulic flushing. To solve the problems on technical parameters that exist in the application of hydraulic flushing technology, the outburst elimination mechanism of hydraulic flushing technology was expatiated, the hydraulic flushing process was introduced, and a field test was performed on the B1 coal seam in Yi’an Coal Mine by using the pressure drop method. Moreover, the effective influence radius of hydraulic flushing measure was determined, and the technical parameters were analyzed. Finally, a series of relationships was obtained, including the relationships between hydraulic pressure and coal output, critical breaking coal pressure and firmness coefficient, flushing time and coal output, drilling hole angle and coal output, and coal output and effective influence radius. Results showed that the effective influence radius of hydraulic flushing in B1 coal seam was 9 m, and the outburst risk of the coal within the influence region was eliminated. In addition, the time of outburst elimination was shortened and the production rate was improved. The research results could provide technical support for the optimization of the technical parameters and the test scheme of hydraulic flushing measures.

  16. Effect of moderate electric field frequency and growth stage on the cell membrane permeability of Lactobacillus acidophilus.

    Loghavi, Laleh; Sastry, Sudhir K; Yousef, Ahmed E


    Changes in growth kinetics and metabolic activity of microorganisms under the presence of a moderate electric field (MEF) have been hypothesized as being due to temporary permeabilization of cell membranes. We investigated herein the effects of frequency and growth stage on cell membrane permeabilization of Lactobacillus acidophilus OSU 133 during MEF fermentation. Cells were stained with two fluorescent nucleic acid stains: the green, nonselective, cell membrane permeable SYTO 9, and the red, cell membrane impermeable propidium iodide (PI). Fluorescence exhibition post-treatment was assessed using fluorescence microscopy. Total plate counting was done to determine whether or not the permeabilized population represented live cells. Fermentation treatments investigated were conventional (control) and MEF (2 V/cm, 45, 60, 1,000, 10,000 Hz) at 30 degrees C. Studies were conducted at 45 Hz for lag, exponential, and stationary phases of growth. Low frequency MEF treated cells exhibited significantly greater numbers of red cell counts than conventional treatments; further, no significant differences existed in viable counts between MEF and conventional treatments, suggesting that the red counts represent permeabilized live cells. MEF treatments at the early stage of bacterial growth at 45 Hz exhibited the maximum permeabilization followed by treatments at 60 Hz. MEF treated samples at frequencies higher than 60 Hz did not exhibit red fluorescence. Cells at lag phase showed the greatest susceptibility to permeabilization followed by those at exponential phase. No evidence of electroporation was observed during the stationary phase. To our knowledge, these observations provide the first evidence that cell membrane permeabilization occurs under the presence of electric fields as low as those under MEF.

  17. Use of 3D Seismic Azimuthal Iso-Frequency Volumes for the Detection and Characterization of High Porosity/Permeability Zones in Carbonate Reservoirs

    Toelle, Brian E.

    Among the most important properties controlling the production from conventional oil and gas reservoirs is the distribution of porosity and permeability within the producing geologic formation. The geometry of the pore space within these reservoirs, and the permeability associated with this pore space geometry, impacts not only where production can occur and at what flow rates but can also have significant influence on many other rock properties. Zones of high matrix porosity can result in an isotropic response for certain reservoir properties whereas aligned porosity/permeability, such as open, natural fracture trends, have been shown to result in reservoirs being anisotropic in many properties. The ability to identify zones within a subsurface reservoir where porosity/permeability is significantly higher and to characterize them according to their geometries would be of great significance when planning where new boreholes, particularly horizontal boreholes, should be drilled. The detection and characterization of these high porosity/permeability zones using their isotropic and anisotropic responses may be possible through the analysis of azimuthal (also referred to as azimuth-limited) 3D seismic volumes. During this study the porosity/permeability systems of a carbonate, pinnacle reef within the northern Michigan Basin undergoing enhanced oil recovery were investigated using selected seismic attributes extracted from azimuthal 3D seismic volumes. Based on the response of these seismic attributes an interpretation of the geometry of the porosity/permeability system within the reef was made. This interpretation was supported by well data that had been obtained during the primary production phase of the field. Additionally, 4D seismic data, obtained as part of the CO2 based EOR project, supported reservoir simulation results that were based on the porosity/permeability interpretation.

  18. MicroRNA-497 impairs the growth of chemoresistant neuroblastoma cells by targeting cell cycle, survival and vascular permeability genes

    Soriano, Aroa; París-Coderch, Laia; Jubierre, Luz; Martínez, Alba; Zhou, Xiangyu; Piskareva, Olga; Bray, Isabella; Vidal, Isaac; Almazán-Moga, Ana; Molist, Carla; Roma, Josep; Bayascas, José R.; Casanovas, Oriol; Stallings, Raymond L.; de Toledo, José Sánchez; Gallego, Soledad; Segura, Miguel F.


    Despite multimodal therapies, a high percentage of high-risk neuroblastoma (NB) become refractory to current treatments, most of which interfere with cell cycle and DNA synthesis or function, activating the DNA damage response (DDR). In cancer, this process is frequently altered by deregulated expression or function of several genes which contribute to multidrug resistance (MDR). MicroRNAs are outstanding candidates for therapy since a single microRNA can modulate the expression of multiple genes of the same or different pathways, thus hindering the development of resistance mechanisms by the tumor. We found several genes implicated in the MDR to be overexpressed in high-risk NB which could be targeted by microRNAs simultaneously. Our functional screening identified several of those microRNAs that reduced proliferation of chemoresistant NB cell lines, the best of which was miR-497. Low expression of miR-497 correlated with poor patient outcome. The overexpression of miR-497 reduced the proliferation of multiple chemoresistant NB cell lines and induced apoptosis in MYCN-amplified cell lines. Moreover, the conditional expression of miR-497 in NB xenografts reduced tumor growth and inhibited vascular permeabilization. MiR-497 targets multiple genes related to the DDR, cell cycle, survival and angiogenesis, which renders this molecule a promising candidate for NB therapy. PMID:26824183

  19. Highly organic natural media as permeable reactive barriers: TCE partitioning and anaerobic degradation profile in eucalyptus mulch and compost.

    Öztürk, Zuhal; Tansel, Berrin; Katsenovich, Yelena; Sukop, Michael; Laha, Shonali


    Batch and column experiments were conducted with eucalyptus mulch and commercial compost to evaluate suitability of highly organic natural media to support anaerobic decomposition of trichloroethylene (TCE) in groundwater. Experimental data for TCE and its dechlorination byproducts were analyzed with Hydrus-1D model to estimate the partitioning and kinetic parameters for the sequential dechlorination reactions during TCE decomposition. The highly organic natural media allowed development of a bioactive zone capable of decomposing TCE under anaerobic conditions. The first order TCE biodecomposition reaction rates were 0.23 and 1.2d(-1) in eucalyptus mulch and compost media, respectively. The retardation factors in the eucalyptus mulch and compost columns for TCE were 35 and 301, respectively. The results showed that natural organic soil amendments can effectively support the anaerobic bioactive zone for remediation of TCE contaminated groundwater. The natural organic media are effective environmentally sustainable materials for use in permeable reactive barriers.

  20. Methods to Measure Water Permeability.

    Solenov, Evgeniy I; Baturina, Galina S; Katkova, Liubov E; Zarogiannis, Sotirios G


    Water permeability is a key feature of the cell plasma membranes and it has seminal importance for a number of cell functions such as cell volume regulation, cell proliferation, cell migration, and angiogenesis to name a few. The transport of water occurs mainly through plasma membrane water channels , the aquaporins, who have very important function in physiological and pathophysiological states. Due to the above the experimental assessment of the water permeability of cells and tissues is necessary. The development of new methodologies of measuring water permeability is a vibrant scientific field that constantly develops during the past three decades along with the advances in imaging mainly. In this chapter we describe and critically assess several methods that have been developed for the measurement of water permeability both in living cells as well as in tissues with a focus in the first category.

  1. High Permeability Ternary Palladium Alloy Membranes with Improved Sulfur and Halide Tolerances

    K. Coulter


    and ternary membranes on a simulated flue gas stream and experienced significant difficulty in mounting and testing the sputter deposited membranes. IdaTech was able to successfully test PdAu and PdAuPt membranes and saw similar sulfur tolerance to what TDA found. The Program met all the deliverables on schedule and on budget. Over ten presentations at national and international conferences were made, four papers were published (two in progress) in technical journals, and three students (2 at GT and 1 at CSM) completed their doctorates using results generated during the course of the program. The three major findings of program were; (1) the DFT modeling was verified as a predictive tool for the permeability of Pd based ternary alloys, (2) while magnetron sputtering is useful in precisely fabricating binary and ternary alloys, the mechanical durability of membranes fabricated using this technique are inferior compared to cold rolled membranes and this preparation method is currently not ready for industrial environments, (3) based on both modeling and experimental verification in pure gas and mixed gas environments PdAu and PdAuPt alloys were found to have the combination of the highest permeability and tolerance to sulfur.

  2. Novel permeability characteristics of red blood cells from sickle cell patients heterozygous for HbS and HbC (HbSC genotype).

    Dalibalta, S; Ellory, J C; Browning, J A; Wilkins, R J; Rees, D C; Gibson, J S


    Individuals heterozygous for HbS and HbC (HbSC) represent about 1/3(rd) of sickle cell disease (SCD) patients. Whilst HbSC disease is generally milder, there is considerable overlap in symptoms with HbSS disease. HbSC patients, as well as HbSS ones, present with the chronic anaemia and panoply of acute vaso-occlusive complications that characterize SCD. However, there are important clinical and haematological differences. Certain complications occur with greater frequency in HbSC patients (like proliferative retinopathy and osteonecrosis) whilst intravascular haemolysis is reduced. Patients with HbSC disease can be considered as a discrete subset of SCD cases. Although much work has been carried out on understanding the pathogenesis of SCD in HbSS homozygotes, including the contribution of altered red blood cell permeability, relatively little pertains directly to HbSC individuals. Results reported in the literature suggest that HbSC cells, and particularly certain subpopulations, present with similar permeability to HbSS cells but there are also important differences - these have not been well characterized. We hypothesise that their unique cell transport properties accounts for the different pattern of disease in HbSC patients and represents a potential chemotherapeutic target not shared in red blood cells from HbSS patients. The distinct pattern of clinical haematology in HbSC disease is emphasised here. We analyse some of the electrophysiological properties of single red blood cells from HbSC patients, comparing them with those from HbSS patients and normal HbAA individuals. We also use the isosmotic haemolysis technique to investigate the behaviour of total red blood cell populations. Whilst both HbSS and HbSC cells show increased monovalent and divalent (Ca(2+)) cation conductance further elevated upon deoxygenation, the distribution of current magnitudes differs, and outward rectification is greatest for HbSC cells. In addition, although Gd(3+) largely

  3. ADME Properties Evaluation in Drug Discovery: Prediction of Caco-2 Cell Permeability Using a Combination of NSGA-II and Boosting.

    Wang, Ning-Ning; Dong, Jie; Deng, Yin-Hua; Zhu, Min-Feng; Wen, Ming; Yao, Zhi-Jiang; Lu, Ai-Ping; Wang, Jian-Bing; Cao, Dong-Sheng


    The Caco-2 cell monolayer model is a popular surrogate in predicting the in vitro human intestinal permeability of a drug due to its morphological and functional similarity with human enterocytes. A quantitative structure-property relationship (QSPR) study was carried out to predict Caco-2 cell permeability of a large data set consisting of 1272 compounds. Four different methods including multivariate linear regression (MLR), partial least-squares (PLS), support vector machine (SVM) regression and Boosting were employed to build prediction models with 30 molecular descriptors selected by nondominated sorting genetic algorithm-II (NSGA-II). The best Boosting model was obtained finally with R(2) = 0.97, RMSEF = 0.12, Q(2) = 0.83, RMSECV = 0.31 for the training set and RT(2) = 0.81, RMSET = 0.31 for the test set. A series of validation methods were used to assess the robustness and predictive ability of our model according to the OECD principles and then define its applicability domain. Compared with the reported QSAR/QSPR models about Caco-2 cell permeability, our model exhibits certain advantage in database size and prediction accuracy to some extent. Finally, we found that the polar volume, the hydrogen bond donor, the surface area and some other descriptors can influence the Caco-2 permeability to some extent. These results suggest that the proposed model is a good tool for predicting the permeability of drug candidates and to perform virtual screening in the early stage of drug development.

  4. Protective effects of l-carnitine and piracetam against mitochondrial permeability transition and PC3 cell necrosis induced by simvastatin.

    Costa, Rute A P; Fernandes, Mariana P; de Souza-Pinto, Nadja C; Vercesi, Aníbal E


    Mitochondrial oxidative stress followed by membrane permeability transition (MPT) has been considered as a possible mechanism for statins cytotoxicity. Statins use has been associated with reduced risk of cancer incidence, especially prostate cancer. Here we investigated the pathways leading to simvastatin-induced prostate cancer cell death as well as the mechanisms of cell death protection by l-carnitine or piracetam. These compounds are known to prevent and/or protect against cell death mediated by oxidative mitochondrial damage induced by a variety of conditions, either in vivo or in vitro. The results provide evidence that simvastatin induced MPT and cell necrosis were sensitive to either l-carnitine or piracetam in a dose-dependent fashion and mediated by additive mechanisms. When combined, l-carnitine and piracetam acted at concentrations significantly lower than they act individually. These results shed new light into both the cytotoxic mechanisms of statins and the mechanisms underlying the protection against MPT and cell death by the compounds l-carnitine and piracetam. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Bax and Bak function as the outer membrane component of the mitochondrial permeability pore in regulating necrotic cell death in mice

    Karch, Jason; Kwong, Jennifer Q; Burr, Adam R; Sargent, Michelle A; Elrod, John W; Peixoto, Pablo M; Martinez-Caballero, Sonia; Osinska, Hanna; Cheng, Emily H-Y; Robbins, Jeffrey; Kinnally, Kathleen W; Molkentin, Jeffery D


    A critical event in ischemia-based cell death is the opening of the mitochondrial permeability transition pore (MPTP). However, the molecular identity of the components of the MPTP remains unknown. Here, we determined that the Bcl-2 family members Bax and Bak, which are central regulators of apoptotic cell death, are also required for mitochondrial pore-dependent necrotic cell death by facilitating outer membrane permeability of the MPTP. Loss of Bax/Bak reduced outer mitochondrial membrane permeability and conductance without altering inner membrane MPTP function, resulting in resistance to mitochondrial calcium overload and necrotic cell death. Reconstitution with mutants of Bax that cannot oligomerize and form apoptotic pores, but still enhance outer membrane permeability, permitted MPTP-dependent mitochondrial swelling and restored necrotic cell death. Our data predict that the MPTP is an inner membrane regulated process, although in the absence of Bax/Bak the outer membrane resists swelling and prevents organelle rupture to prevent cell death. DOI: PMID:23991283

  6. Bax and Bak function as the outer membrane component of the mitochondrial permeability pore in regulating necrotic cell death in mice.

    Karch, Jason; Kwong, Jennifer Q; Burr, Adam R; Sargent, Michelle A; Elrod, John W; Peixoto, Pablo M; Martinez-Caballero, Sonia; Osinska, Hanna; Cheng, Emily H-Y; Robbins, Jeffrey; Kinnally, Kathleen W; Molkentin, Jeffery D


    A critical event in ischemia-based cell death is the opening of the mitochondrial permeability transition pore (MPTP). However, the molecular identity of the components of the MPTP remains unknown. Here, we determined that the Bcl-2 family members Bax and Bak, which are central regulators of apoptotic cell death, are also required for mitochondrial pore-dependent necrotic cell death by facilitating outer membrane permeability of the MPTP. Loss of Bax/Bak reduced outer mitochondrial membrane permeability and conductance without altering inner membrane MPTP function, resulting in resistance to mitochondrial calcium overload and necrotic cell death. Reconstitution with mutants of Bax that cannot oligomerize and form apoptotic pores, but still enhance outer membrane permeability, permitted MPTP-dependent mitochondrial swelling and restored necrotic cell death. Our data predict that the MPTP is an inner membrane regulated process, although in the absence of Bax/Bak the outer membrane resists swelling and prevents organelle rupture to prevent cell death. DOI:

  7. Methane seep in shallow-water permeable sediment harbors high diversity of anaerobic methanotrophic communities, Elba, Italy

    S Emil Ruff


    Full Text Available The anaerobic oxidation of methane (AOM is a key biogeochemical process regulating methane emission from marine sediments into the hydrosphere. AOM is largely mediated by consortia of anaerobic methanotrophic archaea (ANME and sulfate-reducing bacteria (SRB, and has mainly been investigated in deep-sea sediments. Here we studied methane seepage at four spots located at 12 m water depth in coastal, organic-carbon depleted permeable sands off the Island of Elba (Italy. We combined biogeochemical measurements, sequencing-based community analyses and in situ hybridization to investigate the microbial communities of this environment. Increased alkalinity, formation of free sulfide and nearly stoichiometric methane oxidation and sulfate reduction rates up to 200 nmol g-1 day-1 indicated the predominance of sulfate-coupled AOM. With up to 40 cm thickness the zones of AOM activity were unusually large and occurred in deeper sediment horizons (20–50 cm below seafloor as compared to diffusion-dominated deep-sea seeps, which is likely caused by advective flow of pore water due to the shallow water depth and permeability of the sands. Hydrodynamic forces also may be responsible for the substantial phylogenetic and unprecedented morphological diversity of AOM consortia inhabiting these sands, including the clades ANME-1a/b, ANME-2a/b/c, ANME-3 and their partner bacteria SEEP-SRB1a and SEEP-SRB2. High microbial dispersal, the availability of diverse energy sources and high habitat heterogeneity might explain that the emission spots shared few microbial taxa, despite their physical proximity. Although the biogeochemistry of this shallow methane seep was very different to that of deep-sea seeps, their key functional taxa were very closely related, which supports the global dispersal of key taxa and underlines strong selection by methane as the predominant energy source. Mesophilic, methane-fueled ecosystems in shallow-water permeable sediments may comprise

  8. Fabrication and application of a wireless inductance-capacitance coupling microsensor with electroplated high permeability material NiFe

    Chen, Y H; Chang, I N [Ph. D. Program in Electrical and Communications Engineering, Feng-Chia University, Taichung 40724, Taiwan (China); Chang, H C; Lai, C C, E-mail: [Department of Automatic Control Engineering, Feng-Chia University, Taichung 40724, Taiwan (China)


    A fully integrated wireless inductance-capacitance (LC) coupling microsensor was designed and fabricated by MEMS technology. The sensing loop was formed by connecting a deformable parallel-plated capacitor and a planar spiral inductor with a Ni(80)Fe(20) core. Polyimide and PMMA were used to isolate and package the devices. Typical dimension of the sensors was 5 x 5 mm{sup 2} x 0.77 mm. Different electroplated inductive coils (30, 40, and 60 turns) were fabricated to connect with a 4 x 4 mm{sup 2} plate capacitor in series. The LC sensing module for measuring liquid-level induced frequency responses was setup. Experimental results show that frequency response decreased as liquid level increased and sensitivity is about 7.01 kHz/cm with deviation less than 2%. Developed planar spiral inductor with high permeability magnetic core can provide a wide range of frequency variation in LC sensing applications.

  9. Role of mitochondrial permeability transition in human hepatocellular carcinoma Hep-G2 cell death induced by rhein.

    Du, Qiong; Bian, Xiao-Lan; Xu, Xiao-Le; Zhu, Bin; Yu, Bo; Zhai, Qing


    Rhein, a compound found as a glucoside in the root of rhubarb, is currently a subject of interest for its antitumor properties. The apoptosis of tumor cell lines induced by rhein was observed, and the involvement of mitochondria was established; however, the role of mitochondrial permeability transition (MPT) remains unknown. Here we report that MPT plays an important role in the apoptosis of human hepatocellular carcinoma Hep-G2 cells induced by rhein. After adding rhein to the isolated hepatic mitochondria, swelling effects and the leakage of Ca(2+) were observed. These alterations were suppressed by cyclosporin A (CsA), an MPT inhibitor. Furthermore, in Hep-G2 cells, the decrease of ATP production, the loss of mitochondrial transmembrane potential (MTP), the release of cytochrome c (Cyto c), and the activation of caspase 3 were also observed. These toxic effects of rhein can also be attenuated by CsA as well. Moreover, TUNEL assay confirmed that in the presence of CsA, rhein-induced apoptosis was largely inhibited. These results suggest that MPT plays a critical role in the pathogenesis of Hep-G2 cell injury induced by rhein, and imply that MPT may contribute to the anti-cancer activity of rhein. © 2013.

  10. Inhibitory effect of high [Mg2+] on the vasopressin-stimulated hydroosmotic permeability of the isolated perfused cortical collecting duct

    Falkenstein D.


    Full Text Available High magnesium concentration inhibits the effect of arginine vasopressin (AVP on smooth muscle contraction and platelet aggregation and also influences hepatocyte AVP receptor binding. The aim of this study was to determine the role of magnesium concentration [Mg2+] in AVP-stimulated water transport in the kidney collecting duct. The effect of low and high peritubular [Mg2+] on the AVP-stimulated osmotic water permeability coefficient (Pf was evaluated in the isolated perfused rabbit cortical collecting duct (CCD. Control tubules bathed and perfused with standard Ringer bicarbonate solution containing 1 mM Mg2+ presented a Pf of 223.9 ± 27.2 µm/s. When Mg2+ was not added to the bathing solution, an increase in the AVP-stimulated Pf to 363.1 ± 57.2 µm/s (P<0.05 was observed. An elevation of Mg2+ to 5 mM resulted in a decrease in Pf to 202.9 ± 12.6 µm/s (P<0.05. This decrease in the AVP-stimulated Pf at 5 mM Mg2+ persisted when the CCDs were returned to 1 mM Mg2+, Pf = 130.2 ± 20.3 µm/s, and was not normalized by the addition of 8-[4-chlorophenylthio]-adenosine 3',5'-cyclic monophosphate, a cAMP analogue, to the preparation. These data indicate that magnesium may play a modulatory role in the action of AVP on CCD osmotic water permeability, as observed in other tissues.

  11. A novel, cell-permeable, fluorescent probe for ratiometric imaging of zinc ion.

    Maruyama, Satoko; Kikuchi, Kazuya; Hirano, Tomoya; Urano, Yasuteru; Nagano, Tetsuo


    Zn(2+) plays important roles in various biological systems; as a result, the development of tools that can visualize chelatable Zn(2+) has attracted much attention recently. We report here newly synthesized fluorescent sensors for Zn(2+), ZnAF-Rs, whose excitation maximum is shifted by Zn(2+) under physiological conditions. Thus, these sensors enable ratiometric imaging, which is a technique to reduce artifacts by minimizing the influence of extraneous factors on the fluorescence of a probe. Ratiometric measurement can provide precise data, and some probes allow quantitative detection. ZnAF-Rs are the first ratiometric fluorescent sensors for Zn(2+) that enable quantitative analysis under physiological conditions. ZnAF-Rs also possess suitable K(d) for applications, and high selectivity against other biologically relevant cations, especially Ca(2+). Using these probes, changes of intracellular Zn(2+) concentration in cultured cells were monitored successfully. We believe that these probes will be extremely useful in studies on the biological functions of Zn(2+).

  12. Claudin-5 regulates blood-brain barrier permeability by modifying brain microvascular endothelial cell proliferation, migration, and adhesion to prevent lung cancer metastasis.

    Ma, Shun-Chang; Li, Qi; Peng, Jia-Yi; Zhouwen, Jian-Long; Diao, Jin-Fu; Niu, Jian-Xing; Wang, Xi; Guan, Xiu-Dong; Jia, Wang; Jiang, Wen-Guo


    To investigate the roles of Claudin-5 (CLDN5) in regulating the permeability of the blood-brain barrier (BBB) during lung cancer brain metastasis. By silencing and overexpressing the CLDN5 gene in human brain vascular endothelial (hCMEC/D3) cells, we demonstrated the attenuation of cell migration ability and CLDN5's significant positive role in cell proliferation in CLDN5-overexpressing hCMEC/D3 cells and observed the opposite result in the CLDN5 knockdown group. The reinforced CLDN5 expression reduced the paracellular permeability of hCMEC/D3 cells and decreased the invasion of lung adenocarcinoma A549 cells. Overall, 1685 genes were found to be differentially expressed between the CLDN5-overexpressing cells and the control cells using the Affymetrix Human Transcriptome Array 2.0 (HTA 2.0), and the function of these genes was determined by Gene Ontology and pathway analyses. The possible biological functions of the 1685 genes include cell proliferation, adhesion molecules, and the Jak-STAT, PI3K-Akt, Wnt, and Notch signaling pathways. The identified sets of mRNAs that were specific to CLDN5-overexpressing hCMEC/D3 cells were verified by a qRT-PCR experiment. CLDN5 regulates the permeability of BBB by regulating the proliferation, migration, and permeability of hCMEC/D3 cells, especially through the cell adhesion molecule signaling pathway, to enhance the function of the tight junctions, which was involved in reducing the formation of lung cancer brain metastasis. © 2017 John Wiley & Sons Ltd.

  13. Cell Treatment for Stroke in Type Two Diabetic Rats Improves Vascular Permeability Measured by MRI.

    Guangliang Ding

    Full Text Available Treatment of stroke with bone marrow stromal cells (BMSC significantly enhances brain remodeling and improves neurological function in non-diabetic stroke rats. Diabetes is a major risk factor for stroke and induces neurovascular changes which may impact stroke therapy. Thus, it is necessary to test our hypothesis that the treatment of stroke with BMSC has therapeutic efficacy in the most common form of diabetes, type 2 diabetes mellitus (T2DM. T2DM was induced in adult male Wistar rats by administration of a high fat diet in combination with a single intraperitoneal injection (35mg/kg of streptozotocin. These rats were then subjected to 2h of middle cerebral artery occlusion (MCAo. T2DM rats received BMSC (5x106, n = 8 or an equal volume of phosphate-buffered saline (PBS (n = 8 via tail-vein injection at 3 days after MCAo. MRI was performed one day and then weekly for 5 weeks post MCAo for all rats. Compared with vehicle treated control T2DM rats, BMSC treatment of stroke in T2DM rats significantly (p<0.05 decreased blood-brain barrier disruption starting at 1 week post stroke measured using contrast enhanced T1-weighted imaging with gadopentetate, and reduced cerebral hemorrhagic spots starting at 3 weeks post stroke measured using susceptibility weighted imaging, although BMSC treatment did not reduce the ischemic lesion volumes as demarcated by T2 maps. These MRI measurements were consistent with histological data. Thus, BMSC treatment of stroke in T2DM rats initiated at 3 days after stroke significantly reduced ischemic vascular damage, although BMSC treatment did not change infarction volume in T2DM rats, measured by MRI.

  14. Effect of FCCP on tight junction permeability and cellular distribution of ZO-1 protein in epithelial (MDCK) cells.

    Li, C X; Poznansky, M J


    The effect of the uncoupler of oxidative phosphorylation, FCCP (carbonylcyanide p-trifluoromethoxyphenylhydrazone), on the tight junction of Madin-Darby canine kidney cells was examined. FCCP induced an abrupt decrease in the transepithelial electrical resistance of the confluent monolayers over a period of 20 s. When FCCP was withdrawn from the incubation medium, the monolayer resistance recovered to close to the original level in less than 2 h. Staining of the tight junction-associated protein ZO-1 showed that the changes in transepithelial electrical resistance were accompanied by a diffusing of the protein away from cell peripheries and a reconcentration to the tight junction areas following resistance recovery. Intracellular pH was decreased by FCCP on a similar time-scale with no obvious changes in ATP levels over this time-course. These data suggest that the uncoupler FCCP has a profound effect on tight junction permeability and cellular distribution of the tight junction protein ZO-1 in the epithelial cells and that it probably acts by breaking down proton gradients and altering intracellular pH.

  15. SGLT1-mediated transport in Caco-2 cells is highly dependent on cell bank origin

    Steffansen, B; Pedersen, Maria; Laghmoch, A M


    The Caco-2 cell line is a well-established in vitro model for studying transport phenomena for prediction of intestinal nutrient and drug absorption. However, for substances depending on transporters such predictions are complicated due to variable transporter expression and limited knowledge about...... permeability (PSGLT1). Here, the objective was to characterize and compare SGLT1-mediated uptake in Caco-2 cells obtained from different cell banks. SGLT1-mediated uptake of the standard SGLT1 substrate, α-MDG, in Caco-2 cells was shown to be highly dependent on cell bank origin. The most robust and reliable...

  16. Conjugated primary bile salts reduce permeability of endotoxin through intestinal epithelial cells and synergize with phosphatidylcholine in suppression of inflammatory cytokine production

    Parlesak, Alexandr; Schaeckeler, S.; Moser, L.


    : The effect of CPBS (0.5 mM and 1.5 mM), phosphatidylcholine (0.38 mM), and human bile (0.5% vol/vol) on the barrier function was assessed by the measurement of transepithelial electrical resistance, by endotoxin permeability through the intestinal epithelial cell layer, and by basolateral cytokine enzyme...

  17. Conjugated primary bile salts reduce permeability of endotoxin through bacteria-stimulated intestinal epithelial cells and synergize with lecithin in suppression of inflammatory cytokine production

    Parlesak, Alexandr; Schaeckeler, Simone; Moser, Lydia


    : The effect of CPBS (0.5 mM and 1.5 mM), phosphatidylcholine(0.38 mM), and human bile (0.5% vol/vol) on the barrier function was assessed by the measurement of transepithelial electrical resistance, by endotoxin permeability through the intestinal epithelial cell layer, and by basolateral cytokine enzyme...

  18. Preparation of highly permeable BPPO microfiltration membrane with binary porous structures on a colloidal crystal substrate by the breath figure method.

    Yuan, Hua; Yu, Bing; Cong, Hailin; Peng, Qiaohong; Yang, Zhen; Luo, Yongli; Chi, Ming


    A highly permeable brominated poly(phenylene oxide) (BPPO) microfiltration membrane with binary porous structures was fabricated by combination of the breath figure and colloidal crystal template methods. The pore size in the bottom layer of the membrane was adjusted by the diameter of SiO2 microspheres in the colloidal crystal template, while the pore size in the top layer of the membrane was adjusted by varying the BPPO concentration in the casting solution. The permeability of the membrane cast on the colloidal crystal substrate was much higher than that of the membrane cast on a bare silicon wafer. The binary porous BPPO membrane with high permeability and antifouling property was used for microfiltration applications.

  19. Long-term observation of permeability in sedimentary rocks under high-temperature and stress conditions and its interpretation mediated by microstructural investigations

    Yasuhara, Hideaki; Kinoshita, Naoki; Ohfuji, Hiroaki; Takahashi, Manabu; Ito, Kazumasa; Kishida, Kiyoshi


    In this study, a series of long-term, intermittent permeability experiments utilizing Berea sandstone and Horonobe mudstone samples, with and without a single artificial fracture, is conducted for more than 1000 days to examine the evolution of rock permeability under relatively high-temperature and confining pressure conditions. Effluent element concentrations are also measured throughout the experiments. Before and after flow-through experiments, rock samples are prepared for X-ray diffraction, X-ray fluorescence, and scanning electron microscopy coupled with energy dispersive X-ray spectroscopy to examine the mineralogical changes between pre and postexperimental samples, and also for microfocus X-ray CT to evaluate the alteration of the microstructure. Although there are exceptions, the observed, qualitative evolution of permeability is found to be generally consistent in both the intact and the fractured rock samples—the permeability in the intact rock samples increases with time after experiencing no significant changes in permeability for the first several hundred days, while that in the fractured rock samples decreases with time. An evaluation of the Damkohler number and of the net dissolution, using the measured element concentrations, reveals that the increase in permeability can most likely be attributed to the relative dominance of the mineral dissolution in the pore spaces, while the decrease can most likely be attributed to the mineral dissolution/crushing at the propping asperities within the fracture. Taking supplemental observations by microfocus X-ray CT and using the intact sandstone samples, a slight increase in relatively large pore spaces is seen. This supports the increase in permeability observed in the flow-through experiments.

  20. Side population analysis using a violet-excited cell-permeable DNA binding dye.

    Telford, William G; Bradford, Jolene; Godfrey, William; Robey, Robert W; Bates, Susan E


    Hoechst 33342 side population (SP) analysis is a common method for identifying stem cells in mammalian hematopoietic and nonhematopoietic tissues. Although widely employed for stem cell analysis, this method requires an ultraviolet (UV) laser to excite Hoechst 33342. Flow cytometers equipped with UV sources are not common because of the cost of both the laser and optics that can transmit light UV light. Violet laser sources are inexpensive and are now common fixtures on flow cytometers, but have been previously shown to provide insufficient Hoechst dye excitation for consistent resolution of SP cells. One solution to this problem is to identify additional fluorescent substrates with the same pump specificity as Hoechst 33342, but with better violet excitation characteristics. DyeCycle Violet reagent has emission characteristics similar to those of Hoechst 33342, but with a longer wavelength excitation maxima (369 nm). When this dye is loaded into hematopoietic cells, a sharply resolved side population was also observed, similar in appearance to that seen with Hoechst 33342. Unlike Hoechst SP, DCV SP was similar in appearance with both violet and UV excitation. DCV SP could be inhibited fumitremorgin C, and showed the same membrane pump specificity as Hoechst 33342. Simultaneous immunophenotyping with stem cell markers in mouse bone marrow demonstrated that DCV SP was restricted to the stem cell lineage(-) Sca-1(+) c-kit(+) cells population, as is Hoechst SP. Pending confirmation by functional analysis of DCV SP cells, these results suggest that DCV efflux identified approximately the same stem cell population as did Hoechst 33342 efflux. Substituting DCV for Hoechst 33342 in the SP technique may, therefore, allow side population analysis on flow cytometers with violet lasers.

  1. Exogenous arachidonic acid mediates permeability of human brain microvessel endothelial cells through prostaglandin E2 activation of EP3 and EP4 receptors.

    Dalvi, Siddhartha; Nguyen, Hieu H; On, Ngoc; Mitchell, Ryan W; Aukema, Harold M; Miller, Donald W; Hatch, Grant M


    The blood-brain barrier, formed by microvessel endothelial cells, is the restrictive barrier between the brain parenchyma and the circulating blood. Arachidonic acid (ARA; 5,8,11,14-cis-eicosatetraenoic acid) is a conditionally essential polyunsaturated fatty acid [20:4(n-6)] and is a major constituent of brain lipids. The current study examined the transport processes for ARA in confluent monolayers of human brain microvascular endothelial cells (HBMEC). Addition of radioactive ARA to the apical compartment of HBMEC cultured on Transwell(®) inserts resulted in rapid incorporation of radioactivity into the basolateral medium. Knock down of fatty acid transport proteins did not alter ARA passage into the basolateral medium as a result of the rapid generation of prostaglandin E2 (PGE2 ), an eicosanoid known to facilitate opening of the blood-brain barrier. Permeability following ARA or PGE2 exposure was confirmed by an increased movement of fluorescein-labeled dextran from apical to basolateral medium. ARA-mediated permeability was attenuated by specific cyclooxygenase-2 inhibitors. EP3 and EP4 receptor antagonists attenuated the ARA-mediated permeability of HBMEC. The results indicate that ARA increases permeability of HBMEC monolayers likely via increased production of PGE2 which acts upon EP3 and EP4 receptors to mediate permeability. These observations may explain the rapid influx of ARA into the brain previously observed upon plasma infusion with ARA. The blood-brain barrier, formed by microvessel endothelial cells, is a restrictive barrier between the brain parenchyma and the circulating blood. Radiolabeled arachidonic acid (ARA) movement across, and monolayer permeability in the presence of ARA, was examined in confluent monolayers of primary human brain microvessel endothelial cells (HBMECs) cultured on Transwell(®) plates. Incubation of HBMECs with ARA resulted in a rapid increase in HBMEC monolayer permeability. The mechanism was mediated, in part

  2. Can Lucifer Yellow Indicate Correct Permeability of Biological Cell Membrane under An Electric and Magnetic Field?

    Tahereh Pourmirjafari Firoozabadi


    Full Text Available The effect of external magnetic and electric fields, in the range of electroporation and magnetoporation, on Lucifer Yellow (LY fluorescence in the absence of cells is studied. Electric-field-induced quenching and magnetic field-induced increase are observed for fluorescence intensity of LY. Regard to the fact that the variation of field-induced fluorescence, even in the absence of cells, can be observed, the application of LY, as a marker, is debatable in electroporation and magnetoporation techniques.

  3. Rational design of intrinsically ultramicroporous polyimides containing bridgehead-substituted triptycene for highly selective and permeable gas separation membranes

    Swaidan, Raja


    Highly ultramicroporous, solution-processable polyimides bearing 9,10-bridgehead-substituted triptycene demonstrated the highest BET surface area reported for polyimides (840 m2 g-1) and several new highs in gas selectivity and permeability for hydrogen (1630-3980 barrers, H2/CH4 ∼ 38) and air (230-630 barrers, O 2/N2 = 5.5-5.9) separations. Two new dianhydrides bearing 9,10-diethyl- and 9,10-dipropyltriptycenes indicate that the ultramicroporosity is optimized for fast polymeric sieving with the use of short, bulky isopropyl bridgeheads and methyl-substituted diamines (TrMPD, TMPD, and TMBZ) that increase intrachain rigidity. Mechanically, the triptycene-based analogue of a spirobisindane-based polyimide exhibited 50% increases in both tensile strength at break (94 MPa) and elastic modulus (2460 MPa) with corresponding 90% lower elongations at break (6%) likely due to the ability of highly entangled spiro-based chains to unwind. To guide future polyimide design, structure/property relationships are suggested between the geometry of the contortion center, the diamine and bridgehead substituent, and the mechanical, microstructural, and gas transport properties. © 2014 American Chemical Society.

  4. Synthesis and analysis of novel polymers with high permselectivity and permeability in gas separation applications

    Koros, W.J.; Paul, D.R.


    Significant progress was made toward developing advanced materials for gas separation membrane applications and rationalizing molecular structure and efficacy: Synthesized and tested polyarylates based on terephthalic or isophthalic acid or a tertiary butyl derivative of the isophthalic acid with different diols to illustrate the effects of: opening'' the matrix by incorporation of bulky packing inhibiting groups such as the tertiary butyl moiety inhibition of backbone motion via meta connected backbone connections and tightening'' of the matrix by incorporation of polar halogens. Completed high temperature characterization of sorption and transport properties for novel materials. Continued studies of the phenyl-substituted polymers aimed at producing super stable high temperature useful polymers for gas separations. Synthesized a polyarylate based on the spirobiindane diol and bibenzoyl acid chloride to incorporate long flat packable bibenzoyl units between packing disruptive spirobiindane units in an attempt to control the segmental level morphology to produce highly selective bottleneck'' regions between highly open regions.

  5. Autoimmune gastritis and parietal cell reactivity in two children with abnormal intestinal permeability

    Greenwood, Deanne L. V.; Crock, Patricia; Braye, Stephen; Davidson, Patricia; Sentry, John W.


    Autoimmune gastritis is characterised by lymphocytic infiltration of the gastric submucosa, with loss of parietal and chief cells and achlorhydria. Often, gastritis is expressed clinically as cobalamin deficiency with megaloblastic anaemia, which is generally described as a disease of the elderly. H

  6. Autoimmune gastritis and parietal cell reactivity in two children with abnormal intestinal permeability

    Greenwood, Deanne L. V.; Crock, Patricia; Braye, Stephen; Davidson, Patricia; Sentry, John W.

    Autoimmune gastritis is characterised by lymphocytic infiltration of the gastric submucosa, with loss of parietal and chief cells and achlorhydria. Often, gastritis is expressed clinically as cobalamin deficiency with megaloblastic anaemia, which is generally described as a disease of the elderly.

  7. Quantitative Imaging of Cell-Permeable Magnetic Resonance Contrast Agents Using X-Ray Fluorescence

    Paul J. Endres


    Full Text Available The inability to transduce cellular membranes is a limitation of current magnetic resonance imaging probes used in biologic and clinical settings. This constraint confines contrast agents to extracellular and vascular regions of the body, drastically reducing their viability for investigating processes and cycles in developmental biology. Conversely, a contrast agent with the ability to permeate cell membranes could be used in visualizing cell patterning, cell fate mapping, gene therapy, and, eventually, noninvasive cancer diagnosis. Therefore, we describe the synthesis and quantitative imaging of four contrast agents with the capability to cross cell membranes in sufficient quantity for detection. Each agent is based on the conjugation of a Gd(III chelator with a cellular transduction moiety. Specifically, we coupled Gd(III–diethylenetriaminepentaacetic acid DTPA and Gd(III–1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid with an 8–amino acid polyarginine oligomer and an amphipathic stilbene molecule, 4-amino-4'-(N,N-dimethylaminostilbene. The imaging modality that provided the best sensitivity and spatial resolution for direct detection of the contrast agents is synchrotron radiation x-ray fluorescence (SR-XRF. Unlike optical microscopy, SR-XRF provides two-dimensional images with resolution 103 better than 153Gd gamma counting, without altering the agent by organic fluorophore conjugation. The transduction efficiency of the intracellular agents was evaluated by T1 analysis and inductively coupled plasma mass spectrometry to determine the efficacy of each chelate-transporter combination.

  8. Sequence-selective DNA binding with cell-permeable oligoguanidinium-peptide conjugates.

    Mosquera, Jesús; Sánchez, Mateo I; Valero, Julián; de Mendoza, Javier; Vázquez, M Eugenio; Mascareñas, José L


    Conjugation of a short peptide fragment from a bZIP protein to an oligoguanidinium tail results in a DNA-binding miniprotein that selectively interacts with composite sequences containing the peptide-binding site next to an A/T-rich tract. In addition to stabilizing the complex with the target DNA, the oligoguanidinium unit also endows the conjugate with cell internalization properties.

  9. Magnetic ageing study of high and medium permeability nanocrystalline FeSiCuNbB alloys

    Lekdim, Atef; Morel, Laurent; Raulet, Marie-Ange


    increasing the energy efficiency is one of the most important issues in modern power electronic systems. In aircraft applications, the energy efficiency must be associated with a maximum reduction of mass and volume, so a high components compactness. A consequence from this compactness is the increase of operating temperature. Thus, the magnetic materials used in these applications, have to work at high temperature. It raises the question of the thermal ageing problem. The reliability of these components operating at this condition becomes a real problem which deserves serious interest. Our work takes part in this context by studying the magnetic material thermal ageing. The nanocrystalline materials are getting more and more used in power electronic applications. Main advantages of nanocrystalline materials compared to ferrite are: high saturation flux density of almost 1.25 T and low dynamic losses for low and medium frequencies. The nanocrystalline Fe73.5Cu1Nb3Si15.5B7 alloys have been chosen in our aging study. This study is based on monitoring the magnetic characteristics for several continuous thermal ageing (100, 150, 200 and 240 °C). An important experimental work of magnetic characterization is being done following a specific monitoring protocol. Elsewhere, X-Ray Diffraction and magnetostriction measurements were carried out to support the study of the anisotropy energies evolution with ageing. This latter is discussed in this paper to explain and give hypothesis about the ageing phenomena.

  10. Combined T2 and T1 measurements for improved perfusion and permeability studies in high field using dynamic contrast enhancement

    Bazelaire, Cedric de [Saint Louis Hospital, Radiology Department, Paris, Cedex 10 (France); Beth Israel Deaconess Medical Center and Harvard Medical School, Department of Radiology, Cambridge, MA (United States); Rofsky, Neil M.; Duhamel, Guillaume; Zhang, Jingbo; Alsop, David C. [Beth Israel Deaconess Medical Center and Harvard Medical School, Department of Radiology, Cambridge, MA (United States); Michaelson, M.D. [Massachusetts General Hospital, Department of Hematology/Oncology, Boston, MA (United States); George, Daniel [Dana Farber Cancer Institute, Department of Adult Oncology, Boston, MA (United States)


    This study analyzed the T2* effect of extracellularly distributed gadolinium contrast agents in arterial blood during tumor studies using T1-weighted sequences at high field strength. A saturation-prepared dual echo sequence with echo times of 1.5 and 3.5 ms was employed at 3 T to simultaneously characterize T1 and T2* of arterial blood during bolus administration of Gd-DTPA in 28 patients with body tumors. T2* effect and T1 effect of Gd-DTPA on image intensity of whole blood were calibrated in human blood samples with different concentrations of contrast agent. T2* was used to estimate concentration near the peak of the bolus. T1 was used to measure lower concentrations when T2* was not significant. T2* was measurable on calibration curves for Gd-DTPA concentrations higher than 4 mM. This concentration was exceeded in 18 patients. The mean signal intensity reduction because of T2* effect was estimated at 22{+-}14% of the T2* compensated signal. Using T2* measurements reduced underestimations of peak arterial Gd-DTPA concentration (59{+-}38%) and overestimation of permeability K{sup trans} (58%). The T2* effect of gadolinium contrast agents should therefore be accounted for when performing tumors study with T1-weighted sequences at high field strength. (orig.)

  11. Synthesis of ceramic hollow fiber supported zeolitic imidazolate framework-8 (ZIF-8) membranes with high hydrogen permeability

    Pan, Yichang


    Purification and recovery of hydrogen from hydrocarbons in refinery streams in the petrochemical industry is an emerging research field in the study of membrane gas separation. Hollow fiber membrane modules can be easily implemented into separation processes at the industrial scale. In this report, hollow yttria-stabilized zirconia (YSZ) fiber-supported zeolitic imidazole framework-8 (ZIF-8) membranes were successfully prepared using a mild and environmentally friendly seeded growth method. Our single-component permeation studies demonstrated that the membrane had a very high hydrogen permeance (~15×10 -7mol/m 2sPa) and an ideal selectivity of H 2/C 3H 8 of more than 1000 at room temperature. This high membrane permeability and selectivity caused serious concentration polarization in the separation of H 2/C 3H 8 mixtures, which led to almost 50% drop in both the H 2 permeance and the separation factor. Enhanced mixing on the feed side could reduce the effect of the concentration polarization. Our experimental data also indicated that the membranes had excellent reproducibility and long-term stability, indicating that the hollow fiber-supported ZIF-8 membranes developed in this study have great potential in industry-scale separation of hydrogen. © 2012 Elsevier B.V.

  12. Changes in Cell Membrane Permeability in Sunflower Hypocotyls Infected with Sclerotinia sclerotiorum.

    Hancock, J G


    Influx and efflux of water and urea and electrolyte leakage are less for sunflower (Helianthus annuus) hypocotyl sections above lesions caused by Sclerotinia sclerotiorum than for those from healthy plants. Urea uptake by sections above lesions is reduced (celery, squash, and tomato) or unchanged (bean) in other hosts after Sclerotinia infection. Efflux of urea from sunflower hypocotyls is biphasic, suggesting diffusion in series from two cellular compartments (cytoplasm and vacuole). Efflux during the fast phase was 7 to 20 times greater than that during the slow phase. No difference was noted in urea efflux from healthy and diseased tissues during the slow phase. However, efflux during the fast phase from diseased tissues was slower than from healthy tissues, suggesting that the increased resistance to diffusion of urea in host cells above lesions resides in the plasmalemma. Water movement across cell membranes of healthy and diseased sunflower hypocotyls was reduced when tissues were treated with p-hydroxymercuribenzoate.

  13. Water permeability of Na+-K+-2C1- cotransporters in mammalian epithelial cells

    Hammann, Steffen; Herrera-Perez, J.J.; Bundgaard, Magnus


    Water transport properties of the Na+-K+-2Cl- cotransporter (NKCC) were studied in cultures of pigmented epithelial cells (PE) from the ciliary body of the eye. Here, the membrane that faces upwards contains NKCCs and can be subjected to rapid changes in bathing solution composition and osmolarity...... changes of the cotransporter and interaction with Na+, K+ and Cl-. Similar measurements were performed on immortalized cell cultures from the thick ascending limb of the loop of Henle (TALH). Given similar overall transport rates of bumetanide-sensitive 86Rb+, the NKCCs of this tissue did not contribute...... any bumetanide-sensitive Lp. This suggests that the cotransporters of the two tissues are either different isoforms or the same cotransporter but in two different transport modes....

  14. Hydrogen-Rich Medium Attenuated Lipopolysaccharide-Induced Monocyte-Endothelial Cell Adhesion and Vascular Endothelial Permeability via Rho-Associated Coiled-Coil Protein Kinase.

    Xie, Keliang; Wang, Weina; Chen, Hongguang; Han, Huanzhi; Liu, Daquan; Wang, Guolin; Yu, Yonghao


    Sepsis is the leading cause of death in critically ill patients. In recent years, molecular hydrogen, as an effective free radical scavenger, has been shown a selective antioxidant and anti-inflammatory effect, and it is beneficial in the treatment of sepsis. Rho-associated coiled-coil protein kinase (ROCK) participates in junction between normal cells, and regulates vascular endothelial permeability. In this study, we used lipopolysaccharide to stimulate vascular endothelial cells and explored the effects of hydrogen-rich medium on the regulation of adhesion of monocytes to endothelial cells and vascular endothelial permeability. We found that hydrogen-rich medium could inhibit adhesion of monocytes to endothelial cells and decrease levels of adhesion molecules, whereas the levels of transepithelial/endothelial electrical resistance values and the expression of vascular endothelial cadherin were increased after hydrogen-rich medium treatment. Moreover, hydrogen-rich medium could lessen the expression of ROCK, as a similar effect of its inhibitor Y-27632. In addition, hydrogen-rich medium could also inhibit adhesion of polymorphonuclear neutrophils to endothelial cells. In conclusion, hydrogen-rich medium could regulate adhesion of monocytes/polymorphonuclear neutrophils to endothelial cells and vascular endothelial permeability, and this effect might be related to the decreased expression of ROCK protein.

  15. High paracellular nutrient absorption in intact bats is associated with high paracellular permeability in perfused intestinal segments.

    Brun, Antonio; Price, Edwin R; Gontero-Fourcade, Manuel N; Fernandez-Marinone, Guido; Cruz-Neto, Ariovaldo P; Karasov, William H; Caviedes-Vidal, Enrique


    Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular mechanisms. Based on a few previous studies, the capacity for paracellular nutrient absorption seems greater in flying mammals than in nonflying mammals, but there has been little investigation of the mechanisms driving this difference. Therefore, we studied three species each of bats (Artibeus lituratus, Sturnira lilium and Carollia perspicillata) and nonflying mammals (Akodon montensis, Mus musculus and Rattus norvegicus). Using standard pharmacokinetic techniques in intact animals, we confirmed the greater paracellular nutrient absorption in the fliers, comparing one species in each group. Then we conducted in situ intestinal perfusions on individuals of all species. In both approaches, we measured the absorption of 3OMD-glucose, a nonmetabolizable glucose analog absorbed both paracellularly and transcellularly, as well as L-arabinose, which has no mediated transport. Fractional absorption of L-arabinose was three times higher in the bat (S. lilium: 1.2±0.24) than in the rodent (A. montensis: 0.35±0.04), whereas fractional absorption of 3OMD-glucose was complete in both species (1.46±0.4 and 0.97±0.12, respectively). In agreement, bats exhibited two to 12 times higher l-arabinose clearance per square centimeter nominal surface area than rodents in intestinal perfusions. Using L-arabinose, we estimated that the contribution of the paracellular pathway to total glucose absorption was higher in all three bats (109-137%) than in the rodents (13-39%). These findings contribute to an emerging picture that reliance on the paracellular pathway for nutrient absorption is much greater in bats relative to nonflying mammals and that this difference is driven by differences in intestinal permeability to nutrient-sized molecules.

  16. Relaxation and gas drainage boreholes for high performance longwall operations in low permeability coal seams

    Imgrund, Thomas [DMT GmbH und Co. KG, Essen (Germany); Bauer, Frank [Hazemag und EPR GmbH, Duelmen (Germany). Mining


    With an increasing depth of cover, gas emission control and gas outbursts prevention has become an increasingly important issue in coal mining. Deep multi-seam mining often requires operation in an environment characterised by a high gas content and gas pressure. Control of gas related risks has to be realised during heading and close to the longwall by proper risk assessment and flexible drilling schemes. These cover exploration and relaxation drilling, in-seam drilling for pre-drainage and cross measure drilling for drainage of roof and the floor gas emissions. DMT provides comprehensive solutions based on a scientific background. These solutions are engineered considering their technical feasibility. Hazemag Mining offers a large number of complete machinery including tools systems for the implementation of those solutions. (orig.)


    Subramanian, K; Michael Morgan, M


    The Defense Waste Processing Facility (DWPF) is about to process High Level Waste (HLW) Sludge Batch 4 (SB4). This sludge batch is high in alumina and nepheline can crystallize readily depending on the glass composition. Large concentrations of crystallized nepheline can have an adverse effect on HLW glass durability. Several studies have been performed to study the potential for nepheline formation in SB4. The Phase 3 Nepheline Formation study of SB4 glasses examined sixteen different glasses made with four different frits. Melt rate experiments were performed by the Process Science and Engineering Section (PS&E) of the Savannah River National Laboratory (SRNL) using the four frits from the Phase 3 work, plus additional high B2O3/high Fe2O3 frits. Preliminary results from these tests showed the potential for significant improvements in melt rate for SB4 glasses using a higher B2O3-containing frit, particularly Frit 503. The main objective of this study was to investigate the durability of SB4 glasses produced with a high B2O3 frit likely to be recommended for SB4 processing. In addition, a range of waste loadings (WLs) was selected to continue to assess the effectiveness of a nepheline discriminator in predicting concentrations of nepheline crystallization that would be sufficient to influence the durability response of the glass. Five glasses were selected for this study, covering a WL range of 30 to 50 wt% in 5 wt% increments. The Frit 503 glasses were batched and melted. Specimens of each glass were heat-treated to simulate cooling along the centerline of a DWPF-type canister (ccc) to gauge the effects of thermal history on product performance. Visual observations on both quenched and ccc glasses were documented. A representative sample from each glass was submitted to the SRNL Process Science Analytical Laboratory (PSAL) for chemical analysis to confirm that the as-fabricated glasses corresponded to the defined target compositions. The Product Consistency Test

  18. Study of interactions between cells and microbubbles in high speed centrifugation field for biomolecule delivery.

    He, Chuan; Chen, Jie


    Biomolecule delivery has a very wide range of applications in biology and medicine. In this study, a microbubble based delivery method was developed. In a high centrifugation field, cells deform and collide with microbubbles to induce intracellular pathways on cell membranes. As a result, biomaterials can then easily enter cells. Experimental results show that this delivery method can achieve high delivery efficiency. Simulation results showed that cells with more deformed structure experienced higher strain on cell membranes than cells with less deformed structure. The models can help explain how centrifugation affects cell membrane permeability. By controlling cell morphology and its mechanical properties, high biomolecule delivery efficiency can be achieved.

  19. Peanut Allergens Alter Intestinal Barrier Permeability and Tight Junction Localisation in Caco-2 Cell Cultures1

    Dwan B. Price


    Full Text Available Background/Aims: Allergen absorption by epithelia may play an important role in downstream immune responses. Transport mechanisms that can bypass Peyer's patches include transcellular and paracellular transport. The capacity of an allergen to cross via these means can modulate downstream processing of the allergen by the immune system. The aim of this study was to investigate allergen-epithelial interactions of peanut allergens with the human intestinal epithelium. Methods: We achieved this using the human Caco-2 cell culture model, exposed to crude peanut extract. Western and immunofluorescence analysis were used to identify the cellular and molecular changes of peanut extract on the intestinal epithelium. Results: Following exposure of Caco-2 cells to peanut extract, binding of the peanut allergens Ara h 1 and Ara h 2 to the apical cellular membrane and transcytosis across the monolayers were observed. Additionally, the co-localisation of the transmembrane tight junction proteins occludin, JAM-A and claudin-1, with the intracellular adhesion protein ZO-1 was modified. Conclusion: Disruption of Caco-2 barrier integrity through tight junction disruption may enable movement of peanut proteins across the intestinal epithelium. This accounts for peanut's increased allergenicity, compared to other food allergens, and provides an explanation for the potency of peanut allergens in immune response elicitation.

  20. Permeation Dispersal of Treatment Agents for In Situ Remediation in Low Permeability Media: 1. Field Studies in Unconfined Test Cells

    Siegrist, R.L.; Smuin, D.R.; Korte, N.E.; Greene, D.W.; Pickering, D.A.; Lowe, K.S.; Strong-Gunderson, J.


    Chlorocarbons like trichloroethylene (TCE) are common contaminants of concern at US Department of Energy (DOE) facilities and industrial sites across the US and abroad. These contaminants of concern are present in source areas and in soil and ground water plumes as dissolved or sorbed phase constituents as well as dense nonaqueous-phase liquids (DNAPLs). These DNAPL compounds can be released to the environment through a variety of means including leaks in storage tanks and transfer lines, spills during transportation, and land treatment of wastes. When DNAPL compounds are present in low permeability media (LPM) like silt and clay layers or deposits, there are major challenges with assessment of their behavior and implementation of effective in situ remediation technologies. This report describes a field demonstration that was conducted at the Portsmouth Gaseous Diffusion Plant (PORTS) Clean Test Site (CTS) to evaluate the feasibility of permeation and dispersal of reagents into LPM. Various reagents and tracers were injected at seven test cells primarily to evaluate the feasibility of delivery, but also to evaluate the effects of the injected reagents on LPM. The various reagents and tracers were injected at the PORTS CTS using a multi-port injection system (MPIS) developed and provided by Hayward Baker Environmental, Inc.

  1. Increasing the high-frequency magnetic permeability of MnZn ferrite in polyaniline composites by incorporating silver

    Babayan, V.; Kazantseva, N. E.; Sapurina, I.; Moučka, R.; Stejskal, J.; Sáha, P.


    A hybrid composite containing 73 vol% of MnZn ferrite, 21 vol% of polyaniline, and 6 vol% of silver is obtained by oxidative polymerization of aniline with silver nitrate in the presence of ferrite powder. The hybrid composite contains ferrite particles with a size of 40-80 μm coated by an inhomogeneous layer of polyaniline in the conducting emeraldine form. Silver in the form of nano- and submicrometre -size particles is localized both on the surface of ferrite particles and in the bulk of polyaniline coating. The electrical and magnetic properties of the hybrid composite are investigated and compared with the properties of a composite with 71 vol% of MnZn ferrite coated by a conducting polyaniline layer (29 vol%). The hybrid composite containing silver exhibits an increase in the real and imaginary parts of the complex permeability in the radio-frequency band by more than one and a half times compared with those of the MnZn ferrite-polyaniline composite. The high-frequency permittivity of both composites is determined by the properties of core-shell structure: electric properties of shell as well as its composition and uniformity.

  2. Poly-L-arginine-Induced internalization of tight junction proteins increases the paracellular permeability of the Caco-2 cell monolayer to hydrophilic macromolecules.

    Yamaki, Tsutomu; Ohtake, Kazuo; Ichikawa, Keiko; Uchida, Masaki; Uchida, Hiroyuki; Oshima, Shinji; Ohshima, Shinji; Juni, Kazuhiko; Kobayashi, Jun; Morimoto, Yasunori; Natsume, Hideshi


    We investigated whether poly-L-arginine (PLA) enhances the paracellular permeability of the Caco-2 monolayer to hydrophilic macromolecules and clarified the disposition of tight junction (TJ) proteins. The transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC)-dextran (FD-4) permeation were determined after treatment with PLA. TJ proteins were visualized using immunofluorescence microscopy after PLA exposure and depletion, and their expression levels were determined. The barrier function of TJs was also evaluated by measuring the alterations in the TEER and in the localization of TJ proteins. PLA induced an increase in hydrophilic macromolecule, FD-4, permeation through Caco-2 cell monolayers and a decrease in the TEER in a concentration-dependent manner, without any significant impact on the cell viability. This increased paracellular permeability induced by PLA was found to be internalized of claudin-4, ZO-1, tricellulin and mainly occludin from cell-cell junction to the subcellular space. ZO-1 appeared to play an important role in the reconstitution of TJ strand structures following PLA depletion. These results indicate that the PLA led to the internalization of TJ proteins to the subcellular space, subsequently increasing the permeability of the Caco-2 cell monolayer to FD-4 via a paracellular route.

  3. Permeability testing of biomaterial membranes

    Dreesmann, L; Hajosch, R; Nuernberger, J Vaz; Schlosshauer, B [NMI Natural and Medical Sciences Institute at University Tuebingen, Markwiesenstr. 55, D-72770 Reutlingen (Germany); Ahlers, M [GELITA AG, Gammelsbacher Str. 2, D-69412 Eberbach (Germany)], E-mail:


    The permeability characteristics of biomaterials are critical parameters for a variety of implants. To analyse the permeability of membranes made from crosslinked ultrathin gelatin membranes and the transmigration of cells across the membranes, we combined three technical approaches: (1) a two-chamber-based permeability assay, (2) cell culturing with cytochemical analysis and (3) biochemical enzyme electrophoresis (zymography). Based on the diffusion of a coloured marker molecule in conjunction with photometric quantification, permeability data for a gelatin membrane were determined in the presence or absence of gelatin degrading fibroblasts. Cytochemical evaluation after cryosectioning of the membranes was used to ascertain whether fibroblasts had infiltrated the membrane inside. Zymography was used to investigate the potential release of proteases from fibroblasts, which are known to degrade collagen derivatives such as gelatin. Our data show that the diffusion equilibrium of a low molecular weight dye across the selected gelatin membrane is approached after about 6-8 h. Fibroblasts increase the permeability due to cavity formation in the membrane inside without penetrating the membrane for an extended time period (>21 days in vitro). Zymography indicates that cavity formation is most likely due to the secretion of matrix metalloproteinases. In summary, the combination of the depicted methods promises to facilitate a more rational development of biomaterials, because it provides a rapid means of determining permeability characteristics and bridges the gap between descriptive methodology and the mechanistic understanding of permeability alterations due to biological degradation.

  4. Mosquito Saliva Increases Endothelial Permeability in the Skin, Immune Cell Migration, and Dengue Pathogenesis during Antibody-Dependent Enhancement.

    Michael A Schmid


    Full Text Available Dengue remains the most prevalent arthropod-borne viral disease in humans. While probing for blood vessels, Aedes aegypti and Ae. albopictus mosquitoes transmit the four serotypes of dengue virus (DENV1-4 by injecting virus-containing saliva into the skin. Even though arthropod saliva is known to facilitate transmission and modulate host responses to other pathogens, the full impact of mosquito saliva on dengue pathogenesis is still not well understood. Inoculating mice lacking the interferon-α/β receptor intradermally with DENV revealed that mosquito salivary gland extract (SGE exacerbates dengue pathogenesis specifically in the presence of enhancing serotype-cross-reactive antibodies-when individuals already carry an increased risk for severe disease. We further establish that SGE increases viral titers in the skin, boosts antibody-enhanced DENV infection of dendritic cells and macrophages in the dermis, and amplifies dendritic cell migration to skin-draining lymph nodes. We demonstrate that SGE directly disrupts endothelial barrier function in vitro and induces endothelial permeability in vivo in the skin. Finally, we show that surgically removing the site of DENV transmission in the skin after 4 hours rescued mice from disease in the absence of SGE, but no longer prevented lethal antibody-enhanced disease when SGE was present. These results indicate that SGE accelerates the dynamics of dengue pathogenesis after virus transmission in the skin and induces severe antibody-enhanced disease systemically. Our study reveals novel aspects of dengue pathogenesis and suggests that animal models of dengue and pre-clinical testing of dengue vaccines should consider mosquito-derived factors as well as enhancing antibodies.

  5. Mosquito Saliva Increases Endothelial Permeability in the Skin, Immune Cell Migration, and Dengue Pathogenesis during Antibody-Dependent Enhancement.

    Michael A Schmid


    Full Text Available Dengue remains the most prevalent arthropod-borne viral disease in humans. While probing for blood vessels, Aedes aegypti and Ae. albopictus mosquitoes transmit the four serotypes of dengue virus (DENV1-4 by injecting virus-containing saliva into the skin. Even though arthropod saliva is known to facilitate transmission and modulate host responses to other pathogens, the full impact of mosquito saliva on dengue pathogenesis is still not well understood. Inoculating mice lacking the interferon-α/β receptor intradermally with DENV revealed that mosquito salivary gland extract (SGE exacerbates dengue pathogenesis specifically in the presence of enhancing serotype-cross-reactive antibodies-when individuals already carry an increased risk for severe disease. We further establish that SGE increases viral titers in the skin, boosts antibody-enhanced DENV infection of dendritic cells and macrophages in the dermis, and amplifies dendritic cell migration to skin-draining lymph nodes. We demonstrate that SGE directly disrupts endothelial barrier function in vitro and induces endothelial permeability in vivo in the skin. Finally, we show that surgically removing the site of DENV transmission in the skin after 4 hours rescued mice from disease in the absence of SGE, but no longer prevented lethal antibody-enhanced disease when SGE was present. These results indicate that SGE accelerates the dynamics of dengue pathogenesis after virus transmission in the skin and induces severe antibody-enhanced disease systemically. Our study reveals novel aspects of dengue pathogenesis and suggests that animal models of dengue and pre-clinical testing of dengue vaccines should consider mosquito-derived factors as well as enhancing antibodies.

  6. A cell-permeable dominant-negative survivin protein induces apoptosis and sensitizes prostate cancer cells to TNF-α therapy

    Kanwar Jagat R


    Full Text Available Abstract Background Survivin is a member of the inhibitor-of-apoptosis (IAP family which is widely expressed by many different cancers. Overexpression of survivin is associated with drug resistance in cancer cells, and reduced patient survival after chemotherapy and radiotherapy. Agents that antagonize the function of survivin hold promise for treating many forms of cancer. The purpose of this study was to investigate whether a cell-permeable dominant-negative survivin protein would demonstrate bioactivity against prostate and cervical cancer cells grown in three dimensional culture. Results A dominant-negative survivin (C84A protein fused to the cell penetrating peptide poly-arginine (R9 was expressed in E. coli and purified by affinity chromatography. Western blot analysis revealed that dNSurR9-C84A penetrated into 3D-cultured HeLa and DU145 cancer cells, and a cell viability assay revealed it induced cancer cell death. It increased the activities of caspase-9 and caspase-3, and rendered DU145 cells sensitive to TNF-α via by a mechanism involving activation of caspase-8. Conclusions The results demonstrate that antagonism of survivin function triggers the apoptosis of prostate and cervical cancer cells grown in 3D culture. It renders cancer cells sensitive to the proapoptotic affects of TNF-α, suggesting that survivin blocks the extrinsic pathway of apoptosis. Combination of the biologically active dNSurR9-C84A protein or other survivin antagonists with TNF-α therapy warrants consideration as an approach to cancer therapy.

  7. Maintenance measures for preservation and recovery of permeable pavement surface infiltration rate--The effects of street sweeping, vacuum cleaning, high pressure washing, and milling.

    Winston, Ryan J; Al-Rubaei, Ahmed M; Blecken, Godecke T; Viklander, Maria; Hunt, William F


    The surface infiltration rates (SIR) of permeable pavements decline with time as sediment and debris clog pore spaces. Effective maintenance techniques are needed to ensure the hydraulic functionality and water quality benefits of this stormwater control. Eight different small-scale and full-scale maintenance techniques aimed at recovering pavement permeability were evaluated at ten different permeable pavement sites in the USA and Sweden. Maintenance techniques included manual removal of the upper 2 cm of fill material, mechanical street sweeping, regenerative-air street sweeping, vacuum street sweeping, hand-held vacuuming, high pressure washing, and milling of porous asphalt. The removal of the upper 2 cm of clogging material did not significantly improve the SIR of concrete grid paves (CGP) and permeable interlocking concrete pavers (PICP) due to the inclusion of fines in the joint and bedding stone during construction, suggesting routine maintenance cannot overcome improper construction. For porous asphalt maintenance, industrial hand-held vacuum cleaning, pressure washing, and milling were increasingly successful at recovering the SIR. Milling to a depth of 2.5 cm nearly restored the SIR for a 21-year old porous asphalt pavement to like-new conditions. For PICP, street sweepers employing suction were shown to be preferable to mechanical sweepers; additionally, maintenance efforts may become more intensive over time to maintain a threshold SIR, as maintenance was not 100% effective at removing clogging material.

  8. Silver nanoparticles inhibit VEGF-and IL-1β-induced vascular permeability via Src dependent pathway in porcine retinal endothelial cells

    Park Jongsun


    Full Text Available Abstract The aim of this study is to determine the effects of silver nanoparticles (Ag-NP on vascular endothelial growth factor (VEGF-and interleukin-1 beta (IL-1β-induced vascular permeability, and to detect the underlying signaling mechanisms involved in endothelial cells. Porcine retinal endothelial cells (PRECs were exposed to VEGF, IL-1β and Ag-NP at different combinations and endothelial cell permeability was analyzed by measuring the flux of RITC-dextran across the PRECs monolayer. We found that VEGF and IL-1β increase flux of dextran across a PRECs monolayer, and Ag-NP block solute flux induced by both VEGF and IL-1β. To explore the signalling pathway involved VEGF- and IL-1β-induced endothelial alteration, PRECs were treated with Src inhibitor PP2 prior to VEGF and IL-1β treatment, and the effects were recorded. Further, to clarify the possible involvement of the Src pathways in endothelial cell permeability, plasmid encoding dominant negative(DN and constitutively active(CA form of Src kinases were transfected into PRECs, 24 h prior to VEGF and IL-1β exposure and the effects were recorded. Overexpression of DN Src blocked both VEGF-and IL-1β-induced permeability, while overexpression of CA Src rescues the inhibitory action of Ag-NP in the presence or absence of VEGF and IL-1β. Further, an in vitro kinase assay was performed to identify the presence of the Src phosphorylation at Y419. We report that VEGF and IL-1β-stimulate endothelial permeability via Src dependent pathway by increasing the Src phosphorylation and Ag-NP block the VEGF-and IL-1β-induced Src phosphorylation at Y419. These results demonstrate that Ag-NP may inhibit the VEGF-and IL-1β-induced permeability through inactivation of Src kinase pathway and this pathway may represent a potential therapeutic target to inhibit the ocular diseases such as diabetic retinopathy.

  9. Relationship between Passive Permeability and Molecular Polarity Using Block Relevance Analysis.

    Goetz, Gilles H; Shalaeva, Marina; Caron, Giulia; Ermondi, Giuseppe; Philippe, Laurence


    EPSA is an experimental descriptor of molecular polarity obtained from chromatographic retention in supercritical fluid chromatography (SFC) systems, previously shown by Goetz et al. to correlate with passive permeability of cyclic peptides. The present study focuses on EPSA in relation to passive permeability of small molecules. We applied block relevance (BR) analysis to interpret the relative significance of mechanistic forces prevailing in EPSA. The BR analysis is a computational tool that allows the interpretation of the balance of intermolecular interactions governing systems such as the aforementioned chromatographic retention in EPSA. EPSA and passive permeability determined by Ralph Russ canine kidney cells (RRCK) or low efflux Madin Darby canine kidney cells (MDCK-LE) and human epithelial colorectal adenocarcinoma cells (Caco-2), studied on a data set of commercial drugs, indicated that EPSA is relevant in describing permeability of hydrophilic drugs (CLogP value of EPSA < 100 significantly increases the likelihood of high permeability.

  10. Correlation of vascular endothelial growth factor to permeability of blood-brain barrier and brain edema during high-altitude exposure

    Qiquan Zhou; Chang'e Liu; Jing Wang; Yunli Wang; Bo Zhou


    BACKGROUND:Many studies have evaluated the role of vascular endothelial growth factor (VEGF) in traumatic brain edema and hemorrhagic brain edema.OBJECTIVE:To observe the effects of VEGF expression on permeability of the blood-brain barrier (BBB) during high-altitude and hypoxia exposure,and to investigate the correlation between VEGF expression and BBB permeability with regard to Evans blue staining and brain edema during high-altitude exposure.DESIGN,TIME AND SETTING:The randomized,controlled,animal study was performed at the Tanggula Etape,Central Laboratory of Chengdu Medical College,and Central Laboratory of General Hospital of Chengdu Military Area Command of Chinese PLA,China,from July 2003 to November 2004.MATERIALS:Quantitative RT-PCR kit (Sigma,USA),VEGF ELISA kit (Biosource,USA),and Evans blue (Jingchun,China) were acquired for this study.METHODS:A total of 180 Wistar rats were equally and randomly assigned to 15 groups:low-altitude (500 m),middle-altitude (2 880 m),high-altitude (4 200 m),super-high-altitude (5 000 m),1,3,5,7,9,11,13,15,17,19,and 21 days of super high-altitude exposure.Wistar rats were exposed to various altitude gradients to establish a hypoxia model.MAIN OUTCOME MEASURES:Brain water content was calculated according to the wet-to-dry weight ratio.BBB permeability to Evans blue was determined by colorimetric method.VEGF mRNA and protein levels in brain tissues were detected using RT-PCR and double-antibody sandwich ELISA.RESULTS:Brain water content,BBB permeability to Evans blue,and VEGF mRNA and protein levels in brain tissues increased with increasing altitude and prolonged exposure to altitude.The greatest increase was determined on day 9 upon ascending 5 000 m.Simultaneously,VEGF expression positively correlated to BBB permeability of Evans blue and brain water content (r=0.975,0.917,P<0.01).CONCLUSION:Increased VEGF protein and mRNA expression was responsible for increased BBB permeability,which may be an important mechanism

  11. Cell-permeable and plasma-stable peptidomimetic inhibitors of the postsynaptic density-95/N-methyl-D-aspartate receptor interaction

    Bach, Anders*; Eildal, Jonas Nii Nortey*; Stuhr-Hansen, Nicolai


    The protein--protein interaction between the NMDA receptor and its intracellular scaffolding protein, PSD-95, is a potential target for treating ischemic brain diseases, neuropathic pain, and Alzheimer's disease. We have previously demonstrated that N-alkylated tetrapeptides are potent inhibitors...... to identification of N-cyclohexylethyl-ETA(S)V (54) as the most potent, plasma-stable and cell-permeable inhibitor, which is a promising tool in unraveling the therapeutic potential of the PSD-95/NMDA receptor interaction....

  12. Separation of intact proteins on γ-ray-induced polymethacrylate monolithic columns: A highly permeable stationary phase with high peak capacity for capillary high-performance liquid chromatography with high-resolution mass spectrometry.

    Simone, Patrizia; Pierri, Giuseppe; Foglia, Patrizia; Gasparrini, Francesca; Mazzoccanti, Giulia; Capriotti, Anna Laura; Ursini, Ornella; Ciogli, Alessia; Laganà, Aldo


    Polymethacrylate-based monolithic capillary columns, prepared by γ-radiation-induced polymerization, were used to optimize the experimental conditions (nature of the organic modifiers, the content of trifluoroacetic acid and the column temperature) in the separation of nine standard proteins with different hydrophobicities and a wide range of molecular weights. Because of the excellent permeability of the monolithic columns, an ion-pair reversed-phase capillary liquid chromatography with high-resolution mass spectrometry method has been developed by coupling the column directly to the mass spectrometer without a flow-split and using a standard electrospray interface. Additionally, the high working flow and concomitant high efficiency of these columns allowed us to employ a longer column (up to 50 cm) and achieve a peak capacity value superior to 1000. This work is motivated by the need to develop new materials for high-resolution chromatographic separation that combine chemical stability at elevated temperatures (up to 75°C) and a broad pH range, with a high peak capacity value. The advantage of the γ-ray-induced monolithic column lies in the batch-to-batch reproducibility and long-term high-temperature stability. Their proven high loading capacity, recovery, good selectivity and high permeability, moreover, compared well with that of a commercially available poly(styrene-divinylbenzene) monolithic column, which confirms that such monolithic supports might facilitate analysis in proteomics.

  13. Uncaria tomentosa alkaloidal fraction reduces paracellular permeability, IL-8 and NS1 production on human microvascular endothelial cells infected with dengue virus.

    Lima-Junior, Raimundo Sousa; Mello, Cintia da Silva; Siani, Antonio Carlos; Valente, Ligia M Marino; Kubelka, Claire Fernandes


    Dengue is the major Arbovirus in the world, annually causing morbidity and death. Severe dengue is associated with changes in the endothelial barrier function due to the production of inflammatory mediators by immune cells and by the endothelium. Dengue virus (DENV) replicates efficiently in human endothelial cells in vitro and elicits immune responses resulting in endothelial permeability. Uncaria tomentosa (Willd.) DC.(Rubiaceae), known as cat's claw, has been used in folk medicine for the treatment of a wide-array of symptoms, and several scientific studies reported its antiviral, immunomodulatory, anti-inflammatory and antioxidant properties. Here we infected a human lineage of dermal microvascular endothelial cells (HMEC-1) with DENV-2 and treated it with an alkaloidal fraction from U. tomentosa bark (AFUT). We showed antiviral and immunomodulatory activities of U. tomentosa by determining the NS1 antigen and IL-8 in supernatant of DENV-2 infected HMEC-1. Furthermore, by measurement of transendothelial electrical resistance (TEER) we demonstrated, for the first time, that a plant derivative contributed to the reduction of paracellular permeability in DENV-2 infected HMEC-1. We also showed that IL-8 contributed significantly to the induction of permeability. Although further investigations should be conducted before a new drug can be suggested, our in vitro data support evidence that AFUT could be potentially useful in developing a treatment for severe dengue.

  14. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells

    Branco, Patrícia; Monteiro Lomba Viana, Tiago; Albergaria, Helena


    Hi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0pHindicated that this subpopulation was under a viable but non-culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals...... on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability...... by a classical plating method. Results showed that the average pHi of H. guilliermondii cells dropped from 6.5 (healthy cells) to 5.4 (damaged cells) after 20min of exposure to inhibitory concentrations of AMPs, and after 24h 77.0% of the cells completely lost their pH gradient (∆pH=pHi-pHext). After 24h...

  15. Permeability Increase with Deep Borehole Controlled Pre-cracking Blasting Technology in Low Permeability and High Gassy Seam"%低透气性高瓦斯煤层深孔控制预裂爆破增透技术

    弓美疆; 池鹏; 张明杰


    为了解决低透气性高瓦斯煤层难抽采问题,运用RFPA2D数值模拟软件,建立了2种深孔控制预裂爆破力学模型,通过对模拟结果进行对比分析,得出一套适用于鹤煤八矿底板抽采的爆破参数,并进行了现场试验,爆破后煤层透气性系数平均值达到了2.23 m2/(MPa2.d),比爆破前测定的煤层原始透气性平均值提高了近3.05倍,说明该技术可以明显提高钻孔周围煤体裂隙发育程度,煤体透气性增加,达到了提高煤层瓦斯抽采量和抽采率的目的。%In order to solve the difficult gas drainage problem in low permeability and high gassy seam, the RFPA2D numerical simulation software was applied to establish two deep borehole controlled pre-cracking blasting mechanics models. With the comparison and analysis on the simulation results, a set of blasting parameters obtained which were suitable for floor gas drainage in No. 8 mine of Hegang Coal Mining and the site trial with the parameters was conducted. The results showed that after the blasting, the average value of the seam per- meability coefficient reached at 2. 23 m2/ ( MPa2 ·d) and 3.05 times higher than the average value of the seam in-situ permeability be- fore the blasting conducted. Thus the technology could obviously improve the cracking development degree of coal around the borehole and could improve the permeability of the seam. The target to improve the seam gas drainage value and the gas drainage rate could be reached.

  16. TNFα signals via p66(Shc to induce E-Selectin, promote leukocyte transmigration and enhance permeability in human endothelial cells.

    Luigi Laviola

    Full Text Available Endothelial cells participate in inflammatory events leading to atherogenesis by regulating endothelial cell permeability via the expression of VE-Cadherin and β-catenin and leukocyte recruitment via the expression of E-Selectins and other adhesion molecules. The protein p66(Shc acts as a sensor/inducer of oxidative stress and may promote vascular dysfunction. The objective of this study was to investigate the role of p66(Shc in tumor necrosis factor TNFα-induced E-Selectin expression and function in human umbilical vein endothelial cells (HUVEC. Exposure of HUVEC to 50 ng/ml TNFα resulted in increased leukocyte transmigration through the endothelial monolayer and E-Selectin expression, in association with augmented phosphorylation of both p66(Shc on Ser(36 and the stress kinase c-Jun NH2-terminal protein kinase (JNK-1/2, and higher intracellular reactive oxygen species (ROS levels. Overexpression of p66(Shc in HUVEC resulted in enhanced p66(Shc phosphorylation on Ser(36, increased ROS and E-Selectin levels, and amplified endothelial cell permeability and leukocyte transmigration through the HUVEC monolayer. Conversely, overexpression of a phosphorylation-defective p66(Shc protein, in which Ser(36 was replaced by Ala, did not augment ROS and E-Selectin levels, nor modify cell permeability or leukocyte transmigration beyond those found in wild-type cells. Moreover, siRNA-mediated silencing of p66(Shc resulted in marked reduction of E-Selectin expression and leukocyte transmigration. In conclusion, p66(Shc acts as a novel intermediate in the TNFα pathway mediating endothelial dysfunction, and its action requires JNK-dependent phosphorylation of p66(Shc on Ser(36.

  17. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells.

    Branco, Patrícia; Viana, Tiago; Albergaria, Helena; Arneborg, Nils


    Saccharomyces cerevisiae produces antimicrobial peptides (AMPs) during alcoholic fermentation that are active against several wine-related yeasts (e.g. Hanseniaspora guilliermondii) and bacteria (e.g. Oenococcus oeni). In the present study, the physiological changes induced by those AMPs on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability by a classical plating method. Results showed that the average pHi of H. guilliermondii cells dropped from 6.5 (healthy cells) to 5.4 (damaged cells) after 20 min of exposure to inhibitory concentrations of AMPs, and after 24 h 77.0% of the cells completely lost their pH gradient (∆pH=pHi-pHext). After 24h of exposure to AMPs, PI-stained (dead) cells increased from 0% to 77.7% and the number of viable cells fell from 1×10(5) to 10 CFU/ml. This means that virtually all cells (99.99%) became unculturable but that a sub-population of 22.3% of the cells remained viable (as determined by PI staining). Besides, pHi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0pH<3). Taken together, these results indicated that this subpopulation was under a viable but non-culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals that these AMPs compromise the plasma membrane integrity (and possibly also the vacuole membrane) of H. guilliermondii cells, disturbing the pHi homeostasis and inducing a loss of culturability. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Evaluation of different toxicity assays applied to proliferating cells and to stratified epithelium in relation to permeability enhancement with glycocholate

    Eirheim, Heidi Ugelstad; Bundgaard, Christoffer; Nielsen, Hanne Mørck


    exposed to different GC concentrations for 4 h. The MTS/PMS assay and neutral red (NR) retention were performed along with quantitation of ATP, lactate dehydrogenase (LDH) and extracellular protein. The toxicity was calculated as the IC50 value relative to the control. Increase in 3H-mannitol permeability...

  19. Loss of occludin expression and impairment of blood-testis barrier permeability in rats with autoimmune orchitis: effect of interleukin 6 on Sertoli cell tight junctions.

    Pérez, Cecilia Valeria; Sobarzo, Cristian Marcelo; Jacobo, Patricia Verónica; Pellizzari, Eliana Herminia; Cigorraga, Selva Beatriz; Denduchis, Berta; Lustig, Livia


    Inflammation of the male reproductive tract is accepted as being an important etiological factor of infertility. Experimental autoimmune orchitis (EAO) is characterized by interstitial lymphomononuclear cell infiltration and severe damage of seminiferous tubules with germ cells that undergo apoptosis and sloughing. Because the blood-testis barrier (BTB) is relevant for the protection of haploid germ cells against immune attack, the aim of this study was to analyze BTB permeability and the expression of tight junction proteins (occludin, claudin 11, and tight junction protein 1 [TJP1]) in rats during development of autoimmune orchitis. The role of IL6 as modulator of tight junction dynamics was also evaluated because intratesticular content of this cytokine is increased in EAO rats. Orchitis was induced in Sprague-Dawley adult rats by active immunization with testicular homogenate and adjuvants. Control rats (C) were injected with saline solution and adjuvants. Untreated (N) rats were also studied. Concomitant with early signs of germ cell sloughing, a reduced expression of occludin and delocalization of claudin 11 and TJP1 were detected in the testes of rats with EAO compared to C and N groups. The use of tracers showed increased BTB permeability in EAO rats. Intratesticular injection of IL6 induced focal testicular inflammation, which is associated with damaged seminiferous tubules. Rat Sertoli cells cultured in the presence of IL6 exhibited a redistribution of tight junction proteins and reduced transepithelial electrical resistance. These data indicate the possibility that IL6 might be involved in the downregulation of occludin expression and in the modulation of BTB permeability that occur in rats undergoing autoimmune orchitis.

  20. Conductivity and methanol permeability of Nafion-zirconium phosphate composite membranes containing high aspect ratio filler particles

    Bagnasco, G.; Micoli, L.; Turco, M. [Dipartimento di Ingegneria Chimica, Universita di Napoli Federico II, P.le V. Tecchio 80, 80125 - Napoli (Italy); Donnadio, A.; Pica, M.; Sganappa, M. [Dipartimento di Chimica, Universita di Perugia, via Elce di Sotto 8, 06123 - Perugia (Italy); Casciola, M.


    Gels of exfoliated {alpha}-zirconium phosphate (ZrP{sub exf}) in dimethylformamide (DMF) were used to prepare Nafion/ZrP{sub exf} composite membranes with filler loadings up to 7 wt.-% by casting mixtures of Nafion 1100 solutions in DMF and suitable amounts of 2 wt.-% ZrP gels in DMF. TEM pictures showed that the ZrP{sub exf} particles had aspect ratio of at least 20. All samples were characterised by methanol permeability (P) and through-plane ({sigma}{sub thp}) and in-plane ({sigma}{sub inp}) conductivity measurements at 40 C and 100% RH. The methanol permeability of Nafion membranes containing in situ grown ZrP particles with low aspect ratio (Nafion/ZrP{sub isg}) was also determined. The methanol permeability and the swelling behaviour of the composite membranes turned out to be strongly dependent on the filler morphology. As a general trend, both permeability and swelling decreased according to the sequence: Nafion/ZrP{sub isg} > Nafion > Nafion/ZrP{sub exf}. The maximum selectivity ({sigma}{sub thp}/P = 1.4 x 10{sup 5} S cm{sup -3} s) was found for the membrane filled with 1 wt.-% ZrP{sub exf}: this value is seven times higher than that of Nafion. For the Nafion/ZrP{sub exf} membranes, the ratio {sigma}{sub inp}/{sigma}{sub thp} increases with the filler loading, thus indicating that the preferred orientation of the ZrP sheets is parallel to the membrane surface. (Abstract Copyright [2009], Wiley Periodicals, Inc.)

  1. Competitive inhibition of survivin using a cell-permeable recombinant protein induces cancer-specific apoptosis in colon cancer model

    Roy K


    Full Text Available Kislay Roy,1 Rupinder K Kanwar,1 Subramanian Krishnakumar,2,3 Chun Hei Antonio Cheung,4 Jagat R Kanwar1 1Nanomedicine-Laboratory of Immunology and Molecular Biomedical Research (NLIMBR, Molecular and Medical Research (MMR Strategic Research Centre, School of Medicine (SoM, Faculty of Health, Deakin University, Waurn Ponds, VIC, Australia; 2Department of Nanobiotechnology, 3Larsen & Toubro (L&T Ocular Pathology Department, Vision Research Foundation, Kamalnayan Bajaj Institute for Research in Vision and Ophthalmology, Chennai, India; 4Department of Pharmacology, College of Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China Abstract: Endogenous survivin expression has been related with cancer survival, drug resistance, and metastasis. Therapies targeting survivin have been shown to significantly inhibit tumor growth and recurrence. We found out that a cell-permeable dominant negative survivin (SurR9-C84A, referred to as SR9 competitively inhibited endogenous survivin and blocked the cell cycle at the G1/S phase. Nanoencapsulation in mucoadhesive chitosan nanoparticles (CHNP substantially increased the bioavailability and serum stability of SR9. The mechanism of nanoparticle uptake was studied extensively in vitro and in ex vivo models. Our results confirmed that CHNP–SR9 protected primary cells from autophagy and successfully induced tumor-specific apoptosis via both extrinsic and intrinsic apoptotic pathways. CHNP–SR9 significantly reduced the tumor spheroid size (three-dimensional model by nearly 7-fold. Effects of SR9 and CHNP–SR9 were studied on 35 key molecules involved in the apoptotic pathway. Highly significant (4.26-fold, P≤0.005 reduction in tumor volume was observed using an in vivo mouse xenograft colon cancer model. It was also observed that net apoptotic (6.25-fold, P≤0.005 and necrotic indexes (3.5-fold, P≤0.05 were comparatively higher in CHNP–SR9 when compared to void CHNP and CHNP–SR9

  2. The effects of acute oral glutamine supplementation on exercise-induced gastrointestinal permeability and heat shock protein expression in peripheral blood mononuclear cells.

    Zuhl, Micah; Dokladny, Karol; Mermier, Christine; Schneider, Suzanne; Salgado, Roy; Moseley, Pope


    Chronic glutamine supplementation reduces exercise-induced intestinal permeability and inhibits the NF-κB pro-inflammatory pathway in human peripheral blood mononuclear cells. These effects were correlated with activation of HSP70. The purpose of this paper is to test if an acute dose of oral glutamine prior to exercise reduces intestinal permeability along with activation of the heat shock response leading to inhibition of pro-inflammatory markers. Physically active subjects (N = 7) completed baseline and exercise intestinal permeability tests, determined by the percent ratio of urinary lactulose (5 g) to rhamnose (2 g). Exercise included two 60-min treadmill runs at 70 % of VO2max at 30 °C after ingestion of glutamine (Gln) or placebo (Pla). Plasma levels of endotoxin and TNF-α, along with peripheral blood mononuclear cell (PBMC) protein expression of HSP70 and IκBα, were measured pre- and post-exercise and 2 and 4 h post-exercise. Permeability increased in the Pla trial compared to that at rest (0.06 ± 0.01 vs. 0.02 ± 0.018) and did not increase in the Gln trial. Plasma endotoxin was lower at the 4-h time point in the Gln vs. 4 h in the Pla (6.715 ± 0.046 pg/ml vs. 7.952 ± 1.11 pg/ml). TNF-α was lower 4 h post-exercise in the Gln vs. Pla (1.64 ± 0.09 pg/ml vs. 1.87 ± 0.12 pg/ml). PBMC expression of IkBα was higher 4 h post-exercise in the Gln vs. 4 h in the Pla (1.29 ± 0.43 vs. 0.8892 ± 0.040). HSP70 was higher pre-exercise and 2 h post-exercise in the Gln vs. Pla (1.35 ± 0.21 vs. 1.000 ± 0.000 and 1.65 ± 0.21 vs. 1.27 ± 0.40). Acute oral glutamine supplementation prevents an exercise-induced rise in intestinal permeability and suppresses NF-κB activation in peripheral blood mononuclear cells.

  3. Design, synthesis, and biological activities of novel hexahydropyrazino[1,2-a]indole derivatives as potent inhibitors of apoptosis (IAP) proteins antagonists with improved membrane permeability across MDR1 expressing cells.

    Shiokawa, Zenyu; Hashimoto, Kentaro; Saito, Bunnai; Oguro, Yuya; Sumi, Hiroyuki; Yabuki, Masato; Yoshimatsu, Mie; Kosugi, Yohei; Debori, Yasuyuki; Morishita, Nao; Dougan, Douglas R; Snell, Gyorgy P; Yoshida, Sei; Ishikawa, Tomoyasu


    We previously reported octahydropyrrolo[1,2-a]pyrazine derivative 2 (T-3256336) as a potent antagonist for inhibitors of apoptosis (IAP) proteins. Because compound 2 was susceptible to MDR1 mediated efflux, we developed another scaffold, hexahydropyrazino[1,2-a]indole, using structure-based drug design. The fused benzene ring of this scaffold was aimed at increasing the lipophilicity and decreasing the basicity of the scaffold to improve the membrane permeability across MDR1 expressing cells. We established a chiral pool synthetic route to yield the desired tricyclic chiral isomers. Chemical modification of the core scaffold led to a representative compound 50, which showed strong inhibition of IAP binding (X chromosome-linked IAP [XIAP]: IC50 23 nM and cellular IAP [cIAP]: IC50 1.1 nM) and cell growth inhibition (MDA-MB-231 cells: GI50 2.8 nM) with high permeability and low potential of MDR1 substrate.

  4. Effects of 5-hydroxymethyl-2-furfural on the volume and membrane permeability of red blood cells from patients with sickle cell disease.

    Hannemann, Anke; Cytlak, Urszula M; Rees, David C; Tewari, Sanjay; Gibson, John S


    The heterocyclic aldehyde 5-hydroxymethyl-2-furfural (5HMF) interacts allosterically with the abnormal form of haemoglobin (Hb), HbS, in red blood cells (RBCs) from patients with sickle cell disease (SCD), thereby increasing oxygen affinity and decreasing HbS polymerization and RBC sickling during hypoxia. We hypothesized that should 5HMF also inhibit the main cation pathways implicated in the dehydration of RBCs from SCD patients - the deoxygenation-induced cation pathway (Psickle), the Ca(2+)-activated K(+) channel (the Gardos channel) and the K(+)-Cl(-) cotransporter (KCC) - it would have a synergistic effect in protection against sickling, directly through interacting with HbS, and indirectly through maintaining hydration and reducing [HbS]. This study was therefore designed to investigate the effects of 5HMF on RBC volume and K(+) permeability in vitro. 5HMF markedly reduced the deoxygenation-induced dehydration of RBCs whether in response to maintained deoxygenation or to cyclical deoxygenation/re-oxygenation. 5HMF was found to inhibit Psickle, an effect which correlated with its effects on sickling. Deoxygenation-induced activation of the Gardos channel and exposure of phosphatidylserine were also inhibited, probably indirectly via reduced entry of Ca(2+) through the Psickle pathway. Effects of 5HMF on KCC were more modest with a slight inhibition in N-ethylmaleimide (NEM, 1 mm)-treated RBCs and stimulation in RBCs untreated with NEM. These findings support the hypothesis that 5HMF may also be beneficial through effects on RBC ion and water homeostasis.

  5. Cysteamine-based cell-permeable Zn(2+)-specific molecular bioimaging materials: from animal to plant cells.

    Sinha, Sougata; Dey, Gourab; Kumar, Sunil; Mathew, Jomon; Mukherjee, Trinetra; Mukherjee, Subhrakanti; Ghosh, Subrata


    Structure-interaction/fluorescence relationship studies led to the development of a small chemical library of Zn(2+)-specific cysteamine-based molecular probes. The probe L5 with higher excitation/emission wavelengths, which absorbs in the visible region and emits in the green, was chosen as a model imaging material for biological studies. After successful imaging of intracellular zinc in four different kinds of cells including living organisms, plant, and animal cells, in vivo imaging potential of L5 was evaluated using plant systems. In vivo imaging of translocation of zinc through the stem of a small herb with a transparent stem, Peperomia pellucida, confirmed the stability of L5 inside biological systems and the suitability of L5 for real-time analysis. Similarly, fluorescence imaging of zinc in gram sprouts revealed the efficacy of the probe in the detection and localization of zinc in cereal crops. This imaging technique will help in knowing the efficiency of various techniques used for zinc enrichment of cereal crops. Computational analyses were carried out to better understand the structure, the formation of probe-Zn(2+) complexes, and the emission properties of these complexes.

  6. High-frequency magnetic permeability of single- and multilayered (Co41Fe39B20) x (SiO2)100- x nanocomposites

    Tarasova, O. S.; Sitnikov, A. V.; Kalinin, Yu. E.; Starostenko, S. N.; Granovskii, A. B.


    Thin film single-layered (Co41Fe39B20) x (SiO2)100- x nanocomposites at x = 30-80 at % and multilayered nanocomposites composed of 176 pairs of [(Co41Fe39B20)60(SiO2)40]/[(Co41Fe39B20)60(SiO2)40 + O2] have been prepared via ion-beam sputtering of the complex target. The concentration dependences of the magnetic permeability of single-layered films at a frequency of 50 MHz are characterized by maximum losses near x = 60 at %, whereas the percolation threshold with respect to the electric conductivity is x = 50 at %. The high-frequency magnetic permeability of films has been measured by the resonator method in the frequency range of 0.1—10 GHz. As is shown, while the single-layer film passes to the multilayered structure, the ferromagnetic resonance frequency shifts from 1.5 to 2.5 GHz, and the imaginary part of the magnetic permeability attains 200 that is presumably due to the inhibition of the perpendicular magnetic anisotropy component.

  7. Non-Darcian flow in low-permeability media: key issues related to geological disposal of high-level nuclear waste in shale formations

    Liu, Hui-Hai


    In clay or other low-permeability media, water flow becomes non-Darcian and characterized by the non-linear relationship between water flux and hydraulic gradient. This work is devoted to addressing a number of key issues related to geological disposal of high-level nuclear waste in clay/shale formations. It is demonstrated that water flow velocity in the damaged zone (often considered as a potential preferential advection paths in a repository) surrounding the tunnel is extremely small, as a result of non-Darcian flow behavior, such that solute transport is dominated by diffusion, rather than advection. The finding is also consistent with the often-observed existence of persistent abnormal pressures in shale formations. While relative permeability is the key parameter for modeling the unsaturated flow process, without incorporating non-Darcian flow behavior, significant errors can occur in the determination of relative permeability values from traditional measurement methods. An approach for dealing with temperature impact on non-Darcian flow and a formulation to calculate non-Darcian water flux in an anisotropic medium are presented, taking into consideration that a geological repository is subject to temperature evolution in the near field as a result of heat generated by nuclear waste, and that shale formations are generally anisotropic.

  8. Anisotropic permeability in deterministic lateral displacement arrays

    Vernekar, Rohan; Loutherback, Kevin; Morton, Keith; Inglis, David


    We investigate anisotropic permeability of microfluidic deterministic lateral displacement (DLD) arrays. A DLD array can achieve high-resolution bimodal size-based separation of micro-particles, including bioparticles such as cells. Correct operation requires that the fluid flow remains at a fixed angle with respect to the periodic obstacle array. We show via experiments and lattice-Boltzmann simulations that subtle array design features cause anisotropic permeability. The anisotropy, which indicates the array's intrinsic tendency to induce an undesired lateral pressure gradient, can lead to off-axis flows and therefore local changes in the critical separation size. Thus, particle trajectories can become unpredictable and the device useless for the desired separation duty. We show that for circular posts the rotated-square layout, unlike the parallelogram layout, does not suffer from anisotropy and is the preferred geometry. Furthermore, anisotropy becomes severe for arrays with unequal axial and lateral gaps...

  9. High-frequency permeability of electroplated CoNiFe and CoNiFe-C alloys

    Rhen, Fernando M.F.; McCloskey, Paul; O' Donnell, Terence [Tyndall National Institute, Lee Maltings, Prospect Row, Cork (Ireland); Roy, Saibal [Tyndall National Institute, Lee Maltings, Prospect Row, Cork (Ireland)], E-mail:


    We have investigated CoNiFe and CoNiFe-C electrodeposited by pulse reverse plating (PRP) and direct current (DC) techniques. CoNiFe(PRP) films with composition Co{sub 59.4}Fe{sub 27.7}Ni{sub 12.8} show coercivity of 95 A m{sup -1} (1.2 Oe) and magnetization saturation flux ({mu}{sub 0}M{sub s}) of 1.8 T. Resistivity of CoNiFe (PRP) is about 24 {mu}{omega} cm and permeability remains almost constant {mu}{sub r}' {approx}475 up to 30 MHz with a quality factor (Q) larger than 10. Additionally, the permeability spectra analysis shows that CoNiFe exhibits a classical eddy current loss at zero bias field and ferromagnetic resonance (FMR) when biased with 0.05 T. Furthermore, a crossover between eddy current and FMR loss is observed for CoNiFe-PRP when baised with 0.05 T. DC and PRP plated CoNiFe-C, which have resistivity and permeability of 85, 38 {mu}{omega} cm, {mu}{sub r}'=165 and 35 with Q>10 up to 320 MHz, respectively, showed only ferromagnetic resonance losses. The ferromagnetic resonance peaks in CoNiFe and CoNiFe-C are broad and resembles a Gaussian distribution of FMR frequencies. The incorporation of C to CoNiFe reduces eddy current loss, but also reduces the FMR frequency.

  10. Apoptosis Induced by Microbubble-Assisted Acoustic Cavitation in K562 Cells: The Predominant Role of the Cyclosporin A-Dependent Mitochondrial Permeability Transition Pore.

    Zhao, Lu; Feng, Yi; Shi, Aiwei; Zong, Yujin; Wan, Mingxi


    Acoustic cavitation of microbubbles has been described as inducing tumor cell apoptosis that is partly associated with mitochondrial dysfunction; however, the exact mechanisms have not been fully characterized. Here, low-intensity pulsed ultrasound (1 MHz, 0.3-MPa peak negative pressure, 10% duty cycle and 1-kHz pulse repetition frequency) was applied to K562 chronic myelogenous leukemia cells for 1 min with 10% (v/v) SonoVue microbubbles. After ultrasound exposure, the apoptotic index was determined by flow cytometry with annexin V-fluorescein isothiocyanate/propidium iodide. In addition, mitochondrial membrane potential (ΔΨm) was determined with the JC-1 assay. Translocation of apoptosis-associated protein cytochrome c was evaluated by Western blotting. We found that microbubble-assisted acoustic cavitation can increase the cellular apoptotic index, mitochondrial depolarization and cytochrome c release in K562 cells, compared with ultrasound treatment alone. Furthermore, mitochondrial dysfunction and apoptosis were significantly inhibited by cyclosporin A, a classic inhibitor of the mitochondrial permeability transition pore; however, the inhibitor of Bax protein, Bax-inhibiting peptide, could not suppress these effects. Our results suggest that mitochondrial permeability transition pore opening is involved in mitochondrial dysfunction after exposure to microbubble-assisted acoustic cavitation. Moreover, the release of cytochrome c from the mitochondria is dependent on cyclosporin A-sensitive mitochondrial permeability transition pore opening, but not formation of the Bax-voltage dependent anion channel complex or Bax oligomeric pores. These data provide more insight into the mechanisms underlying mitochondrial dysfunction induced by acoustic cavitation and can be used as a basis for therapy. Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  11. Ablation of CD11c(hi) dendritic cells exacerbates Japanese encephalitis by regulating blood-brain barrier permeability and altering tight junction/adhesion molecules.

    Kim, Jin Hyoung; Hossain, Ferdaus Mohd Altaf; Patil, Ajit Mahadev; Choi, Jin Young; Kim, Seong Bum; Uyangaa, Erdenebelig; Park, Sang-Youel; Lee, John-Hwa; Kim, Bumseok; Kim, Koanhoi; Eo, Seong Kug


    Japanese encephalitis (JE), characterized by extensive neuroinflammation following infection with neurotropic JE virus (JEV), is becoming a leading cause of viral encephalitis due to rapid changes in climate and demography. The blood-brain barrier (BBB) plays an important role in restricting neuroinvasion of peripheral leukocytes and virus, thereby regulating the progression of viral encephalitis. In this study, we explored the role of CD11c(hi) dendritic cells (DCs) in regulating BBB integrity and JE progression using a conditional depletion model of CD11c(hi) DCs. Transient ablation of CD11c(hi) DCs resulted in markedly increased susceptibility to JE progression along with highly increased neuro-invasion of JEV. In addition, exacerbated JE progression in CD11c(hi) DC-ablated hosts was closely associated with increased expression of proinflammatory cytokines (IFN-β, IL-6, and TNF-α) and CC chemokines (CCL2, CCL3, CXCL2) in the brain. Moreover, our results revealed that the exacerbation of JE progression in CD11c(hi) DC-ablated hosts was correlated with enhanced BBB permeability and reduced expression of tight junction and adhesion molecules (claudin-5, ZO-1, occluding, JAMs). Ultimately, our data conclude that the ablation of CD11c(hi) DCs provided a subsidiary impact on BBB integrity and the expression of tight junction/adhesion molecules, thereby leading to exacerbated JE progression. These findings provide insight into the secondary role of CD11c(hi) DCs in JE progression through regulation of BBB integrity and the expression of tight junction/adhesion molecules.

  12. Tunable permittivity and permeability of low loss Z + Y-type ferrite composites for ultra-high frequency applications

    Su, Zhijuan; Hu, Bolin; Chen, Yajie, E-mail:; Harris, Vincent G. [Center for Microwave Magnetic Materials and Integrated Circuits, and Department of Electrical and Computer Engineering, Northeastern University, Boston, Massachusetts 02115 (United States); Li, Qifan; Feng, Zekun [School of Optical and Electronic Information, Huazhong University of Science and Technology, Wuhan 430074 (China); Wang, Xian [Center for Microwave Magnetic Materials and Integrated Circuits, and Department of Electrical and Computer Engineering, Northeastern University, Boston, Massachusetts 02115 (United States); School of Optical and Electronic Information, Huazhong University of Science and Technology, Wuhan 430074 (China)


    A series of Z-type and Y-type ferrite composites with various phase fractions were studied for their RF properties including the measurement of permittivity to permeability spectra over a frequency range of 0.1–10 GHz. Phase identification of the ferrite composites' constituents was determined by X-ray diffraction. An effective medium approximation was used to predict the magnetic and dielectric behavior of the composites. The experiments indicated that the composite having 75 vol. % of Z-type ferrite demonstrated a permeability of ∼12 with a nearly equivalent permittivity, yielding a ratio (μ′/ε′) of 0.91 at a frequency range from 0.55 to 0.75 GHz. The dielectric loss (i.e., tan δ{sub ε}) and magnetic loss (i.e., tan δ{sub μ}) were measured to be lower than 0.08 at f = 0.1–1 GHz and 0.29 at f = 0.1–0.7 GHz, respectively. Furthermore, the loss factors, as tan δ{sub ε}/ε′ and tan δ{sub μ}/μ′, were calculated to be 0.003 and 0.02 at 0.65 GHz, respectively.

  13. Can the inflammation markers of patients with high peritoneal permeability on continuous ambulatory peritoneal dialysis be reduced on nocturnal intermittent peritoneal dialysis?

    Cueto-Manzano, Alfonso M; Rojas-Campos, Enrique; Martínez-Ramírez, Héctor R; Valera-González, Isela; Medina, Miguel; Monteón, Francisco; Ruiz, Norma; Becerra, Mauricio; Palomeque, Miguel A; Cortés-Sanabria, Laura


    Patients with high peritoneal permeability have the greatest degree of inflammation on continuous ambulatory peritoneal dialysis (CAPD), which may be associated with their higher mortality. Nocturnal intermittent peritoneal dialysis (NIPD; "dry day") may decrease inflammation by reducing the contact between dialysate and peritoneum and/or providing better fluid overload control. Therefore, the aims of this study were to determine and compare serum and dialysate concentrations of C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) of patients with high or high-average peritoneal transport on CAPD, changed to NIPD, and ultimately to continuous cyclic peritoneal dialysis (CCPD). Crossover clinical trial in 11 randomly selected patients. All subjects had been on CAPD and were changed to NIPD, and ultimately to CCPD (6.4 +/- 3.1 months after initiation of study). All patients used glucose-based dialysate. Evaluations of clinical and biochemical parameters, dialysis adequacy, and serum and dialysis inflammation markers were performed at baseline on CAPD, 7 - 14 days after changing to NIPD, 7 - 14 days after switching to CCPD, and after 1 year of follow-up. All patients used only 1.5% glucose dialysate during evaluation days. CRP was determined by nephelometry, and IL-6 and TNF-alpha by ELISA. Seven patients were high transporters and 4 high average. Ultrafiltration increased (p permeability.

  14. T-type channels become highly permeable to sodium ions using an alternative extracellular turret region (S5-P) outside the selectivity filter.

    Senatore, Adriano; Guan, Wendy; Boone, Adrienne N; Spafford, J David


    T-type (Cav3) channels are categorized as calcium channels, but invertebrate ones can be highly sodium-selective channels. We illustrate that the snail LCav3 T-type channel becomes highly sodium-permeable through exon splicing of an extracellular turret and descending helix in domain II of the four-domain Cav3 channel. Highly sodium-permeable T-type channels are generated without altering the invariant ring of charged residues in the selectivity filter that governs calcium selectivity in calcium channels. The highly sodium-permeant T-type channel expresses in the brain and is the only splice isoform expressed in the snail heart. This unique splicing of turret residues offers T-type channels a capacity to serve as a pacemaking sodium current in the primitive heart and brain in lieu of Nav1-type sodium channels and to substitute for voltage-gated sodium channels lacking in many invertebrates. T-type channels would also contribute substantially to sodium leak conductances at rest in invertebrates because of their large window currents.

  15. The phosphatase PTP-PEST/PTPN12 regulates endothelial cell migration and adhesion, but not permeability, and controls vascular development and embryonic viability.

    Souza, Cleiton Martins; Davidson, Dominique; Rhee, Inmoo; Gratton, Jean-Philippe; Davis, Elaine C; Veillette, André


    Protein-tyrosine phosphatase (PTP)-PEST (PTPN12) is ubiquitously expressed. It is essential for normal embryonic development and embryonic viability in mice. Herein we addressed the involvement of PTP-PEST in endothelial cell functions using a combination of genetic and biochemical approaches. By generating primary endothelial cells from an inducible PTP-PEST-deficient mouse, we found that PTP-PEST is not needed for endothelial cell differentiation and proliferation or for the control of endothelial cell permeability. Nevertheless, it is required for integrin-mediated adhesion and migration of endothelial cells. PTP-PEST-deficient endothelial cells displayed increased tyrosine phosphorylation of Cas, paxillin, and Pyk2, which were previously also implicated in integrin functions. By eliminating PTP-PEST in endothelial cells in vivo, we obtained evidence that expression of PTP-PEST in endothelial cells is required for normal vascular development and embryonic viability. Therefore, PTP-PEST is a key regulator of integrin-mediated functions in endothelial cells seemingly through its capacity to control Cas, paxillin, and Pyk2. This function explains at least in part the essential role of PTP-PEST in embryonic development and viability.

  16. Classical activation of microglia in CD200-deficient mice is a consequence of blood brain barrier permeability and infiltration of peripheral cells.

    Denieffe, Stephanie; Kelly, Ronan J; McDonald, Claire; Lyons, Anthony; Lynch, Marina A


    The interaction between CD200, expressed on several cell types, and its receptor CD200R, expressed on cells of the myeloid lineage, has been shown to be an important factor in modulating inflammation in macrophage function in several conditions including colitis and arthritis. More recently its modulatory effect on microglial activation has been identified and CD200-deficiency has been associated with increased microglial activation accompanied by increased production of inflammatory cytokines. The response of glia prepared from CD200-deficient mice to stimuli like lipopolysaccharide (LPS) is markedly greater than the response of cells prepared from wildtype mice and, consistent with this, is the recent observation that expression of Toll-like receptor (TLR)4 and signalling through NFκB are increased in microglia prepared from CD200-deficient mice. Here we show that glia from CD200-deficient mice are also more responsive to interferon-γ (IFNγ) which triggers classical activation of microglia. We investigated the effects of CD200-deficiency in vivo and report that there is an increase in expression of several markers of microglial activation including tumor necrosis factor (TNF)-α, which is a hallmark of classically-activated microglia. These changes are accompanied by increased IFNγ, and the evidence suggests that this is produced by infiltrating cells including T cells and macrophages. We propose that these cells enter the brain as a consequence of increased blood brain barrier (BBB) permeability in CD200-deficient mice and that infiltration is assisted by increased expression of the chemokines, monocyte chemotactic protein-1 (MCP-1), IFNγ-induced protein-10 (IP-10) and RANTES. This may have implications in neurodegenerative diseases where BBB permeability is compromised.

  17. Neonatal high-permeability pulmonary edema based on serial cytokine profiles and KL-6 in serum: case report.

    Nakamura, Toshihiko; Nakamura, Mari; Takahashi, Naoto


    A newborn male with pulmonary edema was delivered at term by elective Caesarian section. Cytokine profiles of 17 cytokines and KL-6 in cord blood and serial serum values were investigated. The cord blood values of all 17 cytokines and KL-6 were within normal limits. Subsequently, IL-6, IL-8, IL-10, IL-13, IL-17, and IFNγ rapidly elevated during the first several hours after birth and dramatically decreased thereafter, whereas KL-6 rose to 611 U/ml on the 3(rd) day of life and then gradually decreased. These cytokines may induce pulmonary permeability, and KL-6 secreted in lining fluid could result in influx into the bloodstream. This is the first report that we have differentiated neonatal pulmonary edema from TTN by the measurement of serial cytokine profiles and KL-6 in serum.

  18. Study of the Biotransformation of Tongmai Formula by Human Intestinal Flora and Its Intestinal Permeability across the Caco-2 Cell Monolayer

    Shuai Wu


    Full Text Available Tongmai formula (TMF is a well-known Chinese medicinal preparation that contains isoflavones as its major bioactive constituents. As traditional Chinese medicines (TCMs are usually used by oral administration, their fate inside the intestinal lumen, including their biotransformation by human intestinal flora (HIF and intestinal absorption deserves study. In this work TMF extract was incubated with human intestinal bacteria under anaerobic conditions and the changes in the twelve main constituents of TMF were then investigated. Their intestinal permeabilities, i.e., the transport capability across the intestinal brush border were investigated with a human colon carcinoma cell line (Caco­2 cell monolayer model to predict the absorption mechanism. Meanwhile, rapid HPLC-DAD methods were established for the assay. According to the biotransformation curves of the twelve constituents and the permeability coefficients, the intestinal absorption capacity of the typical compounds was elevated from the levels of 10−7 cm/s to 10−5 cm/s from those of the original compounds in TMF. Among them the main isoflavone glycosides puerarin (4, mirificin (6 and daidzin (7 were transformed into the same aglycone, daidzein (10. Therefore it was predicted that the aglycone compounds might be the real active ingredients in TMF. The models used can represent a novel path for the TCM studies.

  19. Study of the Biotransformation of Tongmai Formula by Human Intestinal Flora and Its Intestinal Permeability across the Caco-2 Cell Monolayer.

    Wu, Shuai; Xu, Wei; Wang, Fu-Rong; Yang, Xiu-Wei


    Tongmai formula (TMF) is a well-known Chinese medicinal preparation that contains isoflavones as its major bioactive constituents. As traditional Chinese medicines (TCMs) are usually used by oral administration, their fate inside the intestinal lumen, including their biotransformation by human intestinal flora (HIF) and intestinal absorption deserves study. In this work TMF extract was incubated with human intestinal bacteria under anaerobic conditions and the changes in the twelve main constituents of TMF were then investigated. Their intestinal permeabilities, i.e., the transport capability across the intestinal brush border were investigated with a human colon carcinoma cell line (Caco-2) cell monolayer model to predict the absorption mechanism. Meanwhile, rapid HPLC-DAD methods were established for the assay. According to the biotransformation curves of the twelve constituents and the permeability coefficients, the intestinal absorption capacity of the typical compounds was elevated from the levels of 10(-7) cm/s to 10(-5) cm/s from those of the original compounds in TMF. Among them the main isoflavone glycosides puerarin (4), mirificin (6) and daidzin (7) were transformed into the same aglycone, daidzein (10). Therefore it was predicted that the aglycone compounds might be the real active ingredients in TMF. The models used can represent a novel path for the TCM studies.

  20. Plant actin controls membrane permeability.

    Hohenberger, Petra; Eing, Christian; Straessner, Ralf; Durst, Steffen; Frey, Wolfgang; Nick, Peter


    The biological effects of electric pulses with low rise time, high field strength, and durations in the nanosecond range (nsPEFs) have attracted considerable biotechnological and medical interest. However, the cellular mechanisms causing membrane permeabilization by nanosecond pulsed electric fields are still far from being understood. We investigated the role of actin filaments for membrane permeability in plant cells using cell lines where different degrees of actin bundling had been introduced by genetic engineering. We demonstrate that stabilization of actin increases the stability of the plasma membrane against electric permeabilization recorded by penetration of Trypan Blue into the cytoplasm. By use of a cell line expressing the actin bundling WLIM domain under control of an inducible promotor we can activate membrane stabilization by the glucocorticoid analog dexamethasone. By total internal reflection fluorescence microscopy we can visualize a subset of the cytoskeleton that is directly adjacent to the plasma membrane. We conclude that this submembrane cytoskeleton stabilizes the plasma membrane against permeabilization through electric pulses. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. High Performance Perovskite Solar Cells.

    Tong, Xin; Lin, Feng; Wu, Jiang; Wang, Zhiming M


    Perovskite solar cells fabricated from organometal halide light harvesters have captured significant attention due to their tremendously low device costs as well as unprecedented rapid progress on power conversion efficiency (PCE). A certified PCE of 20.1% was achieved in late 2014 following the first study of long-term stable all-solid-state perovskite solar cell with a PCE of 9.7% in 2012, showing their promising potential towards future cost-effective and high performance solar cells. Here, notable achievements of primary device configuration involving perovskite layer, hole-transporting materials (HTMs) and electron-transporting materials (ETMs) are reviewed. Numerous strategies for enhancing photovoltaic parameters of perovskite solar cells, including morphology and crystallization control of perovskite layer, HTMs design and ETMs modifications are discussed in detail. In addition, perovskite solar cells outside of HTMs and ETMs are mentioned as well, providing guidelines for further simplification of device processing and hence cost reduction.

  2. Estimation of soil permeability

    Amr F. Elhakim


    Full Text Available Soils are permeable materials because of the existence of interconnected voids that allow the flow of fluids when a difference in energy head exists. A good knowledge of soil permeability is needed for estimating the quantity of seepage under dams and dewatering to facilitate underground construction. Soil permeability, also termed hydraulic conductivity, is measured using several methods that include constant and falling head laboratory tests on intact or reconstituted specimens. Alternatively, permeability may be measured in the field using insitu borehole permeability testing (e.g. [2], and field pumping tests. A less attractive method is to empirically deduce the coefficient of permeability from the results of simple laboratory tests such as the grain size distribution. Otherwise, soil permeability has been assessed from the cone/piezocone penetration tests (e.g. [13,14]. In this paper, the coefficient of permeability was measured using field falling head at different depths. Furthermore, the field coefficient of permeability was measured using pumping tests at the same site. The measured permeability values are compared to the values empirically deduced from the cone penetration test for the same location. Likewise, the coefficients of permeability are empirically obtained using correlations based on the index soil properties of the tested sand for comparison with the measured values.

  3. Quantitative Prediction of Structural Fractures in Low Permeability Reservoir

    Zeng Lianbo; Tian Chonglu


    @@ Low -permeability fractured reservoirs will become increasingly prominent along with the enhanced exploration extent and the emerging moderate-high water content in most of the moderate-high permeability reservoirs of the oil fields in eastern China.

  4. High temperature proton exchange membranes based on polybenzimidazoles for fuel cells

    Li, Qingfeng; Jensen, Jens Oluf; Savinell, Robert F;


    in recent years motivated extensive research activities with great progress. This treatise is devoted to updating the development, covering polymer synthesis, membrane casting, physicochemical characterizations and fuel cell technologies. To optimize the membrane properties, high molecular weight polymers...... havebeenmadeincluding spectroscopy,wateruptake and acid doping, thermal and oxidative stability, conductivity, electro-osmoticwater drag, methanol crossover, solubility and permeability of gases, and oxygen reduction kinetics. Related fuel cell technologies such as electrode and MEA fabrication have been developed...

  5. Bentonite Permeability at Elevated Temperature

    Katherine A. Daniels


    Full Text Available Repository designs frequently favour geological disposal of radioactive waste with a backfill material occupying void space around the waste. The backfill material must tolerate the high temperatures produced by decaying radioactive waste to prevent its failure or degradation, leading to increased hydraulic conductivity and reduced sealing performance. The results of four experiments investigating the effect of temperature on the permeability of a bentonite backfill are presented. Bentonite is a clay commonly proposed as the backfill in repository designs because of its high swelling capacity and very low permeability. The experiments were conducted in two sets of purpose-built, temperature controlled apparatus, designed to simulate isotropic pressure and constant volume conditions within the testing range of 4–6 MPa average effective stress. The response of bentonite during thermal loading at temperatures up to 200 °C was investigated, extending the previously considered temperature range. The results provide details of bentonite’s intrinsic permeability, total stress, swelling pressure and porewater pressure during thermal cycles. We find that bentonite’s hydraulic properties are sensitive to thermal loading and the type of imposed boundary condition. However, the permeability change is not large and can mostly be accounted for by water viscosity changes. Thus, under 150 °C, temperature has a minimal impact on bentonite’s hydraulic permeability.

  6. Effects of Lead and Cadmium on Brain Endothelial Cell Survival, Monolayer Permeability, and Crucial Oxidative Stress Markers in an in Vitro Model of the Blood-Brain Barrier

    Shakila Tobwala


    Full Text Available Oxidative stress, which is the loss of balance between antioxidant defense and oxidant production in the cells, is implicated in the molecular mechanism of heavy metal-induced neurotoxicity. Given the key role of lead (Pb and cadmium (Cd in inducing oxidative stress, we investigated their role in disrupting the integrity and function of immortalized human brain microvascular endothelial cells (hCMEC/D3. To study this, hCMEC/D3 cells were exposed to control media or to media containing different concentrations of Pb or Cd. Those exposed to Pb or Cd showed significantly higher oxidative stress than the untreated group, as indicated by cell viability, reactive oxygen species (ROS, glutathione (GSH levels, and catalase enzyme activity. Pb also induced oxidative stress-related disruption of the hCMEC/D3 cell monolayer, as measured by trans-endothelial electrical resistance (TEER, the dextran permeability assay, and the level of tight junction protein, zona occluden protein (ZO-2. However, no significant disruption in the integrity of the endothelial monolayer was seen with cadmium at the concentrations used. Taken together, these results show that Pb and Cd induce cell death and dysfunction in hCMEC/D3 cells and, in the case of Pb, barrier disruption. This suggests blood brain barrier (BBB dysfunction as a contributing mechanism in Pb and Cd neurotoxicities.

  7. The permeation of dynorphin A 1-6 across the blood brain barrier and its effect on bovine brain microvessel endothelial cell monolayer permeability.

    Sloan, Courtney D Kuhnline; Audus, Kenneth L; Aldrich, Jane V; Lunte, Susan M


    Dynorphin A 1-17 (Dyn A 1-17) is an endogenous neuropeptide known to act at the kappa opioid receptor; it has been implicated in a number of neurological disorders, including neuropathic pain, stress, depression, and Alzheimer's and Parkinson's diseases. The investigation of Dyn A 1-17 metabolism at the blood-brain barrier (BBB) is important since the metabolites exhibit unique biological functions compared to the parent compound. In this work, Dyn A 1-6 is identified as a metabolite of Dyn A 1-17 in the presence of bovine brain microvessel endhothelial cells (BBMECs), using LC-MS/MS. The transport of Dyn A 1-6 at the BBB was examined using this in vitro cell culture model of the BBB. Furthermore, the permeation of the BBB by the low molecular weight permeability marker fluorescein was characterized in the presence and absences of Dyn A 1-6.

  8. Self-double-emulsifying drug delivery system (SDEDDS): a new way for oral delivery of drugs with high solubility and low permeability.

    Qi, Xiaole; Wang, Lishuang; Zhu, Jiabi; Hu, Zhenyi; Zhang, Jie


    Water-in-oil-in-water (w/o/w) double emulsions are potential for enhancing oral bioavailability of drugs with high solubility and low permeability, but their industrial application is limited due to the instability. Herein, we developed a novel formulation, self-double-emulsifying drug delivery systems (SDEDDS) by formulating mixtures of hydrophilic surfactants and water-in-oil (w/o) emulsions, which were easier to be stable through formulations optimization. SDEDDS can spontaneously emulsify to water-in-oil-in-water (w/o/w) double emulsions in the mixed aqueous gastrointestinal environment, with drugs encapsulated in the internal water phase of the double emulsions. We employed SDEDDS to improve the oral absorption of pidotimod, a peptide-like drug with high solubility and low permeability. The optimized pidotimod-SDEDDS were found to be stable up to 6 months under 25°C. Plasma concentration-time profiles from pharmacokinetic studies in rats dosed with SDEDDS showed 2.56-fold (p<0.05) increased absorption of pidotimod, compared to the pidotimod solution. Histopathologic studies confirmed that SDEDDS exerted absorption promoting effect without serious local damages. These studies demonstrate that SDEDDS may be a promising strategy for peroral delivery of peptide and peptidomimetic drugs.


    Jensen, Jens Oluf; Qingfeng, Li; He, Ronghuan


    This paper will report recent results from our group on polymer fuel cells (PEMFC) based on the temperature resistant polymer polybenzimidazole (PBI), which allow working temperatures up to 200°C. The membrane has a water drag number near zero and need no water management at all. The high working...

  10. The suitability of an in situ perfusion model for permeability determinations: utility for BCS class I biowaiver requests.

    Kim, Jae-Seung; Mitchell, Stefanie; Kijek, Paul; Tsume, Yasuhiro; Hilfinger, John; Amidon, Gordon L


    The FDA has published recommendations for sponsors who wish to request a waiver of in vivo bioavailability (BA) or bioequivalence (BE) studies for immediate release (IR) solid oral dosage forms based on the Biopharmaceutics Classification System (BCS). Biowaivers can be requested for IR formulations in which the active ingredient is shown to be a BCS class I drug: that is, a drug showing high permeability and high solubility over a pH range of 1-7.5. For permeability determinations, a variety of experimental methods can be used, such as the rat in situ single pass perfusion or Caco-2 cell culture models, once the suitability of the particular method is established. Following the recommended procedure for assessing the suitability of permeability determinations, we determined the permeability of 20 test drugs using the in situ single pass perfusion model in rats. The test compounds were coperfused through jejunal intestinal segments with an internal permeability reference standard (metoprolol) over a 90 min time period. Sample analysis was performed by HPLC, and the ratio of the effective permeability, Peff (cm/s), of test compound to that of metoprolol was determined. To address the question of test drug permeabilities that approach that of the internal standard, we propose that a statistical analysis such as the "0.8-1.25 rule" used for in vivo or in vitro bioequivalence studies provide guidance for permeability classification using the in situ single pass perfusion model. We developed a method using the 90% confidence interval of the permeability ratio of the test to internal reference standard in order to differentiate between high and low permeability compounds. This analysis allowed for the proper permeability classification of all of the test compounds and suggests a robust means for assessing drug permeability classification.

  11. Synthesis and analysis of novel polymers with high permselectivity and permeability in gas separation applications. Final report

    Koros, W.J.; Paul, D.R.


    During the three years of support under this grant, ten novel polymer structures have been synthesized and characterized in detail in terms of sorption and transport properties to test the hypotheses on strategies to develop advanced materials for gas separation membranes. The extremely important O{sub 2}/N{sub 2} and CO{sub 2}/CH{sub 4} systems have been the focus of this work. Data for permeabilities and permselectivities for O{sub 2}/N{sub 2} and CO{sub 2}/CH{sub 4} at 35 C at approximately 2 atm feed pressure for O{sub 2} and N{sub 2} and 10 atm for CO{sub 2} and CH{sub 4} are reported in two tables and will be discussed in two parts, one related to each of the groups of structures in these two tables. For the sake of efficiency, the author will only consider the O{sub 2}/N{sub 2} data; however, similar trends apply for the CO{sub 2}/CH{sub 4} system as well. This gas pair is useful, since solubility selectivity effects are generally of much smaller importance than for the CO{sub 2}/CH{sub 4} pair, so even without detailed solubility and diffusivity data, mobility selectivity based arguments tend to be adequate.

  12. Permeability prediction in chalks

    Alam, Mohammad Monzurul; Fabricius, Ida Lykke; Prasad, Manika


    The velocity of elastic waves is the primary datum available for acquiring information about subsurface characteristics such as lithology and porosity. Cheap and quick (spatial coverage, ease of measurement) information of permeability can be achieved, if sonic velocity is used for permeability....... The relationships between permeability and porosity from core data were first examined using Kozeny’s equation. The data were analyzed for any correlations to the specific surface of the grain, Sg, and to the hydraulic property defined as the flow zone indicator (FZI). These two methods use two different approaches...... to enhance permeability prediction fromKozeny’s equation. The FZI is based on a concept of a tortuous flow path in a granular bed. The Sg concept considers the pore space that is exposed to fluid flow and models permeability resulting from effective flow parallel to pressure drop. The porosity-permeability...

  13. Synthesis of Conformationally Preorganized and Cell-Permeable Guanidine-Based γ-Peptide Nucleic Acids (γGPNAs)

    Sahu, Bichismita; Chenna, Venugopal; Lathrop, Kira L.; Thomas, Sufi M.; Zon, Gerald; Livak, Kenneth J.; Ly, Danith H.


    A general method for preparing optically-pure guanidine-based γ-peptide nucleic acid (γGPNA) monomers for all four natural nucleobases (A, C, G and T) is described. These second-generation γGPNAs differ from the first in that the guanidinium group is installed at the γ-instead of the α-position of the N-(2-aminoethyl)glycine backbone unit. This positional switch enables GPNAs to be synthesized from relatively cheap L- as opposed to D-amino acids. Unlike their α-predecessors, which are randomly-folded, γGPNAs prepared from L-amino acids are preorganized into a right-handed helix and bind to DNA and RNA with exceptionally high affinity and sequence selectivity, and are readily taken up by mammalian cells. PMID:19161276

  14. Development of a cell permeable red-shifted CHEF-based chemosensor for Al3 + ion by controlling PET

    Mukherjee, Manjira; Sen, Buddhadeb; Pal, Siddhartha; Maji, Abhishek; Budhadev, Darshita; Chattopadhyay, Pabitra


    A structurally modified quinazoline derivative (L) acts as highly selective chemosensor for Al3 + ions in DMSO-H2O (1:9, v/v) over the other competitive metal ions. L shows a red shifted fluorescence after the addition of Al3 + ions and later the further fluorescence enhancement is due to chelation enhanced fluorescence (CHEF) through inhibition of photoinduced electron transfer (PET). This probe (L) detects Al3 + ions as low as 9 nM in DMSO-H2O (1:9, v/v) at biological pH. The non-cytotoxic probe (L) can efficiently detect the intercellular distribution of Al3 + ions in living cells under a fluorescence microscope to exhibit its sensible applications in the biological systems.

  15. CI-1010 induced opening of the mitochondrial permeability transition pore precedes oxidative stress and apoptosis in SY5Y neuroblastoma cells.

    Miller, Terry J; Phelka, Amanda D; Tjalkens, Ronald B; Dethloff, Lloyd A; Philbert, Martin A


    The hetero-bifunctional nitroimidazole radiosensitizer CI-1010, R-alpha-[[(2-bromoethyl)-amino]methyl]-2-nitro-1H-imidazole-1-ethanol monohydrobromide, causes selective irreversible apoptotic loss of retinal photoreceptor cells in vivo. The human neuroblastoma cell line, SH-SY5Y, was used as a neuronotypic model of CI-1010-mediated retinal degeneration. Exposure to CI-1010 for 24 h induced apoptosis in neuroblastoma cells, as determined by histopathological and ultrastructural analysis and by TUNEL technique. CI-1010 causes a dose-dependent decrease in cell viability in SY5Y cells, as measured by the reduction of MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. Superoxide dismutase reduced loss of cell viability following CI-1010 treatment suggesting an oxidative stress-mediated mechanism of toxicity. The effects of CI-1010 on mitochondrial membrane potential and intracellular levels of reactive oxygen species were assessed in live SY5Y cells by confocal microscopy using the fluorescent dyes, tetramethylrhodamine methyl ester and 5,6-carboxy-2',7'-dihydrodichlorofluorescein diacetate. CI-1010 caused a rapid depolarization of mitochondria in SY5Y cells followed by an increase in ROS. Both CI-1010-induced mitochondrial depolarization and subsequent increases in ROS were prevented by pretreatment with either the permeability transition pore inhibitor, cyclosporin A (CsA), and by the antioxidant, alpha-tocopherol. However, CsA and alpha-tocopherol were unable to prevent apoptosis in CI-1010-treated cells, suggesting the influence of additional mechanism(s) of CI-1010-induced toxicity. This study evaluates intracellular oxidative stress associated with pore opening prior to apoptosis and provides evidence in support of a mitochondrial mechanism of CI-1010-induced neuronal cell death.

  16. Outer Membrane Permeability of Cyanobacterium Synechocystis sp. Strain PCC 6803: Studies of Passive Diffusion of Small Organic Nutrients Reveal the Absence of Classical Porins and Intrinsically Low Permeability.

    Kowata, Hikaru; Tochigi, Saeko; Takahashi, Hideyuki; Kojima, Seiji


    The outer membrane of heterotrophic Gram-negative bacteria plays the role of a selective permeability barrier that prevents the influx of toxic compounds while allowing the nonspecific passage of small hydrophilic nutrients through porin channels. Compared with heterotrophic Gram-negative bacteria, the outer membrane properties of cyanobacteria, which are Gram-negative photoautotrophs, are not clearly understood. In this study, using small carbohydrates, amino acids, and inorganic ions as permeation probes, we determined the outer membrane permeability of Synechocystis sp. strain PCC 6803 in intact cells and in proteoliposomes reconstituted with outer membrane proteins. The permeability of this cyanobacterium was >20-fold lower than that of Escherichia coli The predominant outer membrane proteins Slr1841, Slr1908, and Slr0042 were not permeable to organic nutrients and allowed only the passage of inorganic ions. Only the less abundant outer membrane protein Slr1270, a homolog of the E. coli export channel TolC, was permeable to organic solutes. The activity of Slr1270 as a channel was verified in a recombinant Slr1270-producing E. coli outer membrane. The lack of putative porins and the low outer membrane permeability appear to suit the cyanobacterial autotrophic lifestyle; the highly impermeable outer membrane would be advantageous to cellular survival by protecting the cell from toxic compounds, especially when the cellular physiology is not dependent on the uptake of organic nutrients.IMPORTANCE Because the outer membrane of Gram-negative bacteria affects the flux rates for various substances into and out of the cell, its permeability is closely associated with cellular physiology. The outer membrane properties of cyanobacteria, which are photoautotrophic Gram-negative bacteria, are not clearly understood. Here, we examined the outer membrane of Synechocystis sp. strain PCC 6803. We revealed that it is relatively permeable to inorganic ions but is markedly less

  17. Formation and Control of Self-Sealing High Permeability Groundwater Mounds in Impermeable Sediment: Implications for SUDS and Sustainable Pressure Mound Management

    David D. J. Antia


    Full Text Available A groundwater mound (or pressure mound is defined as a volume of fluid dominated by viscous flow contained within a sediment volume where the dominant fluid flow is by Knudsen Diffusion. High permeability self-sealing groundwater mounds can be created as part of a sustainable urban drainage scheme (SUDS using infiltration devices. This study considers how they form, and models their expansion and growth as a function of infiltration device recharge. The mounds grow through lateral macropore propagation within a Dupuit envelope. Excess pressure relief is through propagating vertical surge shafts. These surge shafts can, when they intersect the ground surface result, in high volume overland flow. The study considers that the creation of self-sealing groundwater mounds in matrix supported (clayey sediments (intrinsic permeability = 10–8 to 10–30 m3 m–2 s–1 Pa–1 is a low cost, sustainable method which can be used to dispose of large volumes of storm runoff (<20→2,000 m3/24 hr storm/infiltration device and raise groundwater levels. However, the inappropriate location of pressure mounds can result in repeated seepage and ephemeral spring formation associated with substantial volumes of uncontrolled overland flow. The flow rate and flood volume associated with each overland flow event may be substantially larger than the associated recharge to the pressure mound. In some instances, the volume discharged as overland flow in a few hours may exceed the total storm water recharge to the groundwater mound over the previous three weeks. Macropore modeling is used within the context of a pressure mound poro-elastic fluid expulsion model in order to analyze this phenomena and determine (i how this phenomena can be used to extract large volumes of stored filtered storm water (at high flow rates from within a self-sealing high permeability pressure mound and (ii how self-sealing pressure mounds (created using storm water infiltration can be used to

  18. Plasmolysis and Cell Shape Depend on Solute Outer-Membrane Permeability during Hyperosmotic Shock in E. coli

    Pilizota, Teuta; Shaevitz, Joshua W.


    The concentration of chemicals inside the bacterial cytoplasm generates an osmotic pressure, termed turgor, which inflates the cell and is necessary for cell growth and survival. In Escherichia coli, a sudden increase in external concentration causes a pressure drop across the cell envelope that drives changes in cell shape, such as plasmolysis, where the inner and outer membranes separate. Here, we use fluorescence imaging of single cells during hyperosmotic shock with a time resolution on t...

  19. A novel approach to regulate cell membrane permeability for ATP and NADH formation in Saccharomyces cerevisiae induced by air cold plasma

    Xiaoyu, DONG; Tingting, LIU; Yuqin, XIONG


    Air cold plasma has been used as a novel method for enhancing microbial fermentation. The aim of this work was to explore the effect of plasma on membrane permeability and the formation of ATP and NADH in Saccharomyces cerevisiae, so as to provide valuable information for large-scale application of plasma in the fermentation industry. Suspensions of S. cerevisiae cells were exposed to air cold plasma for 0, 1, 2, 3, 4 and 5 min, and then subjected to various analyses prior to fermentation (0 h) and at the 9 and 21 h stages of fermentation. Compared with non-exposed cells, cells exposed to plasma for 1 min exhibited a marked increase in cytoplasmic free Ca2+ concentration as a result of the significant increase in membrane potential prior to fermentation. At the same time, the ATP level in the cell suspension decreased by about 40%, resulting in a reduction of about 60% in NADH prior to culturing. However, the levels of ATP and NADH in the culture at the 9 and 21 h fermentation stages were different from the level at 0 h. Taken together, the results indicated that exposure of S. cerevisiae to air cold plasma could increase its cytoplasmic free Ca2+ concentration by improving the cell membrane potential, consequently leading to changes in ATP and NADH levels. Supported by National Natural Science Foundation of China (Nos. 21246012, 21306015 and 21476032).

  20. High temperature proton exchange membranes based on polybenzimidazole and clay composites for fuel cells

    Plackett, David; Siu, Ana; Li, Qingfeng


    and pyridinium salts with varying polarity and hydrogen-bonding capacity. Clay modification by ion-exchange reactions involving replacement of interlayer inorganic cations was confirmed using X-ray photoelectron and infrared spectroscopy techniques. The cast PBI membranes were characterized by their water uptake......, acid doping and swelling, tensile strength, conductivity and hydrogen permeability as well as by fuel cell tests. For the composite membranes, high acid doping levels were achieved with sufficient mechanical strength and improved dimensional stability or reduced membrane swelling. At an acid doping......-doped pristine PBI membranes. In accordance with the hydrogen permeability measurements, fuel cell tests exhibited high open circuit voltages (i.e., 1.02 V) at room temperature as well as high I–V performance compared with normal PBI membranes....

  1. Implementation of a Permeable Membrane Insert-based Infection System to Study the Effects of Secreted Bacterial Toxins on Mammalian Host Cells.

    Flaherty, Rebecca A; Lee, Shaun W


    Many bacterial pathogens secrete potent toxins to aid in the destruction of host tissue, to initiate signaling changes in host cells or to manipulate immune system responses during the course of infection. Though methods have been developed to successfully purify and produce many of these important virulence factors, there are still many bacterial toxins whose unique structure or extensive post-translational modifications make them difficult to purify and study in in vitro systems. Furthermore, even when pure toxin can be obtained, there are many challenges associated with studying the specific effects of a toxin under relevant physiological conditions. Most in vitro cell culture models designed to assess the effects of secreted bacterial toxins on host cells involve incubating host cells with a one-time dose of toxin. Such methods poorly approximate what host cells actually experience during an infection, where toxin is continually produced by bacterial cells and allowed to accumulate gradually during the course of infection. This protocol describes the design of a permeable membrane insert-based bacterial infection system to study the effects of Streptolysin S, a potent toxin produced by Group A Streptococcus, on human epithelial keratinocytes. This system more closely mimics the natural physiological environment during an infection than methods where pure toxin or bacterial supernatants are directly applied to host cells. Importantly, this method also eliminates the bias of host responses that are due to direct contact between the bacteria and host cells. This system has been utilized to effectively assess the effects of Streptolysin S (SLS) on host membrane integrity, cellular viability, and cellular signaling responses. This technique can be readily applied to the study of other secreted virulence factors on a variety of mammalian host cell types to investigate the specific role of a secreted bacterial factor during the course of infection.

  2. Discovery and Characterization of a Cell-Permeable, Small-Molecule c-Abl Kinase Activator that Binds to the Myristoyl Binding Site

    Yang, Jingsong; Campobasso, Nino; Biju, Mangatt P.; Fisher, Kelly; Pan, Xiao-Qing; Cottom, Josh; Galbraith, Sarah; Ho, Thau; Zhang, Hong; Hong, Xuan; Ward, Paris; Hofmann, Glenn; Siegfried, Brett; Zappacosta, Francesca; Washio, Yoshiaki; Cao, Ping; Qu, Junya; Bertrand, Sophie; Wang, Da-Yuan; Head, Martha S.; Li, Hu; Moores, Sheri; Lai, Zhihong; Johanson, Kyung; Burton, George; Erickson-Miller, Connie; Simpson, Graham; Tummino, Peter; Copeland, Robert A.; Oliff, Allen (GSKPA)


    c-Abl kinase activity is regulated by a unique mechanism involving the formation of an autoinhibited conformation in which the N-terminal myristoyl group binds intramolecularly to the myristoyl binding site on the kinase domain and induces the bending of the {alpha}I helix that creates a docking surface for the SH2 domain. Here, we report a small-molecule c-Abl activator, DPH, that displays potent enzymatic and cellular activity in stimulating c-Abl activation. Structural analyses indicate that DPH binds to the myristoyl binding site and prevents the formation of the bent conformation of the {alpha}I helix through steric hindrance, a mode of action distinct from the previously identified allosteric c-Abl inhibitor, GNF-2, that also binds to the myristoyl binding site. DPH represents the first cell-permeable, small-molecule tool compound for c-Abl activation.

  3. Effect of dexamethasone in feed on intestinal permeability, differential white blood cell counts, and immune organs in broiler chicks.

    Vicuña, E A; Kuttappan, V A; Galarza-Seeber, R; Latorre, J D; Faulkner, O B; Hargis, B M; Tellez, G; Bielke, L R


    We have previously shown that intestinal barrier function can be adversely affected by poorly digested diets or feed restriction, resulting in increased intestinal inflammation-associated permeability. Three experiments were conducted in broilers to evaluate the effect of dexamethasone (DEX) treatment on systemic fluorescein isothiocyanate-dextran (FITC-D; 3-5 kDa) levels, indicative of increased gut epithelial leakage. Experiment 1 compared DEX injections of 1 mg/kg, once per day on d 3, 5, and 9, with feed administration at 0.57, 1.7, or 5.1 ppm d 4 to 10, with FITC-D serum concentrations 2.5 h after gavage with 4.16 mg/kg FITC-D. All DEX treatments resulted in marked (2 to 6X; Psoy agar to evaluate total numbers of aerobic bacteria in the liver as an index of bacterial translocation (BT). In both experiments, FITC-D absorption was significantly enhanced (PBT to the liver, supporting the observation that dietary DEX causes a stress-like inflammatory GI response, which may contribute to subclinical or clinical disease, and may be a useful model for ongoing disease mitigation research related to stress-related diseases of GIT origin. © 2015 Poultry Science Association Inc.

  4. Relationship between permeability and damage in concretes at high temperature; Lien entre la permeabilite et l'endommagement dans les betons a haute temperature

    Dal Pont, St


    Due to its technical and economical advantages, concrete is nowadays the most used building material in civil engineering. Even if its use is known since nearly two centuries, its behavior has not been yet completely explained due to the complexity of its porous microstructure. This fact is quite evident under particular conditions such as, by instance, during an elevation of temperature. This condition can mainly occur in two cases: due to a casualty (e.g. a fire) or in normal use conditions (e.g. storage of nuclear rejects). This work aims at contributing to the study of the phenomena that can be observed in concrete exposed to high temperatures and, in particular, focuses on the study of the evolution of intrinsic permeability. The characterisation of permeability (which is hardly measurable in hot conditions) is necessary for describing and modelling transport phenomena which occur in porous media. An experimental study has been made in collaboration with the CEA. A real-scale hollow cylinder has been instrumented with gauges for studying the evolution of temperature and gas pressure fields inside concrete. Later, the cylinder has been then numerically modelled by means of a thermo-hydro-chemical (THC) and a thermo-hydro-chemo-mechanical (THCM) model. The THC model, implemented by means of the finite volume method, has allowed a first, qualitative study of the behaviour of concrete submitted to high temperature. This model, which, for sake of simplicity, has neglected all mechanical effects, has allowed the description of the main phenomena occurring inside concrete: mass transport, phase changes, microstructure evolution. Later, the modelling has been completed by means of the THCM model using the Hitecosp code, implemented by means of the finite element method at the university of Padua. This code allows a very complete description of the phenomena occurring inside concrete and takes into consideration the mechanical behavior of concrete by means of an

  5. The Mechanism by Which MYCN Amplification Confers an Enhanced Sensitivity to a PCNA-Derived Cell Permeable Peptide in Neuroblastoma Cells

    Long Gu


    Full Text Available Dysregulated expression of MYC family genes is a hallmark of many malignancies. Unfortunately, these proteins are not amenable to blockade by small molecules or protein-based therapeutic agents. Therefore, we must find alternative approaches to target MYC-driven cancers. Amplification of MYCN, a MYC family member, predicts high-risk neuroblastoma (NB disease. We have shown that R9-caPep blocks the interaction of PCNA with its binding partners and selectively kills human NB cells, especially those with MYCN amplification, and we now show the mechanism. We found elevated levels of DNA replication stress in MYCN-amplified NB cells. R9-caPep exacerbated DNA replication stress in MYCN-amplified NB cells and NB cells with an augmented level of MYC by interfering with DNA replication fork extension, leading to Chk1 dependence and susceptibility to Chk1 inhibition. We describe how these effects may be exploited for treating NB.

  6. Preparation of three-layered porous PLA/PEG scaffold: relationship between morphology, mechanical behavior and cell permeability.

    Scaffaro, R; Lopresti, F; Botta, L; Rigogliuso, S; Ghersi, G


    Interface tissue engineering (ITE) is used to repair or regenerate interface living tissue such as for instance bone and cartilage. This kind of tissues present natural different properties from a biological and mechanical point of view. With the aim to imitating the natural gradient occurring in the bone-cartilage tissue, several technologies and methods have been proposed over recent years in order to develop polymeric functionally graded scaffolds (FGS). In this study three-layered scaffolds with a pore size gradient were developed by melt mixing polylactic acid (PLA) and two water-soluble porogen agents: sodium chloride (NaCl) and polyethylene glycol (PEG). Pore dimensions were controlled by NaCl granulometry while PEG solvation created a micropores network within the devices. Scaffolds were characterized from a morphological and mechanical point of view in order to find a correlation between the preparation method, the pore architecture and compressive mechanical behavior. Biological tests were also performed in order to study the effect of pore size gradient on the permeation of different cell lines in co-culture. To imitate the physiological work condition, compressive tests were also performed in phosphate buffered saline (PBS) solution at 37°C. The presented preparation method permitted to prepare three-layered scaffolds with high control of porosity and pore size distribution. Furthermore mechanical behaviors were found to be strongly affected by pore architecture of tested devices as well as the permeation of osteoblast and fibroblast in-vitro.

  7. Accurate determination of characteristic relative permeability curves

    Krause, Michael H.; Benson, Sally M.


    A recently developed technique to accurately characterize sub-core scale heterogeneity is applied to investigate the factors responsible for flowrate-dependent effective relative permeability curves measured on core samples in the laboratory. The dependency of laboratory measured relative permeability on flowrate has long been both supported and challenged by a number of investigators. Studies have shown that this apparent flowrate dependency is a result of both sub-core scale heterogeneity and outlet boundary effects. However this has only been demonstrated numerically for highly simplified models of porous media. In this paper, flowrate dependency of effective relative permeability is demonstrated using two rock cores, a Berea Sandstone and a heterogeneous sandstone from the Otway Basin Pilot Project in Australia. Numerical simulations of steady-state coreflooding experiments are conducted at a number of injection rates using a single set of input characteristic relative permeability curves. Effective relative permeability is then calculated from the simulation data using standard interpretation methods for calculating relative permeability from steady-state tests. Results show that simplified approaches may be used to determine flowrate-independent characteristic relative permeability provided flow rate is sufficiently high, and the core heterogeneity is relatively low. It is also shown that characteristic relative permeability can be determined at any typical flowrate, and even for geologically complex models, when using accurate three-dimensional models.

  8. Enhancement of plume dilution in two-dimensional and three-dimensional porous media by flow focusing in high-permeability inclusions

    Ye, Yu; Chiogna, Gabriele; Cirpka, Olaf A.; Grathwohl, Peter; Rolle, Massimo


    In porous media, lateral mass exchange exerts a significant influence on the dilution of solute plumes in quasi steady state. This process is one of the main mechanisms controlling transport of continuously emitted conservative tracers in groundwater and is fundamental for the understanding of many degradation processes. We investigate the effects of high-permeability inclusions on transverse mixing in three-dimensional versus two-dimensional systems by experimental, theoretical, and numerical analyses. Our results show that mixing enhancement strongly depends on the system dimensionality and on the parameterization used to model transverse dispersion. In particular, no enhancement of transverse mixing would occur in three-dimensional media if the local transverse dispersion coefficient was uniform and flow focusing in both transverse directions was identical, which is fundamentally different from the two-dimensional case. However, the velocity and grain size dependence of the transverse dispersion coefficient and the correlation between hydraulic conductivity and grain size lead to prevailing mixing enhancement within the inclusions, regardless of dimensionality. We perform steady state bench-scale experiments with multiple tracers in three-dimensional and quasi two-dimensional flow-through systems at two different velocities (1 and 5 m/d). We quantify transverse mixing by the flux-related dilution index and compare the experimental results with model simulations. The experiments confirm that, although dilution is larger in three-dimensional systems, the enhancement of transverse mixing due to flow focusing is less effective than in two-dimensional systems. The spatial arrangement of the high-permeability inclusions significantly affects the degree of mixing enhancement. We also observe more pronounced compound-specific effects in the dilution of solute plumes in three-dimensional porous media than in two-dimensional ones.

  9. Comparative NMR studies of diffusional water permeability of red blood cells from different species: XVI Dingo (Canis familiaris dingo) and dog (Canis familiaris).

    Benga, Gheorghe; Chapman, Bogdan E; Matei, Horea; Cox, Guy C; Romeo, Tony; Mironescu, Eugen; Kuchel, Philip W


    As part of a programme of comparative measurements of Pd (diffusional water permeability) the RBCs (red blood cells) from dingo (Canis familiaris dingo) and greyhound dog (Canis familiaris) were studied. The morphologies of the dingo and greyhound RBCs [examined by light and SEM (scanning electron microscopy)] were found to be very similar, with regard to aspect ratio and size; the mean diameters were estimated to be the same (approximately 7.2 microm) for both dingo and greyhound RBCs. The water diffusional permeability was monitored by using an Mn2+-doping 1H NMR technique at 400 MHz. The Pd (cm/s) values of dingo and greyhound RBCs were similar: 6.5 x 10(-3) at 25 degrees C, 7.5 x 10(-3) at 30 degrees C, 10 x 10(-3) at 37 degrees C and 11.5 x 10(-3) at 42 degrees C. The inhibitory effect of a mercury-containing SH (sulfhydryl)-modifying reagent PCMBS (p-chloromercuribenzene sulfonate) was investigated. The maximal inhibition of dingo and greyhound RBCs was reached in 15-30 min at 37 degrees C with 2 mmol/l PCMBS. The values of maximal inhibition were in the range 72-74% when measured at 25 degrees C and 30 degrees C, and approximately 66% at 37 degrees C. The lowest value of Pd (corresponding to the basal permeability to water) was approximately 2-3 x 10(-3) cm/s in the temperature range 25-37 degrees C. The Ea,d (activation energy of water diffusion) was 25 kJ/mol for dingo RBC and 23 kJ/mol for greyhound RBCs. After incubation with PCMBS, the values of Ea,d increased, reaching 46-48 kJ/mol in the condition of maximal inhibition of water exchange. The electrophoretograms of membrane polypeptides of the dingo and greyhound RBCs were compared and seen to be very similar. We postulate that the RBC parameters reported in the present study are characteristic of all canine species and, in particular in the two cases presented here, these parameters have not been changed by the peculiar Australian habitat over the millennia (as in the case of the dingo) or over

  10. High precision measurement of electrical resistance across endothelial cell monolayers.

    Tschugguel, W; Zhegu, Z; Gajdzik, L; Maier, M; Binder, B R; Graf, J


    Effects of vasoactive agonists on endothelial permeability was assessed by measurement of transendothelial electrical resistance (TEER) of human umbilical vein endothelial cells (HUVECs) grown on porous polycarbonate supports. Because of the low values of TEER obtained in this preparation (< 5 omega cm2) a design of an Ussing type recording chamber was chosen that provided for a homogeneous electric field across the monolayer and for proper correction of series resistances. Precision current pulses and appropriate rates of sampling and averaging of the voltage signal allowed for measurement of < 0.1 omega resistance changes of the endothelium on top of a 21 omega series resistance of the support and bathing fluid layers. Histamine (10 microM) and thrombin (10 U/ml) induced an abrupt and substantial decrease of TEER, bradykinin (1 microM) was less effective, PAF (380 nM) and LTC4 (1 microM) had no effect. TEER was also reduced by the calcium ionophore A-23187 (10 microM). The technique allows for measurements of TEER in low resistance monolayer cultures with high precision and time resolution. The results obtained extend previous observations in providing quantitative data on the increase of permeability of HUVECs in response to vasoactive agonists.

  11. Low vanadium ion permeabilities of sulfonated poly(phthalazinone ether ketone)s provide high efficiency and stability for vanadium redox flow batteries

    Chen, Liyun; Zhang, Shouhai; Chen, Yuning; Jian, Xigao


    A series of novel sulfonated poly(phthalazinone ether ketone)s containing pendant phenyl moieties (SPPEK-Ps) are synthesized and thoroughly characterized. The chemical structures of the polymers are confirmed by 1H NMR and FTIR analysis. The physicochemical properties and single cell performance of SPPEK-P membranes are systematically evaluated, revealing that the membranes are thermally, chemically and mechanically stable. The area resistances of SPPEK-P-90 and SPPEK-P-100 are 0.75 Ω cm2 and 0.34 Ω cm2, respectively. SPPEK-P membranes are impermeable to the bulky hydrated VO2+ ion and exhibited low V3+ ion permeability (SPPEK-P-90, 2.53 × 10-5 cm min-1) (Nafion 115 membrane: 9.0 × 10-4 cm min-1). Tests of SPPEK-P-90 in vanadium redox flow batteries (VRFBs) demonstrate a comparable columbic efficiency (CE) and energy efficiency (EE) to that of Nafion 115, where the CE is 98% and the EE is 83% at 60 mA cm-2. Moreover, the SPPEK-P-90 membrane exhibits stable performance in cell over 100 charge-discharge cycles (∼450 h).

  12. A cell-permeable fusion protein based on Clostridium botulinum C2 toxin for delivery of p53 tumorsuppressor into cancer cells.

    Jörg Fahrer

    Full Text Available Genetically engineered bacterial protein toxins are attractive systems for delivery of exogenous proteins into the cytosol of mammalian cells. The binary C2 toxin from C. botulinum has emerged as powerful delivery vehicle, which rests on its binding/translocation component C2IIa and the genetically modified adaptor domain C2IN that act in concert to trigger cell uptake. The p53 tumor suppressor protein has a crucial function in suppressing carcinogenesis and is frequently inactivated by diverse mechanisms in human tumor cells. Therefore, we constructed a C2IN-p53 fusion protein, which is internalized into cancer cells by C2IIa. To this end, the C2IN-p53 fusion construct was overexpressed in E. coli with good solubility, purified by heparin affinity chromatography and protein identity was confirmed by immunoblotting. We demonstrated that the fusion protein is capable of binding to the p53 consensus-DNA with high affinity in a p53-specific manner in vitro. Next, the internalization of C2IN-p53 was monitored in HeLa cells by cell fractionation and immunoblot analysis, which revealed a C2IIa-mediated translocation of the fusion protein into the cytosol. The uptake was also shown in A549 and Saos-2 cells with similar efficiency. These findings were further corroborated by confocal immunofluorescence analyses of C2IN-p53/C2IIa-treated HeLa and A549 cells, displaying predominantly cytoplasmic localization of the fusion construct.


    Jensen, Jens Oluf; Qingfeng, Li; He, Ronghuan


    This paper will report recent results from our group on polymer fuel cells (PEMFC) based on the temperature resistant polymer polybenzimidazole (PBI), which allow working temperatures up to 200°C. The membrane has a water drag number near zero and need no water management at all. The high working...... temperature allows for utilization of the excess heat for fuel processing. Moreover, it provides an excellent CO tolerance of several percent, and the system needs no purification of hydrogen from a reformer. Continuous service for over 6 months at 150°C has been demonstrated....

  14. Synthesis and evaluation of cell-permeable biotinylated PU-H71 derivatives as tumor Hsp90 probes

    Tony Taldone


    Full Text Available The attachment of biotin to a small molecule provides a powerful tool in biology. Here, we present a systematic approach to identify biotinylated analogues of the Hsp90 inhibitor PU-H71 that are capable of permeating cell membranes so as to enable the investigation of Hsp90 complexes in live cells. The identified derivative 2g can isolate Hsp90 through affinity purification and, as we show, represents a unique and useful tool to probe tumor Hsp90 biology in live cells by affinity capture, flow cytometry and confocal microscopy. To our knowledge, 2g is the only reported biotinylated Hsp90 probe to have such combined characteristics.

  15. Synthesis and evaluation of cell-permeable biotinylated PU-H71 derivatives as tumor Hsp90 probes

    DaGama Gomes, Erica M; Riolo, Matthew; Patel, Hardik J; Alonso-Sabadell, Raul; Zatorska, Danuta; Patel, Maulik R; Kishinevsky, Sarah


    Summary The attachment of biotin to a small molecule provides a powerful tool in biology. Here, we present a systematic approach to identify biotinylated analogues of the Hsp90 inhibitor PU-H71 that are capable of permeating cell membranes so as to enable the investigation of Hsp90 complexes in live cells. The identified derivative 2g can isolate Hsp90 through affinity purification and, as we show, represents a unique and useful tool to probe tumor Hsp90 biology in live cells by affinity capture, flow cytometry and confocal microscopy. To our knowledge, 2g is the only reported biotinylated Hsp90 probe to have such combined characteristics. PMID:23616796

  16. Compact rock material gas permeability properties

    Wang, Huanling, E-mail: [Key Laboratory of Coastal Disaster and Defence, Ministry of Education, Hohai University, Nanjing 210098 (China); LML, University of Lille, Cite Scientifique, 59655 Villeneuve d’Ascq (France); Xu, Weiya; Zuo, Jing [Institutes of Geotechnical Engineering, Hohai University, Nanjing 210098 (China)


    Natural compact rocks, such as sandstone, granite, and rock salt, are the main materials and geological environment for storing underground oil, gas, CO{sub 2,} shale gas, and radioactive waste because they have extremely low permeabilities and high mechanical strengths. Using the inert gas argon as the fluid medium, the stress-dependent permeability and porosity of monzonitic granite and granite gneiss from an underground oil storage depot were measured using a permeability and porosity measurement system. Based on the test results, models for describing the relationships among the permeability, porosity, and confining pressure of rock specimens were analyzed and are discussed. A power law is suggested to describe the relationship between the stress-dependent porosity and permeability; for the monzonitic granite and granite gneiss (for monzonitic granite (A-2), the initial porosity is approximately 4.05%, and the permeability is approximately 10{sup −19} m{sup 2}; for the granite gneiss (B-2), the initial porosity is approximately 7.09%, the permeability is approximately 10{sup −17} m{sup 2}; and the porosity-sensitivity exponents that link porosity and permeability are 0.98 and 3.11, respectively). Compared with moderate-porosity and high-porosity rocks, for which φ > 15%, low-porosity rock permeability has a relatively lower sensitivity to stress, but the porosity is more sensitive to stress, and different types of rocks show similar trends. From the test results, it can be inferred that the test rock specimens’ permeability evolution is related to the relative particle movements and microcrack closure.

  17. CDPKs CPK6 and CPK3 function in ABA regulation of guard cell S-type anion- and Ca(2+-permeable channels and stomatal closure.

    Izumi C Mori


    Full Text Available Abscisic acid (ABA signal transduction has been proposed to utilize cytosolic Ca(2+ in guard cell ion channel regulation. However, genetic mutants in Ca(2+ sensors that impair guard cell or plant ion channel signaling responses have not been identified, and whether Ca(2+-independent ABA signaling mechanisms suffice for a full response remains unclear. Calcium-dependent protein kinases (CDPKs have been proposed to contribute to central signal transduction responses in plants. However, no Arabidopsis CDPK gene disruption mutant phenotype has been reported to date, likely due to overlapping redundancies in CDPKs. Two Arabidopsis guard cell-expressed CDPK genes, CPK3 and CPK6, showed gene disruption phenotypes. ABA and Ca(2+ activation of slow-type anion channels and, interestingly, ABA activation of plasma membrane Ca(2+-permeable channels were impaired in independent alleles of single and double cpk3cpk6 mutant guard cells. Furthermore, ABA- and Ca(2+-induced stomatal closing were partially impaired in these cpk3cpk6 mutant alleles. However, rapid-type anion channel current activity was not affected, consistent with the partial stomatal closing response in double mutants via a proposed branched signaling network. Imposed Ca(2+ oscillation experiments revealed that Ca(2+-reactive stomatal closure was reduced in CDPK double mutant plants. However, long-lasting Ca(2+-programmed stomatal closure was not impaired, providing genetic evidence for a functional separation of these two modes of Ca(2+-induced stomatal closing. Our findings show important functions of the CPK6 and CPK3 CDPKs in guard cell ion channel regulation and provide genetic evidence for calcium sensors that transduce stomatal ABA signaling.

  18. Polybenzimidazoles based on high temperature polymer electrolyte fuel cells

    Linares Leon, Jose Joaquin; Camargo, Ana Paula M.; Ashino, Natalia M.; Morgado, Daniella L.; Frollini, Elisabeth; Paganin, Valdecir A.; Gonzalez, Ernesto Rafael [Universidade de Sao Paulo (IQSC/USP), Sao Carlos, SP (Brazil); Bajo, Justo Lobato [University of Castilla-La Mancha, Ciudad Real (Spain). Dept. of Chemical Engineering


    This work presents an interesting approach in order to enhance the performance of Polymer Electrolyte Membrane Fuel Cells (PEMFC) by means of an increase in the operational temperature. For this, two polymeric materials, Poly(2,5-bibenzimidazole) (ABPBI) and Poly[2,2'-(m-phenyl en)-5,5' bib enzimidazol] (PBI), impregnated with phosphoric acid have been utilized. These have shown excellent properties, such as thermal stability above 500 deg C, reasonably high conductivity when impregnated with H{sub 3}PO{sub 4} and a low permeability to alcohols compared to Nafion. Preliminary fuel cells measurements on hydrogen based Polymer Electrolyte Membrane Fuel Cell (PEMFC) displayed an interestingly reasonable good fuel cell performance, a quite reduced loss when the hydrogen stream was polluted with carbon monoxide, and finally, when the system was tested with an ethanol/water (E/W) fuel, it displayed quite promising results that allows placing this system as an attractive option in order to increase the cell performance and deal with the typical limitations of low temperature Nafion-based PEMFC. (author)

  19. cGMP and nitric oxide modulate thrombin-induced endothelial permeability : Regulation via different pathways in human aortic and umbilical vein endothelial cells

    Draijer, R.; Atsma, D.E.; Laarse, A. van der; Hinsbergh, V.W.M. van


    Previous studies have demonstrated that cGMP and cAMP reduce the endothelial permeability for fluids and macromolecules when the endothelial permeability is increased by thrombin. In this study, we have investigated the mechanism by which cGMP improves the endothelial barrier function and examined w

  20. Materials for high-temperature fuel cells

    Jiang, San Ping; Lu, Max


    There are a large number of books available on fuel cells; however, the majority are on specific types of fuel cells such as solid oxide fuel cells, proton exchange membrane fuel cells, or on specific technical aspects of fuel cells, e.g., the system or stack engineering. Thus, there is a need for a book focused on materials requirements in fuel cells. Key Materials in High-Temperature Fuel Cells is a concise source of the most important and key materials and catalysts in high-temperature fuel cells with emphasis on the most important solid oxide fuel cells. A related book will cover key mater

  1. Scientific perspectives on extending the provision for waivers of in vivo bioavailability and bioequivalence studies for drug products containing high solubility-low permeability drugs (BCS-Class 3).

    Stavchansky, Salomon


    Recently, there has been increased interest in extending the provision for waivers of in vivo bioavailability and bioequivalence (BA-BE) studies that appeared in the guidance published by the Food and Drug Administration (FDA) (1) to pharmaceutical products containing Class 3 drugs (High solubility-Low Permeability). The extension of the Biopharmaceutics Classification System (BCS) to Class 3 drugs is meritorious because of its impact on public health policy considerations. The rate limiting step in the absorption of Class 3 drugs is the permeability through the intestinal membrane. This commentary will focus its attention on the scientific considerations which need to be examined to assess the risk and the benefit prior to granting a waiver of in vivo bioavailability and/or bioequivalence studies for Class 3 drugs. It will examine the forces affecting the interconnectivity of the neuronal, immunological and hormonal systems in the gastrointestinal tract that may affect its permeability and functionality. It will also challenge the assumption that in vitro dissolution and in vitro permeability studies in tissue cultures in the presence and absence of excipients are good predictors for in vivo dissolution and in vivo permeability which are at the heart of the BCS.

  2. High Resolution ground penetrating radar (GPR) measurements at the laboratory scale to model porosity and permeability in the Miami Limestone in South Florida.

    Mount, G. J.; Comas, X.


    Subsurface water flow within the Biscayne aquifer is controlled by the heterogeneous distribution of porosity and permeability in the karst Miami Limestone and the presence of numerous dissolution and mega-porous features. The dissolution features and other high porosity areas can create preferential flow paths and direct recharge to the aquifer, which may not be accurately conceptualized in groundwater flow models. As hydrologic conditions are undergoing restoration in the Everglades, understanding the distribution of these high porosity areas within the subsurface would create a better understanding of subsurface flow. This research utilizes ground penetrating radar to estimate the spatial variability of porosity and dielectric permittivity of the Miami Limestone at centimeter scale resolution at the laboratory scale. High frequency GPR antennas were used to measure changes in electromagnetic wave velocity through limestone samples under varying volumetric water contents. The Complex Refractive Index Model (CRIM) was then applied in order to estimate porosity and dielectric permittivity of the solid phase of the limestone. Porosity estimates ranged from 45.2-66.0% from the CRIM model and correspond well with estimates of porosity from analytical and digital image techniques. Dielectric permittivity values of the limestone solid phase ranged from 7.0 and 13.0, which are similar to values in the literature. This research demonstrates the ability of GPR to identify the cm scale spatial variability of aquifer properties that influence subsurface water flow which could have implications for groundwater flow models in the Biscayne and potentially other shallow karst aquifers.

  3. Soils - Mean Permeability

    Kansas Data Access and Support Center — This digital spatial data set provides information on the magnitude and spatial pattern of depth-weighted, mean soil permeability throughout the State of Kansas. The...

  4. Permeable pavement study (Edison)

    U.S. Environmental Protection Agency — While permeable pavement is increasingly being used to control stormwater runoff, field-based, side-by-side investigations on the effects different pavement types...

  5. Highly Permeable AlPO-18 Membranes for N 2 /CH 4 Separation

    Zong, Zhaowang; Elsaidi, Sameh K.; Thallapally, Praveen K.; Carreon, Moises A.


    Herein we demonstrate that AlPO-18 membranes can separate N2/CH4 gas mixtures at unprecedented N2 permeances. The best membranes separated N2/CH4 mixtures with N2 permeances as high as 3076 GPU and separation selectivities as high as 4.6. Gas mixture separation data, N2 and CH4 adsorption isotherms, ideal adsorbed solution theory (IAST), and breakthrough experiments were collected to understand the separation mechanisms. Competitive adsorption and differences in diffusivities were identified as the prevailing separation mechanisms. Differences in diffusivity played a more dominant role than the competitive adsorption, and led to nitrogen selective membranes.

  6. The Determination of Polyethylene Glycol in Untreated Urine Samples by High Performance Liquid Chromatography for Intestinal Permeability Studies

    Larsen, Elfinn; Pedersen, Walther Batsberg; Philipsen, E.


    Polyethylene glycol in urine samples has been investigated by high performance liquid chromatography. The molecular weights ranged from 634 to 1338. The urine samples were applied to the chromatographic system without any pre-treatment. For samples with a concentration of 0.2% polyethylene glycol...

  7. Improved Electrodes for High Temperature Proton Exchange Membrane Fuel Cells using Carbon Nanospheres.

    Zamora, Héctor; Plaza, Jorge; Cañizares, Pablo; Lobato, Justo; Rodrigo, Manuel A


    This work evaluates the use of carbon nanospheres (CNS) in microporous layers (MPL) of high temperature proton exchange membrane fuel cell (HT-PEMFC) electrodes and compares the characteristics and performance with those obtained using conventional MPL based on carbon black. XRD, hydrophobicity, Brunauer-Emmett-Teller theory, and gas permeability of MPL prepared with CNS were the parameters evaluated. In addition, a short life test in a fuel cell was carried out to evaluate performance under accelerated stress conditions. The results demonstrate that CNS is a promising alternative to traditional carbonaceous materials because of its high electrochemical stability and good electrical conductivity, suitable to be used in this technology.

  8. IL-4 and IL-13 induce protection from complement and melittin in endothelial cells despite initial loss of cytoplasmic proteins: membrane resealing impairs quantifying cytotoxicity with the lactate dehydrogenase permeability assay.

    Benson, Barbara A; Vercellotti, Gregory M; Dalmasso, Agustin P


    Endothelial cell activation and injury by the terminal pathway of complement is important in various pathobiological processes, including xenograft rejection. Protection against injury by human complement can be induced in porcine endothelial cells (ECs) with IL-4 and IL-13 through metabolic activation. However, despite this resistance, the complement-treated ECs were found to lose membrane permeability control assessed with the small molecule calcein. Therefore, to define the apparent discrepancy of permeability changes vis-à-vis the protection from killing, we now investigated whether IL-4 and IL-13 influence the release of the large cytoplasmic protein lactate dehydrogenase (LDH) in ECs incubated with complement or the pore-forming protein melittin. Primary cultures of ECs were pre-treated with IL-4 or IL-13 and then incubated with human serum as source of antibody and complement or melittin. Cell death was assessed using neutral red. Membrane permeability was quantitated measuring LDH release. We found that IL-4-/IL-13-induced protection of ECs from killing by complement or melittin despite loss of LDH in amounts similar to control ECs. However, the cytokine-treated ECs that were protected from killing rapidly regained effective control of membrane permeability. Moreover, the viability of the protected ECs was maintained for at least 2 days. We conclude that the protection induced by IL-4/IL-13 in ECs against lethal attack by complement or melittin is effective and durable despite severe initial impairment of membrane permeability. The metabolic changes responsible for protection allow the cells to repair the membrane injury caused by complement or melittin.

  9. Monodomain Design and Permeability Study of High- Q-Factor NiCuZn Ferrites for Near-Field Communication Application

    Liu, Weihu; Yan, Shuoqing; Cheng, Yongzhi; Li, Qifan; Feng, Zekun; Wang, Xian; Gong, Rongzhou; Nie, Yan


    For application in near-field communication (NFC) systems, monodomain NiCuZn ferrite has been prepared by the conventional solid-state reaction method. The results show that the monodomain design is effective; the Q factor of this kind of sample with 0.3 wt.% Co2O3 doping could be as high as 118.19. To obtain guidelines for preparing high-quality materials, the magnetic spectra of monodomain ferrite samples are studied via a numerical fitting method. According to the results of this permeability spectra fitting, the magnetic spectra of our prepared samples are mainly determined by the static spin susceptibility K s, spin resonance frequency ω 0, and relaxation frequency ω r. Specifically, we find that ω r varies with the frequency of the external field f. Furthermore, we demonstrate that Co2O3 influences the relationship between ω r and the frequency of the external field. According to theoretical expectations, a higher Q factor at frequency of 13.56 MHz could be obtained by using an appropriate Co2O3 content.

  10. A controlled wet-spinning and dip-coating process for preparation of high-permeable TiO2 hollow fiber membranes.

    Zhang, Qi; Wang, Hua; Fan, Xinfei; Chen, Shuo; Yu, Hongtao; Quan, Xie


    In order to improve the permeate flux of photocatalytic membranes, we present an approach for coupling TiO2 with ceramic hollow fiber membranes. The ceramic hollow fiber membranes with high permeate flux were fabricated by a controlled wet-spinning process using polyethersulfone (PESf) and ceramic powder as precursors and 1-methyl-2-pyrrolidinone as solvent, and the subsequent TiO2 coating was performed by a dip-coating process using tetra-n-butyl titanate as precursor. It has been found that the PESf/ceramic powder ratio could influence the structure of the membranes. Here the as-prepared TiO2 hollow fiber membranes had a pure water flux of 4,450 L/(m(2)·h). The performance of the TiO2 hollow fiber membrane was evaluated using humic acid (HA) as a test substance. The results demonstrated that this membrane exhibited a higher permeate flux under UV irradiation than in the dark and the HA removal efficiency was enhanced. The approach described here provides an operable route to the development of high-permeable photocatalytic membranes for water treatment.

  11. Tromboembolismo pulmonar masivo de alto riesgo asociado a foramen oval permeable High-risk massive pulmonary thromboembolism associated with patent foramen ovale

    Antonio Miranda


    Full Text Available La alta mortalidad de los pacientes con tromboembolismo pulmonar masivo de alto riesgo amerita un enfoque terapéutico enérgico e invasivo que incluya la embolectomía pulmonar quirúrgica en aquellos pacientes con contraindicación para trombolisis o trombolisis fallida. Describimos un caso de tromboembolismo pulmonar masivo de alto riesgo que recibió tratamiento quirúrgico en vez de trombolisis debido a que al momento del diagnóstico presentaba un trombo móvil a través de un foramen oval permeable con altísima posibilidad de embolismo paradójico arterial.High mortality rate associated with massive pulmonary embolism requires an aggressive invasive approach including surgical pulmonary embolectomy when thrombolytic therapy has failed or is contraindicated. We describe a case of high-risk massive pulmonary embolism who underwent surgical treatment due to the presence of a mobile intracardiac clot in a patent foramen ovale, and the possible risk of paradoxical arterial embolism.

  12. High frequency complex permeability of flake-shaped Fe74Cr2Mo2Sn2P10Si4B4C2 particle composite material


    A composite consisting of flake-shaped Fe74Cr2Mo2Sn2P10Si4B4C2 particles was prepared.The flake-shaped particles included in the composite were prepared by planetary ball milling.The complex permeability of the composite material was measured at frequency range from 10 MHz to 8.5 GHz.The permeability of the composite containing flake-shaped particles was much higher than that of the spherical particles.The permeability of particles was improved by means of heat-treatment.In contrast to the random-spatial-distributed flake-shaped particle composites,the oriented-distributed ones had a higher effective permeability.A model was proposed considering the orientation ratio,and our calculation agreed well with the experimental data.

  13. Hydrogen permeable high-temperature membranes. Development and application in gas separation processes and in hydrogen-producing reactions for process heat utilization. Wasserstoffpermeable Hochtemperaturmembranen. Entwicklung und Einsatz in Gastrennprozessen und in Wasserstoff erzeugenden Reaktionen zur Nutzung von Prozesswaerme

    Weirich, W.


    The thesis deals with hydrogen-permeable, high-temperature membranes consisting of metal-bare or ceramic-base materials, discussing their fabrication and properties as well as the analyses and experimentes performed for studying the applicability of these membranes in hydrogen production processes or in energy conversion processes for the utilization of process heat from HTGR-type reactors. (orig.)

  14. Engineering of a novel tri-functional enzyme with MnSOD, catalase and cell-permeable activities.

    Luangwattananun, Piriya; Yainoy, Sakda; Eiamphungporn, Warawan; Songtawee, Napat; Bülow, Leif; Ayudhya, Chartchalerm Isarankura Na; Prachayasittikul, Virapong


    Cooperative function of superoxide dismutase (SOD) and catalase (CAT), in protection against oxidative stress, is known to be more effective than the action of either single enzyme. Chemical conjugation of the two enzymes resulted in molecules with higher antioxidant activity and therapeutic efficacy. However, chemical methods holds several drawbacks; e.g., loss of enzymatic activity, low homogeneity, time-consuming, and the need of chemical residues removal. Yet, the conjugated enzymes have never been proven to internalize into target cells. In this study, by employing genetic and protein engineering technologies, we reported designing and production of a bi-functional protein with SOD and CAT activities for the first time. To enable cellular internalization, cell penetrating peptide from HIV-1 Tat (TAT) was incorporated. Co-expression of CAT-MnSOD and MnSOD-TAT fusion genes allowed simultaneous self-assembly of the protein sequences into a large protein complex, which is expected to contained one tetrameric structure of CAT, four tetrameric structures of MnSOD and twelve units of TAT. The protein showed cellular internalization and superior protection against paraquat-induced cell death as compared to either complex bi-functional protein without TAT or to native enzymes fused with TAT. This study not only provided an alternative strategy to produce multifunctional protein complex, but also gained an insight into the development of therapeutic agent against oxidative stress-related conditions.

  15. Highly permeable double-skinned forward osmosis membranes for anti-fouling in the emulsified oil-water separation process

    Duong, Hoang Hanh Phuoc


    Forward osmosis (FO) has attracted wide attention in recent years. However, the FO performance may be restricted due to internal concentration polarization (ICP) and fast fouling propensity that occurs in the membrane sublayer. Particularly, these problems significantly affect the membrane performance when treating highly contaminated oily wastewater. Recently, double-skinned flat sheet cellulose acetate (CA) membranes consisting of two selective skins via the phase inversion method have demonstrated less ICP and fouling propensity over typical single-skinned membranes. However, these membranes exhibit low water fluxes of <12 LMH under 2 M NaCl draw solution. Therefore, a novel double-skinned FO membrane with a high water flux has been aimed for in this study for emulsified oil-water treatment. The double-skinned FO membrane comprises a fully porous sublayer sandwiched between (i) a truly dense skin for salt rejection and (ii) a fairly loose dense skin for emulsified oil particle rejection. The former dense skin is a polyamide synthesized via interfacial polymerization, while the latter one is a self-assembled sulfonated pentablock copolymer (Nexar copolymer) layer. The resultant double-skinned membrane exhibits a high water flux of 17.2 LMH and a low reverse salt transport of 4.85 gMH using 0.5 M NaCl as the draw solution and DI water as the feed. The double-skinned membrane outperforms the single-skinned membrane with much lower fouling propensity for emulsified oil-water separation. © 2014 American Chemical Society.

  16. Effective stress law for the permeability of a limestone

    Ghabezloo, Siavash; Guédon, Sylvine; Martineau, François


    The effective stress law for the permeability of a limestone is studied experimentally by performing constant head permeability tests in a triaxial cell with different conditions of confining pressure and pore pressure. Test results have shown that a pore pressure increase and a confining pressure decrease both result in an increase of the permeability, and that the effect of the pore pressure change on the variation of the permeability is more important than the effect of a change of the confining pressure. A power law is proposed for the variation of the permeability with the effective stress. The permeability effective stress coefficient increases linearly with the differential pressure and is greater than one as soon the differential pressure exceeds few bars. The test results are well reproduced using the proposed permeability-effective stress law. A conceptual pore-shell model based on a detailed observation of the microstructure of the studied limestone is proposed. This model is able to explain the ex...

  17. Upscaling of permeability field of fractured rock system: Numerical examples

    Bao, K.


    When the permeability field of a given porous medium domain is heterogeneous by the existence of randomly distributed fractures such that numerical investigation becomes cumbersome, another level of upscaling may be required. That is such complex permeability field could be relaxed (i.e., smoothed) by constructing an effective permeability field. The effective permeability field is an approximation to the real permeability field that preserves certain quantities and provides an overall acceptable description of the flow field. In this work, the effective permeability for a fractured rock system is obtained for different coarsening scenarios starting from very coarse mesh all the way towards the fine mesh simulation. In all these scenarios, the effective permeability as well as the pressure at each cell is obtained. The total flux at the exit boundary is calculated in all these cases, and very good agreement is obtained.

  18. Highly and Stably Water Permeable Thin Film Nanocomposite Membranes Doped with MIL-101 (Cr Nanoparticles for Reverse Osmosis Application

    Yuan Xu


    Full Text Available A hydrophilic, hydrostable porous metal organic framework (MOF material-MIL-101 (Cr was successfully doped into the dense selective polyamide (PA layer on the polysulfone (PS ultrafiltration (UF support to prepare a new thin film nanocomposite (TFN membrane for water desalination. The TFN-MIL-101 (Cr membranes were characterized by SEM, AFM, XPS, wettability measurement and reverse osmosis (RO test. The porous structures of MIL-101 (Cr can establish direct water channels in the dense selective PA layer for water molecules to transport through quickly, leading to the increasing water permeance of membranes. With good compatibility between MIL-101 (Cr nanoparticles and the PA layer, the lab made TFN-MIL-101 (Cr membranes integrated tightly and showed a high NaCl salt rejection. MIL-101 (Cr nanoparticles increased water permeance to 2.2 L/m2·h·bar at 0.05 w/v % concentration, 44% higher than the undoped PA membranes; meanwhile, the NaCl rejection remained higher than 99%. This study experimentally verified the potential use of MIL-101 (Cr in advanced TFN RO membranes, which can be used in the diversified water purification field.

  19. Water permeability in human airway epithelium

    Pedersen, Peter Steen; Procida, Kristina; Larsen, Per Leganger;


    Osmotic water permeability (P(f)) was studied in spheroid-shaped human airway epithelia explants derived from nasal polyps by the use of a new improved tissue collection and isolation procedure. The fluid-filled spheroids were lined with a single cell layer with the ciliated apical cell membrane...

  20. Limitations in the Use of Fluorescein Diacetate/Propidium Iodide (FDA/PI) and Cell Permeable Nucleic Acid Stains for Viability Measurements of Isolated Islets of Langerhans.

    Boyd, Vinc; Cholewa, Olivia Maria; Papas, Klearchos K


    BACKGROUND: A review of current literature shows that the combined use of the cell permeable esterase-substrate fluorescein diacetate (FDA) and the cell impermeant nucleic acid stain propidium iodide (PI) to be one of the most common fluorescence-based methods to assess the viability of isolated islets of Langerhans, and it is currently used for islet product release prior to transplantation in humans. However, results from this assay do not correlate with islet viability and function or islet transplantation success in animals or humans (Eckhard et al. 2004; Ricordi et al. 2001). This may be in part attributed to considerable differences as well as discrepancies in the use of these reagents on islets. We critically surveyed the literature and evaluated the impact of a number of variables associated with the use of FDA/PI to determine their reliability in assessing islet cell viability. In addition, we evaluated other fluorescent stains, such as SYTO(R)13, SYTO(R)24 and SYBR(R)14 as possible alternatives to FDA. RESULTS: We found that the stability of stains in storage and stock solutions, the number of islets stained, concentration of stains, staining incubation time, the buffer/media used, and the method of examining islets were significant in the final scoring of viability. For archival file photos, the exposure time and camera/software settings can also impact interpretation of viability. Although our results show that FDA does detect intracellular esterase activity and staining with PI does assess cell membrane integrity, the results obtained from using these stains did not correlate directly with expected islet function and viability per transplantation into diabetic athymic nude mice (Papas et al. 2007). In addition, the use of two nucleic acid stains, such as SYTO(R)13 and PI, for live/dead scoring exhibited staining anomalies which limit their accuracy in assessing islet viability. CONCLUSIONS: From a review of the literature and from our observations on

  1. Magnetization Response of the Bulk and Supplementary Magnetic Domain Structure in High-Permeability Steel Laminations Visualized In Situ by Neutron Dark-Field Imaging

    Betz, B.; Rauscher, P.; Harti, R. P.; Schäfer, R.; Irastorza-Landa, A.; Van Swygenhoven, H.; Kaestner, A.; Hovind, J.; Pomjakushina, E.; Lehmann, E.; Grünzweig, C.


    Industrial transformer cores are composed of stacked high-permeability steel laminations (HPSLs). The performance and degree of efficiency of transformers are directly determined by the magnetic properties of each HPSL. In this article, we show how the neutron dark-field image (DFI) allows for the in situ visualization of the locally resolved response of the bulk and supplementary magnetic domain structures in HPSLs under the influence of externally applied magnetic fields. In particular, we investigate the domain formation and growth along the initial magnetization curve up to the saturated state. For decreasing field, we visualize the recurrence of the hysteretic domain structure down to the remanent state. Additionally, the DFI allows us to derive a correlation between the grain orientation and the corresponding volume and supplementary domain structure. Furthermore, we visualize the influence of the insulation coating, introducing desired tensile stresses on the domain structures. To compare our DFI findings to traditional methods, we perform complementary surface-sensitive magneto-optical Kerr-microscopy investigations.

  2. Blood flow and permeability in microvessels

    Sugihara-Seki, Masako; Fu, Bingmei M.


    The mechanics of blood flow in microvessels and microvessel permeability are reviewed. In the first part, characteristics of blood flow in vivo and in vitro are described from a fluid-mechanical point of view, and mathematical models for blood flow in microvessels are presented. Possible causes of the increased flow resistance obtained in vivo compared to in vitro are examined, including the effects of irregularities of vessel lumen, the presence of endothelial surface glycocalyx and white blood cells. In the second part, the ultrastructural pathways and mechanisms whereby endothelial cells and the clefts between the cells modulate microvessel permeability to water and solutes are introduced. Previous and current models for microvessel permeability to water and solutes are reviewed. These models examine the role of structural components of interendothelial cleft, such as junction strands and surface glycocalyx, in the determination of water and solute transport across the microvessel walls. Transport models in the tissue space surrounding the microvessel are also described.

  3. Multiple—tube Permeable Element for Combined Blowing Converter

    YANGWen-yuan; XUWei-hua; 等


    The method consisting of cold test and hot simulating has been proved to be reliable for designing permeable ele-ments.The carbon-magnesia multiple-tube permeable ele-ments which are formed by isostatical pressure have higher density,high heat resistance and good thermoshock resis-tance,The brickwork ,maintenance and erosion meha-nism of permeable elements were studied.The tehnology of combined blowing wa established based on the peculiari-ties of semi-steel refining at Panzhihua Irom& Steel Co.(PZISC).The service life of permeable elements reahed 600 heats,that means the permeable elements can work synchronitially with converter lining.

  4. High-Cycle-Life Lithium Cell

    Yen, S. P. S.; Carter, B.; Shen, D.; Somoano, R.


    Lithium-anode electrochemical cell offers increased number of charge/ discharge cycles. Cell uses components selected for compatibility with electrolyte solvent: These materials are wettable and chemically stable. Low vapor pressure and high electrochemical stability of solvent improve cell packaging, handling, and safety. Cell operates at modest temperatures - less than 100 degrees C - and is well suited to automotive, communications, and other applications.

  5. The Membrane Permeability Outcome study.

    Locatelli, Francesco; Cavalli, Andrea; Manzoni, Celestina; Pontoriero, Giuseppe


    Many observational studies have consistently shown that high-flux hemodialysis has positive effects on the survival and morbidity of uremic patients when compared with low-flux hemodialysis. However, the HEMO study, a randomized trial designed to evaluate the effect of membrane permeability on patient survival, showed only an 8% non-statistically significant reduction of mortality, albeit a secondary analysis suggested an advantage for high-flux membranes in certain patient subgroups. The prospective, randomized Membrane Permeability Outcome (MPO) study investigated the impact of membrane permeability on survival in incident hemodialysis patients who had low albumin (≤4 g/dl) and normal albumin ( >4 g/dl) as separate randomization groups. Patients with serum albumin ≤4 g/dl had significantly better survival rates in the high-flux group compared with the low-flux group (p = 0.032). Moreover, a post-hoc secondary analysis showed that high-flux membranes may significantly improve survival in diabetic patients. No difference was found in patients with normal albumin levels. Considering the increasing number of dialysis patients with low serum albumin levels and with diabetes, the relevance of the MPO study led to the publication of a position statement by the European Renal Best Practice Advisory Board. This board strongly recommended that high-flux hemodialysis should be used for high-risk patients and, with a lower degree of evidence, even also for low-risk subjects due to the substantial reduction in β(2)-microglobulin levels observed in the high-flux group. Copyright © 2011 S. Karger AG, Basel.

  6. Biostable glucose permeable polymer


    A new biostable glucose permeable polymer has been developed which is useful, for example, in implantable glucose sensors. This biostable glucose permeable polymer has a number of advantageous characteristics and, for example, does not undergo hydrolytic cleavage and degradation, thereby providing...... a composition that facilitates long term sensor stability in vivo. The versatile characteristics of this polymer allow it to be used in a variety of contexts, for example to form the body of an implantable glucose sensor. The invention includes the polymer composition, sensor systems formed from this polymer...

  7. Lunar electrical conductivity and magnetic permeability

    Dyal, P.; Parkin, C. W.; Daily, W. D.


    Improved analytical techniques are applied to a large Apollo magnetometer data set to yield values of electroconductivity, temperature, magnetic permeability, and iron abundance. Average bulk electroconductivity of the moon is calculated to be .0007 mho/m; a rapid increase with depth to about .003 mho/m within 250 km is indicated. The temperature profile, obtained from the electroconductivity profile for olivine, indicates high lunar temperatures at relatively shallow depths. Magnetic permeability of the moon relative to its environment is calculated to be 1.008 plus or minus .005; a permeability relative to free space of 1.012 plus 0.011, minus 0.008 is obtained. Lunar iron abundances corresponding to this permeability value are 2.5 plus 2.3, minus 1.7 wt% free iron and 5.0-13.5 wt% total iron for a moon composed of a combination of free iron, olivine, and orthopyroxene.

  8. Permeability, diffusivity and solubility of carbon dioxide in fluoropolymers: An experimental and modeling study

    Neela, Vasu; von Solms, Nicolas


    . Using a high-pressure permeation cell, the permeability and diffusivity of carbon dioxide were measured in several polymers used as packing and sealing materials. These were the fluoropolymers PTFE, FKM and TFM, both pure and containing glass, graphite, Ekonol and polysulfone as additives...

  9. Anti-fouling and high water permeable forward osmosis membrane fabricated via layer by layer assembly of chitosan/graphene oxide

    Salehi, Hasan; Rastgar, Masoud; Shakeri, Alireza


    To date, forward osmosis (FO) has received considerable attention due to its potential application in seawater desalination. FO does not require external hydraulic pressure and consequently is believed to have a low fouling propensity. Despite the numerous privileges of FO process, a major challenge ahead for its development is the lack of high performance membranes. In this study, we fabricated a novel highly-efficient FO membrane using layer-by-layer (LbL) assembly of positive chitosan (CS) and negative graphene oxide (GO) nanosheets via electrostatic interaction on a porous support layer. The support layer was prepared by blending hydrophilic sulfonated polyethersulfone (SPES) into polyethersulfone (PES) matrix using wet phase inversion process. Various characterization techniques were used to confirm successful fabrication of LbL membrane. The number of layers formed on the SPES-PES support layer was easily adjusted by repeating the CS and GO deposition cycles. Thin film composite (TFC) membrane was also prepared by the same SPES-PES support layer and polyamide (PA) active layer to compare membranes performances. The water permeability and salt rejection of the fabricated membranes were obtained by two kinds of draw solutions (including Na2SO4 and sucrose) under two different membrane orientations. The results showed that membrane coated by a CS/GO bilayers had water flux of 2-4 orders of magnitude higher than the TFC one. By increasing the number of CS/GO bilayers, the selectivity of the LbL membrane was improved. The novel fabricated LbL membrane showed better fouling resistance than the TFC one in the feed solution containing 200 ppm of sodium alginate as a foulant model.

  10. Production of savinase and population viability of Bacillus clausii during high-cell-density fed-batch cultivations

    Christiansen, Torben; Michaelsen, S.; Wumpelmann, M.


    feed profiles applied and, in addition, there was a time-dependent decrease in specific productivity. The specific glucose uptake rate increased with time for constant specific growth rate indicating that the maintenance requirements increased with time, possibly due to a decreasing K+ concentration......The growth and product formation of a Savinase-producing Bacillus clausii were investigated in high-cell-density fed-batch cultivations with both linear and exponential feed profiles. The highest specific productivity of Savinase was observed shortly after the end of the initial batch phase for all....... The physiological state of the cells was monitored during the cultivations using a flow cytometry assay based on the permeability of the cell membrane to propidium iodide. In the latter parts of the fed-batch cultures with a linear feed profile, a large portion of the cell population was found to have a permeable...

  11. Structural determinants of glomerular permeability.

    Deen, W M; Lazzara, M J; Myers, B D


    Recent progress in relating the functional properties of the glomerular capillary wall to its unique structure is reviewed. The fenestrated endothelium, glomerular basement membrane (GBM), and epithelial filtration slits form a series arrangement in which the flow diverges as it enters the GBM from the fenestrae and converges again at the filtration slits. A hydrodynamic model that combines morphometric findings with water flow data in isolated GBM has predicted overall hydraulic permeabilities that are consistent with measurements in vivo. The resistance of the GBM to water flow, which accounts for roughly half that of the capillary wall, is strongly dependent on the extent to which the GBM surfaces are blocked by cells. The spatial frequency of filtration slits is predicted to be a very important determinant of the overall hydraulic permeability, in keeping with observations in several glomerular diseases in humans. Whereas the hydraulic resistances of the cell layers and GBM are additive, the overall sieving coefficient for a macromolecule (its concentration in Bowman's space divided by that in plasma) is the product of the sieving coefficients for the individual layers. Models for macromolecule filtration reveal that the individual sieving coefficients are influenced by one another and by the filtrate velocity, requiring great care in extrapolating in vitro observations to the living animal. The size selectivity of the glomerular capillary has been shown to be determined largely by the cellular layers, rather than the GBM. Controversial findings concerning glomerular charge selectivity are reviewed, and it is concluded that there is good evidence for a role of charge in restricting the transmural movement of albumin. Also discussed is an effect of albumin that has received little attention, namely, its tendency to increase the sieving coefficients of test macromolecules via steric interactions. Among the unresolved issues are the specific contributions of the

  12. Lymphocyte apoptosis: role of uremia and permeability of dialysis membrane.

    Soriano, Sagrario; Martín-Malo, Alejandro; Carracedo, Julia; Ramírez, Rafael; Rodríguez, Mariano; Aljama, Pedro


    Uremia is associated to host defense mechanism disorders. Lymphocyte apoptosis, which may cause alteration of the immune system, is increased in uremic patients. The aim of the present study was to determine if, in addition to uremia, dialysis membranes with different biocompatibility and permeability have an effect on lymphocyte apoptosis. Cell apoptosis and Fas expression were assessed using flow cytometry in four groups of patients: (1) uremic non-dialyzed (Non-D) patients; (2) hemodialysis (HD) patients on hemophan; (3) low-flux polysulfone, and (4) high-flux polysulfone membrane. Ten healthy volunteers were used as controls. At baseline, lymphocytes from patients on hemophan showed an increase in apoptosis (18.4 +/- 6.9%) as compared with Non-D (7.2 +/- 2.8%; p dialysis patients lymphocyte apoptosis is influenced not only by the biocompatibility but also by the permeability of the dialysis membrane.

  13. Functional characterization of S100A8 and S100A9 in altering monolayer permeability of human umbilical endothelial cells.

    Liqun Wang

    Full Text Available S100A8, S100A9 and S100A8/A9 complexes have been known as important endogenous damage-associated molecular pattern (DAMP proteins. But the pathophysiological roles of S100A8, S100A9 and S100A8/A9 in cardiovascular diseases are incompletely explained. In this present study, the effects of homo S100A8, S100A9 and their hetero-complex S100A8/A9 on endothelial barrier function were tested respectively in cultured human umbilical venous endothelial cells (HUVECs. The involvement of TLR4 and RAGE were observed by using inhibitor of TLR4 and blocking antibody of RAGE. The clarification of different MAPK subtypes in S100A8/A9-induced endothelial response was implemented by using specific inhibitors. The calcium-dependency was detected in the absence of Ca2+ or in the presence of gradient-dose Ca2+. The results showed that S100A8, S100A9 and S100A8/A9 could induce F-actin and ZO-1 disorganization in HUVECs and evoked the increases of HUVEC monolayer permeability in a dose- and time-dependent manner. The effects of S100A8, S100A9 and S100A8/A9 on endothelial barrier function depended on the activation of p38 and ERK1/2 signal pathways through receptors TLR4 and RAGE. Most importantly, we revealed the preference of S100A8 on TLR4 and S100A9 on RAGE in HUVECs. The results also showed the calcium dependency in S100A8- and S100A9-evoked endothelial response, indicating that calcium dependency on formation of S100A8 or A9 dimmers might be the prerequisite for this endothelial functional alteration.

  14. Dopamine enhances duodenal epithelial permeability via the dopamine D5 receptor in rodent.

    Feng, X-Y; Zhang, D-N; Wang, Y-A; Fan, R-F; Hong, F; Zhang, Y; Li, Y; Zhu, J-X


    The intestinal barrier is made up of epithelial cells and intercellular junctional complexes to regulate epithelial ion transport and permeability. Dopamine (DA) is able to promote duodenal epithelial ion transport through D1-like receptors, which includes subtypes of D1 (D1 R) and D5 (D5 R), but whether D1-like receptors influence the duodenal permeability is unclear. FITC-dextran permeability, short-circuit current (ISC ), Western blot, immunohistochemistry and ELISA were used in human D5 R transgenic mice and hyperendogenous enteric DA (HEnD) rats in this study. Dopamine induced a downward deflection in ISC and an increase in FITC-dextran permeability of control rat duodenum, which were inhibited by the D1-like receptor antagonist, SCH-23390. However, DA decreased duodenal transepithelial resistance (TER), an effect also reversed by SCH-23390. A strong immunofluorescence signal for D5 R, but not D1 R, was observed in the duodenum of control rat. In human D5 R knock-in transgenic mice, duodenal mucosa displayed an increased basal ISC with high FITC-dextran permeability and decreased TER with a lowered expression of tight junction proteins, suggesting attenuated duodenal barrier function in these transgenic mice. D5 R knock-down transgenic mice manifested a decreased basal ISC with lowered FITC-dextran permeability. Moreover, an increased FITC-dextran permeability combined with decreased TER and tight junction protein expression in duodenal mucosa were also observed in HEnD rats. This study demonstrates, for the first time, that DA enhances duodenal permeability of control rat via D5 R, which provides new experimental and theoretical evidence for the influence of DA on duodenal epithelial barrier function. © 2016 Scandinavian Physiological Society. Published by John Wiley & Sons Ltd.

  15. The steady-state dipole-flow test for characterization of hydraulic conductivity statistics in a highly permeable aquifer: Horkheimer Insel site, Germany.

    Zlotnik, V A; Zurbuchen, B R; Ptak, T


    Over the last decade the dipole-flow test (DFT) evolved from the general idea of using recirculatory flow to evaluate aquifer properties, to the development of prototype instrumentation and feasibility studies, to a reliable tool for characterization of aquifer heterogeneity. The DFT involves the interpretation of head in recirculatory flow between injection and extraction sections (chambers) in a single well isolated from each other by a multipacker system. In this study, the steady-state dipole flow test (DFT) has been used to characterize the statistics of horizontal hydraulic conductivity (Kr) of the highly permeable, heterogeneous, and thin aquifer at the Horkheimer Insel site, Germany. In previous studies, Kr estimates were based on the steady-state head difference between chambers. A new by-chamber interpretation is proposed that is based on drawdown within each individual chamber. This interpretation yields more detailed information on structure of heterogeneity of the aquifer without introducing complexity into the analysis. The DFT results indicate that Kr ranges from 49 to 6000 m/day (mean ln Kr [(m/s)] approximately -4, and variance of ln Kr [(m/s)] approximately 1-2). Descriptive statistics from the DFT compare well with those from previous field and laboratory tests (pumping, borehole flowmeter, and permeameter tests and grain-size analysis) at this site. It is shown that the role of confining boundaries in the DFT interpretation is negligible even in this case of a thin (< 4 m thick) aquifer. This study demonstrates the flexibility of the DFT and expands the potential application of this method to a wide range of hydrogeologic settings.

  16. Parallel screening of drug-like natural compounds using Caco-2 cell permeability QSAR model with applicability domain, lipophilic ligand efficiency index and shape property: A case study of HIV-1 reverse transcriptase inhibitors

    Patel, Rikin D.; Kumar, Sivakumar Prasanth; Patel, Chirag N.; Shankar, Shetty Shilpa; Pandya, Himanshu A.; Solanki, Hitesh A.


    The traditional drug design strategy centrally focuses on optimizing binding affinity with the receptor target and evaluates pharmacokinetic properties at a later stage which causes high rate of attrition in clinical trials. Alternatively, parallel screening allows evaluation of these properties and affinity simultaneously. In a case study to identify leads from natural compounds with experimental HIV-1 reverse transcriptase (RT) inhibition, we integrated various computational approaches including Caco-2 cell permeability QSAR model with applicability domain (AD) to recognize drug-like natural compounds, molecular docking to study HIV-1 RT interactions and shape similarity analysis with known crystal inhibitors having characteristic butterfly-like model. Further, the lipophilic properties of the compounds refined from the process with best scores were examined using lipophilic ligand efficiency (LLE) index. Seven natural compound hits viz. baicalien, (+)-calanolide A, mniopetal F, fagaronine chloride, 3,5,8-trihydroxy-4-quinolone methyl ether derivative, nitidine chloride and palmatine, were prioritized based on LLE score which demonstrated Caco-2 well absorption labeling, encompassment in AD structural coverage, better receptor affinity, shape adaptation and permissible AlogP value. We showed that this integrative approach is successful in lead exploration of natural compounds targeted against HIV-1 RT enzyme.

  17. Bile acids reduce endocytosis of high-density lipoprotein (HDL) in HepG2 cells.

    Röhrl, Clemens; Eigner, Karin; Fruhwürth, Stefanie; Stangl, Herbert


    High-density lipoprotein (HDL) transports lipids to hepatic cells and the majority of HDL-associated cholesterol is destined for biliary excretion. Cholesterol is excreted into the bile directly or after conversion to bile acids, which are also present in the plasma as they are effectively reabsorbed through the enterohepatic cycle. Here, we provide evidence that bile acids affect HDL endocytosis. Using fluorescent and radiolabeled HDL, we show that HDL endocytosis was reduced in the presence of high concentrations of taurocholate, a natural non-cell-permeable bile acid, in human hepatic HepG2 and HuH7 cells. In contrast, selective cholesteryl-ester (CE) uptake was increased. Taurocholate exerted these effects extracellularly and independently of HDL modification, cell membrane perturbation or blocking of endocytic trafficking. Instead, this reduction of endocytosis and increase in selective uptake was dependent on SR-BI. In addition, cell-permeable bile acids reduced HDL endocytosis by farnesoid X receptor (FXR) activation: chenodeoxycholate and the non-steroidal FXR agonist GW4064 reduced HDL endocytosis, whereas selective CE uptake was unaltered. Reduced HDL endocytosis by FXR activation was independent of SR-BI and was likely mediated by impaired expression of the scavenger receptor cluster of differentiation 36 (CD36). Taken together we have shown that bile acids reduce HDL endocytosis by transcriptional and non-transcriptional mechanisms. Further, we suggest that HDL endocytosis and selective lipid uptake are not necessarily tightly linked to each other.

  18. 深部岩体在高压水头作用下的渗透性状况试验%Experimental examination of permeability of deep rock mass under high water pressure

    黄震; 姜振泉; 孙晓倩; 曹丁涛; 王宗胜; 辛斌


    Water injection experiment is a particularly useful technique to determine the permeability of rock mass in deep geotechnical engineering. To probe the permeability of lower coal seam floor, high borehole pressure water injection experiment has been carried out to analyse the three segments of rock masses in one coal mine in Yanzhou, and numerous data have been obtained. Based on the formula of permeability coefficient as well as the analysis of the data, permeability coefficients of three segments of rock masses have been determined, and a generalized model has been established to describe the rela-tionship between permeability coefficients and pressure. The results show that the test pressure and the water discharge will suddenly change when the hydraulic fracturing occurs. The curves of permeability coefficient and pressure are provided with a significant geometric segmentation, which is stable before mutation and linear after mutation. Fissures will restore to be closed after water injection experiment, and the rock masses still have strong anti-permeability.%现场压水试验是揭示深部岩土工程中岩体渗透性能的可靠方法,为探究深部煤层底板岩体的渗透性,采用钻孔高压压水测试手段,对兖州矿区某矿下组煤底板三段岩体进行了现场压水试验,获得了大量压水测试数据.结合渗透系数计算公式,对试验数据进行了分析,获得了不同深度及不同岩性岩体的渗透系数值及渗透系数与压力关系的概化模式.研究结果表明:岩体发生渗流时,测试水压和流量会发生突变.岩体的渗透系数-压力关系曲线在几何形态表现出明显的分段性,即可分为突变点前的平稳段和突变点后的线性变化段.压水结束后,岩体裂隙会发生闭合,且仍具有较强的抗渗性.

  19. Glycosaminoglycan Binding and Non-Endocytic Membrane Translocation of Cell-Permeable Octaarginine Monitored by Real-Time In-Cell NMR Spectroscopy.

    Takechi-Haraya, Yuki; Aki, Kenzo; Tohyama, Yumi; Harano, Yuichi; Kawakami, Toru; Saito, Hiroyuki; Okamura, Emiko


    Glycosaminoglycans (GAGs), which are covalently-linked membrane proteins at the cell surface have recently been suggested to involve in not only endocytic cellular uptake but also non-endocytic direct cell membrane translocation of arginine-rich cell-penetrating peptides (CPPs). However, in-situ comprehensive observation and the quantitative analysis of the direct membrane translocation processes are challenging, and the mechanism therefore remains still unresolved. In this work, real-time in-cell NMR spectroscopy was applied to investigate the direct membrane translocation of octaarginine (R8) into living cells. By introducing 4-trifluoromethyl-l-phenylalanine to the N terminus of R8, the non-endocytic membrane translocation of (19)F-labeled R8 ((19)F-R8) into a human myeloid leukemia cell line was observed at 4 °C with a time resolution in the order of minutes. (19)F NMR successfully detected real-time R8 translocation: the binding to anionic GAGs at the cell surface, followed by the penetration into the cell membrane, and the entry into cytosol across the membrane. The NMR concentration analysis enabled quantification of how much of R8 was staying in the respective translocation processes with time in situ. Taken together, our in-cell NMR results provide the physicochemical rationale for spontaneous penetration of CPPs in cell membranes.

  20. Carvacrol and 1,8-cineole alone or in combination at sublethal concentrations induce changes in the cell morphology and membrane permeability of Pseudomonas fluorescens in a vegetable-based broth.

    de Sousa, Jossana Pereira; Torres, Rayanne de Araújo; de Azerêdo, Geíza Alves; Figueiredo, Regina Célia Bressan Queiroz; Vasconcelos, Margarida Angélica da Silva; de Souza, Evandro Leite


    This study aimed to investigate the effects of sublethal concentrations of carvacrol (CAR) and 1,8-cineole (CIN) alone and in combination on the morphology, cell viability and membrane permeability of Pseudomonas fluorescens ATCC 11253 cultivated in a vegetable-based broth. Transmission and scanning electron microscopy images of bacterial cells exposed to CAR and CIN alone or in combination showed marked ultrastructural changes after 1h of exposure. These changes included shrunken protoplasm, discontinuity of the outer and cytoplasmic membranes and leakage of the intracellular material. Confocal scanning laser microscopy images corroborated the electron microscopy data, showing a decrease in the number of SYTO-9 cells (intact cells) with a concomitant increase in the number of PI-positive cells (dead cells). All of these morphological changes are indicative of increased membrane permeability and the loss of bacterial envelope integrity, which ultimately lead to cell death. The combination of sublethal concentrations of CAR and CIN could be applied to inhibit the growth of P. fluorescens on vegetables.

  1. High Efficiency Polymer Solar Cells Technologies

    Abdrhman M G; LI Hang-quan; ZHANG Li-ye; ZHOU Bing


    The conjugated polymer-based solar cell is one of the most promising devices in search of sustainable, renewable energy sources in last decade. It is the youngest field in organic solar cell research and also is certainly the fastest growing one at the moment. In addition, the key factor for polymer-based solar cells with high-efficiency is to invent new materials. Organic solar cell has attracted significant researches and commercial interest due to its low cost in fabrication and flexibility in applications. However, they suffer from relatively low conversion efficiency. The summarization of the significance and concept of high efficiency polymer solar cell technologies are presented.

  2. Frequency Dependence of The Properties about High Permeability Ferrite and Transformer%高磁导率铁氧体及其变压器性质的频率依赖关系

    王波; 裴谐第; 廖绍彬


    研究了高起始磁导率(6000~20 000)铁氧体和由该磁芯制成的变压器的电感频率依赖关系.当驱动频率由100Hz增加到300kHz时,电感呈现V形L-f曲线.详细讨论了形成V形曲线的原因.%The research of the high initial permeability(6000~20000)ferrite and the frequency dependence of inductance of the ferrite transformer made by high permeability ferrite core are presented. When the driving frequency increases from 100Hz to 300kHz the inductance shows a Vshape L-f curve. The reasons forming such a V-shape curve are discussed in detail.

  3. High rate, high reliability Li/SO2 cells

    Chireau, R.


    The use of the lithium/sulfur dioxide system for aerospace applications is discussed. The high rate density in the system is compared to some primary systems: mercury zinc, silver zinc, and magnesium oxide. Estimates are provided of the storage life and shelf life of typical lithium sulfur batteries. The design of lithium cells is presented and criteria are given for improving the output of cells in order to achieve high rate and high reliability.

  4. The cell-permeable Aβ1-6A2VTAT(D) peptide reverts synaptopathy induced by Aβ1-42wt.

    Cimini, Sara; Sclip, Alessandra; Mancini, Simona; Colombo, Laura; Messa, Massimo; Cagnotto, Alfredo; Di Fede, Giuseppe; Tagliavini, Fabrizio; Salmona, Mario; Borsello, Tiziana


    Alzheimer disease (AD) is the most prevalent form of dementia. Loss of hippocampal synapses is the first neurodegenerative event in AD. Synaptic loss has been associated with the accumulation in the brain parenchyma of soluble oligomeric forms of amyloid β peptide (Aβ1-42wt). Clinical observations have shown that a mutation in the APP protein (A673V) causes an early onset AD-type dementia in homozygous carriers while heterozygous carriers are unaffected. This mutation leads to the formation of mutated Aβ peptides (Aβ1-42A2V) in homozygous patients, while in heterozygous subjects both Aβ1-42wt and Aβ1-42A2V are present. To better understand the impact of the A673V mutation in AD, we analyzed the synaptotoxic effect of oligomers formed by aggregation of different Aβ peptides (Aβ1-42wt or Aβ1-42A2V) and the combination of the two Aβ1-42MIX (Aβ1-42wt and Aβ1-42A2V) in an in vitro model of synaptic injury. We showed that Aβ1-42A2V oligomers are more toxic than Aβ1-42wt oligomers in hippocampal neurons, confirming the results previously obtained in cell lines. Furthermore, we reported that oligomers obtained by the combination of both wild type and mutated peptides (Aβ1-42MIX) did not exert synaptic toxicity. We concluded that the combination of Aβ1-42wt and Aβ1-42A2V peptides hinders the toxicity of Aβ1-42A2V and counteracts the manifestation of synaptopathy in vitro. Finally we took advantage of this finding to generate a cell-permeable peptide for clinical application, by fusing the first six residues of the Aβ1-42A2V to the TAT cargo sequence (Aβ1-6A2VTAT(D)). Noteworthy, the treatment with Aβ1-6A2VTAT(D) confers neuroprotection against both in vitro and in vivo synaptopathy models. Therefore Aβ1-6A2VTAT(D) may represent an innovative therapeutic tool to prevent synaptic degeneration in AD.

  5. Duration of ultrasound-mediated enhanced plasma membrane permeability

    Lammertink, Bart; Deckers, Roel; Storm, Gert; Moonen, Chrit; Bos, Clemens


    Ultrasound (US) induced cavitation can be used to enhance the intracellular delivery of drugs by transiently increasing the cell membrane permeability. The duration of this increased permeability, termed temporal window, has not been fully elucidated. In this study, the temporal window was investiga

  6. Duration of ultrasound-mediated enhanced plasma membrane permeability

    Lammertink, Bart; Deckers, Roel; Storm, Gerrit; Moonen, Chrit; Bos, Clemens


    Ultrasound (US) induced cavitation can be used to enhance the intracellular delivery of drugs by transiently increasing the cell membrane permeability. The duration of this increased permeability, termed temporal window, has not been fully elucidated. In this study, the temporal window was

  7. Liquid Permeability of Ceramic Foam Filters

    Zhang, Kexu


    This project is in support of the PhD project: ‘Removal of Inclusions from Liquid Aluminium using Electromagnetically Modified Filtration’. The purpose of this project was to measure the tortuosity, and permeability of ~50mm thick: 30, 40, 50 and 80 pores per inch (ppi) commercial alumina ceramic foam filters (CFFs). Measurements have been taken of: cell (pore), window and strut sizes, porosity, tortuosity and liquid permeability. Water velocity from ~0.015-0.77 m/s have been used ...

  8. High Efficiency Reversible Fuel Cell Power Converter

    Pittini, Riccardo

    The large scale integration of renewable energy sources requires suitable energy storage systems to balance energy production and demand in the electrical grid. Bidirectional fuel cells are an attractive technology for energy storage systems due to the high energy density of fuel. Compared...... entitled "High Efficiency Reversible Fuel Cell Power Converter" and it presents the design of a high efficiency dc-dc converter developed and optimized for bidirectional fuel cell applications. First, a brief overview of fuel cell and energy storage technologies is presented. Different system topologies...... to traditional unidirectional fuel cell, bidirectional fuel cells have increased operating voltage and current ranges. These characteristics increase the stresses on dc-dc and dc-ac converters in the electrical system, which require proper design and advanced optimization. This work is part of the PhD project...

  9. The effect of cholesterol and epicholesterol incorporation on the permeability and on the phase transition of intact Acholeplasma laidlawii cell membranes and derived liposomes

    Kruyff, B.; Demel, R.A.; Deenen, L.L.M. van

    1. 1. The effect of incorporated cholesterol and epicholesterol upon the glycerol and erythritol permeability through the membrane of Acholeplasma laidlawii (previously denoted as Mycoplasma laidlawii) is studied. Both sterols, when present in the growth medium, are incorporated to the same extent

  10. Ammonia and urea permeability of mammalian aquaporins

    Litman, Thomas; Søgaard, Rikke; Zeuthen, Thomas


    The human aquaporins,AQP3,AQP7, AQP8,AQP9, and possibly AQP10, are permeable to ammonia, and AQP7, AQP9, and possibly AQP3, are permeable to urea. In humans, these aquaporins supplement the ammonia transport of the Rhesus (Rh) proteins and the urea transporters (UTs). The mechanism by which...... ammonium is transported by aquaporins is not fully resolved. A comparison of transport equations, models, and experimental data shows that ammonia is transported in its neutral form, NH(3). In the presence of NH(3), the aquaporin stimulates H(+) transport. Consequently, this transport of H(+) is only...... significant at alkaline pH. It is debated whether the H(+) ion passes via the aquaporin or by some external route; the investigation of this problem requires the aquaporin-expressing cell to be voltage-clamped. The ammonia-permeable aquaporins differ from other aquaporins by having a less restrictive aromatic...

  11. In vivo human buccal permeability of nicotine

    Adrian, Charlotte L; Olin, Helle B D; Dalhoff, Kim


    The aim was to examine the in vivo buccal pH-dependent permeability of nicotine in humans and furthermore compare the in vivo permeability of nicotine to previous in vitro permeability data. The buccal permeability of nicotine was examined in a three-way cross-over study in eight healthy non......-smokers using a buccal perfusion cell. The disappearance of nicotine from perfusion solutions with pH 6.0, 7.4, and 8.1 was studied for 3h. The apparent permeability of nicotine (P(app)) was determined at each pH value. Parotid saliva was collected in an attempt to assess systemic levels of nicotine....... The disappearance rate of nicotine increased significantly as the pH increased, which resulted in P(app) values of 0.57+/-0.55 x 10(-4), 2.10+/-0.23 x 10(-4), and 3.96+/-0.54 x 10(-4)cms(-1) (mean+/-S.D.) at pH 6.0, 7.4, and 8.1, respectively. A linear relationship (R(2)=0.993) was obtained between the P...

  12. Magnetic core test stand for energy loss and permeability measurements at a high constant magnetization rate and test results for nanocrystalline and ferrite materials.

    Burdt, Russell; Curry, Randy D


    A test stand was developed to measure the energy losses and unsaturated permeability of toroidal magnetic cores, relevant to applications of magnetic switching requiring a constant magnetization rate of the order of 1-10 T/micros. These applications in pulsed power include linear induction accelerators, pulse transformers, and discharge switches. The test stand consists of a coaxial transmission line pulse charged up to 100 kV that is discharged into a magnetic core load. Suitable diagnostics measure the voltage across and the current through a winding on the magnetic core load, from which the energy losses and unsaturated permeability are calculated. The development of the test stand is discussed, and test results for ferrite CN20 and the nanocrystalline material Finemet FT-1HS are compared to demonstrate the unique properties of a nanocrystalline material. The experimental data are compared with published data in a similar parameter space to demonstrate the efficacy of the experimental methods.

  13. Strong coercivity reduction and high initial permeability in NiCoP coated BaFe12O19-polystyrene bilayer composite

    Hamad, Mahmoud A.; El-Sayed, Adly H.; Hemeda, O. M.; Tawfik, A.


    Soft-magnetic NiCoP coated hard-magnetic M-type ferrite BaFe12O19 (BaM)-polystyrene (PS) bilayer composite film was successfully synthesized. X-ray diffraction peaks exhibited no change in the structure of BaM after coating with PS. The NiCoP coated BaM-PS composite exhibited a wasp-waisted magnetic hysteretic loop with remarkable reduction in the coercivity, remanence and squareness with respect to BaM-PS, which is useful for the core of a magnetic switching device to control currents so large that they are unmanageable. Moreover, the initial permeability measurement exhibits initial permeability of around 100 000 and thermal stability up to 558 K, which is good for flux-amplifying components of smaller inductors.

  14. Vascular permeability and drug delivery in cancers

    Sandy eAzzi


    Full Text Available The endothelial barrier strictly maintains vascular and tissue homeostasis, and therefore modulates many physiological processes such as angiogenesis, immune responses, and dynamic exchanges throughout organs. Consequently, alteration of this finely tuned function may have devastating consequences for the organism. This is particularly obvious in cancers, where a disorganized and leaky blood vessel network irrigates solid tumors. In this context, vascular permeability drives tumor-induced angiogenesis, blood flow disturbances, inflammatory cell infiltration, and tumor cell extravasation. This can directly restrain the efficacy of conventional therapies by limiting intravenous drug delivery. Indeed, for more effective anti-angiogenic therapies, it is now accepted that not only should excessive angiogenesis be alleviated, but also that the tumor vasculature needs to be normalized. Recovery of normal state vasculature requires diminishing hyperpermeability, increasing pericyte coverage, and restoring the basement membrane, to subsequently reduce hypoxia and interstitial fluid pressure. In this review, we will introduce how vascular permeability accompanies tumor progression and, as a collateral damage, impacts on efficient drug delivery. The molecular mechanisms involved in tumor-driven vascular permeability will next be detailed, with a particular focus on the main factors produced by tumor cells, especially the emblematic vascular endothelial growth factor (VEGF. Finally, new perspectives in cancer therapy will be presented, centered on the use of anti-permeability factors and normalization agents.

  15. Variability in permeability and integrity of cell membrane and depletion of food reserves in neem (Azadirachta indica) seeds from trees of differ-ent age classes

    Devendra Kumar; Dhruv Kumar Mishra


    We quantified cell membrane permeability (electrical conduc-tivity-EC, water soluble sugar-WSS, and amino acids-AA) and integrity (phospholipids,α-tocopherol and lipid peroxidation) along with food reserve deterioration (total proteins, total sugar, and total starch) of neem seeds collected from various mother tree age classes and stored for 65 days in airtight plastic containers at ambient room temperature (35±5°C). Results show that the activities were higher in fresh seeds (EC 267.56-2950.01 µS/g, WSS 19.96-19.48 mg/g and AA 5.40-5.35 mg/g) and declined with increasing duration of storage period (EC 153.37-195.17 µS/g, WSS 3.13-4.17 mg/g and AA 4.29-4.49 mg/g after 35 days and EC 144.02-161.56 µS/g, WSS 2.06-2.40 mg/g and AA 3.98-4.27 mg/g after 65 days of storage). Phospholipids andα-tocopherol were higher in fresh seed (0.073-0.093 OD at 710 nm and 0.080-0.105 OD, respectively) and declined as storage duration in-creased (0.033-0.042 OD at 710 nm and 00.0010-0.0020 OD, respec-tively). Dead seeds showed reduced amounts of phospholipids and minimum activity ofα- tocopherol (antioxidants). The level of MDA was lower in fresh seeds (0.0066-0.0087 OD at 600-535 nm) and increased as storage duration increased (0.0248-0.0268 OD after 65 days of stor-age). The higher amount of MDA indicated that seeds died due to rancid-ity of the oil inside the seed. Neem seed cake was assessed for deteriora-tion of food reserves (total proteins, total sugar, and total starch), concen-trations of which were higher in fresh seed and declined as storage dura-tion increased. Germination was higher in fresh seeds and after 65 days, no germination was received perhaps due to deterioration of biochemi-cals in seeds. Patterns of seed deterioration were similar across all seed lots.

  16. Quantifying porosity, compressibility and permeability in Shale

    Mbia, Ernest Ncha; Fabricius, Ida Lykke; Frykman, Peter

    (XRD) of shale samples show about 50% silt and high content of kaolinite in the clay fraction when compared with offshore samples from the Central Graben. Porosity measurements from helium porosimetry-mercury immersion (HPMI), mercury injection capillary pressure (MICP) and nuclear magnetic resonance...... (NMR) show that, the MICP porosity is 9-10% points lower than HPMI and NMR porosity. Compressibility result shows that deep shale is stiffer in situ than normally assumed in geotechnical modelling and that static compressibility corresponds with dynamic one only at the begining of unloading stress...... strain data. We found that Kozeny's modelled permeability fall in the same order of magnitude with measured permeability for shale rich in kaolinite but overestimates permeability by two to three orders of magnitudes for shale with high content of smectite. The empirical Yang and Aplin model gives good...

  17. Relative permeability in dual porosity porous media

    Deghmoum, A. [SONATRACH CRD, Boumerdes (Algeria); Tiab, D. [Oklahoma Univ., Norman, OK (United States); Mazouzi, A. [SONATRACH PED (Algeria)


    One of the important factors in the field of reservoir simulation of dual-porosity systems is reliable relative permeability data. Laboratory limitations hinder measurements. The real behaviour of naturally fractures reservoirs is not reflected in the reservoir core samples, which as a rule originate from zones without induced or natural fractures. Therefore, it is commonly assumed that the relative permeability of a naturally fractured system is a straight line, which can cause errors. The authors undertook to conduct special core analyses on Berea outcrop core samples, to simulate fracture opening through the cutting of the samples to get different fracture apertures, to study the effects of dual porosity on the shape of capillary pressure curves, and to evaluate absolute and relative permeability, as they are affected by fracture opening. The correlation obtained between absolute permeability and fracture aperture was good, and capillary pressure curves permitted the observation of the effect of dual porosity. High residual oil saturation was present in the matrix, since the fractures became the easiest route for water flow, and this situation prevented the use of unsteady-state tests to measure relative permeability on the samples. Instead, the centrifuge technique was successfully used. A naturally fractured reservoir (NFR), the Tin Fouye Tabankort (TFT) reservoir in Algeria was selected to extend the findings. The site was principally selected due to the availability of naturally fractured cores and published data. Core observations, well test analysis and borehole imager tools were all TFT natural fracture indicators presented in the paper. Representative data of relative permeability was obtained by conducting a displacement test on a full diameter core to solve the laboratory limitations. The correlation between permeability and fracture opening was used to estimate the aperture of natural fractures in TFT reservoir. 17 refs., 2 tabs., 24 figs.

  18. A novel high reliability CMOS SRAM cell

    Xie Chengmin; Wang Zhongfang; Wu Longsheng; Liu Youbao


    A novel 8T single-event-upset (SEU) hardened and high static noise margin (SNM) SRAM cell is proposed.By adding one transistor paralleled with each access transistor,the drive capability of pull-up PMOS is greater than that of the conventional cell and the read access transistors are weaker than that of the conventional cell.So the hold,read SNM and critical charge increase greatly.The simulation results show that the critical charge is almost three times larger than that of the conventional 6T cell by appropriately sizing the pull-up transistors.The hold and read SNM of the new cell increase by 72% and 141.7%,respectively,compared to the 6T design,but it has a 54% area overhead and read performance penalty.According to these features,this novel cell suits high reliability applications,such as aerospace and military.

  19. Achieving High Performance Perovskite Solar Cells

    Yang, Yang


    Recently, metal halide perovskite based solar cell with the characteristics of rather low raw materials cost, great potential for simple process and scalable production, and extreme high power conversion efficiency (PCE), have been highlighted as one of the most competitive technologies for next generation thin film photovoltaic (PV). In UCLA, we have realized an efficient pathway to achieve high performance pervoskite solar cells, where the findings are beneficial to this unique materials/devices system. Our recent progress lies in perovskite film formation, defect passivation, transport materials design, interface engineering with respect to high performance solar cell, as well as the exploration of its applications beyond photovoltaics. These achievements include: 1) development of vapor assisted solution process (VASP) and moisture assisted solution process, which produces perovskite film with improved conformity, high crystallinity, reduced recombination rate, and the resulting high performance; 2) examination of the defects property of perovskite materials, and demonstration of a self-induced passivation approach to reduce carrier recombination; 3) interface engineering based on design of the carrier transport materials and the electrodes, in combination with high quality perovskite film, which delivers 15 ~ 20% PCEs; 4) a novel integration of bulk heterojunction to perovskite solar cell to achieve better light harvest; 5) fabrication of inverted solar cell device with high efficiency and flexibility and 6) exploration the application of perovskite materials to photodetector. Further development in film, device architecture, and interfaces will lead to continuous improved perovskite solar cells and other organic-inorganic hybrid optoelectronics.

  20. Planar multijunction high voltage solar cells

    Evans, J. C., Jr.; Chai, A. T.; Goradia, C.


    Technical considerations, preliminary results, and fabrication details are discussed for a family of high-voltage planar multi-junction (PMJ) solar cells which combine the attractive features of planar cells with conventional or interdigitated back contacts and the vertical multijunction (VMJ) solar cell. The PMJ solar cell is internally divided into many voltage-generating regions, called unit cells, which are internally connected in series. The key to obtaining reasonable performance from this device was the separation of top surface field regions over each active unit cell. Using existing solar cell fabricating methods, output voltages in excess of 20 volts per linear centimeter are possible. Analysis of the new device is complex, and numerous geometries are being studied which should provide substantial benefits in both normal sunlight usage as well as with concentrators.

  1. Relative permeability through fractures

    Diomampo, Gracel, P.


    The mechanism of two-phase flow through fractures is of importance in understanding many geologic processes. Currently, two-phase flow through fractures is still poorly understood. In this study, nitrogen-water experiments were done on both smooth and rough parallel plates to determine the governing flow mechanism for fractures and the appropriate methodology for data analysis. The experiments were done using a glass plate to allow visualization of flow. Digital video recording allowed instantaneous measurement of pressure, flow rate and saturation. Saturation was computed using image analysis techniques. The experiments showed that gas and liquid phases flow through fractures in nonuniform separate channels. The localized channels change with time as each phase path undergoes continues breaking and reforming due to invasion of the other phase. The stability of the phase paths is dependent on liquid and gas flow rate ratio. This mechanism holds true for over a range of saturation for both smooth and rough fractures. In imbibition for rough-walled fractures, another mechanism similar to wave-like flow in pipes was also observed. The data from the experiments were analyzed using Darcy's law and using the concept of friction factor and equivalent Reynold's number for two-phase flow. For both smooth- and rough-walled fractures a clear relationship between relative permeability and saturation was seen. The calculated relative permeability curves follow Corey-type behavior and can be modeled using Honarpour expressions. The sum of the relative permeabilities is not equal one, indicating phase interference. The equivalent homogeneous single-phase approach did not give satisfactory representation of flow through fractures. The graphs of experimentally derived friction factor with the modified Reynolds number do not reveal a distinctive linear relationship.

  2. Steam-water relative permeability

    Ambusso, W.; Satik, C.; Home, R.N. [Stanford Univ., CA (United States)


    A set of relative permeability relations for simultaneous flow of steam and water in porous media have been measured in steady state experiments conducted under the conditions that eliminate most errors associated with saturation and pressure measurements. These relations show that the relative permeabilities for steam-water flow in porous media vary approximately linearly with saturation. This departure from the nitrogen/water behavior indicates that there are fundamental differences between steam/water and nitrogen/water flows. The saturations in these experiments were measured by using a high resolution X-ray computer tomography (CT) scanner. In addition the pressure gradients were obtained from the measurements of liquid phase pressure over the portions with flat saturation profiles. These two aspects constitute a major improvement in the experimental method compared to those used in the past. Comparison of the saturation profiles measured by the X-ray CT scanner during the experiments shows a good agreement with those predicted by numerical simulations. To obtain results that are applicable to general flow of steam and water in porous media similar experiments will be conducted at higher temperature and with porous rocks of different wetting characteristics and porosity distribution.

  3. Dietary fat and bile juice, but not obesity, are responsible for the increase in small intestinal permeability induced through the suppression of tight junction protein expression in LETO and OLETF rats

    Suzuki Takuya


    Full Text Available Abstract Background An increase in the intestinal permeability is considered to be associated with the inflammatory tone and development in the obesity and diabetes, however, the pathogenesis of the increase in the intestinal permeability is poorly understood. The present study was performed to determine the influence of obesity itself as well as dietary fat on the increase in intestinal permeability. Methods An obese rat strain, Otsuka Long Evans Tokushima Fatty (OLETF, and the lean counter strain, Long Evans Tokushima Otsuka (LETO, were fed standard or high fat diets for 16 weeks. Glucose tolerance, intestinal permeability, intestinal tight junction (TJ proteins expression, plasma bile acids concentration were evaluated. In addition, the effects of rat bile juice and dietary fat, possible mediators of the increase in the intestinal permeability in the obesity, on TJ permeability were explored in human intestinal Caco-2 cells. Results The OLETF rats showed higher glucose intolerance than did the LETO rats, which became more marked with the prolonged feeding of the high fat diet. Intestinal permeability in the OLETF rats evaluated by the urinary excretion of intestinal permeability markers (Cr-EDTA and phenolsulfonphthalein was comparable to that in the LETO rats. Feeding the high fat diet increased intestinal permeability in both the OLETF and LETO rats, and the increases correlated with decreases in TJ proteins (claudin-1, claudin-3, occludin and junctional adhesion molecule-1 expression in the small, but not in the large intestine (cecum or colon. The plasma bile acids concentration was higher in rats fed the high fat diet. Exposure to bile juice and the fat emulsion increased TJ permeability with concomitant reductions in TJ protein expression (claudin-1, claudin-3, and junctional adhesion molecule-1 in the Caco-2 cell monolayers. Conclusion Excessive dietary fat and/or increased levels of luminal bile juice, but not genetic obesity, are

  4. Permeability measuremens of brazilian Eucalyptus

    Marcio Rogério da Silva


    Full Text Available The permeability of Brazilian Eucalyptus grandis and Eucalyptus citriodora wood was measured in a custom build gas analysis chamber in order to determine which species could be successfully treated with preservatives. Liquid permeability was tested using an emulsion of Neen oil and a control of distillated water. Air was used to test the gas phase permeability. For both Eucalyptus grandis and Eucalyptus citriodora, the longitudinal permeability of gas was shown to be about twice as great as the liquid phase permeability. No radial permeability was observed for either wood. The permeability of air and water through the sapwood of Eucalyptus grandis was greater than that through the sapwood of Eucalyptus citriodora. The permeability of neen oil preservative through the sapwood of Eucalyptus grandis was also greater than through the sapwood of E. Citradora, but the difference was not statistically significant. Scanning Electron Microscopy images showed that the distribution and obstruction in the vessels could be correlated with observed permeability properties. Irrespective of the causes of differences in permeability between the species, the fluid phase flux through the sapwood of both species was significant, indicating that both Eucalyptus grandis and Eucalyptus citriodora could be successfully treated with wood preservative.

  5. Very High Efficiency Solar Cell Modules

    Barnett, A.; Kirkpatrick, D.; Honsberg, C.; Moore, D.; Wanlass, M.; Emery, K.; Schwartz, R.; Carlson, D.; Bowden, S.; Aiken, D.; Gray, A.; Kurtz, S.; Kazmerski, L., et al


    The Very High Efficiency Solar Cell (VHESC) program is developing integrated optical system - PV modules for portable applications that operate at greater than 50% efficiency. We are integrating the optical design with the solar cell design, and have entered previously unoccupied design space. Our approach is driven by proven quantitative models for the solar cell design, the optical design, and the integration of these designs. Optical systems efficiency with an optical efficiency of 93% and solar cell device results under ideal dichroic splitting optics summing to 42.7 {+-} 2.5% are described.

  6. Parallel artificial membrane permeability assay for blood-brain permeability determination of illicit drugs and synthetic analogues.

    Clemons, Kristina; Kretsch, Amanda; Verbeck, Guido


    With the number of designer drugs on the streets rampantly on the rise, it's becoming more and more important to be able to rapidly characterize them in a biologically relevant way. Using a parallel artificial membrane permeability assay (PAMPA) to assess the blood brain barrier permeability has shown to be a high throughput way to compare new drugs with currently controlled substances via their effective permeability values. This combined with direct infusion electrospray ionization-mass spectrometry creates a rapid technique for characterization of new designer drugs. PAMPA has successfully determined the effective permeabilities of cocaine, methamphetamine, heroin, MDMA, and several tryptamine derivatives.

  7. The Permeability of Rubble Mound Breakwaters

    Williams, A.F.; Burcharth, H. F.; Adel, H. den


    The results of an extensive series of permeability experiments originally analysed by Shih (1990) are reinterpreted in the light of new experiments. It is proposed that the Forchheimer equation might not fully describe flow at the high Reynolds numbers found in the interior of rubble material....... A new series of tests designed to test for deviations from the Forchheimer equation and investigate the effects of material shape are described. While no evidence can be found to indicate a deviation from the Forchheimer equation a dependency of permeability and the surface roughness the material...

  8. Gyroid Nanoporous Membranes with Tunable Permeability

    Li, Li; Schulte, Lars; Clausen, Lydia D.


    Understanding the relevant permeability properties of ultrafiltration membranes is facilitated by using materials and procedures that allow a high degree of control on morphology and chemical composition. Here we present the first study on diffusion permeability through gyroid nanoporous cross......-sided skin membranes, much faster than expected by a naive resistance-in-series model; the flux through the two-sided skin membranes even increases with the membrane thickness. We propose a model that captures the physics behind the observed phenomena, as confirmed by flow visualization experiments...... the effective diffusion coefficients of a series of antibiotics, proteins, and other biomolecules; solute permeation is discussed in terms of hindered diffusion. The combination of uniform bulk morphology, isotropically percolating porosity, controlled surface chemistry, and tunable permeability is distinctive...

  9. Permeability of Hollow Microspherical Membranes to Helium

    Zinoviev, V. N.; Kazanin, I. V.; Pak, A. Yu.; Vereshchagin, A. S.; Lebiga, V. A.; Fomin, V. M.


    This work is devoted to the study of the sorption characteristics of various hollow microspherical membranes to reveal particles most suitable for application in the membrane-sorption technologies of helium extraction from a natural gas. The permeability of the investigated sorbents to helium and their impermeability to air and methane are shown experimentally. The sorption-desorption dependences of the studied sorbents have been obtained, from which the parameters of their specific permeability to helium are calculated. It has been established that the physicochemical modification of the original particles exerts a great influence on the coefficient of the permeability of a sorbent to helium. Specially treated cenospheres have displayed high efficiency as membranes for selective extraction of helium.

  10. In vitro porcine blood-brain barrier model for permeability studies: pCEL-X software pKa(FLUX) method for aqueous boundary layer correction and detailed data analysis.

    Yusof, Siti R; Avdeef, Alex; Abbott, N Joan


    In vitro blood-brain barrier (BBB) models from primary brain endothelial cells can closely resemble the in vivo BBB, offering valuable models to assay BBB functions and to screen potential central nervous system drugs. We have recently developed an in vitro BBB model using primary porcine brain endothelial cells. The model shows expression of tight junction proteins and high transendothelial electrical resistance, evidence for a restrictive paracellular pathway. Validation studies using small drug-like compounds demonstrated functional uptake and efflux transporters, showing the suitability of the model to assay drug permeability. However, one limitation of in vitro model permeability measurement is the presence of the aqueous boundary layer (ABL) resulting from inefficient stirring during the permeability assay. The ABL can be a rate-limiting step in permeation, particularly for lipophilic compounds, causing underestimation of the permeability. If the ABL effect is ignored, the permeability measured in vitro will not reflect the permeability in vivo. To address the issue, we explored the combination of in vitro permeability measurement using our porcine model with the pKa(FLUX) method in pCEL-X software to correct for the ABL effect and allow a detailed analysis of in vitro (transendothelial) permeability data, Papp. Published Papp using porcine models generated by our group and other groups are also analyzed. From the Papp, intrinsic transcellular permeability (P0) is derived by simultaneous refinement using a weighted nonlinear regression, taking into account permeability through the ABL, paracellular permeability and filter restrictions on permeation. The in vitro P0 derived for 22 compounds (35 measurements) showed good correlation with P0 derived from in situ brain perfusion data (r(2)=0.61). The analysis also gave evidence for carrier-mediated uptake of naloxone, propranolol and vinblastine. The combination of the in vitro porcine model and the software

  11. Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

    Amin Abbaszadeh Banaeiyan


    Full Text Available The possibility to conduct complete cell assays under a precisely controlled environment while consuming minor amounts of chemicals and precious drugs have made microfluidics an interesting candidate for quantitative single-cell studies. Here, we present an application-specific microfluidic device, cellcomb, capable of conducting high-throughput single-cell experiments. The system employs pure hydrodynamic forces for easy cell trapping and is readily fabricated in polydimethylsiloxane (PDMS using soft lithography techniques. The cell-trapping array consists of V-shaped pockets designed to accommodate up to six Saccharomyces cerevisiae (yeast cells with the average diameter of 4 μm. We used this platform to monitor the impact of flow rate modulation on the arsenite (As(III uptake in yeast. Redistribution of a green fluorescent protein (GFP-tagged version of the heat shock protein Hsp104 was followed over time as read out. Results showed a clear reverse correlation between the arsenite uptake and three different adjusted low = 25 nL min−1, moderate = 50 nL min−1, and high = 100 nL min−1 flow rates. We consider the presented device as the first building block of a future integrated application-specific cell-trapping array that can be used to conduct complete single cell experiments on different cell types.

  12. High Radiation Resistance IMM Solar Cell

    Pan, Noren


    Due to high launch costs, weight reduction is a key driver for the development of new solar cell technologies suitable for space applications. This project is developing a unique triple-junction inverted metamorphic multijunction (IMM) technology that enables the manufacture of very lightweight, low-cost InGaAsP-based multijunction solar cells. This IMM technology consists of indium (In) and phosphorous (P) solar cell active materials, which are designed to improve the radiation-resistant properties of the triple-junction solar cell while maintaining high efficiency. The intrinsic radiation hardness of InP materials makes them of great interest for building solar cells suitable for deployment in harsh radiation environments, such as medium Earth orbit and missions to the outer planets. NASA Glenn's recently developed epitaxial lift-off (ELO) process also will be applied to this new structure, which will enable the fabrication of the IMM structure without the substrate.

  13. Test of blockers of AQP1 water permeability by a high-resolution method: no effects of tetraethylammonium ions or acetazolamide

    Søgaard, Rikke; Zeuthen, Thomas


    The effects of putative water channel blockers were tested on AQP1-expressing Xenopus laevis oocytes by a fast optical method with a time resolution of 1 s and a volume resolution of 20 pl. The oocytes were exposed to external hyposmolarity and the osmotic water permeability (Lp) derived from...... will be underestimated. (b) Hyposmotic gradients implemented by dilution with water will entail changes in the ionic strength as well; this may enhance loss of salt from the oocyte. (c) By voltage clamping the AQP1-expressing oocytes during hyposmotic challenges, we found that TEA+-treated oocytes were more electrically...

  14. Hypoxia in high glucose followed by reoxygenation in normal glucose reduces the viability of cortical astrocytes through increased permeability of connexin 43 hemichannels

    Orellana, Juan A.; Hernández, Diego E.; Ezan, Pascal; Velarde, Victoria; Bennett, Michael V. L.; Giaume, Christian; Sáez, Juan C.


    Brain ischemia causes more extensive injury in hyperglycemic than normoglycemic subjects, and the increased damage is to astroglia as well as neurons. In the present work, we found that in cortical astrocytes from rat or mouse, reoxygenation after hypoxia in a medium mimicking interstitial fluid during ischemia increases hemichannel activity and decreases cell-cell communication via gap junctions as indicated by dye uptake and dye coupling, respectively. These effects were potentiated by high glucose during the hypoxia in a concentration-dependent manner (and by zero glucose) and were not observed in connexin 43−/− astrocytes. The responses were transient or persistent after short and long periods of hypoxia, respectively. The persistent responses were associated with a progressive reduction in cell viability that was prevented by La3+ or peptides that block connexin 43 (Cx43) hemichannels or by inhibition of p38 MAP kinase prior to hypoxia-reoxygenation but not by treatments that block pannexin hemichannels. Block of Cx43 hemichannels did not affect the reduction in gap junction mediated dye coupling observed during reoxygenation. Cx43 hemichannels may be a novel therapeutic target to reduce cell death following stroke, particularly in hyperglycemic conditions. PMID:19705457

  15. Temporal and Tight Hepatitis C Virus Gene Activation in Cultured Human Hepatoma Cells Mediated by a Cell-Permeable Cre Recombinase

    Dong XIAO; Kang XU; Ying YUE; Zhong-Min GUO; Bing HUANG; Xin-Yan DENG; Huan TANG; Xi-Gu CHEN


    Conditional gene expression has greatly facilitated the examination of the functions of particular gene products. Using the Cre/lox P switching expression system, we plan to develop efficient conditional transgene activation of hepatitis C virus core protein (HCV-C) cDNA (nucleotide 342-914) in the transgenic mice to overcome "immune tolerance" formed during the embryonic period and "immune escape" against hepatitis virus antigen in our project. To use this system in vivo, the dormant transgenic construct, i.e.,pApoE-SCS-EGFP-HCV-C, was generated using techniques of standard molecular biology. The liverspecific human apoE promoter was here used to target expression of genes of interest (EGFP and HCV-C) to murine liver. Prior to generating the transgenic mice, the availability of Cre/lox P system and construct functionality were successfully verified by a cell-free recombination system and via checking the expression of EGFP and HCV-C in the human hepatoma cells at the mRNA and protein levels. These results suggest that the Cre/lox P system could tightly control expression of EGFP and HCV-C in vitro, which laid a solid foundation to conditionally activate expression of target gene(s) in transgenic mice by Cre-mediated site-specific recombination.

  16. Probability distribution of biofilm thickness and effect of biofilm on the permeability of porous media

    Ye, S.; Sleep, B. E.; Chien, C.


    Probability distribution of biofilm thickness and effect of biofilm on permeability of saturated porous media were investigated in a two-dimensional sand-filled cell (55 cm wide x 45 cm high x 1.28 cm thick) under condition of rich nutrition. Inoculation of the lower portion of the cell with a methanogenic culture and addition of methanol to the bottom of the cell led to biomass growth. Biomass distributions in the water and on the sand in the cell were measured by protein analysis. The biofilm distribution on the sand was observed by confocal laser scanning microscopy (CLSM). Permeability was measured by laboratory hydraulic tests. The biomass levels measured in water and on the sand increased with time, and were highest at the bottom of the cell. The biofilm on the sand at the bottom of the cell was thicker. Biomass distribution on the grain of sand was not uniform. Biofilm thickness was a random variable with a normal distribution by statistical analysis of CLSM images. The results of the hydraulic tests demonstrated that the permeability due to biofilm growth was estimated to be average 12% of the initial value. To investigate the spatial distribution of permeability in the two dimensional cell, three models (Taylor, Seki, and Clement) were used to calculate permeability of porous media with biofilm growth. The results of Taylor's model (Taylor et al., 1990) showed reduction in permeability of 2-5 orders magnitude. The Clement's model (Clement et al., 1996) predicted 3%-98% of the initial value. Seki's model (Seki and Miyazaki, 2001) could not be applied in this study. Conclusively, biofilm growth could obviously decrease the permeability of two dimensional saturated porous media, however, the reduction was much less than that estimated in one dimensional condition. Additionally, under condition of two dimensional saturated porous media with rich nutrition, Seki's model could not be applied, Taylor’s model predicted bigger reductions, and the results of

  17. Selective permeability of PVA membranes. I - Radiation-crosslinked membranes

    Katz, M. G.; Wydeven, T., Jr.


    The water and salt transport properties of ionizing radiation crosslinked poly(vinyl alcohol) (PVA) membranes were investigated. The studied membranes showed high permeabilities and low selectivities for both water and salt. The results were found to be in accord with a modified solution-diffusion model for transport across the membranes, in which pressure-dependent permeability coefficients are employed.

  18. Selective Permeability of PVA Membranes. I: Radiation-Crosslinked Membranes

    Katz, Moshe G.; Wydeven, Theodore, Jr.


    The water and salt transport properties of ionizing radiation crosslinked poly(vinyl alcohol) (PVA) membranes were investigated. The studied membranes showed high permeabilities and low selectivities for both water and salt. The results were found to be in accord with a modified solution-diffusion model for transport across the membranes, in which pressure-dependent permeability coefficients are employed.

  19. Characteristics of permeability in carbonate areas of Korea

    Park, Y.; Lee, J.; Lim, H.; Keehm, Y.


    Permeability (hydraulic conductivity) in carbonate areas is affected by various factors such as fracture, pore and degree of weathering and diagenesis. Also, caves developed in carbonate area are main factors. This study was performed to understand factors controlling the permeability in carbonate areas in Korea. In order to conduct this study, the permeability and well logging data (n=30) were collected from many literatures and rock samples were collected around wells. Vertical profile of the carbonate areas can be classified into soil, weathered carbonate and fresh carbonate zone. They show a different range at each region. Most of the rock samples were hardly weathered. The permeability showed wide ranges (0.009 to 1.1 m/day). The average value of the permeability was 0.159 m/day. However, 80% (n=24) of total data had the permeability valves lower than 0.1 m/day. The permeability values were distinguished according to degree of development of fractures. The permeability showed low values (approximately 0.5 m/day) in highly fractured carbonate. These results mean that fractures are dominant factors controlling the permeability in carbonate areas of Korea than others. This work was supported by Energy Resource R&D program (2009T100200058) under the Ministry of Knowledge Economy, Republic of Korea.

  20. Gut permeability in neonates after a stage 1 Norwood procedure.

    Malagon, Ignacio; Onkenhout, Willem; Klok, Margreet; van der Poel, Petrus F H; Bovill, James G; Hazekamp, Mark G


    Intestinal mucosal ischemia can occur during and after cardiac surgery. Severe decreases in mucosal perfusion may be a causative factor for postoperative mortality or complications such as necrotizing enterocolitis. Mesenteric perfusion is challenged preoperatively due to an imbalance between the systemic and pulmonary circulations and challenged intraoperatively due to hypothermic circulatory arrest. We have investigated gut permeability in seven patients undergoing stage 1 of the Norwood procedure, applying the dual sugar permeability test with L-rhamnose and lactulose. Seven patients with hypoplastic left heart syndrome: clinical presentation, gut permeability findings, and outcome. A 10-bed mixed pediatric intensive care unit in a university hospital. Seven patients admitted for postoperative care after cardiac surgery. Determination of gut permeability with the dual sugar permeability test using lactulose and rhamnose. Intestinal permeability was measured after induction of anesthesia and 12 and 24 hrs later. : All patients had abnormal lactulose/rhamnose ratios. One patient, who had a lactulose/rhamnose ratio 12 hrs after surgery of 2.3 (46-times normal), developed necrotizing enterocolitis postoperatively and died 3 days after surgery. Gut permeability as assessed by the dual sugar permeability test is abnormal in patients with hypoplastic left heart syndrome before and after surgery. Lactulose/rhamnose ratios 46 times the normal value reflect a highly permeable small intestine. This may be a sign of a low output state and may help to identify patients at risk of developing necrotizing enterocolitis.

  1. Low Permeability Polyimide Insulation Project

    National Aeronautics and Space Administration — Resodyn Technologies proposes a new technology that enables the application of polyimide based cryogenic insulation with low hydrogen permeability. This effort...

  2. Low loss factor Co{sub 2}Z ferrite composites with equivalent permittivity and permeability for ultra-high frequency applications

    Su, Zhijuan; Chang, Hong; Sokolov, Alexander S.; Hu, Bolin; Chen, Yajie, E-mail:; Harris, Vincent G. [Center for Microwave Magnetic Materials and Integrated Circuits, Northeastern University, Boston, Massachusetts 02115, USA and the Department of Electrical and Computer Engineering, Northeastern University, Boston, Massachusetts 02115 (United States); Wang, Xian [Center for Microwave Magnetic Materials and Integrated Circuits, Northeastern University, Boston, Massachusetts 02115, USA and the Department of Electrical and Computer Engineering, Northeastern University, Boston, Massachusetts 02115 (United States); School of Optical and Electronic Information, Huazhong University of Science and Technology, Wuhan 430074 (China)


    Ferrite composites of nominal composition Ba{sub 3}Co{sub 2+x}Ir{sub x}Fe{sub 24−2x}O{sub 41} were studied in order to achieve low magnetic and dielectric losses and equivalent permittivity and permeability over a frequency range of 0.3–1 GHz. Crystallographic structure was characterized by X-ray diffraction, which revealed a Z-type phase accompanied by increasing amounts of Y-type phase as the iridium amount was increased. The measured microwave dielectric and magnetic properties showed that the loss tan δ{sub ε} and loss tan δ{sub μ} decreased by 80% and 90% at 0.8 GHz with the addition of iridium of x = 0.12 and 0.15, respectively. An effective medium approximation was adopted to analyze the composite ferrites having mixed phase structures. Moreover, adding Bi{sub 2}O{sub 3} enabled equivalent values of real permittivity and real permeability over the studied frequency range. The resultant data give rise to low loss factors, i.e., tan δ{sub ε}/ε′ = 0.008 and tan δ{sub μ}/μ′ = 0.037 at 0.8 GHz, while characteristic impedance was the same as that of free space.

  3. Heat transparent high intensity high efficiency solar cell

    Evans, J. C., Jr. (Inventor)


    An improved solar cell design is described. A surface of each solar cell has a plurality of grooves. Each groove has a vertical face and a slanted face that is covered by a reflecting metal. Light rays are reflected from the slanted face through the vertical face where they traverse a photovoltaic junction. As the light rays travel to the slanted face of an adjacent groove, they again traverse the junction. The underside of the reflecting coating directs the light rays toward the opposite surface of solar cell as they traverse the junction again. When the light rays travel through the solar cell and reach the saw toothed grooves on the under side, the process of reflection and repeatedly traversing the junction again takes place. The light rays ultimately emerge from the solar cell. These solar cells are particularly useful at very high levels of insolation because the infrared or heat radiation passes through the cells without being appreciably absorbed to heat the cell.

  4. Methodologies for high efficiency perovskite solar cells.

    Park, Nam-Gyu


    Since the report on long-term durable solid-state perovskite solar cell in 2012, perovskite solar cells based on lead halide perovskites having organic cations such as methylammonium CH3NH3PbI3 or formamidinium HC(NH2)2PbI3 have received great attention because of superb photovoltaic performance with power conversion efficiency exceeding 22 %. In this review, emergence of perovskite solar cell is briefly introduced. Since understanding fundamentals of light absorbers is directly related to their photovoltaic performance, opto-electronic properties of organo lead halide perovskites are investigated in order to provide insight into design of higher efficiency perovskite solar cells. Since the conversion efficiency of perovskite solar cell is found to depend significantly on perovskite film quality, methodologies for fabricating high quality perovskite films are particularly emphasized, including various solution-processes and vacuum deposition method.

  5. Methodologies for high efficiency perovskite solar cells

    Park, Nam-Gyu


    Since the report on long-term durable solid-state perovskite solar cell in 2012, perovskite solar cells based on lead halide perovskites having organic cations such as methylammonium CH3NH3PbI3 or formamidinium HC(NH2)2PbI3 have received great attention because of superb photovoltaic performance with power conversion efficiency exceeding 22 %. In this review, emergence of perovskite solar cell is briefly introduced. Since understanding fundamentals of light absorbers is directly related to their photovoltaic performance, opto-electronic properties of organo lead halide perovskites are investigated in order to provide insight into design of higher efficiency perovskite solar cells. Since the conversion efficiency of perovskite solar cell is found to depend significantly on perovskite film quality, methodologies for fabricating high quality perovskite films are particularly emphasized, including various solution-processes and vacuum deposition method.

  6. High Temperature Polymer Electrolyte Fuel Cells

    Fleige, Michael

    This thesis presents the development and application of electrochemical half-cell setups to study the catalytic reactions taking place in High Temperature Polymer Electrolyte Fuel Cells (HTPEM-FCs): (i) a pressurized electrochemical cell with integrated magnetically coupled rotating disk electrode...... (RDE) and (ii) a gas diffusion electrode (GDE) setup designed for experiments in conc. H3PO4. The pressurized cell is demonstrated by tests on polycrystalline platinum electrodes up to 150 ºC. Functionality of the RDE system is proved studying the oxygen reduction reaction (ORR) at temperatures up...... to 140 ºC and oxygen pressures up to ~100 bar at room temperature. The GDE cell is successfully tested at 130 ºC by means of direct oxidation of methanol and ethanol, respectively. In the second part of the thesis, the emphasis is put on the ORR in H3PO4 with particular focus on the mass transport...

  7. High power density carbonate fuel cell

    Yuh, C.; Johnsen, R.; Doyon, J.; Allen, J. [Energy Research Corp., Danbury, CT (United States)


    Carbonate fuel cell is a highly efficient and environmentally clean source of power generation. Many organizations worldwide are actively pursuing the development of the technology. Field demonstration of multi-MW size power plant has been initiated in 1996, a step toward commercialization before the turn of the century, Energy Research Corporation (ERC) is planning to introduce a 2.85MW commercial fuel cell power plant with an efficiency of 58%, which is quite attractive for distributed power generation. However, to further expand competitive edge over alternative systems and to achieve wider market penetration, ERC is exploring advanced carbonate fuel cells having significantly higher power densities. A more compact power plant would also stimulate interest in new markets such as ships and submarines where space limitations exist. The activities focused on reducing cell polarization and internal resistance as well as on advanced thin cell components.

  8. Interferon-gamma regulation of intestinal epithelial permeability.

    Beaurepaire, Cécile; Smyth, David; McKay, Derek M


    The maintenance and regulation of the barrier function of the epithelial lining of the intestine are important homeostatic events, serving to allow selective absorption from the gut lumen while simultaneously limiting the access of bacteria into the mucosa. Interferon-gamma is a pleiotrophic cytokine produced predominantly by natural kill cells and CD4+ T cells that under normal circumstances, and particularly during infection or inflammation, will be a component of the intestinal milieu. Use of colon-derived epithelial cell lines and, to a less extent, murine in vivo analyses, have revealed that interferon-gamma (IFN-gamma) can increase epithelial permeability as gauged by markers of paracellular permeability and bacterial transcytosis, with at least a portion of the bacteria using the transcellular permeation pathway. In this review, we describe the main characteristics of epithelial permeability and then focus on the ability of IFN-gamma to increase epithelial permeability, and the mechanism(s) thereof.

  9. Electrostatically gated membrane permeability in inorganic protocells

    Li, Mei; Harbron, Rachel L.; Weaver, Jonathan V. M.; Binks, Bernard P.; Mann, Stephen


    Although several strategies are now available to produce functional microcompartments analogous to primitive cell-like structures, little progress has been made in generating protocell constructs with self-controlled membrane permeability. Here we describe the preparation of water-dispersible colloidosomes based on silica nanoparticles and delineated by a continuous semipermeable inorganic membrane capable of self-activated, electrostatically gated permeability. We use crosslinking and covalent grafting of a pH-responsive copolymer to generate an ultrathin elastic membrane that exhibits selective release and uptake of small molecules. This behaviour, which depends on the charge of the copolymer coronal layer, serves to trigger enzymatic dephosphorylation reactions specifically within the protocell aqueous interior. This system represents a step towards the design and construction of alternative types of artificial chemical cells and protocell models based on spontaneous processes of inorganic self-organization.

  10. Permeability of cork for water and ethanol.

    Fonseca, Ana Luisa; Brazinha, Carla; Pereira, Helena; Crespo, Joao G; Teodoro, Orlando M N D


    Transport properties of natural (noncompressed) cork were evaluated for water and ethanol in both vapor and liquid phases. The permeability for these permeants has been measured, as well as the sorption and diffusion coefficients. This paper focuses on the differences between the transport of gases' relevant vapors and their liquids (water and ethanol) through cork. A transport mechanism of vapors and liquids is proposed. Experimental evidence shows that both vapors and liquids permeate not only through the small channels across the cells (plasmodesmata), as in the permeation of gases, but also through the walls of cork cells by sorption and diffusion as in dense membranes. The present study also shows that cork permeability for gases was irreversibly and drastically decreased after cork samples were exposed to ethanol or water in liquid phase.

  11. High Temperature Polymer Electrolyte Fuel Cells

    Fleige, Michael

    This thesis presents the development and application of electrochemical half-cell setups to study the catalytic reactions taking place in High Temperature Polymer Electrolyte Fuel Cells (HTPEM-FCs): (i) a pressurized electrochemical cell with integrated magnetically coupled rotating disk electrode...... of dissolved oxygen. A potential step method (hydrodynamic chronocoulometry) is evaluated for simultaneous measurement of diffusivity and solubility of oxygen by means of RDE. Finally, the ORR tests are extended to conc. H3PO4 at more relevant working temperatures and under increased oxygen pressure. Direct...... of platinumphosphoric acid. At room temperature, a relative slow ORR hindering process is active, which requires using a fast method (cyclic voltammetry with high scan rate / hydrodynamic chronocoulometry) to accurately measure the diffusion limited currents, and thus, oxygen diffusivity and solubility. In conc. H3PO4...

  12. Synthesis and characterization of cell-permeable caged phosphates that can be photolyzed by visible light or 800 nm two-photon photolysis.

    Herbivo, Cyril; Omran, Ziad; Revol, Julia; Javot, Hélène; Specht, Alexandre


    We report the synthesis and photolytic properties of caged inorganic phosphates (Pi compounds) based on the 2-(4'-{bis[2-(2-methoxyethoxy)ethyl]amino}-4-nitro-[1,1'-biphenyl]-3-yl)propan-1-ol (EANBP) and 7-(diethylamino)coumarin-4-yl]methyl (DEACM) protecting groups. The EANBP-Pi showed unprecedented photolysis efficiency at 405 nm, with 95 % release of free phosphate and a quantum yield of 0.28. Thanks to the high two-photon sensitivity of the EANBP chromophore, Pi release through two-photon photolysis is also possible, with an action cross section of 20.5 GM at 800 nm. Two bioactivatable acetoxymethyl protection groups were added to the "caged-Pi" compounds. The resulting triesters of phosphoric acid were able to diffuse through the cellular membranes of plant cells. Once inside a cell, the cleavage of these biocleavable motifs by intracellular esterases allows intracellular accumulation of EANBP-Pi. Bis(AM)-EANBP-Pi therefore represents a very attractive tool for study of the Pi signal transduction cascade in living cells.

  13. Membrane permeabilization of mammalian cells using bursts of high magnetic field pulses

    Vitalij Novickij


    Full Text Available Background Cell membrane permeabilization by pulsed electromagnetic fields (PEMF is a novel contactless method which results in effects similar to conventional electroporation. The non-invasiveness of the methodology, independence from the biological object homogeneity and electrical conductance introduce high flexibility and potential applicability of the PEMF in biomedicine, food processing, and biotechnology. The inferior effectiveness of the PEMF permeabilization compared to standard electroporation and the lack of clear description of the induced transmembrane transport are currently of major concern. Methods The PEMF permeabilization experiments have been performed using a 5.5 T, 1.2 J pulse generator with a multilayer inductor as an applicator. We investigated the feasibility to increase membrane permeability of Chinese Hamster Ovary (CHO cells using short microsecond (15 µs pulse bursts (100 or 200 pulses at low frequency (1 Hz and high dB/dt (>106 T/s. The effectiveness of the treatment was evaluated by fluorescence microscopy and flow cytometry using two different fluorescent dyes: propidium iodide (PI and YO-PRO®-1 (YP. The results were compared to conventional electroporation (single pulse, 1.2 kV/cm, 100 µs, i.e., positive control. Results The proposed PEMF protocols (both for 100 and 200 pulses resulted in increased number of permeable cells (70 ± 11% for PI and 67 ± 9% for YP. Both cell permeabilization assays also showed a significant (8 ± 2% for PI and 35 ± 14% for YP increase in fluorescence intensity indicating membrane permeabilization. The survival was not affected. Discussion The obtained results demonstrate the potential of PEMF as a contactless treatment for achieving reversible permeabilization of biological cells. Similar to electroporation, the PEMF permeabilization efficacy is influenced by pulse parameters in a dose-dependent manner.

  14. High temperature polymer electrolyte membrane fuel cell

    K.Scott; M. Mamlouk


    One of the major issues limiting the introduction of polymer electrolyte membrane fuel cells (PEMFCs) is the low temperature of operation which makes platinum-based anode catalysts susceptible to poisoning by the trace amount of CO, inevitably present in reformed fuel. In order to alleviate the problem of CO poisoning and improve the power density of the cell, operating at temperature above 100 ℃ is preferred. Nafion(R) -type perfluorosulfonated polymers have been typically used for PEMFC. However, the conductivity of Nafion(R) -type polymers is not high enough to be used for fuel cell operations at higher temperature ( > 90 ℃) and atmospheric pressure because they dehydrate under these condition.An additional problem which faces the introduction of PEMFC technology is that of supplying or storing hydrogen for cell operation,especially for vehicular applications. Consequently the use of alternative fuels such as methanol and ethanol is of interest, especially if this can be used directly in the fuel cell, without reformation to hydrogen. A limitation of the direct use of alcohol is the lower activity of oxidation in comparison to hydrogen, which means that power densities are considerably lower. Hence to improve activity and power output higher temperatures of operation are preferable. To achieve this goal, requires a new polymer electrolyte membrane which exhibits stability and high conductivity in the absence of liquid water.Experimental data on a polybenzimidazole based PEMFC were presented. A simple steady-state isothermal model of the fuel cell is also used to aid in fuel cell performance optimisation. The governing equations involve the coupling of kinetic, ohmic and mass transport. This paper also considers the advances made in the performance of direct methanol and solid polymer electrolyte fuel cells and considers their limitations in relation to the source and type of fuels to be used.

  15. Study on Permeability Change Rule of Different Rank Coals by Injecting Carbon Dioxide

    Guo Hui


    Full Text Available This paper provides an experimental support for increasing the flow conductivity of coal reservoir by identifying permeability change rule caused by reactions with different minerals in coal after injecting carbon dioxide. Based on measurement of nitrogen adsorption, mineral composition and permeability of medium-high rank coal in Tunlan mine and Sihe mine, it is used to investigate the permeability change rule caused by reactions with different minerals in coal and its improving effect after injecting carbon dioxide. A permeability change model was established by making a nonlinear regression analysis of the initial permeability, the reaction time and the improved permeability. The results showed that as a result of CO2-water-rock interaction, permeability of medium-high rank coal increases at first and then decreases with time going by after injecting carbon dioxide. The permeability of Sihe coal samples reaches maximum value earlier than that of Tunlan coal samples. Improving effect of permeability of Sihe coal samples is better than that of Tunlan coal samples. The initial permeability which is too large or too small is insensitive to the change of permeability, while the medium permeability within 0.1--0.2×10-3μm2 is more favorable. The reliability of the mathematical model is verified by the experiment. The results can also provide a theoretical basis for the analysis of permeability change after injecting carbon dioxide.

  16. Discovery of Sanggenon G as a natural cell-permeable small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP).

    Seiter, Maximilian A; Salcher, Stefan; Rupp, Martina; Hagenbuchner, Judith; Kiechl-Kohlendorfer, Ursula; Mortier, Jérémie; Wolber, Gerhard; Rollinger, Judith M; Obexer, Petra; Ausserlechner, Michael J


    Defects in the regulation of apoptosis are one main cause of cancer development and may result from overexpression of anti-apoptotic proteins such as the X-linked inhibitor of apoptosis protein (XIAP). XIAP is frequently overexpressed in human leukemia and prostate and breast tumors. Inhibition of apoptosis by XIAP is mainly coordinated through direct binding to the initiator caspase-9 via its baculovirus-IAP-repeat-3 (BIR3) domain. XIAP inhibits caspases directly making it to an attractive target for anti-cancer therapy. In the search for novel, non-peptidic XIAP inhibitors in this study we focused on the chemical constituents of sāng bái pí (mulberry root bark). Most promising candidates of this plant were tested biochemically in vitro by a fluorescence polarization (FP) assay and in vivo via protein fragment complementation analysis (PCA). We identified the Diels Alder adduct Sanggenon G (SG1) as a novel, small-molecular weight inhibitor of XIAP. As shown by FP and PCA analyses, SG1 binds specifically to the BIR3 domain of XIAP with a binding affinity of 34.26 μM. Treatment of the transgenic leukemia cell line Molt3/XIAP with SG1 enhances caspase-8, -3 and -9 cleavage, displaces caspase-9 from XIAP as determined by immunoprecipitation experiments and sensitizes these cells to etoposide-induced apoptosis. SG1 not only sensitizes the XIAP-overexpressing leukemia cell line Molt3/XIAP to etoposide treatment but also different neuroblastoma cell lines endogenously expressing high XIAP levels. Taken together, Sanggenon G (SG1) is a novel, natural, non-peptidic, small-molecular inhibitor of XIAP that can serve as a starting point to develop a new class of improved XIAP inhibitors.

  17. Permeability characterization of polymer matrix composites by RTM/VARTM

    Naik, N. K.; Sirisha, M.; Inani, A.


    Cost effective manufacturing of high performance polymer matrix composite structures is an important consideration for the growth of its use. Resin transfer moulding (RTM) and vacuum assisted resin transfer moulding (VARTM) are the efficient processes for the cost effective manufacturing. These processes involve transfer of resin from the tank into the reinforcing preform loaded into a closed mould. Resin flow within the preform and reinforcement wetting can be characterized using the permeability properties. Different reinforcement and resin properties and process parameters affecting the permeability are discussed based on state of art literature review covering experimental studies. General theory for the determination of permeability is presented. Based on the literature review, permeability values for different reinforcement architecture, resin and processing conditions are presented. Further, possible sources of error during experimental determination of permeability and issues involved with reproducibility are discussed.

  18. Oxygen permeable membrane for oxygen enriched combustion

    Asakawa, S. (Matsushita Research Inst., Tokyo, Japan); Saito, Y.; Kawahito, M.; Ito, Y.; Tsuchiya, S.; Sugata, K.


    An oxygen enriched air production system using gas separation membranes has been developed to be used for fuel combustion systems. High oxygen permeable scopolymers, including three dimensional structure, have been synthesized through condensation of polyvinylphenol with some, ..cap alpha.., (diethylamino) polydimethylsiloxanes. The experimental results showed that the oxygen permeability through the copolymer varies as a function of the dimethylsiloxane content of the copolymers. Such composition dependence of the oxygen permeability was explained on the basis of polymer constitution. Typical values of the oxygen permeability, 3.4 x 10/sup -8/ (cc x cm/cm/sup 2/ x sec x cmHg) and ..cap alpha.., 2.1, were obtained at 72% of dimethylsiloxane content. The copolymers are soluble in most common organic solvents and uniform, defect-free membranes as thin as 1000 Angstroms have been formed by spreading solutions of the copolymer on water. Composite membranes fabricated by applying the membranes to porous support materials were used for practical gas separation and 30% oxygen enriched air was produced from ambient air. A new type oxygen enriched combustion system, which is more efficient for energy saving, has been developed by utilizing oxygen enriched air thus produced. 15 references, 15 figures, 1 table.

  19. Tunable permeability of magnetic wires at microwaves

    Panina, L.V., E-mail: [National University of Science and Technology, MISiS, Moscow (Russian Federation); Institute for Design Problems in Microelectronics, RAN, Moscow (Russian Federation); Makhnovskiy, D.P. [School of Computing and Mathematics, University of Plymouth (United Kingdom); Morchenko, A.T.; Kostishin, V.G. [National University of Science and Technology, MISiS, Moscow (Russian Federation)


    This paper presents the analysis into microwave magnetic properties of magnetic microwires and their composites in the context of applications in wireless sensors and tunable microwave materials. It is demonstrated that the intrinsic permeability of wires has a wide frequency dispersion with relatively large values in the GHz band. In the case of a specific magnetic anisotropy this results in a tunable microwave impedance which could be used for distributed wireless sensing networks in functional composites. The other range of applications is related with developing the artificial magnetic dielectrics with large and tunable permeability. The composites with magnetic wires with a circumferential anisotropy have the effective permeability which differs substantially from unity for a relatively low concentration (less than 10%). This can make it possible to design the wire media with a negative and tunable index of refraction utilising natural magnetic properties of wires. - Highlights: • Applications of magnetic microwires for functional composites and distributed sensor networks are proposed. • Diluted composites with magnetic microwires can demonstrate tunable left-handed properties. • Large microwave permeability combined with a specific magnetic structure lead to a large and sensitive microwave magnetoimpedance. • Microwave magnetoimpedance highly sensitive to temperature is demonstrated.

  20. Apoptosis Induced by High Concentrations of Nicotinamide in Tobacco Suspension Cells

    张贵友; 朱瑞宇; 戴尧仁


    As an inhibitor of poly(ADP-ribose) polymerase (PARP), nicotinamide has a restraining effect on apoptosis at certain low concentrations. In our present study, apoptosis induced by high concentrations of nicotinamide was observed in tobacco suspension cells. When cells were preincubated with 250 mmol/L nicotinamide for 24 h, the hallmarks of apoptosis were detected, including DNA fragments increasing in size by multiples of 180-200 bp, the condensation and peripheral distribution of nuclear chromatin, and a positive reaction to the TUNEL assay. At the same time, the degradation of PARP and the reduction in the potential of the inner membrane of mitochondria appeared in apoptotic cells induced by high concentrations of nicotinamide. This result indicates that apoptosis induced by high concentrations of nicotinamide is associated with caspase-3-like activity and with the opening of mitochondrial permeability pores. These results partially support the hypothesis that high concentrations of PARP inhibitor could force cells to enter an apoptotic pathway by delay of DNA repair in replicating cells.

  1. High temperature polymer electrolyte membrane fuel cells

    This book is a comprehensive review of high-temperature polymer electrolyte membrane fuel cells (PEMFCs). PEMFCs are the preferred fuel cells for a variety of applications such as automobiles, cogeneration of heat and power units, emergency power and portable electronics. The first 5 chapters...... of the book describe rationalization and illustration of approaches to high temperature PEM systems. Chapters 6 - 13 are devoted to fabrication, optimization and characterization of phosphoric acid-doped polybenzimidazole membranes, the very first electrolyte system that has demonstrated the concept...... of and motivated extensive research activity in the field. The last 11 chapters summarize the state-of-the-art of technological development of high temperature-PEMFCs based on acid doped PBI membranes including catalysts, electrodes, MEAs, bipolar plates, modelling, stacking, diagnostics and applications....

  2. High performance internal reforming unit for high temperature fuel cells

    Ma, Zhiwen; Venkataraman, Ramakrishnan; Novacco, Lawrence J.


    A fuel reformer having an enclosure with first and second opposing surfaces, a sidewall connecting the first and second opposing surfaces and an inlet port and an outlet port in the sidewall. A plate assembly supporting a catalyst and baffles are also disposed in the enclosure. A main baffle extends into the enclosure from a point of the sidewall between the inlet and outlet ports. The main baffle cooperates with the enclosure and the plate assembly to establish a path for the flow of fuel gas through the reformer from the inlet port to the outlet port. At least a first directing baffle extends in the enclosure from one of the sidewall and the main baffle and cooperates with the plate assembly and the enclosure to alter the gas flow path. Desired graded catalyst loading pattern has been defined for optimized thermal management for the internal reforming high temperature fuel cells so as to achieve high cell performance.

  3. High cell density cultures produced by internal retention: application in continuous ethanol fermentation

    Berta Carola Pérez


    Full Text Available Ethanol has provoked great interest due to its potential as an alternative fuel. Nevertheless, fermentation processes must be developed by increasing the low volumetric productivity achieved in conventional cultures (batch or continuous to make this product become economically competitive. This can be achieved by using techniques leading to high cell concentration and reducing inhibition by the end-product. One of the frequently employed methods involves cell recycling. This work thus developed a membrane reactor incorporating a filtration module with 5 u,m stainless steel tubular units inside a 3L stirred jar fermenter for investigating its application in continuous ethanol production. The effects of cell concentration and transmembrane pressure difference on permeate flux were evaluated for testing the filtration module's performance. The internal cell retention system was operated in Saccharomyces cerevisiae continuous culture derived from sucrose, once fermentation conditions had been selected (30 °C, 1.25 -1.75 vvm, pH 4.5. Filter unit permeability was maintained by applying pulses of air. More than 97% of the grown cells were retained in the fermenter, reaching 51 g/L cell concentration and 8.51 g/L.h average ethanol productivity in culture with internal cell retention; this was twice that obtained in a conventional continuous culture. Key words: Membrane reactor, Saccharomyces cerevisiae, alcoholic fermentation, cell recycling.

  4. High-content screening of drug-induced cardiotoxicity using quantitative single cell imaging cytometry on microfluidic device.

    Kim, Min Jung; Lee, Su Chul; Pal, Sukdeb; Han, Eunyoung; Song, Joon Myong


    Drug-induced cardiotoxicity or cytotoxicity followed by cell death in cardiac muscle is one of the major concerns in drug development. Herein, we report a high-content quantitative multicolor single cell imaging tool for automatic screening of drug-induced cardiotoxicity in an intact cell. A tunable multicolor imaging system coupled with a miniaturized sample platform was destined to elucidate drug-induced cardiotoxicity via simultaneous quantitative monitoring of intracellular sodium ion concentration, potassium ion channel permeability and apoptosis/necrosis in H9c2(2-1) cell line. Cells were treated with cisapride (a human ether-à-go-go-related gene (hERG) channel blocker), digoxin (Na(+)/K(+)-pump blocker), camptothecin (anticancer agent) and a newly synthesized anti-cancer drug candidate (SH-03). Decrease in potassium channel permeability in cisapride-treated cells indicated that it can also inhibit the trafficking of the hERG channel. Digoxin treatment resulted in an increase of intracellular [Na(+)]. However, it did not affect potassium channel permeability. Camptothecin and SH-03 did not show any cytotoxic effect at normal use (≤300 nM and 10 μM, respectively). This result clearly indicates the potential of SH-03 as a new anticancer drug candidate. The developed method was also used to correlate the cell death pathway with alterations in intracellular [Na(+)]. The developed protocol can directly depict and quantitate targeted cellular responses, subsequently enabling an automated, easy to operate tool that is applicable to drug-induced cytotoxicity monitoring with special reference to next generation drug discovery screening. This multicolor imaging based system has great potential as a complementary system to the conventional patch clamp technique and flow cytometric measurement for the screening of drug cardiotoxicity.

  5. Hydrodynamic permeability of aggregates of porous particles with an impermeable core.

    Deo, Satya; Filippov, Anatoly; Tiwari, Ashish; Vasin, Sergey; Starov, Victor


    A hydrodynamic permeability of membranes built up by porous cylindrical or spherical particles with impermeable core is investigated. Different versions of a cell method are used to calculate the hydrodynamic permeability of the membranes. Four known boundary conditions, namely, Happel's, Kuwabara's, Kvashnin's and Cunningham/Mehta-Morse's, are considered on the outer surface of the cell. Comparison of the resulting hydrodynamic permeability is undertaken. A possible jump of a shear stress at the fluid-membrane interface, its impact on the hydrodynamic permeability is also investigated. New results related to the calculated hydrodynamic permeability and the theoretical values of Kozeny constant are reported. Both transversal and normal flows of liquid with respect to the cylindrical fibers that compose the membrane are studied. The deduced theoretical results can be applied for the investigation of the hydrodynamic permeability of colloidal cake layers on the membrane surface, the hydrodynamic permeability of woven materials. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. Highly effective permeability and antifouling performances of polypropylene non-woven fabric membranes modified with graphene oxide by inkjet printing and immersion coating methods.

    Zhao, Chuan-Qi; Xu, Xiao-Chen; Li, Rui-Yun; Chen, Jie; Yang, Feng-Lin


    In the current study, graphene oxide (GO)-modified polypropylene non-woven fabric (PP-NWF) membranes were prepared via inkjet printing and immersion coating methods. Scanning electron microscopy, Fourier transform infrared spectroscopy, contact angle measurements, pure water permeation (JPWP) and protein adsorption were tested to evaluate the impact of the GO nanosheet on the characteristics and performance of modified PP-NWF membranes. The results showed that the exfoliated GO nanosheets uniformly deposited on the membrane surface and firmly embedded into the interlaced fibers, resulting in the improvement of membrane hydrophilicity, permeability and antifouling properties comparing with original PP-NWF membranes. The GO-printed and GO-coated membranes had 113 and 188% higher fluxes, and 70.95 and 75.74% lower protein adsorptions than the original PP-NWF membranes, respectively. After cross-linked treatment, ultrasound processing was conducted to evaluate the stability of the modified PP-NWF membranes. The results demonstrated that there was almost no decrease in permeation after ultrasonic treatment indicating that the cross-linking treatment could enhance the immobilization of the GO nanosheets on and into the modified membranes.


    Goss, W.C.; Gilley, L.F.


    The present invention relates to a high speed camera utilizing a Kerr cell shutter and a novel optical delay system having no moving parts. The camera can selectively photograph at least 6 frames within 9 x 10/sup -8/ seconds during any such time interval of an occurring event. The invention utilizes particularly an optical system which views and transmits 6 images of an event to a multi-channeled optical delay relay system. The delay relay system has optical paths of successively increased length in whole multiples of the first channel optical path length, into which optical paths the 6 images are transmitted. The successively delayed images are accepted from the exit of the delay relay system by an optical image focusing means, which in turn directs the images into a Kerr cell shutter disposed to intercept the image paths. A camera is disposed to simultaneously view and record the 6 images during a single exposure of the Kerr cell shutter. (AEC)

  8. Geothermal Permeability Enhancement - Final Report

    Joe Beall; Mark Walters


    The overall objective is to apply known permeability enhancement techniques to reduce the number of wells needed and demonstrate the applicability of the techniques to other undeveloped or under-developed fields. The Enhanced Geothermal System (EGS) concept presented in this project enhances energy extraction from reduced permeability zones in the super-heated, vapor-dominated Aidlin Field of the The Geysers geothermal reservoir. Numerous geothermal reservoirs worldwide, over a wide temperature range, contain zones of low permeability which limit the development potential and the efficient recovery of heat from these reservoirs. Low permeability results from poorly connected fractures or the lack of fractures. The Enhanced Geothermal System concept presented here expands these technologies by applying and evaluating them in a systematic, integrated program.

  9. Permeability, storage and hydraulic diffusivity controlled by earthquakes

    Brodsky, E. E.; Fulton, P. M.; Xue, L.


    Earthquakes can increase permeability in fractured rocks. In the farfield, such permeability increases are attributed to seismic waves and can last for months after the initial earthquake. Laboratory studies suggest that unclogging of fractures by the transient flow driven by seismic waves is a viable mechanism. These dynamic permeability increases may contribute to permeability enhancement in the seismic clouds accompanying hydraulic fracking. Permeability enhancement by seismic waves could potentially be engineered and the experiments suggest the process will be most effective at a preferred frequency. We have recently observed similar processes inside active fault zones after major earthquakes. A borehole observatory in the fault that generated the M9.0 2011 Tohoku earthquake reveals a sequence of temperature pulses during the secondary aftershock sequence of an M7.3 aftershock. The pulses are attributed to fluid advection by a flow through a zone of transiently increased permeability. Directly after the M7.3 earthquake, the newly damaged fault zone is highly susceptible to further permeability enhancement, but ultimately heals within a month and becomes no longer as sensitive. The observation suggests that the newly damaged fault zone is more prone to fluid pulsing than would be expected based on the long-term permeability structure. Even longer term healing is seen inside the fault zone of the 2008 M7.9 Wenchuan earthquake. The competition between damage and healing (or clogging and unclogging) results in dynamically controlled permeability, storage and hydraulic diffusivity. Recent measurements of in situ fault zone architecture at the 1-10 meter scale suggest that active fault zones often have hydraulic diffusivities near 10-2 m2/s. This uniformity is true even within the damage zone of the San Andreas fault where permeability and storage increases balance each other to achieve this value of diffusivity over a 400 m wide region. We speculate that fault zones

  10. 高渗高强混凝土材料强度特性试验及参数确定%Strength Characteristic Experiment and Parameters Determination of High Permeability-high Strength Concrete Material

    谭仪忠; 刘元雪; 葛增超; 赵吉昌


    High permeability⁃high strength concrete is a new material which is the characterized with high permeability and high strength. When it is used in away⁃from underground space,it can help the around rock to drain the ground water, and it can al⁃so improve the structure strength and give full play to the role of the shock absorption layer. The influence factors of tensile and com⁃pression strength can be confirmed through orthogonal experiment. The results show that the biggest two factors against the tensile and compression strength are water cement ratio and cement aggregate ratio,while the polymeric ash ratio can impact the tensile strength also. Then according to the three⁃dimension stress,the strength parameters of cohesion and internal friction angle which are widely used in geotechnical engineering can be calculated by the test results. It can be found that the relationship between compres⁃sive strength and cohesion are of linear. The results can be used in geotechnical calculation directly.%高渗高强混凝土是一种兼具高渗透性和高强度的新材料,用在离壁式地下工程中既能确保通畅地排除围岩周边地下水,又能提高地下工程结构自身的强度,充分发挥其减震层的作用。通过正交试验得出影响材料单轴抗压强度与抗劈裂强度的4个因素的排序,结果显示:对抗压强度与抗劈裂强度影响最大的2个因素是灰集比与水灰比,聚灰比对抗劈裂强度的影响也较为明显。从材料一点的三向应力状态出发,通过对试验结果的计算得到岩土工程中广泛运用的内摩擦角和黏聚力等强度参数,经回归分析发现材料的黏聚力与抗压强度之间符合线性增长关系,该结果可以直接为岩土工程数值计算中的数据应用提供依据。

  11. Assessing the permeability of engineered capillary networks in a 3D culture.

    Stephanie J Grainger

    Full Text Available Many pathologies are characterized by poor blood vessel growth and reduced nutrient delivery to the surrounding tissue, introducing a need for tissue engineered blood vessels. Our lab has developed a 3D co-culture method to grow interconnected networks of pericyte-invested capillaries, which can anastamose with host vasculature following implantation to restore blood flow to ischemic tissues. However, if the engineered vessels contain endothelial cells (ECs that are misaligned or contain wide junctional gaps, they may function improperly and behave more like the pathologic vessels that nourish tumors. The purpose of this study was to test the resistance to permeability of these networks in vitro, grown with different stromal cell types, as a metric of vessel functionality. A fluorescent dextran tracer was used to visualize transport across the endothelium and the pixel intensity was quantified using a customized MATLAB algorithm. In fibroblast-EC co-cultures, the dextran tracer easily penetrated through the vessel wall and permeability was high through the first 5 days of culture, indicative of vessel immaturity. Beyond day 5, dextran accumulated at the periphery of the vessel, with very little transported across the endothelium. Quantitatively, permeability dropped from initial levels of 61% to 39% after 7 days, and to 7% after 2 weeks. When ECs were co-cultured with bone marrow-derived mesenchymal stem cells (MSCs or adipose-derived stem cells (AdSCs, much tighter control of permeability was achieved. Relative to the EC-fibroblast co-cultures, permeabilities were reduced 41% for the EC-MSC co-cultures and 50% for the EC-AdSC co-cultures after 3 days of culture. By day 14, these permeabilities decreased by 68% and 77% over the EC-fibroblast cultures. Co-cultures containing stem cells exhibit elevated VE-cadherin levels and more prominent EC-EC junctional complexes when compared to cultures containing fibroblasts. These data suggest the stromal

  12. High Aspect Ratio Semiconductor Heterojunction Solar Cells

    Redwing, Joan [Pennsylvania State Univ., University Park, PA (United States). Dept. of Material Science and Engineering; Mallouk, Tom [Pennsylvania State Univ., University Park, PA (United States). Dept. of Chemistry; Mayer, Theresa [Pennsylvania State Univ., University Park, PA (United States). Dept. of Electrical Engineering; Dickey, Elizabeth [Pennsylvania State Univ., University Park, PA (United States). Dept. of Materials Science and Engineering; Wronski, Chris [Pennsylvania State Univ., University Park, PA (United States). Dept. of Electrical Engineering


    The project focused on the development of high aspect ratio silicon heterojunction (HARSH) solar cells. The solar cells developed in this study consisted of high density vertical arrays of radial junction silicon microwires/pillars formed on Si substrates. Prior studies have demonstrated that vertical Si wire/pillar arrays enable reduced reflectivity and improved light trapping characteristics compared to planar solar cells. In addition, the radial junction structure offers the possibility of increased carrier collection in solar cells fabricated using material with short carrier diffusion lengths. However, the high junction and surface area of radial junction Si wire/pillar array devices can be problematic and lead to increased diode leakage and enhanced surface recombination. This study investigated the use of amorphous hydrogenated Si in the form of a heterojunction-intrinsic-thin layer (HIT) structure as a junction formation method for these devices. The HIT layer structure has widely been employed to reduce surface recombination in planar crystalline Si solar cells. Consequently, it was anticipated that it would also provide significant benefits to the performance of radial junction Si wire/pillar array devices. The overall goals of the project were to demonstrate a HARSH cell with a HIT-type structure in the radial junction Si wire/pillar array configuration and to develop potentially low cost pathways to fabricate these devices. Our studies demonstrated that the HIT structure lead to significant improvements in the open circuit voltage (Voc>0.5) of radial junction Si pillar array devices compared to devices fabricated using junctions formed by thermal diffusion or low pressure chemical vapor deposition (LPCVD). In addition, our work experimentally demonstrated that the radial junction structure lead to improvements in efficiency compared to comparable planar devices for devices fabricated using heavily doped Si that had reduced carrier diffusion

  13. Cation Permeability in Soybean Aleurone Layer

    Noda, Hiroko; Fukuda, Mitsuru


    The permeation of water and ions into bean seeds is essential for processing and cooking of beans. The permeability of cations, K, Na, Ca, and Mg ions, into soybean seed tissue, especially aleurone layer, during water uptake was investigated to characterize the ion permeation into soybeans. Aleurone layers and seed coats contained relatively high concentration of endogenous K and Ca ions, and endogenous Ca ion, respectively. The amounts of Ca ion entered seed coats and aleurone layers were gr...

  14. The putative P-gp inhibitor telmisartan does not affect the transcellular permeability and cellular uptake of the calcium channel antagonist verapamil in the P-glycoprotein expressing cell line MDCK II MDR1.

    Saaby, Lasse; Tfelt-Hansen, Peer; Brodin, Birger


    Verapamil is used in high doses for the treatment of cluster headache. Verapamil has been described as a P-glycoprotein (P-gp, ABCB1) substrate. We wished to evaluate in vitro whether co administration of a P-gp inhibitor with verapamil could be a feasible strategy for increasing CNS uptake of verapamil. Fluxes of radiolabelled verapamil across MDCK II MDR1 monolayers were measured in the absence and presence of the putative P-gp inhibitor telmisartan (a clinically approved drug compound). Verapamil displayed a vectorial basolateral-to-apical transepithelial efflux across the MDCK II MDR1 monolayers with a permeability of 5.7 × 10(-5) cm sec(-1) compared to an apical to basolateral permeability of 1.3 × 10(-5) cm sec(-1). The efflux could be inhibited with the P-gp inhibitor zosuquidar. Zosuquidar (0.4 μmol/L) reduced the efflux ratio (PB-A/PA-B) for verapamil 4.6-1.6. The presence of telmisartan, however, only caused a slight reduction in P-gp-mediated verapamil transport to an efflux ratio of 3.4. Overall, the results of the present in vitro approach indicate, that clinical use of telmisartan as a P-gp inhibitor may not be an effective strategy for increasing brain uptake of verapamil by co-administration with telmisartan.

  15. Predicting chemically-induced skin reactions. Part II: QSAR models of skin permeability and the relationships between skin permeability and skin sensitization

    Alves, Vinicius M. [Laboratory of Molecular Modeling and Design, Faculty of Pharmacy, Federal University of Goiás, Goiânia, GO 74605-220 (Brazil); Laboratory for Molecular Modeling, Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599 (United States); Muratov, Eugene [Laboratory for Molecular Modeling, Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599 (United States); Laboratory of Theoretical Chemistry, A.V. Bogatsky Physical–Chemical Institute NAS of Ukraine, Odessa 65080 (Ukraine); Fourches, Denis [Laboratory for Molecular Modeling, Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599 (United States); Strickland, Judy; Kleinstreuer, Nicole [ILS/Contractor supporting the NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM), P.O. Box 13501, Research Triangle Park, NC 27709 (United States); Andrade, Carolina H. [Laboratory of Molecular Modeling and Design, Faculty of Pharmacy, Federal University of Goiás, Goiânia, GO 74605-220 (Brazil); Tropsha, Alexander, E-mail: [Laboratory for Molecular Modeling, Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599 (United States)


    Skin permeability is widely considered to be mechanistically implicated in chemically-induced skin sensitization. Although many chemicals have been identified as skin sensitizers, there have been very few reports analyzing the relationships between molecular structure and skin permeability of sensitizers and non-sensitizers. The goals of this study were to: (i) compile, curate, and integrate the largest publicly available dataset of chemicals studied for their skin permeability; (ii) develop and rigorously validate QSAR models to predict skin permeability; and (iii) explore the complex relationships between skin sensitization and skin permeability. Based on the largest publicly available dataset compiled in this study, we found no overall correlation between skin permeability and skin sensitization. In addition, cross-species correlation coefficient between human and rodent permeability data was found to be as low as R{sup 2} = 0.44. Human skin permeability models based on the random forest method have been developed and validated using OECD-compliant QSAR modeling workflow. Their external accuracy was high (Q{sup 2}{sub ext} = 0.73 for 63% of external compounds inside the applicability domain). The extended analysis using both experimentally-measured and QSAR-imputed data still confirmed the absence of any overall concordance between skin permeability and skin sensitization. This observation suggests that chemical modifications that affect skin permeability should not be presumed a priori to modulate the sensitization potential of chemicals. The models reported herein as well as those developed in the companion paper on skin sensitization suggest that it may be possible to rationally design compounds with the desired high skin permeability but low sensitization potential. - Highlights: • It was compiled the largest publicly-available skin permeability dataset. • Predictive QSAR models were developed for skin permeability. • No concordance between skin

  16. Interaction of wall shear stress magnitude and gradient in the prediction of arterial macromolecular permeability.

    LaMack, Jeffrey A; Himburg, Heather A; Li, Xue-Mei; Friedman, Morton H


    Large spatial shear stress gradients have anecdotally been associated with early atherosclerotic lesion susceptibility in vivo and have been proposed as promoters of endothelial cell dysfunction in vitro. Here, experiments are presented in which several measures of the fluid dynamic shear stress, including its gradient, at the walls of in vivo porcine iliac arteries, are correlated against the transendothelial macromolecular permeability of the vessels. The fluid dynamic measurements are based on postmortem vascular casts, and permeability is measured from Evans blue dye (EBD) uptake. Time-averaged wall shear stress (WSS), as well as a new parameter termed maximum gradient stress (MGS) that describes the spatial shear stress gradient due to flow acceleration at a given point, are mapped for each artery and compared on a point-by-point basis to the corresponding EBD patterns. While there was no apparent relation between MGS and EBD uptake, a composite parameter, WSS(-0.11) MGS(0.044), was highly correlated with permeability. Notwithstanding the small exponents, the parameter varied widely within the region of interest. The results suggest that sites exposed to low wall shear stresses are more likely to exhibit elevated permeability, and that this increase is exacerbated in the presence of large spatial shear stress gradients.

  17. Gas Permeability of Porous Plasma-Sprayed Coatings

    Wittmann-Ténèze, K.; Caron, N.; Alexandre, S.


    For different applications, such as solid oxide fuel cells, there is an interest in understanding the relationship between the microstructure and the gas permeability of plasma-sprayed coatings. Nevertheless, plasma spraying processes allow to elaborate coatings with singular microstructures, depending strongly on the initial material and plasma operating conditions. And so, the evolution of permeability is not directly linked to the porosity. In this work, coatings were manufactured using different initial feedstock and spray parameters to obtain various microporous structures. Measurements of their permeation with the pressure drop method and their open porosity just as the observation of the morphology and the structure by optical microscopy were achieved. The different data show that the evolution of the gas permeability with the open porosity follows the Kozeny-Carman equation. This result correlated with the microstructural observation highlights the relationship between the permeability and the physical properties of porous plasma-sprayed layers.

  18. High Energy Density aluminum/oxygen cell

    Rudd, E. J.; Gibbons, D. W.

    An alternative to a secondary battery as the power source for vehicle propulsion is a fuel cell, an example of which is the metal/air cell using metals such as aluminum, zinc, or iron. Aluminum is a particularly attractive candidate, with high energy and power densities, environmentally acceptable and having a large, established industrial base for production and distribution. An aluminum/oxygen system is currently under development for a prototype unmanned, undersea vehicle (UUV) for the US navy and recent work has focussed upon low corrosion aluminum alloys, and an electrolyte management system for processing the by-products of the energy-producing reactions. This paper summarizes the progress made in both areas. Anode materials capable of providing high utilization factors over current densities ranging from 5 to 150 mA/cm 2 have been identified, such materials being essential to realize mission life for the UUV. With respect to the electrolyte management system, a filter/precipitator unit has been successfully operated for over 250 h in a large scale, half-cell system.

  19. Monitoring pulmonary vascular permeability using radiolabeled transferrin

    Basran, G.S.; Hardy, J.G.


    A simple, noninvasive technique for monitoring pulmonary vascular permeability in patients in critical care units is discussed. High vascular permeability is observed in patients with clinically defined adult respiratory distress syndrome (ARDS) but not in patients with hydrostatic pulmonary edema or in patients with minor pulmonary insults who are considered to be at risk of developing ARDS. The technique has been used in the field of therapeutics and pharmacology to test the effects of the putative antipermeability agents methylprednisolone and terbutaline sulfate. There appears to be a good correlation between the acute inhibitory effect of either drug on transferrin exudation and patient prognosis. Thus, a byproduct of such drug studies may be an index of survival in patients with established ARDS.

  20. Permeability of the peroxisomal membrane: lessons from the glyoxylate cycle

    Markus eKunze


    Full Text Available Glyoxylate serves as intermediate in various metabolic pathways, although high concentrations of this metabolite are toxic to the cell. In many organisms glyoxylate is fed into the glyoxylate cycle. Enzymes participating in this metabolism are located on both sides of the peroxisomal membrane. The permeability of this membrane for small metabolites paves the way for exchange of intermediates between proteins catalyzing consecutive reactions. A model, in which soluble enzymes accumulate in close proximity to both ends of pore-like structures forming a transmembrane metabolon could explain the rapid and targeted exchange of intermediates. The metabolites passing the membrane differ between the three model organisms Saccharomyces cerevisiae, Arabidopsis thaliana and Candida albicans, which reflects the ease of evolutionary adaptation processes whenever specific transporter proteins are not involved. The atypical permeability properties of the peroxisomal membrane together with a flexible structural arrangement ensuring the swift and selective transport across the membrane might represent the molecular basis for the functional versatility of peroxisomes.

  1. Indicaxanthin inhibits NADPH oxidase (NOX)-1 activation and NF-κB-dependent release of inflammatory mediators and prevents the increase of epithelial permeability in IL-1β-exposed Caco-2 cells.

    Tesoriere, L; Attanzio, A; Allegra, M; Gentile, C; Livrea, M A


    Dietary redox-active/antioxidant phytochemicals may help control or mitigate the inflammatory response in chronic inflammatory bowel disease (IBD). In the present study, the anti-inflammatory activity of indicaxanthin (Ind), a pigment from the edible fruit of cactus pear (Opuntia ficus-indica, L.), was shown in an IBD model consisting of a human intestinal epithelial cell line (Caco-2 cells) stimulated by IL-1β, a cytokine known to play a major role in the initiation and amplification of inflammatory activity in IBD. The exposure of Caco-2 cells to IL-1β brought about the activation of NADPH oxidase (NOX-1) and the generation of reactive oxygen species (ROS) to activate intracellular signalling leading to the activation of NF-κB, with the over-expression of inflammatory enzymes and release of pro-inflammatory mediators. The co-incubation of the cells with Ind, at a nutritionally relevant concentration (5-25 μM), and IL-1β prevented the release of the pro-inflammatory cytokines IL-6 and IL-8, PGE2 and NO, the formation of ROS and the loss of thiols in a dose-dependent manner. The co-incubation of the cells with Ind and IL-1β also prevented the IL-1β-induced increase of epithelial permeability. It was also shown that the activation of NOX-1 and NF-κB was prevented by Ind and the expression of COX-2 and inducible NO synthase was reduced. The uptake of Ind in Caco-2 cell monolayers appeared to be unaffected by the inflamed state of the cells. In conclusion, our findings suggest that the dietary pigment Ind may have the potential to modulate inflammatory processes at the intestinal level.

  2. Probing permeability and microstructure: Unravelling the role of a low-permeability dome on the explosivity of Merapi (Indonesia)

    Kushnir, Alexandra R. L.; Martel, Caroline; Bourdier, Jean-Louis; Heap, Michael J.; Reuschlé, Thierry; Erdmann, Saskia; Komorowski, Jean-Christophe; Cholik, Noer


    permeability of these rocks is more likely associated with their lower fracture density. We propose that diktytaxitic textures may arise from late-stage gas filter pressing of a silica-rich melt phase, which leaves behind a microlite-supported groundmass and cristobalite in neighbouring vesicles. Due to the ubiquity of the Type 3 rocks in all Merapi eruptions, we do not invoke the emplacement of a low-permeability cap as having favoured a particularly high pressurization and subsequent high explosivity of the 2010 eruption. The debate as to the reasons for the highly explosive 2010 eruption rages on.

  3. Development of Low Permeability Oilfields in Shengli Oil Province

    Wang Binhai; Fan Naifu


    @@ Shengli oil province,complicated in geology and rich in resources, is the second largest oil production basin of China. Except for some big/medium sized oil/gas field with high and medium permeability which have already been put into production. There are 11oil fields at great depth with low permeability of less than 50 ×10-3 μm2 have been discovered with 12. 2% of the total proved original oil in place.

  4. Caco-2 Permeability Studies and In Vitro hERG Liability Assessment of Tryptanthrin and Indolinone.

    Jähne, Evelyn A; Eigenmann, Daniela E; Moradi-Afrapoli, Fahimeh; Verjee, Sheela; Butterweck, Veronika; Hebeisen, Simon; Hettich, Timm; Schlotterbeck, Götz; Smieško, Martin; Hamburger, Matthias; Oufir, Mouhssin


    Tryptanthrin and (E,Z)-3-(4-hydroxy-3,5-dimethoxybenzylidene)indolinone (indolinone) were recently isolated from Isatis tinctoria as potent anti-inflammatory and antiallergic alkaloids, and shown to inhibit COX-2, 5-LOX catalyzed leukotriene synthesis, and mast cell degranulation at low µM to nM concentrations. To assess their suitability for oral administration, we screened the compounds in an in vitro intestinal permeability assay using human colonic adenocarcinoma cells. For exact quantification of the compounds, validated UPLC-MS/MS methods were used. Tryptanthrin displayed high permeability (apparent permeability coefficient > 32.0 × 10(-6) cm/s) across the cell monolayer. The efflux ratio below 2 ( 10 µM) and indolinone (IC50 of 24.96 µM). The analysis of compounds using various in silico methods confirmed favorable pharmacokinetic properties, as well as a slight inhibition of the human ether-a-go-go-related gene potassium channel at micromolar concentrations.